Structural stability and sustained release of protein from a multilayer nanofiber/nanoparticle composite.

PubMed

Vakilian, Saeid; Mashayekhan, Shohreh; Shabani, Iman; Khorashadizadeh, Mohsen; Fallah, Ali; Soleimani, Masoud

2015-04-01

The cellular microenvironment can be engineered through the utilization of various nano-patterns and matrix-loaded bioactive molecules. In this study, a multilayer system of electrospun scaffold containing chitosan nanoparticles was introduced to overcome the common problems of instability and burst release of proteins from nanofibrous scaffolds. Bovine serum albumin (BSA)-loaded chitosan nanoparticles was fabricated based on ionic gelation interaction between chitosan and sodium tripolyphosphate. Suspension electrospinning was employed to fabricate poly-ɛ-caprolacton (PCL) containing protein-loaded chitosan nanoparticles with a core-shell structure. To obtain the desired scaffold mechanical properties with enough elasticity for expansion and contraction, a hybrid mono and multilayer electrospun scaffold was fabricated using PCL containing protein-loaded chitosan nanoparticles and poly-L-lactic acid (PLLA). According to the BSA release profile, the multi-layered structure of nanofibers with two barrier layers provided a programmable release pattern of the loaded protein. Moreover, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and circular dichroism spectra results showed that the electrospinning process had no significant effect on the primary and secondary structure of the protein. The results indicated a desirable biocompatibility and mechanical cues of the multilayer nanofibrous scaffolds supporting structural stability and controlled release of the protein, which can offer diverse applications in hollow organ tissue engineering. Copyright © 2015 Elsevier B.V. All rights reserved.

Towards ultraporous poly(L-lactide) scaffolds from quaternary immiscible polymer blends.

PubMed

Virgilio, N; Sarazin, P; Favis, B D

2010-08-01

Ultraporous poly(l-lactide) (PLLA) scaffolds were prepared by melt-processing quaternary ethylene propylene diene rubber/poly(epsilon-caprolactone)/polystyrene/poly(l-lactide) (EPDM/PCL/PS/PLLA) 45/45/5/5 %vol. polymer blends modified with a PS-b-PLLA diblock copolymer. The morphology consists of a PS+PLLA+copolymer sub-blend layer forming at the interface of the EPDM and PCL phases. Quiescent annealing and interfacial modification using the block copolymer are used to control the blend microstructure. The ultraporous structure is subsequently obtained by selectively extracting the EPDM, PS and PCL phases. The PLLA scaffolds modified with the PS-b-PLLA copolymer present themselves as fully interconnected porous networks with asymmetric channel walls, one side being smooth while the other is covered with an array of submicron-sized PLLA droplets. They are prepared with a high degree of control over the pore size, with averages ranging from 5microm to over 100microm and a specific surface from 9.1 to 23.1m(2)/g of PLLA, as annealing is carried out from 0 to 60min. The void volume reaches values as high as 95% and in all cases the shape and dimensions of the scaffolds are maintained with a high level of integrity. The proposed method represents a comprehensive approach towards the design and generation of porous PLLA scaffolds based on complex morphologies from melt-processed multiphase polymer systems. Copyright 2010 Elsevier Ltd. All rights reserved.

PLLA scaffolds surface-engineered via poly (propylene imine) dendrimers for improvement on its biocompatibility/controlled pH biodegradability

NASA Astrophysics Data System (ADS)

Ganjalinia, Atiyeah.; Akbari, Somaye.; Solouk, Atefeh.

2017-02-01

Novel aminolyzed Poly (L) Lactic Acid (PLLA) films and electrospun nanofibrous scaffolds were fabricated and characterized as potential substrates for tissue engineering. The second generation polypropylene imine dendrimer (PPI-G2) was used as the aminolysis agent to functionalize the inert surface of PLLA substrates directly without any pre-modification process. The effect of the solvent type, G2 concentration, reaction temperature and time were studied by following weight reduction percentage, FTIR and contact angle measurements due to determined optimum conditions. In addition, the modified scaffolds abbreviated by PLLA/G2 were analyzed using mechanical properties, SEM images and dye assays as host-guest modeling. The results indicate that under the 0.5 (wt.%) G2 concentration, ethanol as the solvent, room temperature and 4 h of treatment, the optimum conditions were obtained. It was shown that the hydrophilic properties of PLLA/G2 were greatly enhanced. Also, pH value analysis revealed that after 4 weeks, the biodegradation of PLLA caused massive immune cells infusion and inflammation in the medium through increasing the acidic rate by secretion the lactic acid, whereas the PLLA/G2 scaffolds greatly reduced and stabilize the acidic rate through aminolysis reaction. Finally, promoted cell adhesion and viability underlined the favorable properties of PLLA/G2 scaffolds as a biodegradable biomaterial for biomedical implants.

Synthesis, characterization and foaming of PHEA-PLLA, a new graft copolymer for biomedical engineering.

PubMed

Carfì Pavia, Francesco; La Carrubba, Vincenzo; Brucato, Valerio; Palumbo, Fabio Salvatore; Giammona, Gaetano

2014-08-01

In this study a chemical grafting procedure was set up in order to link high molecular weight poly L-lactic acid (PLLA) chains to the hydrophilic α,β-poly(N-2-hydroxyethyl)-DL-aspartamide (PHEA) backbone. A graft copolymer named PHEA-g-PLLA (or simply PHEA-PLLA) was obtained bearing a degree of derivatization of 1.0 mol.% of PLLA as grafted chain. This new hybrid derivative offers both the opportune crystallinity necessary for the production of scaffolds trough a thermally induced phase separation (TIPS) technique and the proper chemical reactivity to perform further functionalizations with bio-effectors and drugs. PHEA-PLLA porous scaffolds for tissue engineering applications were successfully obtained via TIPS and characterized. Structures with an open porosity and a good level of interconnection were detected. As the applicability of the scaffold is mainly dependent on its pore size, preliminary studies about the mechanisms governing scaffold's pore diameter were carried out. Copyright © 2014 Elsevier B.V. All rights reserved.

Anterior cruciate ligament regeneration using braided biodegradable scaffolds: in vitro optimization studies.

PubMed

Lu, Helen H; Cooper, James A; Manuel, Sharron; Freeman, Joseph W; Attawia, Mohammed A; Ko, Frank K; Laurencin, Cato T

2005-08-01

The anterior cruciate ligament (ACL) is the most commonly injured intra-articular ligament of the knee, and limitations in existing reconstruction grafts have prompted an interest in tissue engineered solutions. Previously, we reported on a tissue-engineered ACL scaffold fabricated using a novel, three-dimensional braiding technology. A critical factor in determining cellular response to such a graft is material selection. The objective of this in vitro study was to optimize the braided scaffold, focusing on material composition and the identification of an appropriate polymer. The selection criteria are based on cellular response, construct degradation, and the associated mechanical properties. Three compositions of poly-alpha-hydroxyester fibers, namely polyglycolic acid (PGA), poly-L-lactic acid (PLLA), and polylactic-co-glycolic acid 82:18 (PLAGA) were examined. The effects of polymer composition on scaffold mechanical properties and degradation were evaluated in physiologically relevant solutions. Prior to culturing with primary rabbit ACL cells, scaffolds were pre-coated with fibronectin (Fn, PGA-Fn, PLAGA-Fn, PLLA-Fn), an important protein which is upregulated during ligament healing. Cell attachment and growth were examined as a function of time and polymer composition. While PGA scaffolds measured the highest tensile strength followed by PLLA and PLAGA, its rapid degradation in vitro resulted in matrix disruption and cell death over time. PLLA-based scaffolds maintained their structural integrity and exhibited superior mechanical properties over time. The response of ACL cells was found to be dependent on polymer composition, with the highest cell number measured on PLLA-Fn scaffolds. Surface modification of polymer scaffolds with Fn improved cell attachment efficiency and effected the long-term matrix production by ACL cells on PLLA and PLAGA scaffolds. Therefore based on the overall cellular response and its temporal mechanical and degradation properties in vitro, the PLLA braided scaffold pre-coated with Fn was found to be the most suitable substrate for ACL tissue engineering.

Multiplicity of morphologies in poly (l-lactide) bioresorbable vascular scaffolds

PubMed Central

Ailianou, Artemis; Ramachandran, Karthik; Kossuth, Mary Beth; Oberhauser, James Paul; Kornfield, Julia A.

2016-01-01

Poly(l-lactide) (PLLA) is the structural material of the first clinically approved bioresorbable vascular scaffold (BVS), a promising alternative to permanent metal stents for treatment of coronary heart disease. BVSs are transient implants that support the occluded artery for 6 mo and are completely resorbed in 2 y. Clinical trials of BVSs report restoration of arterial vasomotion and elimination of serious complications such as late stent thrombosis. It is remarkable that a scaffold made from PLLA, known as a brittle polymer, does not fracture when crimped onto a balloon catheter or during deployment in the artery. We used X-ray microdiffraction to discover how PLLA acquired ductile character and found that the crimping process creates localized regions of extreme anisotropy; PLLA chains in the scaffold change orientation from the hoop direction to the radial direction on micrometer-scale distances. This multiplicity of morphologies in the crimped scaffold works in tandem to enable a low-stress response during deployment, which avoids fracture of the PLLA hoops and leaves them with the strength needed to support the artery. Thus, the transformations of the semicrystalline PLLA microstructure during crimping explain the unexpected strength and ductility of the current BVS and point the way to thinner resorbable scaffolds in the future. PMID:27671659

A one-step method to fabricate PLLA scaffolds with deposition of bioactive hydroxyapatite and collagen using ice-based microporogens

PubMed Central

Li, Jiashen; Chen, Yun; Mak, Arthur F.T.; Tuan, Rocky S.; Li, Lin; Li, Yi

2010-01-01

Porous poly(L-lactic acid) (PLLA) scaffolds with bioactive coatings were prepared by a novel one-step method. In this process, ice-based microporogens containing bioactive molecules, such as hydroxyapatite (HA) and collagen, served as both porogens to form the porous structure and vehicles to transfer the bioactive molecules to the inside of PLLA scaffolds in a single step. Based on scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) analysis, the bioactive components were found to be transferred successfully from the porogens to PLLA scaffolds evenly. Osteoblast cells were used to evaluate the cellular behaviors of the composite scaffolds. After 8 days culturing, MTT assay and alkaline phosphatase (ALP) activity results suggested that HA/collagen could improve the interactions between osteoblast cells and the polymeric scaffold. PMID:20004261

Growth on poly(L-lactic acid) porous scaffold preserves CD73 and CD90 immunophenotype markers of rat bone marrow mesenchymal stromal cells.

PubMed

Zamparelli, Alessandra; Zini, Nicoletta; Cattini, Luca; Spaletta, Giulia; Dallatana, Davide; Bassi, Elena; Barbaro, Fulvio; Iafisco, Michele; Mosca, Salvatore; Parrilli, Annapaola; Fini, Milena; Giardino, Roberto; Sandri, Monica; Sprio, Simone; Tampieri, Anna; Maraldi, Nadir M; Toni, Roberto

2014-10-01

Few data are available on the effect of biomaterials on surface antigens of mammalian bone marrow-derived, adult mesenchymal stromal cells (MSCs). Since poly(L-lactic acid) or PLLA is largely used in tissue engineering of human bones, and we are developing a reverse engineering program to prototype with biomaterials the vascular architecture of bones for their bioartificial reconstruction, both in humans and animal models, we have studied the effect of porous, flat and smooth PLLA scaffolds on the immunophenotype of in vitro grown, rat MSCs in the absence of any coating, co-polymeric enrichment, and differentiation stimuli. Similar to controls on plastic, we show that our PLLA scaffold does not modify the distribution of some surface markers in rat MSCs. In particular, the maintained expression of CD73 and CD90 on two different subpopulations (small and large cells) is consistent with their adhesion to the PLLA scaffold through specialized appendages, and to their prominent content in actin. In addition, our PLLA scaffold favours retention of the intermediate filament desmin, believed a putative marker of undifferentiated state. Finally, it preserves all rat MSCs morphotypes, and allows for their survival, adhesion to the substrate, and replication. Remarkably, a subpopulation of rat MSCs grown on our PLLA scaffold exhibited formation of membrane protrusions of uncertain significance, although in a size range and morphology compatible with either motility blebs or shedding vesicles. In summary, our PLLA scaffold has no detrimental effect on a number of features of rat MSCs, primarily the expression of CD73 and CD90.

Comparative Study of the Physical, Topographical and Biological Properties of Electrospinning PCL, PLLA, their Blend and Copolymer Scaffolds

NASA Astrophysics Data System (ADS)

Bolbasov, E.; Goreninskii, S.; Tverdokhlebov, S.; Mishanin, A.; Viknianshchuk, A.; Bezuidenhout, D.; Golovkin, A.

2018-05-01

Biodegradable polymers (blends, copolymers) could be the ideal materials for manufacturing of scaffolds for small diameter vascular graft. Such material characteristics as mechanical properties, chemical structure, nano- and micro topography, surface charge, porosity, wettability etc. are becoming the most important aspects for effectiveness of prosthesis biofunctionalization because of their great impact on cell adhesion, spreading, cell proliferation, differentiation and cell function. The aim of the study is to compare physical, topographical and biological properties of polycaprolactone (PCL), poly-L-lactic acid (PLLA), polycaprolactone + poly-L-lactic acid blend (PCL PLLA), L-lactide/Caprolactone copolymer (PLC7015) scaffolds fabricated with the same fiber thickness using electrospun technology. PCL PLLA scaffolds had the highest average pore area (p<0.01) and the lowest strength (p<0.01). PLC7015 scaffolds had the significantly lower average pore area (p=0.03) but the highest elastic deformation (p<0.01). Biological testing with MMSC (multipotent mesenchyme stem cells) demonstrated that after 72 hours of co-cultivation only on PCL and PLLA scaffolds cells entered to the active phase of adhesion process. We propose that physical and topographical properties of PCL, PLLA, their blend and copolymer are of a great dependence of chemical structure but could be changed during the manufacturing process that will lead to changes in biological properties.

Modification of bone graft by blending with lecithin to improve hydrophilicity and biocompatibility.

PubMed

Wang, Y; Cui, F Z; Jiao, Y P; Hu, K; Fan, D D

2008-03-01

Lecithin was blended to improve the hydrophilicity and biocompatibility of bone graft containing poly(l-lactic acid) (PLLA). Solution blending and freeze drying were used to fabricate symmetrical scaffolds containing different percentages of lecithin (lecithin: PLLA = 0, 5, 10 wt%). Scanning electron microscopy showed that the scaffolds maintained the three-dimensional porous structure. A water uptake experiment proved the significant improvement of hydrophilicity of the blend scaffold. With the addition of lecithin, the compressive strength and compressive modulus decreased. When the weight ratio of lecithin to PLLA was up to 10%, the compressive strength was still more than the lower limit of natural cancellous bone. To test the biocompatibility of the scaffolds, cell culture in vitro and subcutaneous implantation in vivo were performed. MC3T3-E1 preosteoblastic cells were cultured on the scaffolds for 7 days. Methylthiazol tetrazolium assay and laser scanning confocal microscopy were used to exhibit proliferation and morphology of the cells. The subcutaneous implantation in rats tested inflammatory response to the scaffolds. The results proved the better biocompatibility and milder inflammatory reactions of the blend scaffold (lecithin: PLLA = 5%) compared with the scaffold without lecithin. The modified scaffold containing lecithin is promising for bone tissue engineering.

Polymeric vs hydroxyapatite-based scaffolds on dental pulp stem cell proliferation and differentiation

PubMed Central

Khojasteh, Arash; Motamedian, Saeed Reza; Rad, Maryam Rezai; Shahriari, Mehrnoosh Hasan; Nadjmi, Nasser

2015-01-01

AIM: To evaluate adhesion, proliferation and differentiation of human dental pulp stem cells (hDPSCs) on four commercially available scaffold biomaterials. METHODS: hDPSCs were isolated from human dental pulp tissues of extracted wisdom teeth and established in stem cell growth medium. hDPSCs at passage 3-5 were seeded on four commercially available scaffold biomaterials, SureOss (Allograft), Cerabone (Xenograft), PLLA (Synthetic), and OSTEON II Collagen (Composite), for 7 and 14 d in osteogenic medium. Cell adhesion and morphology to the scaffolds were evaluated by scanning electron microscopy (SEM). Cell proliferation and differentiation into osteogenic lineage were evaluated using DNA counting and alkaline phosphatase (ALP) activity assay, respectively. RESULTS: All scaffold biomaterials except SureOss (Allograft) supported hDPSC adhesion, proliferation and differentiation. hDPSCs seeded on PLLA (Synthetic) scaffold showed the highest cell proliferation and attachment as indicated with both SEM and DNA counting assay. Evaluating the osteogenic differentiation capability of hDPSCs on different scaffold biomaterials with ALP activity assay showed high level of ALP activity on cells cultured on PLLA (Synthetic) and OSTEON II Collagen (Composite) scaffolds. SEM micrographs also showed that in the presence of Cerabone (Xenograft) and OSTEON II Collagen (Composite) scaffolds, the hDPSCs demonstrated the fibroblastic phenotype with several cytoplasmic extension, while the cells on PLLA scaffold showed the osteoblastic-like morphology, round-like shape. CONCLUSION: PLLA scaffold supports adhesion, proliferation and osteogenic differentiation of hDPSCs. Hence, it may be useful in combination with hDPSCs for cell-based reconstructive therapy. PMID:26640621

Doxorubicin-loaded mesoporous silica nanoparticle composite nanofibers for long-term adjustments of tumor apoptosis

NASA Astrophysics Data System (ADS)

Yuan, Ziming; Pan, Yue; Cheng, Ruoyu; Sheng, Lulu; Wu, Wei; Pan, Guoqing; Feng, Qiming; Cui, Wenguo

2016-06-01

There is a high local recurrence (LR) rate in breast-conserving therapy (BCT) and enhancement of the local treatment is promising as a way to improve this. Thus we propose a drug delivery system using doxorubicin (DOX)-loaded mesoporous silica nanoparticle composite nanofibers which can release anti-tumor drugs in two phases—burst release in the early stage and sustained release at a later stage—to reduce the LR of BCT. In the present study, we designed a novel composite nanofibrous scaffold to realize the efficient release of drugs by loading both DOX and DOX-loaded mesoporous silica nanoparticles into an electrospun PLLA nanofibrous scaffold. In vitro results demonstrated that this kind of nanomaterial can release DOX in two phases, and the results of in vivo experiments showed that this hybrid nanomaterial significantly inhibited the tumor growth in a solid tumor model. Histopathological examination demonstrated that the apoptosis of tumor cells in the treated group over a 10 week period was significant. The anti-cancer effects were also accompanied with decreased expression of Bcl-2 and TNF-α, along with up-regulation of Bax, Fas and the activation of caspase-3 levels. The present study illustrates that the mesoporous silica nanoparticle composite nanofibrous scaffold could have anti-tumor properties and could be further developed as adjuvant therapeutic protocols for the treatment of cancer.

Enhanced bone regeneration composite scaffolds of PLLA/β-TCP matrix grafted with gelatin and HAp.

PubMed

Wang, Jie-Lin; Chen, Qian; Du, Bei-Bei; Cao, Lu; Lin, Hong; Fan, Zhong-Yong; Dong, Jian

2018-06-01

The composite polylactide PLLA/β-TCP scaffolds were fabricated by solution casting and were coated with gelatin/hydroxyapatite (Gel/HAp) to improve the biological properties of the composite scaffolds. The Gel/HAp mixture was prepared using an in situ reaction, and a grafting-coating method was used to increase the efficiency of coating the PLLA/β-TCP matrix with Gel/HAp. First, free amino groups were introduced by 1,6-hexanediamine to aminolyze the PLLA/β-TCP matrix surface. Second, glutaraldehyde was coupled to Gel/HAp as a crosslinking agent. The structure and properties of Gel/HAp-modified PLLA/β-TCP films were characterized by Fourier transform infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and water contact angle measurements (WCA). The experimental results show that 23 wt% HAp was uniformly dispersed in the gelatin coating by in situ synthesis. The Gel/HAp composite coating was successfully immobilized on the aminolyzed PLLA/β-TCP surface via a chemical grafting method, which promoted a lower degradation rate and was more hydrophilic than a physical grafting method. The Gel/HAp composite coating adhered tightly and homogeneously to the hydrophobic PLLA/β-TCP surface. Moreover, mouse embryo osteoblast precursor (MC3T3-E1) cells grown on the scaffolds were behaviorally and morphologically characterized. The results indicated that the Gel/HAp composite coating was favorable for the attachment and proliferation of preosteoblasts and that Gel/HAp-NH-PLLA/β-TCP would be a candidate scaffold for bone repair. Copyright © 2018 Elsevier B.V. All rights reserved.

Wetspun poly-L-(lactic acid)-borosilicate bioactive glass scaffolds for guided bone regeneration.

PubMed

Fernandes, João S; Reis, Rui L; Pires, Ricardo A

2017-02-01

We developed a porous poly-L-lactic acid (PLLA) scaffold compounded with borosilicate bioactive glasses (BBGs) endowing it with bioactive properties. Porous PLLA-BBG fibre mesh scaffolds were successfully prepared by the combination of wet spinning and fibre bonding techniques. Micro-computed tomography (μCT) confirmed that the PLLA-BBG scaffolds containing ≈25% of BBGs (w/w) exhibited randomly interconnected porous (58 to 62% of interconnectivity and 53 to 67% of porosity) with mean pore diameters higher that 100μm. Bioactivity and degradation studies were performed by immersing the scaffolds in simulated body fluid (SBF) and ultrapure water, respectively. The PLLA-BBG scaffolds presented a faster degradation rate with a constant release of inorganic species, which are capable to produce calcium phosphate structures at the surface of the material after 7days of immersion in SBF (Ca/P ratio of ~1.7). Cellular in vitro studies with human osteosarcoma cell line (Saos-2) and human adipose-derived stem cells (hASCs) showed that PLLA-BBGs are not cytotoxic to cells, while demonstrating their capacity to promote cell adhesion and proliferation. Overall, we showed that the proposed scaffolds present a tailored kinetics on the release of inorganic species and controlled biological response under conditions that mimic the bone physiological environment. Copyright © 2016 Elsevier B.V. All rights reserved.

Developing bioactive composite scaffolds for bone tissue engineering

NASA Astrophysics Data System (ADS)

Chen, Yun

Poly(L-lactic acid) (PLLA) films were fabricated using the method of dissolving and evaporation. PLLA scaffold was prepared by solid-liquid phase separation of polymer solutions and subsequent sublimation of solvent. Bonelike apatite coating was formed on PLLA films, PLLA scaffolds and poly(glycolic acid) (PGA) scaffolds in 24 hours through an accelerated biomimetic process. The ion concentrations in the simulated body fluid (SBF) were nearly 5 times of those in human blood plasma. The apatite formed was characterized using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The apatite formed in 5SBF was similar in morphology and composition to that formed in the classical biomimetic process employing SBF or 1.5SBF, and similar to that of natural bone. This indicated that the biomimetic apatite coating process could be accelerated by using concentrated simulated body fluid at 37°C. Besides saving time, the accelerated biomimetic process is particularly significant to biodegradable polymers. Some polymers which degrade too fast to be coated with apatite by a classical biomimetic process, for example PGA, could be coated with bone-like apatite in an accelerated biomimetic process. Collagen and apatite were co-precipitated as a composite coating on poly(L-lactic acid) (PLLA) in an accelerated biomimetic process. The incubation solution contained collagen (1g/L) and simulated body fluid (SBF) with 5 times inorganic ionic concentrations as human blood plasma. The coating formed on PLLA films and scaffolds after 24 hours incubation was characterized using EDX, XRD, FTIR, and SEM. It was shown that the coating contained carbonated bone-like apatite and collagen, the primary constituents of natural bone. SEM showed a complex composite coating of submicron bone-like apatite particulates combined with collagen fibrils. This work provided an efficient process to obtain bone-like apatite/collagen composite coating. Saos-2 osteoblast-like cells were used to evaluate the cellular behaviors on these biomimetic coatings. Cell morphologies on the surfaces of PLLA films and scaffolds, PLLA films and scaffolds with apatite coating, and PLLA films and scaffolds with apatite/collagen composite coating were studied by SEM. Cell viability was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrasodium bromide (MTT) assay. In addition, differentiated cell function was assessed by measuring alkaline phosphatase activity. These results suggested that the apatite coating and apatite/collagen composite coating fabricated through the accelerated biomimetic processes could improve the interactions between osteoblasts and PLLA. The composite coating was more effective than apatite coating in improving such interactions. PLLA scaffolds coated with submicron collagen fibrils and submicron apatite paticulates are expected to be one of the promising 3D substrates for bone tissue engineering. To facilitate coating into scaffolds, the flowing condition was introduced into the accelerated biomimetic process. The apatite formed in the different sites in the scaffold was characterized using SEM. It was found that the accelerated biomimetic process performed in the flowing condition yielded more uniform spatial distribution of apatite particles than that in the regular shaking condition. This work provides a novel condition for obtaining uniform spatial distribution of bone-like apatite within the scaffolds in a timely manner, which is expected to facilitate uniform distribution of attached cells within the scaffoldsin vitro and in vivo.

Large-scale and highly efficient synthesis of micro- and nano-fibers with controlled fiber morphology by centrifugal jet spinning for tissue regeneration

NASA Astrophysics Data System (ADS)

Ren, Liyun; Pandit, Vaibhav; Elkin, Joshua; Denman, Tyler; Cooper, James A.; Kotha, Shiva P.

2013-02-01

PLLA fibrous tissue scaffolds with controlled fiber nanoscale surface roughness are fabricated with a novel centrifugal jet spinning process. The centrifugal jet spinning technique is a highly efficient synthesis method for micron- to nano-sized fibers with a production rate up to 0.5 g min-1. During the centrifugal jet spinning process, a polymer solution jet is stretched by the centrifugal force of a rotating chamber. By engineering the rheological properties of the polymer solution, solvent evaporation rate and centrifugal force that are applied on the solution jet, polyvinylpyrrolidone (PVP) and poly(l-lactic acid) (PLLA) composite fibers with various diameters are fabricated. Viscosity measurements of polymer solutions allowed us to determine critical polymer chain entanglement limits that allow the generation of continuous fiber as opposed to beads or beaded fibers. Above a critical concentration at which polymer chains are partially or fully entangled, lower polymer concentrations and higher centrifugal forces resulted in thinner fibers. Etching of PVP from the PLLA-PVP composite fibers doped with increasing PVP concentrations yielded PLLA fibers with increasing nano-scale surface roughness and porosity, which increased the fiber hydrophilicity dramatically. Scanning electron micrographs of the etched composite fibers suggest that PVP and PLLA were co-contiguously phase separated within the composite fibers during spinning and nano-scale roughness features were created after the partial etching of PVP. To study the tissue regeneration efficacy of the engineered PLLA fiber matrix, human dermal fibroblasts are used to simulate partial skin graft. Fibers with increased PLLA surface roughness and porosity demonstrated a trend towards higher cell attachment and proliferation.PLLA fibrous tissue scaffolds with controlled fiber nanoscale surface roughness are fabricated with a novel centrifugal jet spinning process. The centrifugal jet spinning technique is a highly efficient synthesis method for micron- to nano-sized fibers with a production rate up to 0.5 g min-1. During the centrifugal jet spinning process, a polymer solution jet is stretched by the centrifugal force of a rotating chamber. By engineering the rheological properties of the polymer solution, solvent evaporation rate and centrifugal force that are applied on the solution jet, polyvinylpyrrolidone (PVP) and poly(l-lactic acid) (PLLA) composite fibers with various diameters are fabricated. Viscosity measurements of polymer solutions allowed us to determine critical polymer chain entanglement limits that allow the generation of continuous fiber as opposed to beads or beaded fibers. Above a critical concentration at which polymer chains are partially or fully entangled, lower polymer concentrations and higher centrifugal forces resulted in thinner fibers. Etching of PVP from the PLLA-PVP composite fibers doped with increasing PVP concentrations yielded PLLA fibers with increasing nano-scale surface roughness and porosity, which increased the fiber hydrophilicity dramatically. Scanning electron micrographs of the etched composite fibers suggest that PVP and PLLA were co-contiguously phase separated within the composite fibers during spinning and nano-scale roughness features were created after the partial etching of PVP. To study the tissue regeneration efficacy of the engineered PLLA fiber matrix, human dermal fibroblasts are used to simulate partial skin graft. Fibers with increased PLLA surface roughness and porosity demonstrated a trend towards higher cell attachment and proliferation. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr33423f

A space network structure constructed by tetraneedlelike ZnO whiskers supporting boron nitride nanosheets to enhance comprehensive properties of poly(L-lacti acid) scaffolds

PubMed Central

Feng, Pei; Peng, Shuping; Wu, Ping; Gao, Chengde; Huang, Wei; Deng, Youwen; Shuai, Cijun

2016-01-01

In this study, the mechanical strength and modulus of poly(L-lacti acid) (PLLA) scaffolds were enhanced with the mechanical properties of boron nitride nanosheets (BNNSs) and tetraneedlelike ZnO whiskers (T-ZnOw). The adhesion and proliferation of cells were improved as well as osteogenic differentiation of stem cells was increased. Their dispersion statues in PLLA matrix were improved through a space network structure constructed by three-dimensional T-ZnOw supporting two-dimensional BNNSs. The results showed that the compressive strength, modulus and Vickers hardness of the scaffolds with incorporation of 1 wt% BNNSs and 7 wt% T-ZnOw together were about 96.15%, 32.86% and 357.19% higher than that of the PLLA scaffolds, respectively. This might be due to the effect of the pull out and bridging of BNNSs and T-ZnOw as well as the crack deflection, facilitating the formation of effective stress transfer between the reinforcement phases and the matrix. Furthermore, incorporation of BNNSs and T-ZnOw together into PLLA scaffolds was beneficial for attachment and viability of MG-63 cells. More importantly, the scaffolds significantly increased proliferation and promoted osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs). The enhanced mechanical and biological properties provide the potentials of PLLA/BNNSs/T-ZnOw scaffolds for the application into bone tissue engineering. PMID:27629058

Electrospun Poly(L-lactide)/Poly(ε-caprolactone) Blend Nanofibrous Scaffold: Characterization and Biocompatibility with Human Adipose-Derived Stem Cells

PubMed Central

Liao, Guiying; Peng, Ejun; Wu, Bolin; Wang, Yuxi; Zeng, Xiaoyong; Xie, Xiaolin

2013-01-01

The essence of tissue engineering is the fabrication of autologous cells or induced stem cells in naturally derived or synthetic scaffolds to form specific tissues. Polymer is thought as an appealing source of cell-seeded scaffold owing to the diversity of its physicochemical property and can be electrospun into nano-size to mimic natural structure. Poly (L-lactic acid) (PLLA) and poly (ε-caprolactone) (PCL) are both excellent aliphatic polyester with almost “opposite” characteristics. The controlling combination of PLLA and PCL provides varying properties and makes diverse applications. Compared with the copolymers of the same components, PLLA/PCL blend demonstrates its potential in regenerative medicine as a simple, efficient and scalable alternative. In this study, we electrospun PLLA/PCL blends of different weight ratios into nanofibrous scaffolds (NFS) and their properties were detected including morphology, porosity, degradation, ATR-FTIR analysis, stress-stain assay, and inflammatory reaction. To explore the biocompatibility of the NFS we synthesized, human adipose-derived stem cells (hASCs) were used to evaluate proliferation, attachment, viability and multi-lineage differentiation. In conclusion, the electrospun PLLA/PCL blend nanofibrous scaffold with the indicated weight ratios all supported hASCs well. However, the NFS of 1/1 weight ratio showed better properties and cellular responses in all assessments, implying it a biocompatible scaffold for tissue engineering. PMID:23990941

Plasma treatment induces internal surface modifications of electrospun poly(L-lactic) acid scaffold to enhance protein coating

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jin Seo, Hyok; Hee Lee, Mi; Kwon, Byeong-Ju

2013-08-21

Advanced biomaterials should also be bioactive with regard to desirable cellular responses, such as selective protein adsorption and cell attachment, proliferation, and differentiation. To enhance cell-material interactions, surface modifications have commonly been performed. Among the various surface modification approaches, atmospheric pressure glow discharge plasma has been used to change a hydrophobic polymer surface to a hydrophilic surface. Poly(L-lactic acid) (PLLA)-derived scaffolds lack cell recognition signals and the hydrophobic nature of PLLA hinders cell seeding. To make PLLA surfaces more conducive to cell attachment and spreading, surface modifications may be used to create cell-biomaterial interfaces that elicit controlled cell adhesion andmore » maintain differentiated phenotypes. In this study, (He) gaseous atmospheric plasma glow discharge was used to change the characteristics of a 3D-type polymeric scaffold from hydrophobic to hydrophilic on both the outer and inner surfaces of the scaffold and the penetration efficiency with fibronectin was investigated. Field-emission scanning electron microscope images showed that some grooves were formed on the PLLA fibers after plasma treatment. X-ray photoelectron spectroscopy data also showed chemical changes in the PLLA structure. After plasma treatment, -CN (285.76 eV) was increased in C1s and -NH{sub 2} (399.70 eV) was increased significantly and –N=CH (400.80 eV) and –NH{sub 3}{sup +} (402.05 eV) were newly appeared in N1s. These changes allowed fibronectin to penetrate into the PLLA scaffold; this could be observed by confocal microscopy. In conclusion, helium atmospheric pressure plasma treatment was effective in modifying the polymeric scaffold, making it hydrophilic, and this treatment can also be used in tissue engineering research as needed to make polymers hydrophilic.« less

Relevance of fiber integrated gelatin-nanohydroxyapatite composite scaffold for bone tissue regeneration

NASA Astrophysics Data System (ADS)

Halima Shamaz, Bibi; Anitha, A.; Vijayamohan, Manju; Kuttappan, Shruthy; Nair, Shantikumar; Nair, Manitha B.

2015-10-01

Porous nanohydroxyapatite (nanoHA) is a promising bone substitute, but it is brittle, which limits its utility for load bearing applications. To address this issue, herein, biodegradable electrospun microfibrous sheets of poly(L-lactic acid)-(PLLA)-polyvinyl alcohol (PVA) were incorporated into a gelatin-nanoHA matrix which was investigated for its mechanical properties, the physical integration of the fibers with the matrix, cell infiltration, osteogenic differentiation and bone regeneration. The inclusion of sacrificial fibers like PVA along with PLLA and leaching resulted in improved cellular infiltration towards the center of the scaffold. Furthermore, the treatment of PLLA fibers with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide enhanced their hydrophilicity, ensuring firm anchorage between the fibers and the gelatin-HA matrix. The incorporation of PLLA microfibers within the gelatin-nanoHA matrix reduced the brittleness of the scaffolds, the effect being proportional to the number of layers of fibrous sheets in the matrix. The proliferation and osteogenic differentiation of human adipose-derived mesenchymal stem cells was augmented on the fibrous scaffolds in comparison to those scaffolds devoid of fibers. Finally, the scaffold could promote cell infiltration, together with bone regeneration, upon implantation in a rabbit femoral cortical defect within 4 weeks. The bone regeneration potential was significantly higher when compared to commercially available HA (Surgiwear™). Thus, this biomimetic, porous, 3D composite scaffold could be offered as a promising candidate for bone regeneration in orthopedics.

Heparin-binding peptide amphiphile supramolecular architectures as platforms for angiogenesis and drug delivery

NASA Astrophysics Data System (ADS)

Chow, Lesleyann W.

A fascinating phenomenon in nature is the self-assembly of molecules into a functional, hierarchical structure. In the past decade, the Stupp Laboratory has developed several classes of self-assembling biomaterials, one of which is the synthetic peptide amphiphile (PA). Self-assembling PAs are attractive and versatile biomolecules that can be customized for specific applications in regenerative medicine. In particular, a heparin-binding peptide amphiphile (HBPA) containing a specific heparin-binding peptide sequence was used here to induce angiogenesis and serve as a delivery vehicle for growth factors and small hydrophobic molecules. Throughout this dissertation, the HBPA/heparin system is used in different architectures for a variety of regenerative medicine applications. In one aspect of this work, hybrid scaffolds made from HBPA/heparin gelled on a poly(L-lactic acid) (PLLA) fiber mesh were used to promote angiogenesis to facilitate pancreatic islet transplantation for the treatment of type 1 diabetes. Delivery of growth factors with HBPA/PLLA scafflolds increased vessel density in vivo and correlated with improved transplant outcomes in a streptozotocin-induced diabetic mouse model. Soluble HBPA nanofiber architectures were also useful for islet transplantation applications. These nanofibers were used at concentrations below gelation to deliver growth factors into the dense islet cell aggregate, promoting cell survival and angiogenesis in vitro. The nanostructures infiltrated the islets and promoted the retention of heparin and growth factors within the islet. Another interesting growth factor release system discussed here is the HBPA membrane structure. HBPA was found to self-assemble with hyaluronic acid, a large biopolymer found in the body, into macroscopic, hierarchically-ordered membranes. Heparin was incorporated into these membranes and affected the membrane's mechanical properties and growth factor release. Human mesenchymal stem cells were also shown to attach and maintain viability on these membranes. Finally, HBPA nanofibers were used to control the release of small hydrophobic molecules. HBPA nanofiber gels released nitric oxide (NO) to inhibit neointimal hyperplasia, a major cause for vascular graft or stent failure. HBPA/heparin gels were shown to prolong the release of NO generated from NO donors, significantly reducing neointimal hyperplasia in injured carotid arteries in vivo.

Biocompatibility of MG-63 cells on collagen, poly-L-lactic acid, hydroxyapatite scaffolds with different parameters.

PubMed

Cecen, Berivan; Kozaci, Didem; Yuksel, Mithat; Erdemli, Diler; Bagriyanik, Alper; Havitcioglu, Hasan

2015-03-18

In this study, osteoblast-like MG-63 cells were cultured on 3 different scaffold types composed of (a) collagen + poly-L-lactic acid (PLLA), (b) collagen + hydroxyapatite (HA; 30ºC) or (c) collagen + hydroxyapatite (HA; 37ºC) and produced with different porosities. Biomechanical properties of the scaffolds were characterized by tensile strength measurements. Properties of the cell-seeded scaffolds were evaluated with scanning electron microscopy (SEM). Cell adhesion and proliferation capacities were evaluated. Alkaline phosphatase (ALP) levels in media were measured. Transmission electron microscopy (TEM) and histological analyses were used to assess morphological characteristics. Our results showed that collagen-based PLLA and HA scaffolds have good cell biocompatibility. MTT test showed that the scaffolds exhibited no cytotoxicity. According to the force and displacement data, collagen + HA at 37ºC showed the highest mechanical strength and displacement. The results suggest that collagen-based PLLA and HA scaffolds might improve osteoblastic growth in vitro and have biomaterial integration potential in possible therapeutic approaches for future clinical studies.

Preparation of conductive gold nanowires in confined environment of gold-filled polymer nanotubes.

PubMed

Mitschang, Fabian; Langner, Markus; Vieker, Henning; Beyer, André; Greiner, Andreas

2015-02-01

Continuous conductive gold nanofibers are prepared via the "tubes by fiber templates" process. First, poly(l-lactide) (PLLA)-stabilized gold nanoparticles (AuNP) with over 60 wt% gold are synthesized and characterized, including gel permeation chromatography coupled with a diode array detector. Subsequent electrospinning of these AuNP with template PLLA results in composite nanofibers featuring a high gold content of 57 wt%. Highly homogeneous gold nanowires are obtained after chemical vapor deposition of 345 nm of poly(p-xylylene) (PPX) onto the composite fibers followed by pyrolysis of the polymers at 1050 °C. The corresponding heat-induced transition from continuous gold-loaded polymer tubes to smooth gold nanofibers is studied by transmission electron microscopy and helium ion microscopy using both secondary electrons and Rutherford backscattered ions. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Development of porous lamellar poly(L-lactic acid) scaffolds by conventional injection molding process.

PubMed

Ghosh, Satyabrata; Viana, Júlio C; Reis, Rui L; Mano, João F

2008-07-01

A novel fabrication technique is proposed for the preparation of unidirectionally oriented, porous scaffolds by selective polymer leaching from lamellar structures created by conventional injection molding. The proof of the concept is implemented using a 50/50 wt.% poly(L-lactic acid)/poly(ethylene oxide) (PLLA/PEO) blend. With this composition, the PLLA and PEO blend is biphasic, containing a homogeneous PLLA/PEO phase and a PEO-rich phase. The two phases were structured using injection molding into well-defined alternating layers of homogeneous PLLA/PEO phase and PEO-rich phase. Leaching of water-soluble PEO from the PEO-rich phase produces macropores, and leaching of phase-separated PEO from the initially homogeneous PLLA/PEO phase produces micropores in the lamellae. Thus, scaffolds with a macroporous lamellar architecture with microporous walls can be produced. The lamellae are continuous along the flow direction, and a continuous lamellar thickness of less than 1 microm could be achieved. Porosities of 57-74% and pore sizes of around 50-100 microm can be obtained using this process. The tensile elastic moduli of the porous constructs were between 580 and 800 MPa. We propose that this organic-solvent-free method of preparing lamellar scaffolds with good mechanical properties, and the reproducibility associated with the injection molding technique, holds promise for a wide range of guided tissue engineering applications.

A structural model for the flexural mechanics of nonwoven tissue engineering scaffolds.

PubMed

Engelmayr, George C; Sacks, Michael S

2006-08-01

The development of methods to predict the strength and stiffness of biomaterials used in tissue engineering is critical for load-bearing applications in which the essential functional requirements are primarily mechanical. We previously quantified changes in the effective stiffness (E) of needled nonwoven polyglycolic acid (PGA) and poly-L-lactic acid (PLLA) scaffolds due to tissue formation and scaffold degradation under three-point bending. Toward predicting these changes, we present a structural model for E of a needled nonwoven scaffold in flexure. The model accounted for the number and orientation of fibers within a representative volume element of the scaffold demarcated by the needling process. The spring-like effective stiffness of the curved fibers was calculated using the sinusoidal fiber shapes. Structural and mechanical properties of PGA and PLLA fibers and PGA, PLLA, and 50:50 PGA/PLLA scaffolds were measured and compared with model predictions. To verify the general predictive capability, the predicted dependence of E on fiber diameter was compared with experimental measurements. Needled nonwoven scaffolds were found to exhibit distinct preferred (PD) and cross-preferred (XD) fiber directions, with an E ratio (PD/XD) of approximately 3:1. The good agreement between the predicted and experimental dependence of E on fiber diameter (R2 = 0.987) suggests that the structural model can be used to design scaffolds with E values more similar to native soft tissues. A comparison with previous results for cell-seeded scaffolds (Engelmayr, G. C., Jr., et al., 2005, Biomaterials, 26(2), pp. 175-187) suggests, for the first time, that the primary mechanical effect of collagen deposition is an increase in the number of fiber-fiber bond points yielding effectively stiffer scaffold fibers. This finding indicated that the effects of tissue deposition on needled nonwoven scaffold mechanics do not follow a rule-of-mixtures behavior. These important results underscore the need for structural approaches in modeling the effects of engineered tissue formation on nonwoven scaffolds, and their potential utility in scaffold design.

Poly(L-lactic acid) nanofibers containing Cissus quadrangularis induced osteogenic differentiation in vitro.

PubMed

Parvathi, K; Krishnan, Amit G; Anitha, A; Jayakumar, R; Nair, Manitha B

2018-04-15

Cissus quadrangularis (CQ) is known as "bone setter" in Ayurvedic Medicine because of its ability to promote fracture healing. Polymers incorporated with CQ at lower concentration have shown to enhance osteogenic differentiation of mesenchymal stem cells (MSCs) in vitro. However, for the healing of clinically relevant critical sized bone defects, large amount of CQ would be required. Based on this perception, a herbal fibrous sheet containing high weight percentage of CQ [20,40 and 60wt/wt% in poly (L-lactic acid) (PLLA)] was fabricated through electrospinning. The solution concentration, flow rate, voltage and tip-target distance was optimized to obtain nanofibers. The hydrophobicity of PLLA fibers was reduced through CQ incorporation. There was considerable increase in the adhesion, proliferation and osteogenic differentiation of MSCs on herbal fibers than normal fibers, mainly on P-Q20 and P-CQ40. MSCs were differentiated into osteoblasts without providing any osteogenic supplements in the medium, indicating its osteoinductive capability. The herbal sheet also could promote mineralization when immersed in simulated body fluid for 14days. These studies specify that PLLA nanofibers loaded with 20 and 40wt% of CQ could serve as a potential candidate for bone tissue engineering applications. Copyright © 2017 Elsevier B.V. All rights reserved.

Impact of polymer structure and composition on fully resorbable endovascular scaffold performance

PubMed Central

Ferdous, Jahid; Kolachalama, Vijaya B.; Shazly, Tarek

2014-01-01

Fully erodible endovascular scaffolds are being increasingly considered for the treatment of obstructive arterial disease owing to their potential to mitigate long-term risks associated with permanent alternatives. While complete scaffold erosion facilitates vessel healing, generation and release of material degradation by-products from candidate materials such as poly-l-lactide (PLLA) may elicit local inflammatory responses that limit implant efficacy. We developed a computational framework to quantify how the compositional and structural parameters of PLLA-based fully erodible endovascular scaffolds affect degradation kinetics, erosion kinetics and the transient accumulation of material by-products within the arterial wall. Parametric studies reveal that, while some material properties have similar effects on these critical processes, others induce qualitatively opposing responses. For example, scaffold degradation is only mildly responsive to changes in either PLLA polydispersity or the initial degree of crystallinity, while the erosion kinetics is comparatively sensitive to crystallinity. Moreover, lactide doping can effectively tune both scaffold degradation and erosion, but a concomitant increase in local byproduct accumulation raises concerns about implant safety. Optimized erodible endovascular scaffolds must precisely balance therapeutic function and biological response over the implant lifetime, where compositional and structural parameters will have differential effects on implant performance. PMID:23261926

Functional Maturation of Induced Pluripotent Stem Cell Hepatocytes in Extracellular Matrix—A Comparative Analysis of Bioartificial Liver Microenvironments

PubMed Central

Wang, Bo; Jakus, Adam E.; Baptista, Pedro M.; Soker, Shay; Soto-Gutierrez, Alejandro; Abecassis, Michael M.; Shah, Ramille N.

2016-01-01

Induced pluripotent stem cells (iPSCs) are new diagnostic and potentially therapeutic tools to model disease and assess the toxicity of pharmaceutical medications. A common limitation of cell lineages derived from iPSCs is a blunted phenotype compared with fully developed, endogenous cells. We examined the influence of novel three-dimensional bioartificial microenvironments on function and maturation of hepatocyte-like cells differentiated from iPSCs and grown within an acellular, liver-derived extracellular matrix (ECM) scaffold. In parallel, we also compared a bioplotted poly-l-lactic acid (PLLA) scaffold that allows for cell growth in three dimensions and formation of cell-cell contacts but is infused with type I collagen (PLLA-collagen scaffold) alone as a “deconstructed” control scaffold with narrowed biological diversity. iPSC-derived hepatocytes cultured within both scaffolds remained viable, became polarized, and formed bile canaliculi-like structures; however, cells grown within ECM scaffolds had significantly higher P450 (CYP2C9, CYP3A4, CYP1A2) mRNA levels and metabolic enzyme activity compared with iPSC hepatocytes grown in either bioplotted PLLA collagen or Matrigel sandwich control culture. Additionally, the rate of albumin synthesis approached the level of primary cryopreserved hepatocytes with lower transcription of fetal-specific genes, α-fetoprotein and CYP3A7, compared with either PLLA-collagen scaffolds or sandwich culture. These studies show that two acellular, three-dimensional culture systems increase the function of iPSC-derived hepatocytes. However, scaffolds derived from ECM alone induced further hepatocyte maturation compared with bioplotted PLLA-collagen scaffolds. This effect is likely mediated by the complex composition of ECM scaffolds in contrast to bioplotted scaffolds, suggesting their utility for in vitro hepatocyte assays or drug discovery. Significance Through the use of novel technology to develop three-dimensional (3D) scaffolds, the present study demonstrated that hepatocyte-like cells derived via induced pluripotent stem cell (iPSC) technology mature on 3D extracellular matrix scaffolds as a result of 3D matrix structure and scaffold biology. The result is an improved hepatic phenotype with increased synthetic and catalytic potency, an improvement on the blunted phenotype of iPSC-derived hepatocytes, a critical limitation of iPSC technology. These findings provide insight into the influence of 3D microenvironments on the viability, proliferation, and function of iPSC hepatocytes to yield a more mature population of cells for cell toxicity studies and disease modeling. PMID:27421950

Improving effects of chitosan nanofiber scaffolds on osteoblast proliferation and maturation

PubMed Central

Ho, Ming-Hua; Liao, Mei-Hsiu; Lin, Yi-Ling; Lai, Chien-Hao; Lin, Pei-I; Chen, Ruei-Ming

2014-01-01

Osteoblast maturation plays a key role in regulating osteogenesis. Electrospun nanofibrous products were reported to possess a high surface area and porosity. In this study, we developed chitosan nanofibers and examined the effects of nanofibrous scaffolds on osteoblast maturation and the possible mechanisms. Macro- and micro observations of the chitosan nanofibers revealed that these nanoproducts had a flat surface and well-distributed fibers with nanoscale diameters. Mouse osteoblasts were able to attach onto the chitosan nanofiber scaffolds, and the scaffolds degraded in a time-dependent manner. Analysis by scanning electron microscopy further showed mouse osteoblasts adhered onto the scaffolds along the nanofibers, and cell–cell communication was also detected. Mouse osteoblasts grew much better on chitosan nanofiber scaffolds than on chitosan films. In addition, human osteoblasts were able to adhere and grow on the chitosan nanofiber scaffolds. Interestingly, culturing human osteoblasts on chitosan nanofiber scaffolds time-dependently increased DNA replication and cell proliferation. In parallel, administration of human osteoblasts onto chitosan nanofibers significantly induced osteopontin, osteocalcin, and alkaline phosphatase (ALP) messenger (m)RNA expression. As to the mechanism, chitosan nanofibers triggered runt-related transcription factor 2 mRNA and protein syntheses. Consequently, results of ALP-, alizarin red-, and von Kossa-staining analyses showed that chitosan nanofibers improved osteoblast mineralization. Taken together, results of this study demonstrate that chitosan nanofibers can stimulate osteoblast proliferation and maturation via runt-related transcription factor 2-mediated regulation of osteoblast-associated osteopontin, osteocalcin, and ALP gene expression. PMID:25246786

A Comparison of Degradable Synthetic Polymer Fibers for Anterior Cruciate Ligament Reconstruction

PubMed Central

Tovar, Nick; Bourke, Sharon; Jaffe, Michael; Murthy, N. Sanjeeva; Kohn, Joachim; Gatt, Charles; Dunn, Michael G.

2009-01-01

We compared mechanical properties, degradation rates, and cellular compatibilities of two synthetic polymer fibers potentially useful as ACL reconstruction scaffolds: poly(desaminotyrosyl-tyrosine dodecyl dodecanedioate)(12,10), p(DTD DD) and poly(L-lactic acid), PLLA. The yield stress of ethylene oxide (ETO) sterilized wet fibers was 150 ± 22 MPa and 87 ± 12 MPa for p(DTD DD) and PLLA, respectively, with moduli of 1.7 ± 0.1 MPa and 4.4 ± 0.43 MPa. Strength and molecular weight retention were determined after incubation under physiological conditions at varying times. After 64 weeks strength decreased to 20 and 37% of the initial sterile fiber values and MW decreased to 41% and 36% of the initial values for p(DTD DD) and PLLA, respectively. ETO sterilization had no significant effect on mechanical properties. Differences in mechanical behavior may be due to the semicrystalline nature of PLLA and the small degree of crystallinity induced by mesogenic ordering in p(DTD DD) suggested by DSC analysis. Fibroblast growth was similar on 50-fiber scaffolds of both polymers through 16 days in vitro. These data suggest that p(DTD DD) fibers, with higher strength, lower stiffness, favorable degradation rate and cellular compatibility, may be a superior alternative to PLLA fibers for development of ACL reconstruction scaffolds. PMID:19623532

* Hierarchically Structured Electrospun Scaffolds with Chemically Conjugated Growth Factor for Ligament Tissue Engineering.

PubMed

Pauly, Hannah M; Sathy, Binulal N; Olvera, Dinorath; McCarthy, Helen O; Kelly, Daniel J; Popat, Ketul C; Dunne, Nicholas J; Haut Donahue, Tammy Lynn

2017-08-01

The anterior cruciate ligament (ACL) of the knee is vital for proper joint function and is commonly ruptured during sports injuries or car accidents. Due to a lack of intrinsic healing capacity and drawbacks with allografts and autografts, there is a need for a tissue-engineered ACL replacement. Our group has previously used aligned sheets of electrospun polycaprolactone nanofibers to develop solid cylindrical bundles of longitudinally aligned nanofibers. We have shown that these nanofiber bundles support cell proliferation and elongation and the hierarchical structure and material properties are similar to the native human ACL. It is possible to combine multiple nanofiber bundles to create a scaffold that attempts to mimic the macroscale structure of the ACL. The goal of this work was to develop a hierarchical bioactive scaffold for ligament tissue engineering using connective tissue growth factor (CTGF)-conjugated nanofiber bundles and evaluate the behavior of mesenchymal stem cells (MSCs) on these scaffolds in vitro and in vivo. CTGF was immobilized onto the surface of individual nanofiber bundles or scaffolds consisting of multiple nanofiber bundles. The conjugation efficiency and the release of conjugated CTGF were assessed using X-ray photoelectron spectroscopy, assays, and immunofluorescence staining. Scaffolds were seeded with MSCs and maintained in vitro for 7 days (individual nanofiber bundles), in vitro for 21 days (scaled-up scaffolds of 20 nanofiber bundles), or in vivo for 6 weeks (small scaffolds of 4 nanofiber bundles), and ligament-specific tissue formation was assessed in comparison to non-CTGF-conjugated control scaffolds. Results showed that CTGF conjugation encouraged cell proliferation and ligament-specific tissue formation in vitro and in vivo. The results suggest that hierarchical electrospun nanofiber bundles conjugated with CTGF are a scalable and bioactive scaffold for ACL tissue engineering.

Chitosan nanofiber scaffold improves bone healing via stimulating trabecular bone production due to upregulation of the Runx2/osteocalcin/alkaline phosphatase signaling pathway

PubMed Central

Ho, Ming-Hua; Yao, Chih-Jung; Liao, Mei-Hsiu; Lin, Pei-I; Liu, Shing-Hwa; Chen, Ruei-Ming

2015-01-01

Osteoblasts play critical roles in bone formation. Our previous study showed that chitosan nanofibers can stimulate osteoblast proliferation and maturation. This translational study used an animal model of bone defects to evaluate the effects of chitosan nanofiber scaffolds on bone healing and the possible mechanisms. In this study, we produced uniform chitosan nanofibers with fiber diameters of approximately 200 nm. A bone defect was surgically created in the proximal femurs of male C57LB/6 mice, and then the left femur was implanted with chitosan nanofiber scaffolds for 21 days and compared with the right femur, which served as a control. Histological analyses revealed that implantation of chitosan nanofiber scaffolds did not lead to hepatotoxicity or nephrotoxicity. Instead, imaging analyses by X-ray transmission and microcomputed tomography showed that implantation of chitosan nanofiber scaffolds improved bone healing compared with the control group. In parallel, microcomputed tomography and bone histomorphometric assays further demonstrated augmentation of the production of new trabecular bone in the chitosan nanofiber-treated group. Furthermore, implantation of chitosan nanofiber scaffolds led to a significant increase in the trabecular bone thickness but a reduction in the trabecular parameter factor. As to the mechanisms, analysis by confocal microscopy showed that implantation of chitosan nanofiber scaffolds increased levels of Runt-related transcription factor 2 (Runx2), a key transcription factor that regulates osteogenesis, in the bone defect sites. Successively, amounts of alkaline phosphatase and osteocalcin, two typical biomarkers that can simulate bone maturation, were augmented following implantation of chitosan nanofiber scaffolds. Taken together, this translational study showed a beneficial effect of chitosan nanofiber scaffolds on bone healing through stimulating trabecular bone production due to upregulation of Runx2-mediated alkaline phosphatase and osteocalcin gene expressions. Our results suggest the potential of chitosan nanofiber scaffolds for therapy of bone diseases, including bone defects and bone fractures. PMID:26451104

Surface modification of bioactive glass nanoparticles and the mechanical and biological properties of poly(L-lactide) composites.

PubMed

Liu, Aixue; Hong, Zhongkui; Zhuang, Xiuli; Chen, Xuesi; Cui, Yang; Liu, Yi; Jing, Xiabin

2008-07-01

Novel bioactive glass (BG) nanoparticles/poly(L-lactide) (PLLA) composites were prepared as promising bone-repairing materials. The BG nanoparticles (Si:P:Ca=29:13:58 weight ratio) of about 40nm diameter were prepared via the sol-gel method. In order to improve the phase compatibility between the polymer and the inorganic phase, PLLA (M(n)=9700Da) was linked to the surface of the BG particles by diisocyanate. The grafting ratio of PLLA was in the vicinity of 20 wt.%. The grafting modification could improve the tensile strength, tensile modulus and impact energy of the composites by increasing the phase compatibility. When the filler loading reached around 4 wt.%, the tensile strength of the composite increased from 56.7 to 69.2MPa for the pure PLLA, and the impact strength energy increased from 15.8 to 18.0 kJ m(-2). The morphology of the tensile fracture surface of the composite showed surface-grafted bioactive glass particles (g-BG) to be dispersed homogeneously in the PLLA matrix. An in vitro bioactivity test showed that, compared to pure PLLA scaffold, the BG/PLLA nanocomposite demonstrated a greater capability to induce the formation of an apatite layer on the scaffold surface. The results of marrow stromal cell culture revealed that the composites containing either BG or g-BG particles have much better biocompatibility compared to pure PLLA material.

Poly(hydroxybutyrate)/cellulose acetate blend nanofiber scaffolds: Preparation, characterization and cytocompatibility.

PubMed

Zhijiang, Cai; Yi, Xu; Haizheng, Yang; Jia, Jianru; Liu, Yuanpei

2016-01-01

Poly(hydroxybutyrate) (PHB)/cellulose acetate (CA) blend nanofiber scaffolds were fabricated by electrospinning using the blends of chloroform and DMF as solvent. The blend nanofiber scaffolds were characterized by SEM, FTIR, XRD, DSC, contact angle and tensile test. The blend nanofibers exhibited cylindrical, uniform, bead-free and random orientation with the diameter ranged from 80-680 nm. The scaffolds had very well interconnected porous fibrous network structure and large aspect surface areas. It was found that the presence of CA affected the crystallization of PHB due to formation of intermolecular hydrogen bonds, which restricted the preferential orientation of PHB molecules. The DSC result showed that the PHB and CA were miscible in the blend nanofiber. An increase in the glass transition temperature was observed with increasing CA content. Additionally, the mechanical properties of blend nanofiber scaffolds were largely influenced by the weight ratio of PHB/CA. The tensile strength, yield strength and elongation at break of the blend nanofiber scaffolds increased from 3.3 ± 0.35 MPa, 2.8 ± 0.26 MPa, and 8 ± 0.77% to 5.05 ± 0.52 MPa, 4.6 ± 0.82 MPa, and 17.6 ± 1.24% by increasing PHB content from 60% to 90%, respectively. The water contact angle of blend nanofiber scaffolds decreased about 50% from 112 ± 2.1° to 60 ± 0.75°. The biodegradability was evaluated by in vitro degradation test and the results revealed that the blend nanofiber scaffolds showed much higher degradation rates than the neat PHB. The cytocompatibility of the blend nanofiber scaffolds was preliminarily evaluated by cell adhesion studies. The cells incubated with PHB/CA blend nanofiber scaffold for 48 h were capable of forming cell adhesion and proliferation. It showed much better biocompatibility than pure PHB film. Thus, the prepared PHB/CA blend nanofiber scaffolds are bioactive and may be more suitable for cell proliferation suggesting that these scaffolds can be used for wound dressing or tissue-engineering scaffolds. Copyright © 2015 Elsevier B.V. All rights reserved.

Effects of hydrostatic pressure on leporine meniscus cell-seeded PLLA scaffolds.

PubMed

Gunja, Najmuddin J; Athanasiou, Kyriacos A

2010-03-01

Hydrostatic pressure (HP) is an important component of the loading environment of the knee joint. Studies with articular chondrocytes and TMJ disc fibrochondrocytes have identified certain benefits of HP for tissue engineering purposes. However, similar studies with meniscus cells are lacking. Thus, in this experiment, the effects of applying 10 MPa of HP at three different frequencies (0, 0.1, and 1 Hz) to leporine meniscus cell-seeded PLLA scaffolds were examined. HP was applied once every 3 days for 1 h for a period of 28 days. Constructs were analyzed for cellular, biochemical, and biomechanical properties. At t = 4 weeks, total collagen/scaffold was found to be significantly higher in the 10 MPa, 0 Hz group when compared with other groups. This despite the fact that the cell numbers/scaffold were found to be lower in all HP groups when compared with the culture control. Additionally, the total GAG/scaffold, instantaneous modulus, and relaxation modulus were significantly increased in the 10 MPa, 0 Hz group when compared with the culture control. In summary, this experiment provides evidence for the benefit of a 10 MPa, 0 Hz stimulus, on both biochemical and biomechanical aspects, for the purposes of meniscus tissue engineering using PLLA scaffolds. (c) 2009 Wiley Periodicals, Inc.

Dynamic reassembly of peptide RADA16 nanofiber scaffold

NASA Astrophysics Data System (ADS)

Yokoi, Hidenori; Kinoshita, Takatoshi; Zhang, Shuguang

2005-06-01

Nanofiber structures of some peptides and proteins as biological materials have been studied extensively, but their molecular mechanism of self-assembly and reassembly still remains unclear. We report here the reassembly of an ionic self-complementary peptide RADARADARADARADA (RADA16-I) that forms a well defined nanofiber scaffold. The 16-residue peptide forms stable -sheet structure and undergoes molecular self-assembly into nanofibers and eventually a scaffold hydrogel consisting of >99.5% water. In this study, the nanofiber scaffold was sonicated into smaller fragments. Circular dichroism, atomic force microscopy, and rheology were used to follow the kinetics of the reassembly. These sonicated fragments not only quickly reassemble into nanofibers that were indistinguishable from the original material, but their reassembly also correlated with the rheological analyses showing an increase of scaffold rigidity as a function of nanofiber length. The disassembly and reassembly processes were repeated four times and, each time, the reassembly reached the original length. We proposed a plausible sliding diffusion model to interpret the reassembly involving complementary nanofiber cohesive ends. This reassembly process is important for fabrication of new scaffolds for 3D cell culture, tissue repair, and regenerative medicine. atomic force microscopy | circular dichroism | dynamic behaviors | ionic self-complementary peptides | nanofiber hydrogels

Enhanced human bone marrow mesenchymal stem cell functions in novel 3D cartilage scaffolds with hydrogen treated multi-walled carbon nanotubes.

PubMed

Holmes, Benjamin; Castro, Nathan J; Li, Jian; Keidar, Michael; Zhang, Lijie Grace

2013-09-13

Cartilage tissue is a nanostructured tissue which is notoriously hard to regenerate due to its extremely poor inherent regenerative capacity and complex stratified architecture. Current treatment methods are highly invasive and may have many complications. Thus, the goal of this work is to use nanomaterials and nano/microfabrication methods to create novel biologically inspired tissue engineered cartilage scaffolds to facilitate human bone marrow mesenchymal stem cell (MSC) chondrogenesis. To this end we utilized electrospinning to design and fabricate a series of novel 3D biomimetic nanostructured scaffolds based on hydrogen (H2) treated multi-walled carbon nanotubes (MWCNTs) and biocompatible poly(L-lactic acid) (PLLA) polymers. Specifically, a series of electrospun fibrous PLLA scaffolds with controlled fiber dimension were fabricated in this study. In vitro MSC studies showed that stem cells prefer to attach in the scaffolds with smaller fiber diameter. More importantly, the MWCNT embedded scaffolds showed a drastic increase in mechanical strength and a compressive Young's modulus matching to natural cartilage. Furthermore, our MSC differentiation results demonstrated that incorporation of the H2 treated carbon nanotubes and poly-L-lysine coating can induce more chondrogenic differentiations of MSCs than controls. After two weeks of culture, PLLA scaffolds with H2 treated MWCNTs and poly-L-lysine can achieve the highest glycosaminoglycan synthesis, making them promising for further exploration for cartilage regeneration.

Enhanced human bone marrow mesenchymal stem cell functions in novel 3D cartilage scaffolds with hydrogen treated multi-walled carbon nanotubes

NASA Astrophysics Data System (ADS)

Holmes, Benjamin; Castro, Nathan J.; Li, Jian; Keidar, Michael; Zhang, Lijie Grace

2013-09-01

Cartilage tissue is a nanostructured tissue which is notoriously hard to regenerate due to its extremely poor inherent regenerative capacity and complex stratified architecture. Current treatment methods are highly invasive and may have many complications. Thus, the goal of this work is to use nanomaterials and nano/microfabrication methods to create novel biologically inspired tissue engineered cartilage scaffolds to facilitate human bone marrow mesenchymal stem cell (MSC) chondrogenesis. To this end we utilized electrospinning to design and fabricate a series of novel 3D biomimetic nanostructured scaffolds based on hydrogen (H2) treated multi-walled carbon nanotubes (MWCNTs) and biocompatible poly(L-lactic acid) (PLLA) polymers. Specifically, a series of electrospun fibrous PLLA scaffolds with controlled fiber dimension were fabricated in this study. In vitro MSC studies showed that stem cells prefer to attach in the scaffolds with smaller fiber diameter. More importantly, the MWCNT embedded scaffolds showed a drastic increase in mechanical strength and a compressive Young’s modulus matching to natural cartilage. Furthermore, our MSC differentiation results demonstrated that incorporation of the H2 treated carbon nanotubes and poly-L-lysine coating can induce more chondrogenic differentiations of MSCs than controls. After two weeks of culture, PLLA scaffolds with H2 treated MWCNTs and poly-L-lysine can achieve the highest glycosaminoglycan synthesis, making them promising for further exploration for cartilage regeneration.

Carbon nanotube-coating accelerated cell adhesion and proliferation on poly (L-lactide)

NASA Astrophysics Data System (ADS)

Hirata, Eri; Akasaka, Tsukasa; Uo, Motohiro; Takita, Hiroko; Watari, Fumio; Yokoyama, Atsuro

2012-12-01

The surface of a polylactic acid (PLLA) was coated multiwalled carbon nanotubes (MWCNTs) in order to improve the surface properties. In addition, its surface characteristics and cell culturing properties were examined. Whole surface of PLLA was homogeneously covered by MWCNTs maintained a unique tubular structure. MWCNT-coated PLLA showed remarkable higher wettability than uncoated PLLA. Human osteosarcoma cell line (Saos2) adhered well on the CNT-coated PLLA whereas there are few cells attached on the uncoated PLLA at 2 h after seeding. The number of the cells on uncoated PLLA was still smaller than on the MWCNT-coated PLLA at 1 and 3 days. Moreover, The DNA content in the cells attached to the MWCNT-coated PLLA was significantly higher than that on the uncoated PLLA (p < 0.05) at 1 and 3 days. There was no significant difference between the scaffolds for ALP activity normalized by DNA content at both term (p > 0.1). Therefore MWCNT-coating on PLLA improved the surface wettability and initial cell attachment at early stage.

A switchable positive and negative air pressure device for efficient and gentle handling of nanofiber scaffolds

NASA Astrophysics Data System (ADS)

Hotaling, Nathan A.; Khristov, Vladimir; Maminishkis, Arvydas; Bharti, Kapil; Simon, Carl G.

2017-10-01

A scaffold handling device (SHD) has been designed that can switch from gentle suction to positive pressure to lift and place nanofiber scaffolds. In tissue engineering laboratories, delicate fibrous scaffolds, such as electrospun nanofiber scaffolds, are often used as substrates for cell culture. Typical scaffold handling procedures include lifting the scaffolds, moving them from one container to another, sterilization, and loading scaffolds into cell culture plates. Using tweezers to handle the scaffolds can be slow, can damage the scaffolds, and can cause them to wrinkle or fold. Scaffolds may also acquire a static charge which makes them difficult to put down as they cling to tweezers. An SHD has been designed that enables more efficient, gentle lifting, and placement of delicate scaffolds. Most of the parts to make the SHD can be purchased, except for the tip which can be 3D-printed. The SHD enables more reliable handling of nanofiber scaffolds that may improve the consistency of biomanufacturing processes.

Preparation and characterization of electrospun in-situ cross-linked gelatin-graphite oxide nanofibers.

PubMed

Zhan, Jianchao; Morsi, Yosry; Ei-Hamshary, Hany; Al-Deyab, Salem S; Mo, Xiumei

2016-01-01

Electrospun gelatin(Gel) nanofibers scaffold has such defects as poor mechanical property and quick degradation due to high solubility. In this study, the in situ cross-linked electrospinning technique was used for the production of gelatin nanofibers. Deionized water was chosen as the spinning solvent and graphite oxide (GO) was chosen as the enhancer. The morphological structure, porosity, thermal property, moisture absorption, and moisture retention performance, hydrolysis resistance, mechanical property, and biocompatibility of the produced nanofibers were investigated. Compared with in situ cross-linked gelatin nanofibers scaffold, in situ cross-linked Gel-GO nanofibers scaffold has the following features: (1) the hydrophilicity, moisture absorption, and moisture retention performance slightly reduce, while the hydrolysis resistance is improved; (2) the breaking strength, breaking elongation, and Young's modulus are significantly improved; (3) the porosity slightly reduces while the biocompatibility considerably increases. The in situ cross-linked Gel-GO nanofibers scaffold is likely to be applied in such fields as drug delivery and scaffold for skin tissue engineering.

Biomineralized poly (l-lactic-co-glycolic acid)-tussah silk fibroin nanofiber fabric with hierarchical architecture as a scaffold for bone tissue engineering.

PubMed

Gao, Yanfei; Shao, Weili; Qian, Wang; He, Jianxin; Zhou, Yuman; Qi, Kun; Wang, Lidan; Cui, Shizhong; Wang, Rui

2018-03-01

In bone tissue engineering, the fabrication of a scaffold with a hierarchical architecture, excellent mechanical properties, and good biocompatibility remains a challenge. Here, a solution of polylactic acid (PLA) and Tussah silk fibroin (TSF) was electrospun into nanofiber yarns and woven into multilayer fabrics. Then, composite scaffolds were obtained by mineralization in simulated body fluid (SBF) using the multilayer fabrics as a template. The structure and related properties of the composite scaffolds were characterized using different techniques. PLA/TSF (mass ratio, 9:1) nanofiber yarns with uniform diameters of 72±9μm were obtained by conjugated electrospinning; the presence of 10wt% TSF accelerated the nucleation and growth of hydroxyapatite on the surface of the composite scaffolds in SBF. Furthermore, the compressive mechanical properties of the PLA/TSF multilayer nanofiber fabrics were improved after mineralization; the compressive modulus and stress of the mineralized composite scaffolds were 32.8 and 3.0 times higher than that of the composite scaffolds without mineralization, respectively. Interestingly, these values were higher than those of scaffolds containing random nanofibers. Biological assay results showed that the mineralization and multilayer fabric structure of the composite nanofiber scaffolds significantly increased cell adhesion and proliferation and enhanced the mesenchymal stem cell differentiation toward osteoblasts. Our results indicated that the mineralized nanofiber scaffolds with multilayer fabrics possessed excellent cytocompatibility and good osteogenic activity, making them versatile biocompatible scaffolds for bone tissue engineering. Copyright © 2017 Elsevier B.V. All rights reserved.

Highlighting the Importance of Surface Grafting in Combination with a Layer-by-Layer Approach for Fabricating Advanced 3D Poly(l-lactide) Microsphere Scaffolds

PubMed Central

2016-01-01

A combined surface treatment (i.e., surface grafting and a layer-by-layer (LbL) approach) is presented to create advanced biomaterials, i.e., 3D poly(l-lactide) (PLLA) microsphere scaffolds, at room temperature. The grafted surface plays a crucial role in assembling polyelectrolyte multilayers (PEMs) onto the surface of the microspheres, thus improving the physicochemical properties of the 3D microsphere scaffolds. The grafted surface of the PLLA microspheres demonstrates much better PEM adsorption, improved surface coverage at low pH, and smoother surfaces at high pH compared with those of nongrafted surfaces of PLLA microspheres during the assembly of PEMs. They induce more swelling than nongrafted surfaces after the assembly of the PEMs and exhibit blue emission after functionalization of the microsphere surface with a fluorescent dye molecule. The 3D scaffolds functionalized with and without nanosheets not only exhibit good mechanical performance similar to the compressive modulus of cancellous bone but also exhibit the porosity required for cancellous bone regeneration. The magnetic nanoparticle-functionalized 3D scaffolds result in an electrical conductivity in the high range of semiconducting materials (i.e., 1–250 S cm–1). Thus, these 3D microsphere scaffolds fabricated by surface grafting and the LbL approach are promising candidates for bone tissue engineering. PMID:29503506

The regulation of focal adhesion complex formation and salivary gland epithelial cell organization by nanofibrous PLGA scaffolds

PubMed Central

Sequeira, Sharon J.; Soscia, David A.; Oztan, Basak; Mosier, Aaron P.; Jean-Gilles, Riffard; Gadre, Anand; Cady, Nathaniel C.; Yener, Bülent; Castracane, James; Larsen, Melinda

2012-01-01

Nanofiber scaffolds have been useful for engineering tissues derived from mesenchymal cells, but few studies have investigated their applicability for epithelial cell-derived tissues. In this study, we generated nanofiber (250 nm) or microfiber (1200 nm) scaffolds via electrospinning from the polymer, poly-L-lactic-co-glycolic acid (PLGA). Cell-scaffold contacts were visualized using fluorescent immunocytochemistry and laser scanning confocal microscopy. Focal adhesion (FA) proteins, such as phosphorylated FAK (Tyr397), paxillin (Tyr118), talin and vinculin were localized to FA complexes in adult cells grown on planar surfaces but were reduced and diffusely localized in cells grown on nanofiber surfaces, similar to the pattern observed in adult mouse salivary gland tissues. Significant differences in epithelial cell morphology and cell clustering were also observed and quantified, using image segmentation and computational cell-graph analyses. No statistically significant differences in scaffold stiffness between planar PLGA film controls compared to nanofibers scaffolds were detected using nanoindentation with atomic force microscopy, indicating that scaffold topography rather than mechanical properties accounts for changes in cell attachments and cell structure. Finally, PLGA nanofiber scaffolds could support the spontaneous self-organization and branching of dissociated embryonic salivary gland cells. Nanofiber scaffolds may therefore have applicability in the future for engineering an artificial salivary gland. PMID:22285464

Bone regeneration by means of a three-dimensional printed scaffold in a rat cranial defect.

PubMed

Kwon, Doo Yeon; Park, Ji Hoon; Jang, So Hee; Park, Joon Yeong; Jang, Ju Woong; Min, Byoung Hyun; Kim, Wan-Doo; Lee, Hai Bang; Lee, Junhee; Kim, Moon Suk

2018-02-01

Recently, computer-designed three-dimensional (3D) printing techniques have emerged as an active research area with almost unlimited possibilities. In this study, we used a computer-designed 3D scaffold to drive new bone formation in a bone defect. Poly-L-lactide (PLLA) and bioactive β-tricalcium phosphate (TCP) were simply mixed to prepare ink. PLLA + TCP showed good printability from the micronozzle and solidification within few seconds, indicating that it was indeed printable ink for layer-by-layer printing. In the images, TCP on the surface of (and/or inside) PLLA in the printed PLLA + TCP scaffold looked dispersed. MG-63 cells (human osteoblastoma) adhered to and proliferated well on the printed PLLA + TCP scaffold. To assess new bone formation in vivo, the printed PLLA + TCP scaffold was implanted into a full-thickness cranial bone defect in rats. The new bone formation was monitored by microcomputed tomography and histological analysis of the in vivo PLLA + TCP scaffold with or without MG-63 cells. The bone defect was gradually spontaneously replaced with new bone tissues when we used both bioactive TCP and MG-63 cells in the PLLA scaffold. Bone formation driven by the PLLA + TCP30 scaffold with MG-63 cells was significantly greater than that in other experimental groups. Furthermore, the PLLA + TCP scaffold gradually degraded and matched well the extent of the gradual new bone formation on microcomputed tomography. In conclusion, the printed PLLA + TCP scaffold effectively supports new bone formation in a cranial bone defect. Copyright © 2017 John Wiley & Sons, Ltd.

PCL and PCL-gelatin nanofibers as esophageal tissue scaffolds: optimization, characterization and cell-matrix interactions.

PubMed

Kuppan, Purushothaman; Sethuraman, Swaminathan; Krishnan, Uma Maheswari

2013-09-01

Nanofiber based scaffolds offer great promise in regeneration of various tissues including esophagus. Diseases of the esophagus such as malignancy and strictures require surgical intervention to repair the affected region using an appropriate substitute. Long gap esophageal atresia poses a clinical challenge to bridge the gap. In this study, nanofibrous scaffolds made of PCL and PCL-gelatin were fabricated through electrospinning. The average diameter of PCL and PCL-gelatin nanofibers were found to be 324 +/- 50 nm and 242 +/- 30 nm respectively. PCL-gelatin nanofibers was characterized using FTIR, DSC, UTM, Goniometer, suture retention strength and in vitro degradation and the results were compared with the PCL nanofibers. PCL nanofiber characterization results showed that it exhibited higher tensile strength, suture retention strength, contact angle and slower degradation when compared with the PCL-gelatin nanofibers. Further, the interaction of human esophageal epithelial cells with PCL and PCL-gelatin nanofibrous scaffold was determined by cell adhesion, proliferation and gene expression studies. Our results demonstrated that the epithelial cells adhered and proliferated well on both PCL and PCL-gelatin nanofibrous scaffolds and also exhibited the characteristic cobblestone morphology. Cell proliferation on PCL-gelatin nanofibrous scaffold was significantly higher than the PCL nanofibrous scaffold (*p <0.05). Therefore, these scaffolds could be explored as potential candidates for regeneration of functional esophagus.

Rational design of nanofiber scaffolds for orthopedic tissue repair and regeneration

PubMed Central

Ma, Bing; Xie, Jingwei; Jiang, Jiang; Shuler, Franklin D; Bartlett, David E

2013-01-01

This article reviews recent significant advances in the design of nanofiber scaffolds for orthopedic tissue repair and regeneration. It begins with a brief introduction on the limitations of current approaches for orthopedic tissue repair and regeneration. It then illustrates that rationally designed scaffolds made up of electrospun nanofibers could be a promising solution to overcome the problems that current approaches encounter. The article also discusses the intriguing properties of electrospun nanofibers, including control of composition, structures, orders, alignments and mechanical properties, use as carriers for topical drug and/or gene sustained delivery, and serving as substrates for the regulation of cell behaviors, which could benefit musculoskeletal tissue repair and regeneration. It further highlights a few of the many recent applications of electrospun nanofiber scaffolds in repairing and regenerating various orthopedic tissues. Finally, the article concludes with perspectives on the challenges and future directions for better design, fabrication and utilization of nanofiber scaffolds for orthopedic tissue engineering. PMID:23987110

A mesoporous silica composite scaffold: Cell behaviors, biomineralization and mechanical properties

NASA Astrophysics Data System (ADS)

Xu, Yong; Gao, Dan; Feng, Pei; Gao, Chengde; Peng, Shuping; Ma, HaoTian; Yang, Sheng; Shuai, Cijun

2017-11-01

Mesoporous structure is beneficial to cellular response due to the large specific surface area and high pore volume. In this study, mesoporous silica (SBA15) was incorporated into poly-L-lactic acid (PLLA) to construct composite scaffold by selective laser sintering. The results showed that SBA15 facilitated cells proliferation, which was mainly attributed to its unique intrinsic mesoporous structure and the released bioactive silicon. Moreover, the hydrolyzate of soluble mesoporous silica can adsorb ions to form nucleation sites that promote biomineralization, leading to improve biological activity of the composite scaffold. In addition, the compressive strength, compressive modulus and Vickers hardness of the scaffold were increased by 47.6%, 35.5% and 29.53% respectively with 1.5 wt.% SBA15. It was found that the particle enhancement of uniform distributed SBA15 accounted for the mechanic reinforcement of the composite scaffold. It indicated that the PLLA-SBA15 composite scaffold had potential applications in bone tissue engineering.

Double emulsion electrospun nanofibers as a growth factor delivery vehicle for salivary gland regeneration

NASA Astrophysics Data System (ADS)

Foraida, Zahraa I.; Sharikova, Anna; Peerzada, Lubna N.; Khmaladze, Alexander; Larsen, Melinda; Castracane, James

2017-08-01

Sustained delivery of growth factors, proteins, drugs and other biologically active molecules is necessary for tissue engineering applications. Electrospun fibers are attractive tissue engineering scaffolds as they partially mimic the topography of the extracellular matrix (ECM). However, they do not provide continuous nourishment to the tissue. In search of a biomimetic scaffold for salivary gland tissue regeneration, we previously developed a blend nanofiber scaffold composed of the protein elastin and the synthetic polymer polylactic-co-glycolic acid (PLGA). The nanofiber scaffold promoted in vivo-like salivary epithelial cell tissue organization and apicobasal polarization. However, in order to enhance the salivary cell proliferation and biomimetic character of the scaffold, sustained growth factor delivery is needed. The composite nanofiber scaffold was optimized to act as a growth factor delivery system using epidermal growth factor (EGF) as a model protein. The nanofiber/EGF hybrid nanofibers were synthesized by double emulsion electrospinning where EGF is emulsified within a water/oil/water (w/o/w) double emulsion system. Successful incorporation of EGF was confirmed using Raman spectroscopy. EGF release profile was characterized using enzyme-linked immunosorbent assay (ELIZA) of the EGF content. Double emulsion electrospinning resulted in slower release of EGF. We demonstrated the potential of the proposed double emulsion electrospun nanofiber scaffold for the delivery of growth factors and/or drugs for tissue engineering and pharmaceutical applications.

The effects of lactate and acid on articular chondrocytes function: Implications for polymeric cartilage scaffold design.

PubMed

Zhang, Xiaolei; Wu, Yan; Pan, Zongyou; Sun, Heng; Wang, Junjuan; Yu, Dongsheng; Zhu, Shouan; Dai, Jun; Chen, Yishan; Tian, Naifeng; Heng, Boon Chin; Coen, Noelle D; Xu, Huazi; Ouyang, Hongwei

2016-09-15

Poly (lactic-co-glycolic acid) (PLGA) and poly-l-lactate acid (PLLA) are biodegradable polymers widely utilized as scaffold materials for cartilage tissue engineering. Their acid degradation products have been widely recognized as being detrimental to cell function. However, the biological effects of lactate, rather than lactic acid, on chondrocytes have never been investigated. This is the major focus of this study. The amounts of lactate and the pH value (acid) of the PLGA and PLLA degradation medium were measured. The effects of PLGA and PLLA degradation medium, as well as different lactate concentrations and timing of exposure on chondrocytes proliferation and cartilage-specific matrix synthesis were investigated by various techniques including global gene expression profiling and gene knockdown experiments. It was shown that PLGA and PLLA degradation medium differentially regulated chondrocyte proliferation and matrix synthesis. Acidic pH caused by lactate inhibited chondrocyte proliferation and matrix synthesis. The effect of lactate on chondrocyte matrix synthesis was both time and dose dependent. A lactate concentration of 100mM and exposure duration of 8h significantly enhanced matrix synthesis. Lactate could also inhibit expression of cartilage matrix degradation genes in osteoarthritic chondrocytes, such as the major aggrecanase ADAMTS5, whilst promoting matrix synthesis simultaneously. Pulsed addition of lactate was shown to be more efficient in promoting COL2A1 expression. Global gene expression data and gene knock down experiments demonstrated that lactate promote matrix synthesis through up-regulation of HIF1A. These observed differential biological effects of lactate on chondrocytes would have implications for the future design of polymeric cartilage scaffolds. Lactic acid is a widely used substrate for polymers synthesis, PLGA and PLLA in particular. Although physical and biological modifications have been made on these polymers to make them be better cartilage scaffolds, little concern has been given on the biological effect of lactic acid, the main degradation product of these polymers, on chondrocytes. Our finding illustrates the differential biological function of lactate and acid on chondrocytes matrix synthesis. These results can facilitate future design of lactate polymers-based cartilage scaffolds. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Fabrication of novel high performance ductile poly(lactic acid) nanofiber scaffold coated with poly(vinyl alcohol) for tissue engineering applications.

PubMed

Abdal-Hay, Abdalla; Hussein, Kamal Hany; Casettari, Luca; Khalil, Khalil Abdelrazek; Hamdy, Abdel Salam

2016-03-01

Poly(lactic acid) (PLA) nanofiber scaffold has received increasing interest as a promising material for potential application in the field of regenerative medicine. However, the low hydrophilicity and poor ductility restrict its practical application. Integration of hydrophilic elastic polymer onto the surface of the nanofiber scaffold may help to overcome the drawbacks of PLA material. Herein, we successfully optimized the parameters for in situ deposition of poly(vinyl alcohol), (PVA) onto post-electrospun PLA nanofibers using a simple hydrothermal approach. Our results showed that the average fiber diameter of coated nanofiber mat is about 1265±222 nm, which is remarkably higher than its pristine counterpart (650±180 nm). The hydrophilicity of PLA nanofiber scaffold coated with a PVA thin layer improved dramatically (36.11±1.5°) compared to that of pristine PLA (119.7±1.5°) scaffold. The mechanical testing showed that the PLA nanofiber scaffold could be converted from rigid to ductile with enhanced tensile strength, due to maximizing the hydrogen bond interaction during the heat treatment and in the presence of PVA. Cytocompatibility performance of the pristine and coated PLA fibers with PVA was observed through an in vitro experiment based on cell attachment and the MTT assay by EA.hy926 human endothelial cells. The cytocompatibility results showed that human cells induced more favorable attachment and proliferation behavior on hydrophilic PLA composite scaffold than that of pristine PLA. Hence, PVA coating resulted in an increase in initial human cell attachment and proliferation. We believe that the novel PVA-coated PLA nanofiber scaffold developed in this study, could be a promising high performance biomaterial in regeneration medicine. Copyright © 2015. Published by Elsevier B.V.

Fabricating poly(1,8-octanediol citrate) elastomer based fibrous mats via electrospinning for soft tissue engineering scaffold.

PubMed

Zhu, Lei; Zhang, Yuanzheng; Ji, Yali

2017-06-01

Poly(1,8-octanediol citrate) (POC) is a recently developed biodegradable crosslinked elastomer that possesses good cytocompatibility and matchable mechanical properties to soft tissues. However, the thermosetting characteristic reveals a big challenge to manufacture its porous scaffold. Herein, POC elastomer was electrospun into fiber mat using poly(L-lactic acid) (PLLA) as a spinnable carrier. The obtained POC/PLLA fiber mats were characterized by scanning electron microscopy (SEM), dynamic mechanical analysis (DMA), uniaxial tensile test, static-water-contact-angle, thermal analysis, in vitro degradation and biocompatibility test. It was found that the fibrous structure could be formed so long as the POC pre-polymer's content was no more than 50 wt%. The presence of elastic POC component not only strengthened the fiber mats but also toughened the fiber mats. The hydrophilicity of 50/50 fiber mat significantly improved. In vitro degradation rate of POC based fiber mats was much faster than that of pure PLLA. Cyto- and histo-compatibility tests confirmed that the POC/PLLA fiber mats had good biocompatibility for potential applications in soft tissue engineering.

Development of Semicrystalline Morphology of Poly(L-lactic acid) During Processing of a Vascular Scaffold

NASA Astrophysics Data System (ADS)

Ailianou, Artemis

New and promising treatments for coronary heart disease are enabled by vascular scaffolds made of poly(L-lactic acid) (PLLA), as demonstrated by Abbott Vascular's bioresorbable vascular scaffold. PLLA is a semicrystalline polymer whose degree of crystallinity and crystalline microstructure depend on the thermal and deformation history during processing. In turn, the semicrystalline morphology determines scaffold strength and biodegradation time. However, spatially-resolved information about the resulting material structure (crystallinity and crystal orientation) is needed to interpret in vivo observations. The first manufacturing step of the scaffold is tube expansion in a process similar to injection blow molding. Spatial uniformity of the tube microstructure is essential for the consistent production and performance of the final scaffold. For implantation into the artery, solid-state deformation below the glass transition temperature is imposed on a laser-cut subassembly to crimp it into a small diameter. Regions of localized strain during crimping are implicated in deployment behavior. To examine the semicrystalline microstructure development of the scaffold, we employed complementary techniques of scanning electron and polarized light microscopy, wide-angle X-ray scattering, and X-ray microdiffraction. These techniques enabled us to assess the microstructure at the micro and nano length scale. The results show that the expanded tube is very uniform in the azimuthal and axial directions and that radial variations are more pronounced. The crimping step dramatically changes the microstructure of the subassembly by imposing extreme elongation and compression. Spatial information on the degree and direction of chain orientation from X-ray microdiffraction data gives insight into the mechanism by which the PLLA dissipates the stresses during crimping, without fracture. Finally, analysis of the microstructure after deployment shows that it is inherited from the crimping step and contributes to the scaffold's successful implantation in vivo.

Ozone Gas as a Benign Sterilization Treatment for PLGA Nanofiber Scaffolds

PubMed Central

de Jesus Andreoli Pinto, Terezinha; Bou-Chacra, Nadia Araci; Galante, Raquel; de Araújo, Gabriel Lima Barros; do Nascimento Pedrosa, Tatiana; Maria-Engler, Silvya Stuchi

2016-01-01

The use of electrospun nanofibers for tissue engineering and regenerative medicine applications is a growing trend as they provide improved support for cell proliferation and survival due, in part, to their morphology mimicking that of the extracellular matrix. Sterilization is a critical step in the fabrication process of implantable biomaterial scaffolds for clinical use, but many of the existing methods used to date can negatively affect scaffold properties and performance. Poly(lactic-co-glycolic acid) (PLGA) has been widely used as a biodegradable polymer for 3D scaffolds and can be significantly affected by current sterilization techniques. The aim of this study was to investigate pulsed ozone gas as an alternative method for sterilizing PLGA nanofibers. The morphology, mechanical properties, physicochemical properties, and response of cells to PLGA nanofiber scaffolds were assessed following different degrees of ozone gas sterilization. This treatment killed Geobacillus stearothermophilus spores, the most common biological indicator used for validation of sterilization processes. In addition, the method preserved all of the characteristics of nonsterilized PLGA nanofibers at all degrees of sterilization tested. These findings suggest that ozone gas can be applied as an alternative method for sterilizing electrospun PLGA nanofiber scaffolds without detrimental effects. PMID:26757850

Ozone Gas as a Benign Sterilization Treatment for PLGA Nanofiber Scaffolds.

PubMed

Rediguieri, Carolina Fracalossi; Pinto, Terezinha de Jesus Andreoli; Bou-Chacra, Nadia Araci; Galante, Raquel; de Araújo, Gabriel Lima Barros; Pedrosa, Tatiana do Nascimento; Maria-Engler, Silvya Stuchi; De Bank, Paul A

2016-04-01

The use of electrospun nanofibers for tissue engineering and regenerative medicine applications is a growing trend as they provide improved support for cell proliferation and survival due, in part, to their morphology mimicking that of the extracellular matrix. Sterilization is a critical step in the fabrication process of implantable biomaterial scaffolds for clinical use, but many of the existing methods used to date can negatively affect scaffold properties and performance. Poly(lactic-co-glycolic acid) (PLGA) has been widely used as a biodegradable polymer for 3D scaffolds and can be significantly affected by current sterilization techniques. The aim of this study was to investigate pulsed ozone gas as an alternative method for sterilizing PLGA nanofibers. The morphology, mechanical properties, physicochemical properties, and response of cells to PLGA nanofiber scaffolds were assessed following different degrees of ozone gas sterilization. This treatment killed Geobacillus stearothermophilus spores, the most common biological indicator used for validation of sterilization processes. In addition, the method preserved all of the characteristics of nonsterilized PLGA nanofibers at all degrees of sterilization tested. These findings suggest that ozone gas can be applied as an alternative method for sterilizing electrospun PLGA nanofiber scaffolds without detrimental effects.

Electrospun nanofibers for neural tissue engineering

NASA Astrophysics Data System (ADS)

Xie, Jingwei; MacEwan, Matthew R.; Schwartz, Andrea G.; Xia, Younan

2010-01-01

Biodegradable nanofibers produced by electrospinning represent a new class of promising scaffolds to support nerve regeneration. We begin with a brief discussion on the electrospinning of nanofibers and methods for controlling the structure, porosity, and alignment of the electrospun nanofibers. The methods include control of the nanoscale morphology and microscale alignment of the nanofibers, as well as the fabrication of macroscale, three-dimensional tubular structures. We then highlight recent studies that utilize electrospun nanofibers to manipulate biological processes relevant to nervous tissue regeneration, including stem cell differentiation, guidance of neurite extension, and peripheral nerve injury treatments. The main objective of this feature article is to provide valuable insights into methods for investigating the mechanisms of neurite growth on novel nanofibrous scaffolds and optimization of the nanofiber scaffolds and conduits for repairing peripheral nerve injuries.

Nanofibers and their applications in tissue engineering

PubMed Central

Vasita, Rajesh; Katti, Dhirendra S

2006-01-01

Developing scaffolds that mimic the architecture of tissue at the nanoscale is one of the major challenges in the field of tissue engineering. The development of nanofibers has greatly enhanced the scope for fabricating scaffolds that can potentially meet this challenge. Currently, there are three techniques available for the synthesis of nanofibers: electrospinning, self-assembly, and phase separation. Of these techniques, electrospinning is the most widely studied technique and has also demonstrated the most promising results in terms of tissue engineering applications. The availability of a wide range of natural and synthetic biomaterials has broadened the scope for development of nanofibrous scaffolds, especially using the electrospinning technique. The three dimensional synthetic biodegradable scaffolds designed using nanofibers serve as an excellent framework for cell adhesion, proliferation, and differentiation. Therefore, nanofibers, irrespective of their method of synthesis, have been used as scaffolds for musculoskeletal tissue engineering (including bone, cartilage, ligament, and skeletal muscle), skin tissue engineering, vascular tissue engineering, neural tissue engineering, and as carriers for the controlled delivery of drugs, proteins, and DNA. This review summarizes the currently available techniques for nanofiber synthesis and discusses the use of nanofibers in tissue engineering and drug delivery applications. PMID:17722259

Suppressing Mesenchymal Stem Cell Hypertrophy and Endochondral Ossification in 3D Cartilage Regeneration with Nanofibrous Poly(l-Lactic Acid) Scaffold and Matrilin-3.

PubMed

Liu, Qihai; Wang, Jun; Chen, Yupeng; Zhang, Zhanpeng; Saunders, Laura; Schipani, Ernestina; Chen, Qian; Ma, Peter X

2018-06-22

Articular cartilage has a very limited ability to self-heal after injury or degeneration due to its low cellularity, poor proliferative activity, and avascular nature. Current clinical options are able to alleviate patient suffering, but cannot sufficiently regenerate the lost tissue. Biomimetic scaffolds that recapitulate the important features of the extracellular matrix (ECM) of cartilage are hypothesized to be advantageous in supporting cell growth, chondrogenic differentiation, and integration of regenerated cartilage with native cartilage, ultimately restoring the injured tissue to its normal function. It's a challenge to support and maintain articular cartilage regenerated by bone marrow-derived mesenchymal stem cells (BMSCs), which are prone to hypertrophy and endochondral ossification after implanted in vivo. In the present work, a nanofibrous poly(l-lactic acid) (NF PLLA) scaffold developed by our group was utilized because of the desired highly porous structure, high interconnectivity, collagen-like NF architecture to support rabbit BMSCs for articular cartilage regeneration. We further hypothesized that Matrilin-3 (MATN3), a non-collagenous, cartilage-specific ECM protein, would enhance the microenvironment of the NF PLLA scaffold for cartilage regeneration and maintaining its property. To test this hypothesis, we seeded BMSCs on the NF PLLA scaffold with or without MATN3. We found that MATN3 suppresses hypertrophy in this 3D culture system in vitro. Subcutaneous implantation of the chondrogenic cell/scaffold constructs in a nude mouse model showed that pretreatment with MATN3 was able to maintain chondrogenesis and prevent hypertrophy and endochondral ossification in vivo. These results demonstrate that the porous NF PLLA scaffold treated with MATN3 represents an advantageous 3D microenvironment for cartilage regeneration and phenotype maintenance, and is a promising strategy for articular cartilage repair. Articular cartilage defects, caused by trauma, inflammation, or joint instability, may ultimately lead to debilitating pain and disability. Bone marrow-derived mesenchymal stem cells (BMSCs) are an attractive cell source for articular cartilage tissue engineering. However, chondrogenic induction of BMSCs is often accompanied by undesired hypertrophy, which can lead to calcification and ultimately damage the cartilage. Therefore, a therapy to prevent hypertrophy and endochondral ossification is of paramount importance to adequately regenerate articular cartilage. We hypothesized that MATN3 (a non-collagenous ECM protein expressed exclusively in cartilage) may improve regeneration of articular cartilage with BMSCs by maintaining chondrogenesis and preventing hypertrophic transition in an ECM mimicking nanofibrous scaffold. Our results showed that the administration of MATN3 to the cell/nanofibrous scaffold constructs favorably maintained chondrogenesis and prevented hypertrophy/endochondral ossification in the chondrogenic constructs in vitro and in vivo. The combination of nanofibrous PLLA scaffolds and MATN3 treatment provides a very promising strategy to generate chondrogenic grafts with phenotypic stability for articular cartilage repair. Copyright © 2018. Published by Elsevier Ltd.

Tyrosinase-Mediated Construction of a Silk Fibroin/Elastin Nanofiber Bioscaffold.

PubMed

Hong, Yanqing; Zhu, Xueke; Wang, Ping; Fu, Haitian; Deng, Chao; Cui, Li; Wang, Qiang; Fan, Xuerong

2016-04-01

Elastin has characteristics of elasticity, biological activity, and mechanical stability. In the present work, tyrosinase-mediated construction of a bioscaffold with silk fibroin and elastin was carried out, aiming at developing a novel medical biomaterial. The efficiency of enzymatic oxidation of silk fibroin and the covalent reaction between fibroin and elastin were examined by spectrophotometry, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and size exclusion chromatography (SEC). The properties of composite air-dried and nanofiber scaffolds were investigated. The results reveal that elastin was successfully bonded to silk fibroins, resulting in an increase in molecular weight of fibroin proteins. ATR-FTIR spectra indicated that tyrosinase treatment impacted the conformational structure of fibroin-based membrane. The thermal behaviors and mechanical properties of the tyrosinase-treated scaffolds were also improved compared with the untreated group. NIH/3T3 cells exhibited optimum densities when grown on the nanofiber scaffold, implying that the nanofiber scaffold has enhanced biocompatibility compared to the air-dried scaffold. A biological nanofiber scaffold constructed from tyrosinase-treated fibroin and elastin could potentially be utilized in biomedical applications.

Heterofunctional nanosheet controlling cell adhesion properties by collagen coating.

PubMed

Niwa, Daisuke; Fujie, Toshinori; Lang, Thorsten; Goda, Nobuhito; Takeoka, Shinji

2012-08-01

Recently, biomaterials have been widely used in a variety of medical applications. We previously reported that a poly-l-lactic acid (PLLA) nanosheet shows anti-adhesive properties and constitutes a useful biomaterial for preventing unwanted wound adhesion in surgical operations. In this article, we examine whether the PLLA nanosheet can be specifically modified with biomacromolecules on one surface only. Such an approach would endow each side of the nanosheet with discrete functions, that is anti-adhesive and pro-healing properties. We fabricated two distinct PLLA nanosheets: (i) collagen cast on the surface of a PLLA nanosheet (Col-Cast-PLLA) and (ii) collagen spin-coated on the nanosheet (Col-Spin-PLLA). In the Col-Spin-PLLA nanosheet, the collagen layer had a thickness of 5-10 nm on the PLLA surface and displayed increased hydrophilicity compared to both PLLA and Col-Cast-PLLA nanosheets. In addition, atomic force microscopy showed disorganized collagen fibril formation on the PLLA layer when covered using the spin-coating method, while apparent bundle formations of collagen were formed in the Col-Cast-PLLA nanosheet. The Col-Spin-PLLA nanosheet provided a microenvironment for cells to adhere and spread. By contrast, the Col-Cast-PLLA nanosheet displayed reduced cell adhesion compared to the Col-Spin-PLLA nanosheet. Consistent with these findings, immunocytochemical analysis clearly showed fine networks of actin filaments in cells cultured on the Col-Spin-PLLA, but not the Col-Cast-PLLA nanosheet. Therefore, the Col-Spin-PLLA nanosheet was shown to be more suitable for acting as a scaffold. In conclusion, we have succeeded in developing a heterofunctional nanosheet comprising a collagen modified side, which has the ability to rapidly adhere cells, and an unmodified side, which acts as an adhesion barrier, by using a spin-coating technique.

Hybrid microfabrication of nanofiber-based sheets and rods for tissue engineering applications.

PubMed

Park, Suk-Hee; Kim, Min Sung; Lee, Dasom; Choi, Yong Whan; Kim, Deok-Ho; Suh, Kahp-Yang

2013-12-01

Electrospun nanofibers have been developed into a variety of forms for tissue engineering scaffolds to regulate the cellular functions guided by nanotopographical cues. Here, we have successfully fabricated nanofiber-based scaffold complexes of rod and sheet type by combining the three microfabrication techniques of electrospinning, spin coating, and polymer melt deposition. It was demonstrated that this hybrid fabrication could produce uniaxially aligned nanofiber scaffolds supported by a thin film, allowing for a mechanically enforced substrate for cell culture as well as facile scaffold manipulation. The results of cell analysis indicated that nanofibers on spin-coated films could provide contact guidance effects on cells and retain them even after manipulation. As an application of the cell-laden nanofiber film, we built a rod-type structure by rolling up the film around a mechanically supporting core microfiber, which was incorporated by polymer melt deposition. A biocompatible and biodegradable polymer, polycaprolactone, was used throughout the processes and thus could be used as a directly implantable substitute in tissue regeneration.

Intravenous injections of soluble drag-reducing polymers reduce foreign body reaction to implants.

PubMed

Marascalco, Philip J; Blair, Harry C; Nieponice, Alejandro; Robinson, Lisa J; Kameneva, Marina V

2009-01-01

We tested whether soluble viscoelastic drag-reducing polymers (DRPs), which modify blood flow in the macro- and microcirculation, affect host response to implanted biomaterials and control biodegradation and tissue ingrowth processes. Porous poly(L-lactate) (PLLA) implants, which are naturally hydrolyzed by foreign body giant cells, were used to evaluate differences in host response. Intravenous DRPs, high-molecular weight poly(ethylene oxide) (PEO) or poly(mannose) (PMNN), were given biweekly at 0.3-0.4 nM in saline (equivalent volumes of saline in controls) to rats with subcutaneous PLLA implants. After 7 weeks, there was no difference in weight gain or behavior between control and DRP-injected groups. Implanted PLLA scaffolds in controls were almost totally degraded and replaced by giant cell granulomas. On the contrary, PEO- or PMNN-treated animals retained a significant part of the implanted scaffold (p < 0.0001 vs. controls). The foreign body reaction was markedly decreased, and there was an increase in well-oriented collagen deposition within the implanted scaffold area in the animals treated with DRPs. The DRP-mediated effects observed in this study potentially reflect alteration in inflammatory events in response to implanted bioengineered materials, and, thus, warrant further investigation.

Osteoinductive peptide-functionalized nanofibers with highly ordered structure as biomimetic scaffolds for bone tissue engineering.

PubMed

Gao, Xiang; Zhang, Xiaohong; Song, Jinlin; Xu, Xiao; Xu, Anxiu; Wang, Mengke; Xie, Bingwu; Huang, Enyi; Deng, Feng; Wei, Shicheng

2015-01-01

The construction of functional biomimetic scaffolds that recapitulate the topographical and biochemical features of bone tissue extracellular matrix is now of topical interest in bone tissue engineering. In this study, a novel surface-functionalized electrospun polycaprolactone (PCL) nanofiber scaffold with highly ordered structure was developed to simulate the critical features of native bone tissue via a single step of catechol chemistry. Specially, under slightly alkaline aqueous solution, polydopamine (pDA) was coated on the surface of aligned PCL nanofibers after electrospinning, followed by covalent immobilization of bone morphogenetic protein-7-derived peptides onto the pDA-coated nanofiber surface. Contact angle measurement, Raman spectroscopy, and X-ray photoelectron spectroscopy confirmed the presence of pDA and peptides on PCL nanofiber surface. Our results demonstrated that surface modification with osteoinductive peptides could improve cytocompatibility of nanofibers in terms of cell adhesion, spreading, and proliferation. Most importantly, Alizarin Red S staining, quantitative real-time polymerase chain reaction, immunostaining, and Western blot revealed that human mesenchymal stem cells cultured on aligned nanofibers with osteoinductive peptides exhibited enhanced osteogenic differentiation potential than cells on randomly oriented nanofibers. Furthermore, the aligned nanofibers with osteoinductive peptides could direct osteogenic differentiation of human mesenchymal stem cells even in the absence of osteoinducting factors, suggesting superior osteogenic efficacy of biomimetic design that combines the advantages of osteoinductive peptide signal and highly ordered nanofibers on cell fate decision. The presented peptide-decorated bone-mimic nanofiber scaffolds hold a promising potential in the context of bone tissue engineering.

Effect of nanofiber content on bone regeneration of silk fibroin/poly(ε-caprolactone) nano/microfibrous composite scaffolds

PubMed Central

Kim, Beom Su; Park, Ko Eun; Kim, Min Hee; You, Hyung Keun; Lee, Jun; Park, Won Ho

2015-01-01

The broad application of electrospun nanofibrous scaffolds in tissue engineering is limited by their small pore size, which has a negative influence on cell migration. This disadvantage could be significantly improved through the combination of nano- and microfibrous structure. To accomplish this, different nano/microfibrous scaffolds were produced by hybrid electrospinning, combining solution electrospinning with melt electrospinning, while varying the content of the nanofiber. The morphology of the silk fibroin (SF)/poly(ε-caprolactone) (PCL) nano/microfibrous composite scaffolds was investigated with field-emission scanning electron microscopy, while the mechanical and pore properties were assessed by measurement of tensile strength and mercury porosimetry. To assay cell proliferation, cell viability, and infiltration ability, human mesenchymal stem cells were seeded on the SF/PCL nano/microfibrous composite scaffolds. From in vivo tests, it was found that the bone-regenerating ability of SF/PCL nano/microfibrous composite scaffolds was closely associated with the nanofiber content in the composite scaffolds. In conclusion, this approach of controlling the nanofiber content in SF/PCL nano/microfibrous composite scaffolds could be useful in the design of novel scaffolds for tissue engineering. PMID:25624762

PLGA/nHA hybrid nanofiber scaffold as a nanocargo carrier of insulin for accelerating bone tissue regeneration

NASA Astrophysics Data System (ADS)

Haider, Adnan; Gupta, Kailash Chandra; Kang, Inn-Kyu

2014-06-01

The development of tissue engineering in the field of orthopedic surgery is booming. Two fields of research in particular have emerged: approaches for tailoring the surface properties of implantable materials with osteoinductive factors as well as evaluation of the response of osteogenic cells to these fabricated implanted materials (hybrid material). In the present study, we chemically grafted insulin onto the surface of hydroxyapatite nanorods (nHA). The insulin-grafted nHAs (nHA-I) were dispersed into poly(lactide-co-glycolide) (PLGA) polymer solution, which was electrospun to prepare PLGA/nHA-I composite nanofiber scaffolds. The morphology of the electrospun nanofiber scaffolds was assessed by field emission scanning electron microscopy (FESEM). After extensive characterization of the PLGA/nHA-I and PLGA/nHA composite nanofiber scaffolds by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction spectroscopy (XRD), X-ray photoelectron spectroscopy (XPS), energy-dispersive X-ray spectrometry (EDS), and transmission electron microscopy (TEM), the PLGA/nHA-I and PLGA/nHA (used as control) composite nanofiber scaffolds were subjected to cell studies. The results obtained from cell adhesion, alizarin red staining, and Von Kossa assay suggested that the PLGA/nHA-I composite nanofiber scaffold has enhanced osteoblastic cell growth, as more cells were proliferated and differentiated. The fact that insulin enhanced osteoblastic cell proliferation will open new possibilities for the development of artificial scaffolds for bone tissue regeneration.

Shish-kebab-structured poly(ε-caprolactone) nanofibers hierarchically decorated with chitosan-poly(ε-caprolactone) copolymers for bone tissue engineering.

PubMed

Jing, Xin; Mi, Hao-Yang; Wang, Xin-Chao; Peng, Xiang-Fang; Turng, Lih-Sheng

2015-04-01

In this work, scaffolds with a shish-kebab (SK) structure formed by poly(ε-caprolactone) (PCL) nanofibers and chitosan-PCL (CS-PCL) copolymers were prepared via electrospinning and subsequent crystallization for bone tissue engineering applications. The aim of this study was to introduce nanosized topography and the high biocompatibility of chitosan onto PCL nanofibers to enhance cell affinity to PCL scaffolds. CS-PCL copolymers with various ratios were synthesized, and then spontaneously crystallized as kebabs onto the electrospun PCL fibers, which acted as shishes. Scanning electron microscopy (SEM) results demonstrated that the copolymer with PCL to chitosan ratio of 8.8 could hierarchically decorate the PCL nanofibers and formed well-shaped kebabs on the PCL nanofiber surface. Water contact angle tests and biomimetic activity experiments revealed that the shish-kebab scaffolds with CS-PCL kebabs (PCL-SK(CS-PCL(8.8))) showed enhanced hydrophilicity and mineralization ability compared with smooth PCL and PCL-SK(PCL) shish-kebab scaffolds. Osteoblast-like MG63 cells cultured on the PCL-SK(CS-PCL(8.8)) scaffolds showed optimizing cell attachment, cell viability, and metabolic activity, demonstrating that this kind of scaffold has potential applications in bone tissue engineering.

Electrospun Nanofiber Scaffolds with Gradations in Fiber Organization

PubMed Central

Khandalavala, Karl; Jiang, Jiang; Shuler, Franklin D.; Xie, Jingwei

2015-01-01

The goal of this protocol is to report a simple method for generating nanofiber scaffolds with gradations in fiber organization and test their possible applications in controlling cell morphology/orientation. Nanofiber organization is controlled with a new fabrication apparatus that enables the gradual decrease of fiber organization in a scaffold. Changing the alignment of fibers is achieved through decreasing deposition time of random electrospun fibers on a uniaxially aligned fiber mat. By covering the collector with a moving barrier/mask, along the same axis as fiber deposition, the organizational structure is easily controlled. For tissue engineering purposes, adipose-derived stem cells can be seeded to these scaffolds. Stem cells undergo morphological changes as a result of their position on the varied organizational structure, and can potentially differentiate into different cell types depending on their locations. Additionally, the graded organization of fibers enhances the biomimicry of nanofiber scaffolds so they more closely resemble the natural orientations of collagen nanofibers at tendon-to-bone insertion site compared to traditional scaffolds. Through nanoencapsulation, the gradated fibers also afford the possibility to construct chemical gradients in fiber scaffolds, and thereby further strengthen their potential applications in fast screening of cell-materials interaction and interfacial tissue regeneration. This technique enables the production of continuous gradient scaffolds, but it also can potentially produce fibers in discrete steps by controlling the movement of the moving barrier/mask in a discrete fashion. PMID:25938562

Continuing differentiation of human mesenchymal stem cells and induced chondrogenic and osteogenic lineages in electrospun PLGA nanofiber scaffold

PubMed Central

Xin, Xuejun; Hussain, Mohammad; Mao, Jeremy J.

2010-01-01

Nanofibers have recently gained substantial interest for potential applications in tissue engineering. The objective of this study was to determine whether electrospun nanofibers accommodate the viability, growth, and differentiation of human mesenchymal stem cells (hMSCs) as well as their osteogenic (hMSC-Ob) and chondrogenic (hMSC-Ch) derivatives. Poly(D,L-lactide-co-glycolide) (PLGA) beads with a PLA:PGA ratio of 85:15 were electrospun into non-woven fibers with an average diameter of 760±210 nm. The average Young’s modulus of electrospun PLGA nanofibers was 42±26 kPa, per nanoindentation with atomic force microscopy (AFM). Human MSCs were seeded 1–4 weeks at a density of 2×106 cells/mL in PLGA nanofiber sheets. After 2 week culture on PLGA nanofiber scaffold, hMSCs remained as precursors upon immunoblotting with hKL12 antibody. SEM taken up to 7 days after cell seeding revealed that hMSCs, hMSC-Ob and hMSC-Ch apparently attached to PLGA nanofibers. The overwhelming majority of hMSCs was viable and proliferating in PLGA nanofiber scaffolds up to the tested 14 days, as assayed live/dead tests, DNA assay and BrdU. In a separate experiment, hMSCs seeded in PLGA nanofiber scaffolds were differentiated into chodrogenic and osteogenic cells. Histological assays revealed that hMSCs continuously differentiated into chondrogenic cells and osteogenic cells after 2 week incubation in PLGA nanofibers. Taken together, these data represent an original investigation of continuous differentiation of hMSCs into chondrogenic and osteogenic cells in PLGA nanofiber scaffold. Consistent with previous work, these findings also suggest that nanofibers may serve as accommodative milieu for not only hMSCs, but also as a 3D carrier vehicle for lineage specific cells. PMID:17010425

Spiral-structured, nanofibrous, 3D scaffolds for bone tissue engineering.

PubMed

Wang, Junping; Valmikinathan, Chandra M; Liu, Wei; Laurencin, Cato T; Yu, Xiaojun

2010-05-01

Polymeric nanofiber matrices have already been widely used in tissue engineering. However, the fabrication of nanofibers into complex three-dimensional (3D) structures is restricted due to current manufacturing techniques. To overcome this limitation, we have incorporated nanofibers onto spiral-structured 3D scaffolds made of poly (epsilon-caprolactone) (PCL). The spiral structure with open geometries, large surface areas, and porosity will be helpful for improving nutrient transport and cell penetration into the scaffolds, which are otherwise limited in conventional tissue-engineered scaffolds for large bone defects repair. To investigate the effect of structure and fiber coating on the performance of the scaffolds, three groups of scaffolds including cylindrical PCL scaffolds, spiral PCL scaffolds (without fiber coating), and spiral-structured fibrous PCL scaffolds (with fiber coating) have been prepared. The morphology, porosity, and mechanical properties of the scaffolds have been characterized. Furthermore, human osteoblast cells are seeded on these scaffolds, and the cell attachment, proliferation, differentiation, and mineralized matrix deposition on the scaffolds are evaluated. The results indicated that the spiral scaffolds possess porosities within the range of human trabecular bone and an appropriate pore structure for cell growth, and significantly lower compressive modulus and strength than cylindrical scaffolds. When compared with the cylindrical scaffolds, the spiral-structured scaffolds demonstrated enhanced cell proliferation, differentiation, and mineralization and allowed better cellular growth and penetration. The incorporation of nanofibers onto spiral scaffolds further enhanced cell attachment, proliferation, and differentiation. These studies suggest that spiral-structured nanofibrous scaffolds may serve as promising alternatives for bone tissue engineering applications. Copyright 2009 Wiley Periodicals, Inc.

Electrically Conductive Chitosan/Carbon Scaffolds for Cardiac Tissue Engineering

PubMed Central

2015-01-01

In this work, carbon nanofibers were used as doping material to develop a highly conductive chitosan-based composite. Scaffolds based on chitosan only and chitosan/carbon composites were prepared by precipitation. Carbon nanofibers were homogeneously dispersed throughout the chitosan matrix, and the composite scaffold was highly porous with fully interconnected pores. Chitosan/carbon scaffolds had an elastic modulus of 28.1 ± 3.3 KPa, similar to that measured for rat myocardium, and excellent electrical properties, with a conductivity of 0.25 ± 0.09 S/m. The scaffolds were seeded with neonatal rat heart cells and cultured for up to 14 days, without electrical stimulation. After 14 days of culture, the scaffold pores throughout the construct volume were filled with cells. The metabolic activity of cells in chitosan/carbon constructs was significantly higher as compared to cells in chitosan scaffolds. The incorporation of carbon nanofibers also led to increased expression of cardiac-specific genes involved in muscle contraction and electrical coupling. This study demonstrates that the incorporation of carbon nanofibers into porous chitosan scaffolds improved the properties of cardiac tissue constructs, presumably through enhanced transmission of electrical signals between the cells. PMID:24417502

Sustained-release of naproxen sodium from electrospun-aligned PLLA-PCL scaffolds.

PubMed

Lui, Yuan Siang; Lewis, Mark P; Loo, Say Chye Joachim

2017-04-01

Spontaneous tendon healing may result in reduced tissue functionality. In view of this, tissue engineering (TE) emerges as a promising approach in promoting proper tendon regeneration. However, unfavourable post-surgical adhesion formations restrict adequate tendon healing through the TE approach. Naproxen sodium (NPS), a non-steroidal anti-inflammatory drug (NSAID), has been demonstrated to prevent adhesions by inhibiting the inflammatory response. Therefore, in this study, various factors, such as polymer composition, i.e. different poly-l-lactic acid (PLLA):polycaprolactone (PCL) ratios, and percentage of water:hexafluoroisopropanol (HFIP; as co-solvent) ratios, were investigated to understand how these can influence the release of NPS from electrospun scaffolds. By adjusting the amount of water as the co-solvent, NPS could be released sustainably for as long as 2 weeks. Scaffold breaking strength was also enhanced with the addition of water as the co-solvent. This NPS-loaded scaffold showed no significant cytotoxicity, and L929 murine fibroblasts cultured on the scaffolds were able to proliferate and align along the fibre orientation. These scaffolds with desirable tendon TE characteristics would be promising candidates in achieving better tendon regeneration in vivo. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

Nanobiocomposite of poly(lactide-co-glycolide)/chitosan electrospun scaffold can promote proliferation and transdifferentiation of Schwann-like cells from human adipose-derived stem cells.

PubMed

Razavi, Shahnaz; Zarkesh-Esfahani, Hamid; Morshed, Mohammad; Vaezifar, Sedigheh; Karbasi, Saeed; Golozar, Mohammad Ali

2015-08-01

The transdifferentiation of human adipose-derived stem cells (ADSCs) into Schwann-like cells on biocomposite scaffolds may be a critical issue in nerve regeneration medicine. In this study, tissue-engineered scaffold with chitosan (CS) nanopowders and poly(lactide-co-glycolide) (PLGA) was investigated for its potential Schwann cells (SCs) transdifferentiation. The differentiation of human ADSCs into S-like cells was induced with different CS content and direction of nanofibers on PLGA/CS scaffolds. Cell morphology and proliferation of differentiated cells were investigated by scanning electron microscopy and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay respectively. For assessment efficiency of transdifferentiation, the expression of SC markers (glial fibrillary acidic protein and S100), and myelinogenic marker (myelin basic protein) was investigated in different nanochitosan content and direction of nanofibers scaffolds, using immunocytochemistry technique. The nanochitosan can significantly promote cell proliferation of differentiated cells (p < 0.05). The mean percentage of S-like cells on greater CS content nanofibers scaffold was significantly higher than others (p < 0.05). In addition, the align orientation of nanofibers in scaffolds guided the differentiation of ADSCs toward myelinating S-like cells on the constructs. Overall, we found that high CS content and aligned-orientation of nanofibers in biocomposite scaffold (70/30A) can promote differentiation and myelinogenic capacity of S-like cells induced from human ADSCs. © 2015 Wiley Periodicals, Inc.

Elastin-PLGA hybrid electrospun nanofiber scaffolds for salivary epithelial cell self-organization and polarization.

PubMed

Foraida, Zahraa I; Kamaldinov, Tim; Nelson, Deirdre A; Larsen, Melinda; Castracane, James

2017-10-15

Development of electrospun nanofibers that mimic the structural, mechanical and biochemical properties of natural extracellular matrices (ECMs) is a promising approach for tissue regeneration. Electrospun fibers of synthetic polymers partially mimic the topography of the ECM, however, their high stiffness, poor hydrophilicity and lack of in vivo-like biochemical cues is not optimal for epithelial cell self-organization and function. In search of a biomimetic scaffold for salivary gland tissue regeneration, we investigated the potential of elastin, an ECM protein, to generate elastin hybrid nanofibers that have favorable physical and biochemical properties for regeneration of the salivary glands. Elastin was introduced to our previously developed poly-lactic-co-glycolic acid (PLGA) nanofiber scaffolds by two methods, blend electrospinning (EP-blend) and covalent conjugation (EP-covalent). Both methods for elastin incorporation into the nanofibers improved the wettability of the scaffolds while only blend electrospinning of elastin-PLGA nanofibers and not surface conjugation of elastin to PLGA fibers, conferred increased elasticity to the nanofibers measured by Young's modulus. After two days, only the blend electrospun nanofiber scaffolds facilitated epithelial cell self-organization into cell clusters, assessed with nuclear area and nearest neighbor distance measurements, leading to the apicobasal polarization of salivary gland epithelial cells after six days, which is vital for cell function. This study suggests that elastin electrospun nanofiber scaffolds have potential application in regenerative therapies for salivary glands and other epithelial organs. Regenerating the salivary glands by mimicking the extracellular matrix (ECM) is a promising approach for long term treatment of salivary gland damage. Despite their topographic similarity to the ECM, electrospun fibers of synthetic polymers lack the biochemical complexity, elasticity and hydrophilicity of the ECM. Elastin is an ECM protein abundant in the salivary glands and responsible for tissue elasticity. Although it's widely used for tissue regeneration of other organs, little is known about its utility in regenerating the salivary tissue. This study describes the use of elastin to improve the elasticity, hydrophilicity and biochemical complexity of synthetic nanofibers and its potential in directing in vivo-like organization of epithelial salivary cells which helps the design of efficient salivary gland regeneration scaffolds. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

A multi-scaled hybrid orthopedic implant: bone ECM-shaped Sr-HA nanofibers on the microporous walls of a macroporous titanium scaffold.

PubMed

Han, Yong; Zhou, Jianhong; Zhang, Lan; Xu, Kewei

2011-07-08

We report here, for the first time, a novel multi-scaled hybrid orthopedic implant material consisting of a macroporous Ti scaffold, whose macropores' walls have a microporous titania layer which is fully covered with nanofibers of Sr-doped hydroxyapatite (Sr-HA). The microporous titania layer is formed on and within the Ti scaffold by micro-arc oxidation, which firmly binds to the Ti substrate and contains Ca2+, Sr2+ and PO4(3-) ions. It is then hydrothermally treated to form Sr-HA nanofibers. During the hydrothermal treatment, Sr-HA nanoprisms nucleate from Ca0.5Sr0.5TiO3 pre-formed on the TiO2 and grow in length to nanofibers at the expense of Ca2+, Sr2+ and PO4(3-) ions that migrate from the TiO2. These Sr-HA nanofibers construct a network structure similar to the hierarchical organization of bone extracellular matrix (ECM), and the resulting nanofibrous surface displays a firm adhesion to substrate, superhydrophilicity and apatite-inducing ability. The induced apatite prefers to nucleate on the basal-faceted surfaces of Sr-HA nanofibers. The nanofiber-walled scaffold has a great potential for load-bearing orthotopic use.

A multi-scaled hybrid orthopedic implant: bone ECM-shaped Sr-HA nanofibers on the microporous walls of a macroporous titanium scaffold

NASA Astrophysics Data System (ADS)

Han, Yong; Zhou, Jianhong; Zhang, Lan; Xu, Kewei

2011-07-01

We report here, for the first time, a novel multi-scaled hybrid orthopedic implant material consisting of a macroporous Ti scaffold, whose macropores' walls have a microporous titania layer which is fully covered with nanofibers of Sr-doped hydroxyapatite (Sr-HA). The microporous titania layer is formed on and within the Ti scaffold by micro-arc oxidation, which firmly binds to the Ti substrate and contains Ca2 + , Sr2 + and PO43 - ions. It is then hydrothermally treated to form Sr-HA nanofibers. During the hydrothermal treatment, Sr-HA nanoprisms nucleate from Ca0.5Sr0.5TiO3 pre-formed on the TiO2 and grow in length to nanofibers at the expense of Ca2 + , Sr2 + and PO43 - ions that migrate from the TiO2. These Sr-HA nanofibers construct a network structure similar to the hierarchical organization of bone extracellular matrix (ECM), and the resulting nanofibrous surface displays a firm adhesion to substrate, superhydrophilicity and apatite-inducing ability. The induced apatite prefers to nucleate on the basal-faceted surfaces of Sr-HA nanofibers. The nanofiber-walled scaffold has a great potential for load-bearing orthotopic use.

Electrospun Nanofiber Scaffolds and Their Hydrogel Composites for the Engineering and Regeneration of Soft Tissues.

PubMed

Manoukian, Ohan S; Matta, Rita; Letendre, Justin; Collins, Paige; Mazzocca, Augustus D; Kumbar, Sangamesh G

2017-01-01

Electrospinning has emerged as a simple, elegant, and scalable technique that can be used to fabricate polymeric nanofibers. Pure polymers as well as blends and composites of both natural and synthetic ones have been successfully electrospun into nanofiber matrices for many biomedical applications. Tissue-engineered medical implants, such as polymeric nanofiber scaffolds, are potential alternatives to autografts and allografts, which are short in supply and carry risks of disease transmission. These scaffolds have been used to engineer various soft tissues, including connective tissues, such as skin, ligament, and tendon, as well as nonconnective ones, such as vascular, muscle, and neural tissue. Electrospun nanofiber matrices show morphological similarities to the natural extracellular matrix (ECM), characterized by ultrafine continuous fibers, high surface-to-volume ratios, high porosities, and variable pore-size distributions. The physiochemical properties of nanofiber matrices can be controlled by manipulating electrospinning parameters so that they meet the requirements of a specific application.Nanostructured implants show improved biological performance over bulk materials in aspects of cellular infiltration and in vivo integration, taking advantage of unique quantum, physical, and atomic properties. Furthermore, the topographies of such scaffolds has been shown to dictate cellular attachment, migration, proliferation, and differentiation, which are critical in engineering complex functional tissues with improved biocompatibility and functional performance. This chapter discusses the use of the electrospinning technique in the fabrication of polymer nanofiber scaffolds utilized for the regeneration of soft tissues. Selected scaffolds will be seeded with human mesenchymal stem cells (hMSCs), imaged using scanning electron and confocal microscopy, and then evaluated for their mechanical properties as well as their abilities to promote cell adhesion, proliferation , migration, and differentiation.

Development and molecular characterization of polymeric micro-nanofibrous scaffold of a defined 3-D niche for in vitro chemosensitivity analysis against acute myeloid leukemia cells

PubMed Central

Nair, Maya S; Mony, Ullas; Menon, Deepthy; Koyakutty, Manzoor; Sidharthan, Neeraj; Pavithran, Keechilat; Nair, Shantikumar V; Menon, Krishnakumar N

2015-01-01

Standard in vitro drug testing employs 2-D tissue culture plate systems to test anti-leukemic drugs against cell adhesion-mediated drug-resistant leukemic cells that harbor in 3-D bone marrow microenvironments. This drawback necessitates the fabrication of 3-D scaffolds that have cell adhesion-mediated drug-resistant properties similar to in vivo niches. We therefore aimed at exploiting the known property of polyurethane (PU)/poly-l-lactic acid (PLLA) in forming a micro-nanofibrous structure to fabricate unique, not presented before, as far as we are aware, 3-D micro-nanofibrous scaffold composites using a thermally induced phase separation technique. Among the different combinations of PU/PLLA composites generated, the unique PU/PLLA 60:40 composite displayed micro-nanofibrous morphology similar to decellularized bone marrow with increased protein and fibronectin adsorption. Culturing of acute myeloid leukemia (AML) KG1a cells in FN-coated PU/PLLA 60:40 shows increased cell adhesion and cell adhesion-mediated drug resistance to the drugs cytarabine and daunorubicin without changing the original CD34+/CD38−/CD33− phenotype for 168 hours compared to fibronectin tissue culture plate systems. Molecularly, as seen in vivo, increased chemoresistance is associated with the upregulation of anti-apoptotic Bcl2 and the cell cycle regulatory protein p27Kip1 leading to cell growth arrest. Abrogation of Bcl2 activity by the Bcl2-specific inhibitor ABT 737 led to cell death in the presence of both cytarabine and daunorubicin, demonstrating that the cell adhesion-mediated drug resistance induced by Bcl2 and p27Kip1 in the scaffold was similar to that seen in vivo. These results thus show the utility of a platform technology, wherein drug testing can be performed before administering to patients without the necessity for stromal cells. PMID:26028971

The potential of nanofibers in tissue engineering and stem cell therapy.

PubMed

Gholizadeh-Ghaleh Aziz, Shiva; Gholizadeh-Ghaleh Aziz, Sara; Akbarzadeh, Abolfazl

2016-08-01

Electrospinning is a technique in which materials in solution are shaped into continuous nano- and micro-sized fibers. Combining stem cells with biomaterial scaffolds and nanofibers affords a favorable approach for bone tissue engineering, stem cell growth and transfer, ocular surface reconstruction, and treatment of congenital corneal diseases. This review seeks to describe the current examples of the use of scaffolds in stem cell therapy. Stem cells are classified as adult or embryonic stem (ES) cells, and the advantages and drawbacks of each group are detailed. The nanofibers and scaffolds are further classified in Tables I and II , which describe specific examples from the literature. Finally, the current applications of biomaterial scaffolds containing stem cells for tissue engineering applications are presented. Overall, this review seeks to give an overview of the biomaterials available for use in combination with stem cells, and the application of nanofibers in stem cell therapy.

Biomimetic and bioactive nanofibrous scaffolds from electrospun composite nanofibers

PubMed Central

Zhang, YZ; Su, B; Venugopal, J; Ramakrishna, S; Lim, CT

2007-01-01

Electrospinning is an enabling technology that can architecturally (in terms of geometry, morphology or topography) and biochemically fabricate engineered cellular scaffolds that mimic the native extracellular matrix (ECM). This is especially important and forms one of the essential paradigms in the area of tissue engineering. While biomimesis of the physical dimensions of native ECM’s major constituents (eg, collagen) is no longer a fabrication-related challenge in tissue engineering research, conveying bioactivity to electrospun nanofibrous structures will determine the efficiency of utilizing electrospun nanofibers for regenerating biologically functional tissues. This can certainly be achieved through developing composite nanofibers. This article gives a brief overview on the current development and application status of employing electrospun composite nanofibers for constructing biomimetic and bioactive tissue scaffolds. Considering that composites consist of at least two material components and phases, this review details three different configurations of nanofibrous composite structures by using hybridizing basic binary material systems as example. These are components blended composite nanofiber, core-shell structured composite nanofiber, and nanofibrous mingled structure. PMID:18203429

Layer-by-layer 3-dimensional nanofiber tissue scaffold with controlled gap by electrospinning

NASA Astrophysics Data System (ADS)

Lin, Sai-Jun; Xue, Ya-Ping; Chang, Guoqing; Han, Qiao-Ling; Chen, Li-Fang; Jia, Yan-Bo; Zheng, Yu-Guo

2018-02-01

The development of three-dimensional (3D) nanofiber structures by electrospinning has drawn considerable attention in the field of tissue scaffolds. However, the generation of two dimensional mats using the conventional method limits electrospinning, the electrical charging of polymer liquids, as a means of nanofiber fabrication. In this study, we established a facile method of fabrication of layer-by-layer 3D polycaprolactone (PCL) nanofiber structures by utilizing a booklet collector with controlled morphology. Meanwhile, we explore the application of the manufactured 3D architectures in the field of tissue scaffolds. The approximately 20 μm layer-to-layer distance enhanced the ability of cells to migrate freely into tissues and induce cells in an ordered arrangement.

microRNA regulatory mechanism by which PLLA aligned nanofibers influence PC12 cell differentiation

NASA Astrophysics Data System (ADS)

Yu, Yadong; Lü, Xiaoying; Ding, Fei

2015-08-01

Objective. Aligned nanofibers (AFs) are regarded as promising biomaterials in nerve tissue engineering. However, a full understanding of the biocompatibility of AFs at the molecular level is still challenging. Therefore, the present study focused on identifying the microRNA (miRNA)-mediated regulatory mechanism by which poly-L-lactic acid (PLLA) AFs influence PC12 cell differentiation. Approach. Firstly, the effects of PLLA random nanofibers (RFs)/AFs and PLLA films (control) on the biological responses of PC12 cells that are associated with neuronal differentiation were examined. Then, SOLiD sequencing and cDNA microarray were employed to profile the expressions of miRNAs and mRNAs. The target genes of the misregulated miRNAs were predicted and compared with the mRNA profile data. Functions of the matched target genes (the intersection between the predicted target genes and the experimentally-determined, misregulated genes) were analyzed. Main results. The results revealed that neurites spread in various directions in control and RF groups. In the AF group, most neurites extended in parallel with each other. The glucose consumption and lactic acid production in the RF and AF groups were higher than those in the control group. Compared with the control group, 42 and 94 miRNAs were significantly dysregulated in the RF and AF groups, respectively. By comparing the predicted target genes with the mRNA profile data, five and 87 matched target genes were found in the RF and AF groups, respectively. Three of the matched target genes in the AF group were found to be associated with neuronal differentiation, whereas none had this association in the RF group. The PLLA AFs induced the dysregulation of miRNAs that regulate many biological functions, including axonal guidance, lipid metabolism and long-term potentiation. In particular, two miRNA-matched target gene-biological function modules associated with neuronal differentiation were identified as follows: (1) miR-23b, miR-18a, miR-107 and miR-103 regulate the Rras2 and Nf1 gene and thereby, affect cytoskeleton regulation and MAPK pathway; (2) miR-92a, miR-339-5p, miR-25, miR-125a-5p, miR-351 and miR-19b co-regulate the Pafah1b1 gene, affecting PC12 cell migration and differentiation. Significance. This work demonstrates a bioinformatic approach to accomplish miRNA-mRNA profile integrative analysis and provides more insights for understanding the regulatory mechanism of miRNA in AFs affecting neuronal differentiation. These findings will be greatly beneficial for the application and design of AFs in nerve tissue engineering.

Multilayer cell-seeded polymer nanofiber constructs for soft-tissue reconstruction.

PubMed

Barker, Daniel A; Bowers, Daniel T; Hughley, Brian; Chance, Elizabeth W; Klembczyk, Kevin J; Brayman, Kenneth L; Park, Stephen S; Botchwey, Edward A

2013-09-01

Cell seeding throughout the thickness of a nanofiber construct allows for patient-specific implant alternatives with long-lasting effects, earlier integration, and reduced inflammation when compared with traditional implants. Cell seeding may improve implant integration with host tissue; however, the effect of cell seeding on thick nanofiber constructs has not been studied. To use a novel cell-preseeded nanofiber tissue engineering technique to create a 3-dimensional biocompatible implant alternative to decellularized extracellular matrix. Animal study with mammalian cell culture to study tissue engineered scaffolds. Academic research laboratory. Thirty-six Sprague-Dawley rats. The rats each received 4 implant types. The grafts included rat primary (enhanced green fluorescent protein-positive [eGFP+]) fibroblast-seeded polycaprolactone (PCL)/collagen nanofiber scaffold, PCL/collagen cell-free nanofiber scaffold, acellular human cadaveric dermis (AlloDerm), and acellular porcine dermis (ENDURAGen). Rats were monitored postoperatively and received enrofloxacin in the drinking water for 4 days prophylactically and buprenorphine (0.2-0.5 mg/kg administered subcutaneously twice a day postoperatively for pain for 48 hours). The viability of NIH/3T3 fibroblasts cultured on PCL electrospun nanofibers was evaluated using fluorescence microscopy. Soft-tissue remodeling was examined histologically and with novel ex vivo volume determinations of implants using micro-computed tomography of cell-seeded implants relative to nanofibers without cells and commonly used dermal grafts of porcine and human origin (ENDURAGen and AlloDerm, respectively). The fate and distribution of eGFP+ seeded donor fibroblasts were assessed using immunohistochemistry. Fibroblasts migrated across nanofiber layers within 12 hours and remained viable on a single layer for up to 14 days. Scanning electron microscopy confirmed a nanoscale structure with a mean (SD) diameter of 158 (72) nm. Low extrusion rates demonstrated the excellent biocompatibility in vivo. Histological examination of the scaffolds demonstrated minimal inflammation. Cell seeding encouraged rapid vascularization of the nanofiber implants. Cells of donor origin (eGFP+) declined with the duration of implantation. Implant volume was not significantly affected for up to 8 weeks by the preseeding of cells (P > .05). Polymer nanofiber-based scaffolds mimic natural extracellular matrix. Preseeding the nanofiber construct with cells improved vascularization without notable effects on volume. An effect of cell preseeding on scaffold vascularization was evident beyond the presence of preseeded cells. This 3-dimensional, multilayer method of cell seeding throughout a 1-mm-thick construct is simple and feasible for clinical application. Further development of this technique may affect the clinical practice of facial plastic and reconstructive surgeons.

Electrospun Gelatin/β-TCP Composite Nanofibers Enhance Osteogenic Differentiation of BMSCs and In Vivo Bone Formation by Activating Ca (2+) -Sensing Receptor Signaling.

PubMed

Zhang, Xuehui; Meng, Song; Huang, Ying; Xu, Mingming; He, Ying; Lin, Hong; Han, Jianmin; Chai, Yuan; Wei, Yan; Deng, Xuliang

2015-01-01

Calcium phosphate- (CaP-) based composite scaffolds have been used extensively for the bone regeneration in bone tissue engineering. Previously, we developed a biomimetic composite nanofibrous membrane of gelatin/β-tricalcium phosphate (TCP) and confirmed their biological activity in vitro and bone regeneration in vivo. However, how these composite nanofibers promote the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) is unknown. Here, gelatin/β-TCP composite nanofibers were fabricated by incorporating 20 wt% β-TCP nanoparticles into electrospun gelatin nanofibers. Electron microscopy showed that the composite β-TCP nanofibers had a nonwoven structure with a porous network and a rough surface. Spectral analyses confirmed the presence and chemical stability of the β-TCP and gelatin components. Compared with pure gelatin nanofibers, gelatin/β-TCP composite nanofibers caused increased cell attachment, proliferation, alkaline phosphatase activity, and osteogenic gene expression in rat BMSCs. Interestingly, the expression level of the calcium-sensing receptor (CaSR) was significantly higher on the composite nanofibrous scaffolds than on pure gelatin. For rat calvarial critical sized defects, more extensive osteogenesis and neovascularization occurred in the composite scaffolds group compared with the gelatin group. Thus, gelatin/β-TCP composite scaffolds promote osteogenic differentiation of BMSCs in vitro and bone regeneration in vivo by activating Ca(2+)-sensing receptor signaling.

Nanofiber alignment of a small diameter elastic electrospun scaffold

NASA Astrophysics Data System (ADS)

Patel, Jignesh

Cardiovascular disease is the leading cause of death in western countries with coronary heart disease making up 50% of these deaths. As a treatment option, tissue engineered grafts have great potential. Elastic scaffolds that mimic arterial extracellular matrix (ECM) may hold the key to creating viable vascular grafts. Electrospinning is a widely used scaffold fabrication technique to engineer tubular scaffolds. In this study, we investigated how the collector rotation speed altered the nanofiber alignment which may improve mechanical characteristics making the scaffold more suitable for arterial grafts. The scaffold was fabricated from a blend of PCL/Elastin. 2D Fast Fourier Transform (FFT) image processing tool and MatLab were used to quantitatively analyze nanofiber orientation at different collector speeds (13500 to 15500 rpm). Both Image J and MatLab showed graphical peaks indicating predominant fiber orientation angles. A collector speed of 15000 rpm was found to produce the best nanofiber alignment with narrow peaks at 90 and 270 degrees, and a relative amplitude of 200. This indicates a narrow distribution of circumferentially aligned nanofibers. Collector speeds below and above 15000 rpm caused a decrease in fiber alignment with a broader orientation distribution. Uniformity of fiber diameter was also measured. Of 600 measures from the 15000 rpm scaffolds, the fiber diameter range from 500 nm to 899 nm was most prevalent. This diameter range was slightly larger than native ECM which ranges from 50 nm to 500 nm. The second most prevalent diameter range had an average of 404 nm which is within the diameter range of collagen. This study concluded that with proper electrospinning technique and collector speed, it is possible to fabricate highly aligned small diameter elastic scaffolds. Image J 2D FFT results confirmed MatLab findings for the analyses of circumferentially aligned nanofibers. In addition, MatLab analyses simplified the FFT orientation data providing an accurate, user friendly orientation measurement tool.

Scanning probe recognition microscopy investigation of tissue scaffold properties

PubMed Central

Fan, Yuan; Chen, Qian; Ayres, Virginia M; Baczewski, Andrew D; Udpa, Lalita; Kumar, Shiva

2007-01-01

Scanning probe recognition microscopy is a new scanning probe microscopy technique which enables selective scanning along individual nanofibers within a tissue scaffold. Statistically significant data for multiple properties can be collected by repetitively fine-scanning an identical region of interest. The results of a scanning probe recognition microscopy investigation of the surface roughness and elasticity of a series of tissue scaffolds are presented. Deconvolution and statistical methods were developed and used for data accuracy along curved nanofiber surfaces. Nanofiber features were also independently analyzed using transmission electron microscopy, with results that supported the scanning probe recognition microscopy-based analysis. PMID:18203431

Scanning probe recognition microscopy investigation of tissue scaffold properties.

PubMed

Fan, Yuan; Chen, Qian; Ayres, Virginia M; Baczewski, Andrew D; Udpa, Lalita; Kumar, Shiva

2007-01-01

Scanning probe recognition microscopy is a new scanning probe microscopy technique which enables selective scanning along individual nanofibers within a tissue scaffold. Statistically significant data for multiple properties can be collected by repetitively fine-scanning an identical region of interest. The results of a scanning probe recognition microscopy investigation of the surface roughness and elasticity of a series of tissue scaffolds are presented. Deconvolution and statistical methods were developed and used for data accuracy along curved nanofiber surfaces. Nanofiber features were also independently analyzed using transmission electron microscopy, with results that supported the scanning probe recognition microscopy-based analysis.

Fabrication of conductive polymer-based nanofiber scaffolds for tissue engineering applications.

PubMed

Gu, Bon Kang; Kim, Min Sup; Kang, Chang Mo; Kim, Jong-Ll; Park, Sang Jun; Kim, Chun-Ho

2014-10-01

Natural and synthetic polymers, in particular those that are conductive, are of great interest in the field of tissue engineering and the pursuit of biomimetic extracellular matrix (ECM) structures for adhesion, proliferation, and differentiation of cells. In the present study, natural chitin and conductive polyaniline (PANi) blended solutions were electrospun to produce biodegradable and conductive biomimetic nanostructured scaffolds. The chitin/PANi (Chi-PANi) nanofibrous materials were characterized using field emission scanning electron microscopy, Fourier transform-infrared spectroscopy, wettability analysis, mechanical testing, and electrical conductivity measurements using a 4-point probe method. The calculated electrical conductivities of the PANi-containing nanofiber scaffolds significantly increased as the amount of PANi increased, reaching 5.21 ± 0.28 x 10(-3) S/cm for 0.3 wt% content of the conducting polymer. In addition, the viability of human mesenchymal stem cells (hMSCs) cultured on the Chi-PANi nanofiber scaffolds in vitro was found to be excellent. These results suggest that the Chi-PANi nanofiber scaffolds have great potential for use in tissue engineering applications that involve electrical stimulation.

Designer Self-Assembling Peptide Nanofiber Scaffolds Containing Link Protein N-Terminal Peptide Induce Chondrogenesis of Rabbit Bone Marrow Stem Cells

PubMed Central

Wang, Baichuan; Sun, Caixia; Shao, Zengwu; Yang, Shuhua; Che, Biao; Wu, Qiang; Liu, Jianxiang

2014-01-01

Designer self-assembling peptide nanofiber hydrogel scaffolds have been considered as promising biomaterials for tissue engineering because of their excellent biocompatibility and biofunctionality. Our previous studies have shown that a novel designer functionalized self-assembling peptide nanofiber hydrogel scaffold (RLN/RADA16, LN-NS) containing N-terminal peptide sequence of link protein (link N) can promote nucleus pulposus cells (NPCs) adhesion and three-dimensional (3D) migration and stimulate biosynthesis of type II collagen and aggrecan by NPCs in vitro. The present study has extended these investigations to determine the effects of this functionalized LN-NS on bone marrow stem cells (BMSCs), a potential cell source for NP regeneration. Although the functionalized LN-NS cannot promote BMSCs proliferation, it significantly promotes BMSCs adhesion compared with that of the pure RADA16 hydrogel scaffold. Moreover, the functionalized LN-NS remarkably stimulates biosynthesis and deposition of type II collagen and aggrecan. These data demonstrate that the functionalized peptide nanofiber hydrogel scaffold containing link N peptide as a potential matrix substrate will be very useful in the NP tissue regeneration. PMID:25243141

Cell infiltration and growth in a low density, uncompressed three-dimensional electrospun nanofibrous scaffold.

PubMed

Blakeney, Bryan A; Tambralli, Ajay; Anderson, Joel M; Andukuri, Adinarayana; Lim, Dong-Jin; Dean, Derrick R; Jun, Ho-Wook

2011-02-01

A limiting factor of traditional electrospinning is that the electrospun scaffolds consist entirely of tightly packed nanofiber layers that only provide a superficial porous structure due to the sheet-like assembly process. This unavoidable characteristic hinders cell infiltration and growth throughout the nanofibrous scaffolds. Numerous strategies have been tried to overcome this challenge, including the incorporation of nanoparticles, using larger microfibers, or removing embedded salt or water-soluble fibers to increase porosity. However, these methods still produce sheet-like nanofibrous scaffolds, failing to create a porous three-dimensional scaffold with good structural integrity. Thus, we have developed a three-dimensional cotton ball-like electrospun scaffold that consists of an accumulation of nanofibers in a low density and uncompressed manner. Instead of a traditional flat-plate collector, a grounded spherical dish and an array of needle-like probes were used to create a Focused, Low density, Uncompressed nanoFiber (FLUF) mesh scaffold. Scanning electron microscopy showed that the cotton ball-like scaffold consisted of electrospun nanofibers with a similar diameter but larger pores and less-dense structure compared to the traditional electrospun scaffolds. In addition, laser confocal microscopy demonstrated an open porosity and loosely packed structure throughout the depth of the cotton ball-like scaffold, contrasting the superficially porous and tightly packed structure of the traditional electrospun scaffold. Cells seeded on the cotton ball-like scaffold infiltrated into the scaffold after 7 days of growth, compared to no penetrating growth for the traditional electrospun scaffold. Quantitative analysis showed approximately a 40% higher growth rate for cells on the cotton ball-like scaffold over a 7 day period, possibly due to the increased space for in-growth within the three-dimensional scaffolds. Overall, this method assembles a nanofibrous scaffold that is more advantageous for highly porous interconnectivity and demonstrates great potential for tackling current challenges of electrospun scaffolds. 2010 Elsevier Ltd. All rights reserved.

Four-year polymer biocompatibility and vascular healing profile of a novel ultrahigh molecular weight amorphous PLLA bioresorbable vascular scaffold: an OCT study in healthy porcine coronary arteries.

PubMed

Vahl, Torsten P; Gasior, Pawel; Gongora, Carlos A; Ramzipoor, Kamal; Lee, Chang; Cheng, Yanping; McGregor, Jenn; Shibuya, Masahiko; Estrada, Edward A; Conditt, Gerard B; Kaluza, Greg L; Granada, Juan F

2016-12-20

The vascular healing profile of polymers used in bioresorbable vascular scaffolds (BRS) has not been fully characterised in the absence of antiproliferative drugs. In this study, we aimed to compare the polymer biocompatibility profile and vascular healing response of a novel ultrahigh molecular weight amorphous PLLA BRS (FORTITUDE®; Amaranth Medical, Mountain View, CA, USA) against bare metal stent (BMS) controls in porcine coronary arteries. Following device implantation, optical coherence tomography (OCT) evaluation was performed at 0 and 28 days, and at one, two, three and four years. A second group of animals underwent histomorphometric evaluation at 28 and 90 days. At four years, both lumen (BRS 13.19±1.50 mm2 vs. BMS 7.69±2.41 mm2) and scaffold areas (BRS 15.62±1.95 mm2 vs. BMS 8.65±2.37 mm2) were significantly greater for BRS than BMS controls. The degree of neointimal proliferation was comparable between groups. Histology up to 90 days showed comparable healing and inflammation profiles for both devices. At four years, the novel PLLA BRS elicited a vascular healing response comparable to BMS in healthy pigs. Expansive vascular remodelling was evident only in the BRS group, a biological phenomenon that appears to be independent of the presence of antiproliferative drugs.

The use of chitosan/PLA nano-fibers by emulsion eletrospinning for periodontal tissue engineering.

PubMed

Shen, Renze; Xu, Weihong; Xue, Yanxiang; Chen, Luyuan; Ye, Haicheng; Zhong, Enyi; Ye, Zhanchao; Gao, Jie; Yan, Yurong

2018-04-16

In this study, nanofibrous scaffolds base on pure polylactic acid (PLA) and chitosan/PLA blends were fabricated by emulsion eletrospinning. By modulating their mechanical and biological properties, cell-compatible and biodegradable scaffolds were developed for periodontal bone regeneration. Pure PLA and different weight ratios of chitosan nano-particle/PLA nano-fibers were fabricated by emulsion eletrospinning. Scanning electron microscope (SEM) was performed to observe the morphology of nano-fibers. Mechanical properties of nano-fibers were tested by single fiber strength tester. Hydrophilic/hydrophobic nature of the nano-fibers was observed by stereomicroscope. In vitro degradation was also tested. Cells were seeded on nano-fibers scaffolds. Changes in cell adhesion, proliferation and osteogenic differentiation were tested by MTT assay and Alizarin Red S staining. Reverse transcription-polymerase chain reaction (RT-PCR) assay was used to evaluate the expression of (Toll-like receptor 4) TLR4, IL-6, IL-8, IL-1β, OPG, RUNX2 mRNA. It is shown that the mean diameter of nano-fibers is about 200 nm. The mean diameter of chitosan nano-particles is about 50 nm. The combination of chitosan nano-particles enhanced the mechanical properties of pure PLA nano-fibers. By adding a certain amount of chitosan nano-particles, it promoted cell adhesion. It also promoted the osteogenic differentiation of bone marrow stem cells (BMSCs) by elevating the expression of osteogenic marker genes such as BSP, Ocn, collagen I, and OPN and enhanced ECM mineralization. Nonetheless, it caused higher expression of inflammatory mediators and TLR4 of human periodontal ligament cells (hPDLCs). The combination of chitosan nano-particles enhanced the mechanical properties of pure PLA nano-fibers and increased its hydrophilicity. Pure PLA nano-fibers scaffold facilitated BMSCs proliferation. Adding an appropriate amount of chitosan nano-particles may promote its properties of cell proliferation and osteogenic differentiation. The higher expression of inflammatory mediators caused by nano-fibers may be regulated via TLR4 pathway.

Bladder tissue engineering using biocompatible nanofibrous electrospun constructs: feasibility and safety investigation.

PubMed

Shakhssalim, Nasser; Dehghan, Mohammad Mehdi; Moghadasali, Reza; Soltani, Mohammad Hossein; Shabani, Iman; Soleimani, Masoud

2012-01-01

To investigate the feasibility and safety of using biocompatible, nanofibrous electrospun polycaprolactone (PCL) and combination of polylactic acid (PLLA) and PCL mats in a canine model. Plasma-treated electrospun unseeded mats were implanted in three dogs. The first dog was sacrificed after 3 months and the second and third ones after 4 months, and then, the graft was examined macroscopically with subsequent morphological and histochemical evaluation. Both films showed high levels of cell infiltration and tissue formation, but body response to PLLA/PCL mat in comparison to PCL mat was very low. All three implantation models showed the same light microscopic morphology, immunohistochemistry, and scanning electron microscopy results; nevertheless, only the PCL/PLLA model showed favorable clinical results. Based on these data, nanofibrous PLLA/PCL scaffolding could be a suitable material for the bladder tissue engineering; however, it deserves further investigations.

Repair of osteochondral defects with hyaluronan- and polyester-based scaffolds.

PubMed

Solchaga, Luis A; Temenoff, Johnna S; Gao, Jizong; Mikos, Antonios G; Caplan, Arnold I; Goldberg, Victor M

2005-04-01

The natural repair of osteochondral defects can be enhanced with biocompatible, biodegradable materials that support the repair process. It is our hypothesis that hyaluronan-based scaffolds are superior to synthetic scaffolds because they provide biological cues. We tested this thesis by comparing two hyaluronan-based scaffolds [auto cross-linked polysaccharide polymer (ACP) and HYAFF-11] to polyester-based scaffolds [poly(DL-lactic-co-glycolic acid) (PLGA) and poly(L-lactic acid) (PLLA)] with similar pore size, porosity and degradation times. Fifty-four rabbits received bilateral osteochondral defects. One defect received a hyaluronan-based scaffold and the contralateral defect received the corresponding polyester-based scaffold. Rabbits were euthanized 4, 12 and 20 weeks after surgery and the condyles dissected and processed for histology. Only ACP-treated defects presented bone at the base of the defect at 4 weeks. At 12 weeks, only defects treated with rapidly dissolving implants (ACP and PLGA) presented bone reconstitution consistently, while bone was present in only one third of those treated with slowly dissolving scaffolds (HYAFF-11 and PLLA). After 20 weeks, the articular surface of PLGA-treated defects presented fibrillation more frequently than in ACP-treated defects. The surface of defects treated with slowly dissolving scaffolds presented more cracks and fissures. The degradation rate of the scaffolds is critical for the repair process. Slowly dissolving scaffolds sustain thicker cartilage at the surface but, it frequently presents cracks and discontinuities. These scaffolds also delay bone formation at the base of the defects. Hyaluronan-based scaffolds appear to allow faster cell infiltration leading to faster tissue formation. The degradation of ACP leads to rapid bone formation while the slow degradation of HYAFF-11 prolongs the presence of cartilage and delays endochondral bone formation.

Preparation, process optimization and characterization of core-shell polyurethane/chitosan nanofibers as a potential platform for bioactive scaffolds.

PubMed

Maleknia, Laleh; Dilamian, Mandana; Pilehrood, Mohammad Kazemi; Sadeghi-Aliabadi, Hojjat; Hekmati, Amir Houshang

2018-06-01

In this paper, polyurethane (PU), chitosan (Cs)/polyethylene oxide (PEO), and core-shell PU/Cs nanofibers were produced at the optimal processing conditions using electrospinning technique. Several methods including SEM, TEM, FTIR, XRD, DSC, TGA and image analysis were utilized to characterize these nanofibrous structures. SEM images exhibited that the core-shell PU/Cs nanofibers were spun without any structural imperfections at the optimized processing conditions. TEM image confirmed the PU/Cs core-shell nanofibers were formed apparently. It that seems the inclusion of Cs/PEO to the shell, did not induce the significant variations in the crystallinity in the core-shell nanofibers. DSC analysis showed that the inclusion of Cs/PEO led to the glass temperature of the composition increased significantly compared to those of neat PU nanofibers. The thermal degradation of core-shell PU/Cs was similar to PU nanofibers degradation due to the higher PU concentration compared to other components. It was hypothesized that the core-shell PU/Cs nanofibers can be used as a potential platform for the bioactive scaffolds in tissue engineering. Further biological tests should be conducted to evaluate this platform as a three dimensional scaffold with the capabilities of releasing the bioactive molecules in a sustained manner.

A novel gellan-PVA nanofibrous scaffold for skin tissue regeneration: Fabrication and characterization.

PubMed

Vashisth, Priya; Nikhil, Kumar; Roy, Partha; Pruthi, Parul A; Singh, Rajesh P; Pruthi, Vikas

2016-01-20

In this investigation, we have introduced novel electrospun gellan based nanofibers as a hydrophilic scaffolding material for skin tissue regeneration. These nanofibers were fabricated using a blend mixture of gellan with polyvinyl alcohol (PVA). PVA reduced the repulsive force of resulting solution and lead to formation of uniform fibers with improved nanostructure. Field emission scanning electron microscopy (FESEM) confirmed the average diameter of nanofibers down to 50 nm. The infrared spectra (IR), differential scanning calorimetry (DSC) and X-ray diffraction (XRD) analysis evaluated the crosslinking, thermal stability and highly crystalline nature of gellan-PVA nanofibers, respectively. Furthermore, the cell culture studies using human dermal fibroblast (3T3L1) cells established that these gellan based nanofibrous scaffold could induce improved cell adhesion and enhanced cell growth than conventionally proposed gellan based hydrogels and dry films. Importantly, the nanofibrous scaffold are biodegradable and could be potentially used as a temporary substrate/or biomedical graft to induce skin tissue regeneration. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enhanced bone formation in electrospun poly(L-lactic-co-glycolic acid)-tussah silk fibroin ultrafine nanofiber scaffolds incorporated with graphene oxide.

PubMed

Shao, Weili; He, Jianxin; Sang, Feng; Wang, Qian; Chen, Li; Cui, Shizhong; Ding, Bin

2016-05-01

To engineer bone tissue, it is necessary to provide a biocompatible, mechanically robust scaffold. In this study, we fabricated an ultrafine nanofiber scaffold by electrospinning a blend of poly(L-lactic-co-glycolic acid), tussah silk fibroin, and graphene oxide (GO) and characterized its morphology, biocompatibility, mechanical properties, and biological activity. The data indicate that incorporation of 10 wt.% tussah silk and 1 wt.% graphene oxide into poly(L-lactic-co-glycolic acid) nanofibers significantly decreased the fiber diameter from 280 to 130 nm. Furthermore, tussah silk and graphene oxide boosted the Young's modulus and tensile strength by nearly 4-fold and 3-fold, respectively, and significantly enhanced adhesion, proliferation in mouse mesenchymal stem cells and functionally promoted biomineralization-relevant alkaline phosphatase (ALP) and mineral deposition. The results indicate that composite nanofibers could be excellent and versatile scaffolds for bone tissue engineering. Copyright © 2016 Elsevier B.V. All rights reserved.

Electroactive BaTiO3 nanoparticle-functionalized fibrous scaffolds enhance osteogenic differentiation of mesenchymal stem cells

PubMed Central

Li, Yiping; Dai, Xiaohan; Bai, Yunyang; Liu, Yun; Wang, Yuehong; Liu, Ousheng; Yan, Fei; Tang, Zhangui; Zhang, Xuehui; Deng, Xuliang

2017-01-01

It has been proven that the surface topographic cues of fiber arrangement can induce osteogenic differentiation of mesenchymal stem cells. However, this effect alone is weak and insufficient to meet the needs of regenerative medicine. In this work, electroactivity concept was introduced to enhance the osteoinductivity of fibrous scaffolds. The randomly oriented and aligned electroactive fibrous scaffolds of poly-(l-lactic acid) (PLLA) with incorporation of ferroelectric ceramic BaTiO3 (BTO) nanoparticles (NPs) were fabricated by electrospinning. Physicochemical properties, including fiber morphology, microstructure, composition, thermal stability, surface roughness, and surface wettability, of these fibrous scaffolds were studied. The dielectric properties of the scaffolds were evaluated. The results showed that the randomly oriented BTO/PLLA composite fibrous scaffolds had the highest dielectric permittivity of 1.19, which is of the same order of magnitude as the natural bone. The combined effects of fiber orientation and electrical activity on the osteogenic responses of bone marrow mesenchymal stem cells (BM-MSCs) were specifically investigated. Randomly oriented composite fibrous scaffolds significantly promoted polygonal spreading and encouraged early osteogenic differentiation in BM-MSCs, whereas aligned composite fibrous scaffolds promoted cell elongation and discouraged osteogenic differentiation. These results evidenced that randomly fiber orientation and biomimetic electric activity have combining effects on osteogenic differentiation of BM-MSCs. Our findings indicate that coupling effects of multi-physical properties should be paid more attention to mimic the microenvironment for enhancing osteogenic differentiation of BM-MSCs. PMID:28603415

Injectable biomimetic liquid crystalline scaffolds enhance muscle stem cell transplantation

PubMed Central

Sleep, Eduard; McClendon, Mark T.; Preslar, Adam T.; Chen, Charlotte H.; Sangji, M. Hussain; Pérez, Charles M. Rubert; Haynes, Russell D.; Meade, Thomas J.; Blau, Helen M.; Stupp, Samuel I.

2017-01-01

Muscle stem cells are a potent cell population dedicated to efficacious skeletal muscle regeneration, but their therapeutic utility is currently limited by mode of delivery. We developed a cell delivery strategy based on a supramolecular liquid crystal formed by peptide amphiphiles (PAs) that encapsulates cells and growth factors within a muscle-like unidirectionally ordered environment of nanofibers. The stiffness of the PA scaffolds, dependent on amino acid sequence, was found to determine the macroscopic degree of cell alignment templated by the nanofibers in vitro. Furthermore, these PA scaffolds support myogenic progenitor cell survival and proliferation and they can be optimized to induce cell differentiation and maturation. We engineered an in vivo delivery system to assemble scaffolds by injection of a PA solution that enabled coalignment of scaffold nanofibers with endogenous myofibers. These scaffolds locally retained growth factors, displayed degradation rates matching the time course of muscle tissue regeneration, and markedly enhanced the engraftment of muscle stem cells in injured and noninjured muscles in mice. PMID:28874575

Highly porous 3D nanofiber scaffold using an electrospinning technique.

PubMed

Kim, Geunhyung; Kim, WanDoo

2007-04-01

A successful 3D tissue-engineering scaffold must have a highly porous structure and good mechanical stability. High porosity and optimally designed pore size provide structural space for cell accommodation and migration and enable the exchange of nutrients between the scaffold and environment. Poly(epsilon-carprolactone) fibers were electrospun using an auxiliary electrode and chemical blowing agent (BA), and characterized according to porosity, pore size, and their mechanical properties. We also investigated the effect of the BA on the electrospinning processability. The growth characteristic of human dermal fibroblasts cells cultured in the webs showed the good adhesion with the blown web relative to a normal electrospun mat. The blown nanofiber web had good tensile properties and high porosity compared to a typical electrospun nanofiber scaffold. (c) 2006 Wiley Periodicals, Inc.

Lycium barbarum polysaccharide encapsulated Poly lactic-co-glycolic acid Nanofibers: cost effective herbal medicine for potential application in peripheral nerve tissue engineering.

PubMed

Wang, Jing; Tian, Lingling; He, Liumin; Chen, Nuan; Ramakrishna, Seeram; So, Kwok-Fai; Mo, Xiumei

2018-06-06

Nerve regeneration is a serious clinical challenge following peripheral nerve injury. Lycium barbarum polysaccharide (LBP) is the major component of wolfberry extract, which has been shown to be neuroprotective and promising in nerve recovery in many studies. Electrospun nanofibers, especially core-shell structured nanofibers being capable of serving as both drug delivery system and tissue engineering scaffolds, are well known to be suitable scaffolds for regeneration of peripheral nerve applications. In this study, LBP was incorporated into core-shell structured nanofibrous scaffolds via coaxial electrospinning. Alamar blue assays were performed to investigate the proliferation of both PC12 and Schwann cells cultured on the scaffolds. The neuronal differentiation of PC12 cells was evaluated by NF200 expression with immunostaining and morphology changes observed by SEM. The results indicated that the released LBP dramatically enhanced both proliferation and neuronal differentiation of PC12 cells induced by NGF. Additionally, the promotion of Schwann cells myelination and neurite outgrowth of DRG neurons were also observed on LBP loaded scaffolds by LSCM with immunostaining. In summary, LBP, as a drug with neuroprotection, encapsulated into electrospun nanofibers could be a potential candidate as tissue engineered scaffold for peripheral nerve regeneration.

Porous starch/cellulose nanofibers composite prepared by salt leaching technique for tissue engineering.

PubMed

Nasri-Nasrabadi, Bijan; Mehrasa, Mohammad; Rafienia, Mohammad; Bonakdar, Shahin; Behzad, Tayebeh; Gavanji, Shahin

2014-08-08

Starch/cellulose nanofibers composites with proper porosity pore size, mechanical strength, and biodegradability for cartilage tissue engineering have been reported in this study. The porous thermoplastic starch-based composites were prepared by combining film casting, salt leaching, and freeze drying methods. The diameter of 70% nanofibers was in the range of 40-90 nm. All samples had interconnected porous morphology; however an increase in pore interconnectivity was observed when the sodium chloride ratio was increased in the salt leaching. Scaffolds with the total porogen content of 70 wt% exhibited adequate mechanical properties for cartilage tissue engineering applications. The water uptake ratio of nanocomposites was remarkably enhanced by adding 10% cellulose nanofibers. The scaffolds were partially destroyed due to low in vitro degradation rate after more than 20 weeks. Cultivation of isolated rabbit chondrocytes on the fabricated scaffold proved that the incorporation of nanofibers in starch structure improves cell attachment and proliferation. Copyright © 2014 Elsevier Ltd. All rights reserved.

Controlled drug delivery from composites of nanostructured porous silicon and poly(L-lactide).

PubMed

McInnes, Steven J P; Irani, Yazad; Williams, Keryn A; Voelcker, Nicolas H

2012-07-01

Porous silicon (pSi) and poly(L-lactide) (PLLA) both display good biocompatibility and tunable degradation behavior, suggesting that composites of both materials are suitable candidates as biomaterials for localized drug delivery into the human body. The combination of a pliable and soft polymeric material with a hard inorganic porous material of high drug loading capacity may engender improved control over degradation and drug release profiles and be beneficial for the preparation of advanced drug delivery devices and biodegradable implants or scaffolds. In this work, three different pSi and PLLA composite formats were prepared. The first format involved grafting PLLA from pSi films via surface-initiated ring-opening polymerization (pSi-PLLA [grafted]). The second format involved spin coating a PLLA solution onto oxidized pSi films (pSi-PLLA [spin-coated]) and the third format consisted of a melt-cast PLLA monolith containing dispersed pSi microparticles (pSi-PLLA [monoliths]). The surface characterization of these composites was performed via infrared spectroscopy, scanning electron microscopy, atomic force microscopy and water contact angle measurements. The composite materials were loaded with a model cytotoxic drug, camptothecin (CPT). Drug release from the composites was monitored via fluorimetry and the release profiles of CPT showed distinct characteristics for each of the composites studied. In some cases, controlled CPT release was observed for more than 5 days. The PLLA spin coat on pSi and the PLLA monolith containing pSi microparticles both released a CPT payload in accordance with the Higuchi and Ritger-Peppas release models. Composite materials were also brought into contact with human lens epithelial cells to determine the extent of cytotoxicity. We observed that all the CPT containing materials were highly efficient at releasing bioactive CPT, based on the cytotoxicity data.

Phage nanofibers induce vascularized osteogenesis in 3D printed bone scaffolds.

PubMed

Wang, Jianglin; Yang, Mingying; Zhu, Ye; Wang, Lin; Tomsia, Antoni P; Mao, Chuanbin

2014-08-06

A virus-activated matrix is developed to overcome the challenge of forming vascularized bone tissue. It is generated by filling a 3D printed bioceramic scaffold with phage nanofibers displaying high-density RGD peptide. After it is seeded with mesenchymal stem cells (MSCs) and implanted into a bone defect, the phage nanofibers induce osteogenesis and angiogenesis by activating endothelialization and osteogenic differentiation of MSCs. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Controlled release of antibiotics encapsulated in the electrospinning polylactide nanofibrous scaffold and their antibacterial and biocompatible properties

NASA Astrophysics Data System (ADS)

Wang, Shu-Dong; Zhang, Sheng-Zhong; Liu, Hua; Zhang, You-Zhu

2014-04-01

In this research, the drug loaded polylactide nanofibers are fabricated by electrospinning. Morphology, microstructure and mechanical properties are characterized. Properties and mechanism of the controlled release of the nanofibers are investigated. The results show that the drug loaded polylactide nanofibers do not show dispersed phase, and there is a good compatibility between polylactide and drugs. FTIR spectra show that drugs are encapsulated inside the polylactide nanofibers, and drugs do not break the structure of polylcatide. Flexibility of drug loaded polylactide scaffolds is higher than that of the pure polylactide nanofibers. Release rate of the drug loaded nanofibers is significantly slower than that of the drug powder. Release rate increases with the increase of the drugs’ concentration. The research mechanism suggests a typical diffusion-controlled release of the three loaded drugs. Antibacterial and cell culture show that drug loaded nanofibers possess effective antibacterial activity and biocompatible properties.

Gelatine/PLLA sponge-like scaffolds: morphological and biological characterization.

PubMed

Lazzeri, Luigi; Cascone, Maria Grazia; Danti, Serena; Serino, Lorenzo Pio; Moscato, Stefania; Bernardini, Nunzia

2007-07-01

Biodegradable synthetic polymers such as poly(lactic acid) (PLA) are widely used to prepare scaffolds for cell transplantation and tissue growth, using different techniques set up for the purpose. However the poor hydrophilicity of these polymers represents the main limitation to their use as scaffolds because it causes a low affinity for the cells. An effective way to solve this problem could be represented by the addition of biopolymers that are in general highly hydrophilic. The present work concerns porous biodegradable sponge-like systems based on poly(L-lactic acid) (PLLA) and gelatine. Morphology and porosity characteristics of the sponges were studied by scanning electron microscopy and mercury intrusion porosimetry respectively. Blood compatibility was investigated by bovine plasma fibrinogen (BPF) adsorption test and platelet adhesion test (PAT). The cell culture method was used in order to evaluate the ability of the matrices to work as scaffolds for tissue regeneration. The obtained results indicate that the sponges have interesting porous characteristics, good blood compatibility and above all good ability to support cell adhesion and growth. In fact viable and metabolically active animal cells were found inside the sponges after 8 weeks in culture. On this basis the systems produced seem to be good candidates as scaffolds for tissue regeneration.

Formation of Nanofibrous Matrices, Three-Dimensional Scaffolds, and Microspheres: From Theory to Practice

PubMed Central

Ma, Chi

2017-01-01

Nanofibrous architecture presents unique biophysical cues to facilitate cellular responses and is considered an indispensable feature of a biomimetic three-dimensional (3D) scaffold and cell carrier. While electrospinning is a widely used method to prepare natural extracellular matrix-like nanofibers, it faces significant challenges to incorporate nanofibrous architecture into well-defined macroporous 3D scaffolds or injectable microspheres. Here we report a nonelectrospinning approach that is effective at generating nanofibers from a variety of synthetic and natural biodegradable polymers and integrating these nanofibers into (1) 3D scaffolds with constructive geometry and designed internal macropore structures; and (2) injectable microspheres. Our approach to generating polymer nanofibers is based on the control of polymer–solvent interaction parameter χp-s. We obtained the χp-s and solvent composition phase diagrams of different temperatures according to the Flory–Huggins classic lattice model and the Hildebrand-Scott solubility parameter equation. A critical polymer–solvent interaction parameter χcrit was introduced as a criterion to predict phase separation and nanofiber formation. To test the effectiveness of our approach, a total of 15 widely used biodegradable polymers were selected and successfully fabricated into nanofibrous matrices. Furthermore, macroporous nanofibrous 3D scaffolds with complex architecture and nanofibrous injectable microspheres were generated from those biodegradable polymers by combining our method with other processes. Our approach is universally effective to fabricate nanofibrous matrices from any polymeric materials. This work, therefore, greatly expands our ability to design appropriate biomimetic 3D scaffolds and injectable cell carriers for advanced regenerative therapies. PMID:27923327

First in human evaluation of the vascular biocompatibility and biomechanical performance of a novel ultra high molecular weight amorphous PLLA bioresorbable scaffold in the absence of anti-proliferative drugs: Two-year imaging results in humans.

PubMed

Moncada, Miguel; Delgado, Juan A; Colombo, Antonio; Gasior, Pawel; Ramzipoor, Kamal; Estrada, Alex; Lee, Chang; Dokko, Danny; Granada, Juan F

2017-12-15

In this first-in-human study, we prospectively studied the vascular compatibility and mechanical performance of a novel bare ultra-high molecular weight amorphous PLLA bioresorbable scaffold (BRS, FORTITUDE®, Amaranth Medical, Mountain View, California) up to two years after implantation using multimodality imaging techniques. The vascular biocompatibility of polymers used in BRS has not been fully characterized in the absence of anti-proliferative drugs. A total of 10 patients undergoing single scaffold implantation were included in the final analysis and were followed up using optical coherence tomography (OCT) at 2-years. All devices were implanted under angiographic and intravascular ultrasound (IVUS) guidance. Angiographic and IVUS follow up was performed at 6 months. Additionally, angiography and OCT imaging were performed at 2-years. At 6 months, mean intra-scaffold angiographic MLD slightly decreased from baseline procedural values. However, at 2 years, mean angiographic MLD increased (post procedure; 2.9 [2.7, 3.1] mm vs. 6 months; 2.1 [1.6, 2.5] vs. 2 years; 2.4 [2.1, 2.6], P = .001). Also, angiographic percent diameter stenosis decreased and late lumen gain increased between 6 months and 2 years follow up. Mean neointimal hyperplasia volume assessed by IVUS at 6 months was 26% [15.2, 29.3]. At 2 years OCT follow up neointimal hyperplasia volume was 24.2% [19.4, 28.9]. No presence of neoatherosclerosis was identified in any of the analyzed cases. At 2 years, this novel PLLA-based BRS induced expansive vascular remodeling from 6 to 24 months, a biological phenomenon that appears to be independent of the presence of anti-proliferative drugs. © 2017 Wiley Periodicals, Inc.

Tissue Extracellular Matrix Nanoparticle Presentation in Electrospun Nanofibers

PubMed Central

Gibson, Matt; Mao, Hai-Quan; Elisseeff, Jennifer

2014-01-01

Biomaterials derived from the decellularization of mature tissues retain biological and architectural features that profoundly influence cellular activity. However, the clinical utility of such materials remains limited as the shape and physical properties are difficult to control. In contrast, scaffolds based on synthetic polymers can be engineered to exhibit specific physical properties, yet often suffer from limited biological functionality. This study characterizes composite materials that present decellularized extracellular matrix (DECM) particles in combination with synthetic nanofibers and examines the ability of these materials to influence stem cell differentiation. Mechanical processing of decellularized tissues yielded particles with diameters ranging from 71 to 334 nm. Nanofiber scaffolds containing up to 10% DECM particles (wt/wt) derived from six different tissues were engineered and evaluated to confirm DECM particle incorporation and to measure bioactivity. Scaffolds containing bone, cartilage, and fat promoted osteogenesis at 1 and 3 weeks compared to controls. In contrast, spleen and lung DECM significantly reduced osteogenic outcomes compared to controls. These findings highlight the potential to incorporate appropriate source DECM nanoparticles within nanofiber composites to design a scaffold with bioactivity targeted to specific applications. PMID:24971329

Designer bFGF-incorporated d-form self-assembly peptide nanofiber scaffolds to promote bone repair.

PubMed

He, Bin; Ou, Yunsheng; Chen, Shuo; Zhao, Weikang; Zhou, Ao; Zhao, Jinqiu; Li, Hong; Jiang, Dianming; Zhu, Yong

2017-05-01

d-Form and l-form peptide nanofiber scaffolds can spontaneously form stable β-sheet secondary structures and nanofiber hydrogel scaffolds, and hold some promise in hemostasis and wound healing. We report here on the synthetic self-assembling peptide d-RADA16 and l-RADA16 are both found to produce stable β-sheet secondary structure and nanofiber hydrogel scaffolds based on circular dichroism (CD) spectroscopy, transmission electron microscopy (TEM) and rheology analysis etc. d-RADA16 hydrogel and l-RADA16 hydrogel can enhance obvious bone repair in femoral condyle defects of the Sprague-Dawley (SD) rat model compared to PBS treatment. Based on micro-computed tomography (CT), it was revealed that d-RADA16 hydrogel and l-RADA16 hydrogel were capable to obtain the extensive bone healing. Histological evaluation also found that these two hydrogels facilitate the presence of more mature bone tissue within the femoral condyle defects. Additionally, d-RADA16 hydrogel showed some potential in storing and releasing basic-fibroblast growth factor (bFGF) which was able to further promote bone regeneration based on micro-CT analysis. These results indicate that d-form peptide nanofiber hydrogel have some special capacity for bone repair. Copyright © 2016 Elsevier B.V. All rights reserved.

Electrospun nanofibers: Work for medicine?

NASA Astrophysics Data System (ADS)

Liao, Susan; Chan, Casey K.; Ramakrishna, S.

2010-03-01

Attempts have been made to fabricate nanofibrous scaffolds to mimic the chemical composition and structural properties of the extracellular matrix (ECM) for tissue/organ replacement. Nanofiber scaffolds with various patterns have been successfully produced from synthetic and natural polymers through a relatively simple technique of electrospinning. The resulting patterns can mimic some of the diverse tissue-specific orientation and three-dimensional (3D) fibrous structures. Studies on cell-nanofiber interactions, including studies on stem cells, have revealed the importance of nanotopography on cell adhesion, proliferation and differentiation. Furthermore, clinical application of electrospun nanofibers including wound healing, tissue regeneration, drug delivery and stem cell therapy are highly feasible due to the ease and flexibility of fabrication of making nanofiber with this cost-effective method using electrospinning. In this review, we have highlighted the current state of the art and provided future perspectives on electrospun nanofiber in medical applications.

Neural Differentiation of Mesenchymal Stem Cells on Scaffolds for Nerve Tissue Engineering Applications.

PubMed

Quintiliano, Kerlin; Crestani, Thayane; Silveira, Davi; Helfer, Virginia Etges; Rosa, Annelise; Balbueno, Eduardo; Steffens, Daniela; Jotz, Geraldo Pereira; Pilger, Diogo André; Pranke, Patricia

2016-11-01

Scaffolds produced by electrospinning act as supports for cell proliferation and differentiation, improved through the release of neurotrophic factors. The objective of this study was to develop aligned and random nanofiber scaffolds with and without nerve growth factor to evaluate the potential of mesenchymal stem cells (MSCs) for neural differentiation. Nanofiber morphology, diameter, degradability, cell morphology, adhesion, proliferation, viability, cytotoxicity, and neural differentiation were performed to characterize the scaffolds. The expression for nestin, β-III tubulin, and neuron-specific enolase was also evaluated. The scaffolds demonstrated a satisfactory environment for MSC growth, being nontoxic. The MSCs cultivated on the scaffolds were able to adhere and proliferate. The evaluation of neural differentiation indicated that in all groups of scaffolds the MSCs were able to upregulate neural gene expression.

Thermal Annealing to Modulate the Shape Memory Behavior of a Biobased and Biocompatible Triblock Copolymer Scaffold in the Human Body Temperature Range.

PubMed

Merlettini, Andrea; Gigli, Matteo; Ramella, Martina; Gualandi, Chiara; Soccio, Michelina; Boccafoschi, Francesca; Munari, Andrea; Lotti, Nadia; Focarete, Maria Letizia

2017-08-14

A biodegradable and biocompatible electrospun scaffold with shape memory behavior in the physiological temperature range is here presented. It was obtained starting from a specifically designed, biobased PLLA-based triblock copolymer, where the central block is poly(propylene azelate-co-propylene sebacate) (P(PAz60PSeb40)) random copolymer. Shape memory properties are determined by the contemporary presence of the low melting crystals of the P(PAz60PSeb40) block, acting as switching segment, and of the high melting crystal phase of PLLA blocks, acting as physical network. It is demonstrated that a straightforward annealing process applied to the crystal phase of the switching element gives the possibility to tune the shape recovery temperature from about 25 to 50 °C, without the need of varying the copolymer's chemical structure. The thermal annealing approach here presented can be thus considered a powerful strategy for "ad hoc" programming the same material for applications requiring different recovery temperatures. Fibroblast culture experiments demonstrated scaffold biocompatibility.

Electrospinning of poly(glycerol sebacate)-based nanofibers for nerve tissue engineering.

PubMed

Hu, Jue; Kai, Dan; Ye, Hongye; Tian, Lingling; Ding, Xin; Ramakrishna, Seeram; Loh, Xian Jun

2017-01-01

Nerve tissue engineering (TE) requires biomimetic scaffolds providing essential chemical and topographical cues for nerve regeneration. Poly(glycerol sebacate) (PGS) is a biodegradable and elastic polymer that has gained great interest as a TE scaffolding biomaterial. However, uncured PGS is difficult to be electrospun into nanofibers. PGS would, therefore, require the addition of electrospinning agents. In this study, we modified PGS by using atom transfer radical polymerization (ATRP) to synthesize PGS-based copolymers with methyl methacrylate (MMA). The synthesized PGS-PMMA copolymer showed a molecular weight of 82kDa and a glass transition temperature of 115°C. More importantly, the PGS-PMMA could be easily electrospun into nanofiber with a fiber diameter of 167±33nm. Blending gelatin into PGS-PMMA nanofibers was found to increase its hydrophilicity and biocompatibility. Rat PC12 cells were seeded onto the PGS-PMMA/gelatin nanofibers to investigate their potential for nerve regeneration. It was found that gelatin-containing PGS-based nanofibers promoted cell proliferation. The elongated cell morphology observed on such nanofibers indicated that the scaffolds could induce the neurite outgrowth of the nerve stem cells. Overall, our study suggested that the synthesis of PGS-based copolymers might be a promising approach to enhance their processability, and therefore advancing bioscaffold engineering for various TE applications. Copyright © 2016 Elsevier B.V. All rights reserved.

HB-EGF embedded in PGA/PLLA scaffolds via subcritical CO2 augments the production of tissue engineered intestine.

PubMed

Liu, Yanchun; Nelson, Tyler; Cromeens, Barrett; Rager, Terrence; Lannutti, John; Johnson, Jed; Besner, Gail E

2016-10-01

The ability to deliver sustained-release, biologically active growth factors through custom designed tissue engineering scaffolds at sites of tissue regeneration offers great therapeutic opportunity. Due to the short in vivo half-lives of most growth factors, it is challenging to deliver these proteins to sites of interest where they may be used before being degraded. The application of subcritical CO2 uses gas-phase CO2 at subcritical pressures ranging from 41 to 62 bar (595-913 PSI) which avoids foaming by reducing the amount of CO2 dissolved in the polymer and maintains completely reversible plasticization. In the current study, heparin-binding EGF-like growth factor (HB-EGF) was embedded into polyglycolic acid (PGA)/Poly-l-latic acid (PLLA) scaffolds via subcritical CO2 exposure for the production of tissue engineered intestine (TEI). PGA fiber morphology after subcritical CO2 exposure was examined by scanning electron microscopy (SEM) and the distribution of HB-EGF embedded in the scaffold fibers was detected by HB-EGF immunofluorescent staining. In vivo implantation of HB-EGF-embedded scaffolds confirmed significantly improved TEI structure as a result of local delivery of the trophic growth factor. These findings may be critical for the production of TEI in the treatment of patients with short bowel syndrome in the future. Copyright © 2016 Elsevier Ltd. All rights reserved.

Incorporation of mesoporous silica nanoparticles into random electrospun PLGA and PLGA/gelatin nanofibrous scaffolds enhances mechanical and cell proliferation properties.

PubMed

Mehrasa, Mohammad; Asadollahi, Mohammad Ali; Nasri-Nasrabadi, Bijan; Ghaedi, Kamran; Salehi, Hossein; Dolatshahi-Pirouz, Alireza; Arpanaei, Ayyoob

2016-09-01

Poly(lactic-co-glycolic acid) (PLGA) and PLGA/gelatin random nanofibrous scaffolds embedded with different amounts of mesoporous silica nanoparticles (MSNPs) were fabricated using electrospinning method. To evaluate the effects of nanoparticles on the scaffolds, physical, chemical, and mechanical properties as well as in vitro degradation behavior of scaffolds were investigated. The mean diameters of nanofibers were 974±68nm for the pure PLGA scaffolds vs 832±70, 764±80, and 486±64 for the PLGA/gelatin, PLGA/10wt% MSNPs, and the PLGA/gelatin/10wt% MSNPs scaffolds, respectively. The results suggested that the incorporation of gelatin and MSNPs into PLGA-based scaffolds enhances the hydrophilicity of scaffolds due to an increase of hydrophilic functional groups on the surface of nanofibers. With porosity examination, it was concluded that the incorporation of MSNPs and gelatin decrease the porosity of scaffolds. Nanoparticles also improved the tensile mechanical properties of scaffolds. Using in vitro degradation analysis, it was shown that the addition of nanoparticles to the nanofibers matrix increases the weight loss percentage of PLGA-based samples, whereas it decreases the weight loss percentage in the PLGA/gelatin composites. Cultivation of rat pheochromocytoma cell line (PC12), as precursor cells of dopaminergic neural cells, on the scaffolds demonstrated that the introduction of MSNPs into PLGA and PLGA/gelatin matrix leads to improved cell attachment and proliferation and enhances cellular processes. Copyright © 2016 Elsevier B.V. All rights reserved.

Biosilica-loaded poly(ϵ-caprolactone) nanofibers mats provide a morphogenetically active surface scaffold for the growth and mineralization of the osteoclast-related SaOS-2 cells.

PubMed

Müller, Werner E G; Tolba, Emad; Schröder, Heinz C; Diehl-Seifert, Bärbel; Link, Thorben; Wang, Xiaohong

2014-10-01

Bioprinting/3D cell printing procedures for the preparation of scaffolds/implants have the potential to revolutionize regenerative medicine. Besides biocompatibility and biodegradability, the hardness of the scaffold material is of critical importance to allow sufficient mechanical protection and, to the same extent, allow migration, cell-cell, and cell-substrate contact formation of the matrix-embedded cells. In the present study, we present a strategy to encase a bioprinted, cell-containing, and soft scaffold with an electrospun mat. The electrospun poly(ϵ-caprolactone) (PCL) nanofibers mats, containing tetraethyl orthosilicate (TEOS), were subsequently incubated with silicatein. Silicatein synthesizes polymeric biosilica by polycondensation of ortho-silicate that is formed from prehydrolyzed TEOS. Biosilica provides a morphogenetically active matrix for the growth and mineralization of osteoblast-related SaOS-2 cells in vitro. Analysis of the microstructure of the 300-700 nm thick PCL/TEOS nanofibers, incubated with silicatein and prehydrolyzed TEOS, displayed biosilica deposits on the mats formed by the nanofibers. We conclude and propose that electrospun PCL nanofibers mats, coated with biosilica, may represent a morphogenetically active and protective cover for bioprinted cell/tissue-like units with a suitable mechanical stability, even if the cells are embedded in a softer matrix. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Braided and Stacked Electrospun Nanofibrous Scaffolds for Tendon and Ligament Tissue Engineering

PubMed Central

Rothrauff, Benjamin B.; Lauro, Brian B.; Yang, Guang; Debski, Richard E.; Musahl, Volker

2017-01-01

Tendon and ligament injuries are a persistent orthopedic challenge given their poor innate healing capacity. Nonwoven electrospun nanofibrous scaffolds composed of polyesters have been used to mimic the mechanics and topographical cues of native tendons and ligaments. However, nonwoven nanofibers have several limitations that prevent broader clinical application, including poor cell infiltration, as well as tensile and suture-retention strengths that are inferior to native tissues. In this study, multilayered scaffolds of aligned electrospun nanofibers of two designs–stacked or braided–were fabricated. Mechanical properties, including structural and mechanical properties and suture-retention strength, were determined using acellular scaffolds. Human bone marrow-derived mesenchymal stem cells (MSCs) were seeded on scaffolds for up to 28 days, and assays for tenogenic differentiation, histology, and biochemical composition were performed. Braided scaffolds exhibited improved tensile and suture-retention strengths, but reduced moduli. Both scaffold designs supported expression of tenogenic markers, although the effect was greater on braided scaffolds. Conversely, cell infiltration was superior in stacked constructs, resulting in enhanced cell number, total collagen content, and total sulfated glycosaminoglycan content. However, when normalized against cell number, both designs modulated extracellular matrix protein deposition to a similar degree. Taken together, this study demonstrates that multilayered scaffolds of aligned electrospun nanofibers supported tenogenic differentiation of seeded MSCs, but the macroarchitecture is an important consideration for applications of tendon and ligament tissue engineering. PMID:28071988

Braided and Stacked Electrospun Nanofibrous Scaffolds for Tendon and Ligament Tissue Engineering.

PubMed

Rothrauff, Benjamin B; Lauro, Brian B; Yang, Guang; Debski, Richard E; Musahl, Volker; Tuan, Rocky S

2017-05-01

Tendon and ligament injuries are a persistent orthopedic challenge given their poor innate healing capacity. Nonwoven electrospun nanofibrous scaffolds composed of polyesters have been used to mimic the mechanics and topographical cues of native tendons and ligaments. However, nonwoven nanofibers have several limitations that prevent broader clinical application, including poor cell infiltration, as well as tensile and suture-retention strengths that are inferior to native tissues. In this study, multilayered scaffolds of aligned electrospun nanofibers of two designs-stacked or braided-were fabricated. Mechanical properties, including structural and mechanical properties and suture-retention strength, were determined using acellular scaffolds. Human bone marrow-derived mesenchymal stem cells (MSCs) were seeded on scaffolds for up to 28 days, and assays for tenogenic differentiation, histology, and biochemical composition were performed. Braided scaffolds exhibited improved tensile and suture-retention strengths, but reduced moduli. Both scaffold designs supported expression of tenogenic markers, although the effect was greater on braided scaffolds. Conversely, cell infiltration was superior in stacked constructs, resulting in enhanced cell number, total collagen content, and total sulfated glycosaminoglycan content. However, when normalized against cell number, both designs modulated extracellular matrix protein deposition to a similar degree. Taken together, this study demonstrates that multilayered scaffolds of aligned electrospun nanofibers supported tenogenic differentiation of seeded MSCs, but the macroarchitecture is an important consideration for applications of tendon and ligament tissue engineering.

Encapsulation of β-Sitosterol in Polyurethane by Sol-Gel Electrospinning.

PubMed

Amina, Musarat; Amna, Touseef; Al-Musayeib, Nawal; Zabin, Sami A; Hassan, M Shamshi; Khil, Myung-Seob

2017-06-01

Pristine β-sitosterol or in combination with other phytosterols is utilized in an array of enriched commercial foods. Considering the presence of β-sitosterol in different functional foods and its potential role in prevention and cure of neurodegenerative diseases, the aims of our investigation were to encapsulate β-sitosterol in nanofibers and to estimate influence of β-sitosterol on proliferation of fibroblasts. Electrospun nanofibers have widely been used as scaffolds to mimic natural extracellular matrix. Herein, our group for the first time establishes an innovative scaffold based on β-sitosterol and polyurethane using electrospinning. β-Sitosterol promotes epithelialization and possesses anti-oxidant and anti-inflammatory activities, whereas polyurethane, besides possessing biomedical uses, also enhances epithelial growth. We optimized the concentration (5%) of β-sitosterol in polyurethane to obtain homogenous solution, which can be spun without difficulty for the synthesis of β-sitosterol amalgamated scaffold. The resulted twisted nanofibers have been characterized via scanning electron microscopy and Fourier transform infrared spectroscopy. The viability of cells on twisted scaffold was examined using NIH 3T3 fibroblasts as model cell line. Incorporation of β-sitosterol in polyurethane changed the structure and size of nanofibers, and the twisted scaffolds were non-cytotoxic. Thus, the twisted nanoribbons, which contain anti-inflammatory β-sitosterol, can be utilized as a promising future material, which will help to ease inflammation and also aid in wound healing. In conclusion, the outcome of the preliminary research evidently points out the potential of twisted scaffold in biomedical applications.

Nanofiber Scaffold-Based Tissue-Engineered Retinal Pigment Epithelium to Treat Degenerative Eye Diseases

PubMed Central

Khristov, Vladimir; Wan, Qin; Sharma, Ruchi; Jha, Balendu Shekhar; Lotfi, Mostafa; Maminishkis, Arvydas; Simon, Carl G.

2016-01-01

Abstract Clinical-grade manufacturing of a functional retinal pigment epithelium (RPE) monolayer requires reproducing, as closely as possible, the natural environment in which RPE grows. In vitro, this can be achieved by a tissue engineering approach, in which the RPE is grown on a nanofibrous biological or synthetic scaffold. Recent research has shown that nanofiber scaffolds perform better for cell growth and transplantability compared with their membrane counterparts and that the success of the scaffold in promoting cell growth/function is not heavily material dependent. With these strides, the field has advanced enough to begin to consider implementation of one, or a combination, of the tissue engineering strategies discussed herein. In this study, we review the current state of tissue engineering research for in vitro culture of RPE/scaffolds and the parameters for optimal scaffold design that have been uncovered during this research. Next, we discuss production methods and manufacturers that are capable of producing the nanofiber scaffolds in such a way that would be biologically, regulatory, clinically, and commercially viable. Then, a discussion of how the scaffolds could be characterized, both morphologically and mechanically, to develop a testing process that is viable for regulatory screening is performed. Finally, an example of a tissue-engineered RPE/scaffold construct is given to provide the reader a framework for understanding how these pieces could fit together to develop a tissue-engineered RPE/scaffold construct that could pass regulatory scrutiny and can be commercially successful. PMID:27110730

Long-term liver-specific functions of hepatocytes in electrospun chitosan nanofiber scaffolds coated with fibronectin.

PubMed

Rajendran, Divya; Hussain, Ali; Yip, Derek; Parekh, Amit; Shrirao, Anil; Cho, Cheul H

2017-08-01

In this study, a new 3D liver model was developed using biomimetic nanofiber scaffolds and co-culture system consisting of hepatocytes and fibroblasts for the maintenance of long-term liver functions. The chitosan nanofiber scaffolds were fabricated by the electrospinning technique. To enhance cellular adhesion and spreading, the surfaces of the chitosan scaffolds were coated with fibronectin (FN) by adsorption and evaluated for various cell types. Cellular phenotype, protein expression, and liver-specific functions were extensively characterized by immunofluorescent and histochemical stainings, albumin enzyme-linked immunosorbent assay and Cytochrome p450 detoxification assays, and scanning electron microscopy. The electrospun chitosan scaffolds exhibited a highly porous and randomly oriented nanofibrous structure. The FN coating on the surface of the chitosan nanofibers significantly enhanced cell attachment and spreading, as expected, as surface modification with this cell adhesion molecule on the chitosan surface is important for focal adhesion formation and integrin binding. Comparison of hepatocyte mono-cultures and co-cultures in 3D culture systems indicated that the hepatocytes in co-cultures formed colonies and maintained their morphologies and functions for prolonged periods of time. The 3D liver tissue model developed in this study will provide useful tools toward the development of engineered liver tissues for drug screening and tissue engineering applications. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2119-2128, 2017. © 2017 Wiley Periodicals, Inc.

Combined effects of chemical priming and mechanical stimulation on mesenchymal stem cell differentiation on nanofiber scaffolds

PubMed Central

Subramony, Siddarth D.; Su, Amanda; Yeager, Keith; Lu, Helen H.

2014-01-01

Functional tissue engineering of connective tissues such as the anterior cruciate ligament (ACL) remains a significant clinical challenge, largely due to the need for mechanically competent scaffold systems for grafting, as well as a reliable cell source for tissue formation. We have designed an aligned, polylactide-co-glycolide (PLGA) nanofiber-based scaffold with physiologically relevant mechanical properties for ligament regeneration. The objective of this study is to identify optimal tissue engineering strategies for fibroblastic induction of human mesenchymal stem cells (hMSC), testing the hypothesis that basic fibroblast growth factor (bFGF) priming coupled with tensile loading will enhance hMSC-mediated ligament regeneration. It was observed that compared to the unloaded, as well as growth factor-primed but unloaded controls, bFGF stimulation followed by physiologically relevant tensile loading enhanced hMSC proliferation, collagen production and subsequent differentiation into ligament fibroblast-like cells, upregulating the expression of types I and III collagen, as well as tenasin-C and tenomodulin. The results of this study suggest that bFGF priming increases cell proliferation, while mechanical stimulation of the hMSCs on the aligned nanofiber scaffold promotes fibroblastic induction of these cells. In addition to demonstrating the potential of nanofiber scaffolds for hMSC-mediated functional ligament tissue engineering, this study yields new insights into the interactive effects of chemical and mechanical stimuli on stem cell differentiation. PMID:24267271

Self-assembly of silk-elastinlike protein polymers into three-dimensional scaffolds for biomedical applications

NASA Astrophysics Data System (ADS)

Zeng, Like

Production of brand new protein-based materials with precise control over the amino acid sequences at single residue level has been made possible by genetic engineering, through which artificial genes can be developed that encode protein-based materials with desired features. As an example, silk-elastinlike protein polymers (SELPs), composed of tandem repeats of amino acid sequence motifs from Bombyx mori (silkworm) silk and mammalian elastin, have been produced in this approach. SELPs have been studied extensively in the past two decades, however, the fundamental mechanism governing the self-assembly process to date still remains largely unresolved. Further, regardless of the unprecedented success when exploited in areas including drug delivery, gene therapy, and tissue augmentation, SELPs scaffolds as a three-dimensional cell culture model system are complicated by the inability of SELPs to provide the embedded tissue cells with appropriate biochemical stimuli essential for cell survival and function. In this dissertation, it is reported that the self-assembly of silk-elastinlike protein polymers (SELPs) into nanofibers in aqueous solutions can be modulated by tuning the curing temperature, the size of the silk blocks, and the charge of the elastin blocks. A core-sheath model was proposed for nanofiber formation, with the silk blocks in the cores and the hydrated elastin blocks in the sheaths. The folding of the silk blocks into stable cores -- affected by the size of the silk blocks and the charge of the elastin blocks -- plays a critical role in the assembly of silk-elastin nanofibers. The assembled nanofibers further form nanofiber clusters on the microscale, and the nanofiber clusters then coalesce into nanofiber micro-assemblies, interconnection of which eventually leads to the formation of three-dimensional scaffolds with distinct nanoscale and microscale features. SELP-Collagen hybrid scaffolds were also fabricated to enable independent control over the scaffolds' biochemical input and matrix stiffness. It is reported herein that in the hybrid scaffolds, collagen provides essential biochemical cues needed to promote cell attachment and function while SELP imparts matrix stiffness tunability. To obtain tissue-specificity in matrix stiffness that spans over several orders of magnitude covering from soft brain to stiff cartilage, the hybrid SELP-Collagen scaffolds were crosslinked by transglutaminase at physiological conditions compatible for simultaneous cell encapsulation. The effect of the increase in matrix stiffness induced by such enzymatic crosslinking on cellular viability and proliferation was also evaluated using in vitro cell assays.

Fabrication and characterization of anisotropic nanofiber scaffolds for advanced drug delivery systems

PubMed Central

Jalani, Ghulam; Jung, Chan Woo; Lee, Jae Sang; Lim, Dong Woo

2014-01-01

Stimuli-responsive, polymer-based nanostructures with anisotropic compartments are of great interest as advanced materials because they are capable of switching their shape via environmentally-triggered conformational changes, while maintaining discrete compartments. In this study, a new class of stimuli-responsive, anisotropic nanofiber scaffolds with physically and chemically distinct compartments was prepared via electrohydrodynamic cojetting with side-by-side needle geometry. These nanofibers have a thermally responsive, physically-crosslinked compartment, and a chemically-crosslinked compartment at the nanoscale. The thermally responsive compartment is composed of physically crosslinkable poly(N-isopropylacrylamide) poly(NIPAM) copolymers, and poly(NIPAM-co-stearyl acrylate) poly(NIPAM-co-SA), while the thermally-unresponsive compartment is composed of polyethylene glycol dimethacrylates. The two distinct compartments were physically crosslinked by the hydrophobic interaction of the stearyl chains of poly(NIPAM-co-SA) or chemically stabilized via ultraviolet irradiation, and were swollen in physiologically relevant buffers due to their hydrophilic polymer networks. Bicompartmental nanofibers with the physically-crosslinked network of the poly(NIPAM-co-SA) compartment showed a thermally-triggered shape change due to thermally-induced aggregation of poly(NIPAM-co-SA). Furthermore, when bovine serum albumin and dexamethasone phosphate were separately loaded into each compartment, the bicompartmental nanofibers with anisotropic actuation exhibited decoupled, controlled release profiles of both drugs in response to a temperature. A new class of multicompartmental nanofibers could be useful for advanced nanofiber scaffolds with two or more drugs released with different kinetics in response to environmental stimuli. PMID:24872702

Synthesis and Fabrication of Collagen-Coated Ostholamide Electrospun Nanofiber Scaffold for Wound Healing.

PubMed

Kandhasamy, Subramani; Perumal, Sathiamurthi; Madhan, Balaraman; Umamaheswari, Narayanan; Banday, Javid Ahmad; Perumal, Paramasivan Thirumalai; Santhanakrishnan, Vichangal Pridiuldi

2017-03-15

A novel scaffold for effective wound healing treatment was developed utilizing natural product bearing collagen-based biocompatible electrospun nanofibers. Initially, ostholamide (OSA) was synthesized from osthole (a natural coumarin), characterized by 1 H, 13 C, DEPT-135 NMR, ESI-MS, and FT-IR spectroscopy analysis. OSA was incorporated into polyhydroxybutyrate (PHB) and gelatin (GEL), which serve as templates for electrospun nanofibers. The coating of OSA-PHB-GEL nanofibers with collagen resulted in PHB-GEL-OSA-COL nanofibrous scaffold which mimics extracellular matrix and serves as an effective biomaterial for tissue engineering applications, especially for wound healing. PHB-GEL-OSA-COL, along with PHB-GEL-OSA and collagen film (COLF), was characterized in vitro and in vivo to determine its efficacy. The developed PHB-GEL-OSA-COL nanofibers posed an impressive mechanical stability, an essential requirement for wound healing. The presence of OSA had contributed to antimicrobial efficacy. These scaffolds exhibited efficient antibacterial activity against common wound pathogens, Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus). The zones of inhibition were observed to be 14 ± 22 and 10 ± 2 mm, respectively. It was observed that nanofibrous scaffold had the ability to release OSA in a controlled manner, and hence, OSA would be present at the site of application and exhibit bioactivity in a sustained manner. PHB-GEL-OSA-COL nanofiber was determined to be stable against enzymatic degradation, which is the most important parameter for promoting proliferation of cells contributing to repair and remodeling of tissues during wound healing applications. As hypothesized, PHB-GEL-OSA-COL was observed to imbibe excellent cytocompatibility, which was determined using NIH 3T3 fibroblast cell proliferation studies. PHB-GEL-OSA-COL exhibited excellent wound healing efficacy which was confirmed using full thickness excision wound model in Wistar rats. The rats treated with PHB-GEL-OSA-COL nanofibrous scaffold displayed enhanced healing when compared to untreated control. Both in vitro and in vivo analysis of PHB-GEL-OSA-COL presents a strong case of therapeutic biomaterial suiting wound repair and regeneration.

Fabrication and Characterization of Magnesium Ferrite-Based PCL/Aloe Vera Nanofibers

PubMed Central

Thompson, Zanshe; Rahman, Shekh; Yarmolenko, Sergey; Sankar, Jagannathan; Kumar, Dhananjay

2017-01-01

Composite nanofibers of biopolymers and inorganic materials have been widely explored as tissue engineering scaffolds because of their superior structural, mechanical and biological properties. In this study, magnesium ferrite (Mg-ferrite) based composite nanofibers were synthesized using an electrospinning technique. Mg-ferrite nanoparticles were first synthesized using the reverse micelle method, and then blended in a mixture of polycaprolactone (PCL), a synthetic polymer, and Aloe vera, a natural polymer, to create magnetic nanofibers by electrospinning. The morphology, structural and magnetic properties, and cellular compatibility of the magnetic nanofibers were analyzed. Mg-ferrite/PCL/Aloe vera nanofibers showed good uniformity in fiber morphology, retained their structural integrity, and displayed magnetic strength. Experimental results, using cell viability assay and scanning electron microscopy imaging showed that magnetic nanofibers supported 3T3 cell viability. We believe that the new composite nanofibrous membranes developed in this study have the ability to mimic the physical structure and function of tissue extracellular matrix, as well as provide the magnetic and soluble metal ion attributes in the scaffolds with enhanced cell attachment, and thus improve tissue regeneration. PMID:28800071

Nanofiber scaffold gradients for interfacial tissue engineering.

PubMed

Ramalingam, Murugan; Young, Marian F; Thomas, Vinoy; Sun, Limin; Chow, Laurence C; Tison, Christopher K; Chatterjee, Kaushik; Miles, William C; Simon, Carl G

2013-02-01

We have designed a 2-spinnerette device that can directly electrospin nanofiber scaffolds containing a gradient in composition that can be used to engineer interfacial tissues such as ligament and tendon. Two types of nanofibers are simultaneously electrospun in an overlapping pattern to create a nonwoven mat of nanofibers containing a composition gradient. The approach is an advance over previous methods due to its versatility - gradients can be formed from any materials that can be electrospun. A dye was used to characterize the 2-spinnerette approach and applicability to tissue engineering was demonstrated by fabricating nanofibers with gradients in amorphous calcium phosphate nanoparticles (nACP). Adhesion and proliferation of osteogenic cells (MC3T3-E1 murine pre-osteoblasts) on gradients was enhanced on the regions of the gradients that contained higher nACP content yielding a graded osteoblast response. Since increases in soluble calcium and phosphate ions stimulate osteoblast function, we measured their release and observed significant release from nanofibers containing nACP. The nanofiber-nACP gradients fabricated herein can be applied to generate tissues with osteoblast gradients such as ligaments or tendons. In conclusion, these results introduce a versatile approach for fabricating nanofiber gradients that can have application for engineering graded tissues.

Cell-Adhesive Matrices Composed of RGD Peptide-Displaying M13 Bacteriophage/Poly(lactic-co-glycolic acid) Nanofibers Beneficial to Myoblast Differentiation.

PubMed

Shin, Yong Cheol; Lee, Jong Ho; Jin, Linhua; Kim, Min Jeong; Kim, Chuntae; Hong, Suck Won; Oh, Jin Woo; Han, Dong-Wook

2015-10-01

Recently, there has been considerable effort to develop suitable scaffolds for tissue engineering applications. Cell adhesion is a prerequisite for cells to survive. In nature, the extracellular matrix (ECM) plays this role. Therefore, an ideal scaffold should be structurally similar to the natural ECM and have biocompatibility and biodegradability. In addition, the scaffold should have biofunctionality, which provides the potent ability to enhance the cellular behaviors, such as adhesion, proliferation and differentiation. This study concentrates on fabricating cell-adhesive matrices composed of RGD peptide-displaying M13 bacteriophage (RGD-M13 phage) and poly(lactic-co-glycolic acid, PLGA) nanofibers. Long rod-shaped M13 bacteriophages are non-toxic and can express many desired proteins on their surface. A genetically engineered M13 phage was constructed to display RGD peptides on its surface. PLGA is a biodegradable polymer with excellent biocompatibility and suitable physicochemical property for adhesive matrices. In this study, RGD-M13 phage/PLGA hybrid nanofiber matrices were fabricated by electrospinning. The physicochemical properties of these matrices were characterized by scanning electron microscopy, atomic force microscopy, Raman spectroscopy, and contact angle measurement. In addition, the cellular behaviors, such as the initial attachment, proliferation and differentiation, were analyzed by a CCK-8 assay and immunofluorescence staining to evaluate the potential application of these matrices to tissue engineering scaffolds. The RGD-M13 phage/PLGA nanofiber matrices could enhance the cellular behaviors and promote the differentiation of C2C12 myoblasts. These results suggest that the RGD-M13 phage/PLGA nanofiber matrices are beneficial to myoblast differentiation and can serve as effective tissue engineering scaffolds.

Guiding the orientation of smooth muscle cells on random and aligned polyurethane/collagen nanofibers.

PubMed

Jia, Lin; Prabhakaran, Molamma P; Qin, Xiaohong; Ramakrishna, Seeram

2014-09-01

Fabricating scaffolds that can simulate the architecture and functionality of native extracellular matrix is a huge challenge in vascular tissue engineering. Various kinds of materials are engineered via nano-technological approaches to meet the current challenges in vascular tissue regeneration. During this study, nanofibers from pure polyurethane and hybrid polyurethane/collagen in two different morphologies (random and aligned) and in three different ratios of polyurethane:collagen (75:25; 50:50; 25:75) are fabricated by electrospinning. The fiber diameters of the nanofibrous scaffolds are in the range of 174-453 nm and 145-419 for random and aligned fibers, respectively, where they closely mimic the nanoscale dimensions of native extracellular matrix. The aligned polyurethane/collagen nanofibers expressed anisotropic wettability with mechanical properties which is suitable for regeneration of the artery. After 12 days of human aortic smooth muscle cells culture on different scaffolds, the proliferation of smooth muscle cells on hybrid polyurethane/collagen (3:1) nanofibers was 173% and 212% higher than on pure polyurethane scaffolds for random and aligned scaffolds, respectively. The results of cell morphology and protein staining showed that the aligned polyurethane/collagen (3:1) scaffold promote smooth muscle cells alignment through contact guidance, while the random polyurethane/collagen (3:1) also guided cell orientation most probably due to the inherent biochemical composition. Our studies demonstrate the potential of aligned and random polyurethane/collagen (3:1) as promising substrates for vascular tissue regeneration. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

Superabsorbent biphasic system based on poly(lactic acid) and poly(acrylic acid)

NASA Astrophysics Data System (ADS)

Sartore, Luciana; Pandini, Stefano; Baldi, Francesco; Bignotti, Fabio

2016-05-01

In this research work, biocomposites based on crosslinked particles of poly(acrylic acid), commonly used as superabsorbent polymer (SAP), and poly-L-lactic acid (PLLA) were developed to elucidate the role of the filler (i.e., polymeric crosslinked particles) on the overall physico-mechanical behavior and to obtain superabsorbent thermoplastic products. Samples prepared by melt-blending of components in different ratios showed a biphasic system with a regular distribution of particles, with diameter ranging from 5 to 10 μm, within the PLLA polymeric matrix. The polymeric biphasic system, coded PLASA i.e. superabsorbent poly(lactic acid), showed excellent swelling properties, demonstrating that cross-linked particles retain their superabsorbent ability, as in their free counterparts, even if distributed in a thermoplastic polymeric matrix. The thermal characteristics of the biocomposites evidence enhanced thermal stability in comparison with neat PLLA and also mechanical properties are markedly modified by addition of crosslinked particles which induce regular stiffening effect. Furthermore, in aqueous environments the particles swell and are leached from PLLA matrix generating very high porosity. These new open-pore PLLA foams, produced in absence of organic solvents and chemical foaming agents, with good physico-mechanical properties appear very promising for several applications, for instance in tissue engineering for scaffold production.

Aligned ovine diaphragmatic myoblasts overexpressing human connexin-43 seeded on poly (L-lactic acid) scaffolds for potential use in cardiac regeneration.

PubMed

Giménez, Carlos Sebastián; Locatelli, Paola; Montini Ballarin, Florencia; Orlowski, Alejandro; Dewey, Ricardo A; Pena, Milagros; Abraham, Gustavo Abel; Aiello, Ernesto Alejandro; Bauzá, María Del Rosario; Cuniberti, Luis; Olea, Fernanda Daniela; Crottogini, Alberto

2018-04-01

Diaphragmatic myoblasts (DMs) are precursors of type-1 muscle cells displaying high exhaustion threshold on account that they contract and relax 20 times/min over a lifespan, making them potentially useful in cardiac regeneration strategies. Besides, it has been shown that biomaterials for stem cell delivery improve cell retention and viability in the target organ. In the present study, we aimed at developing a novel approach based on the use of poly (L-lactic acid) (PLLA) scaffolds seeded with DMs overexpressing connexin-43 (cx43), a gap junction protein that promotes inter-cell connectivity. DMs isolated from ovine diaphragm biopsies were characterized by immunohistochemistry and ability to differentiate into myotubes (MTs) and transduced with a lentiviral vector encoding cx43. After confirming cx43 expression (RT-qPCR and Western blot) and its effect on inter-cell connectivity (fluorescence recovery after photobleaching), DMs were grown on fiber-aligned or random PLLA scaffolds. DMs were successfully isolated and characterized. Cx43 mRNA and protein were overexpressed and favored inter-cell connectivity. Alignment of the scaffold fibers not only aligned but also elongated the cells, increasing the contact surface between them. This novel approach is feasible and combines the advantages of bioresorbable scaffolds as delivery method and a cell type that on account of its features may be suitable for cardiac regeneration. Future studies on animal models of myocardial infarction are needed to establish its usefulness on scar reduction and cardiac function.

Comparative evaluation of Chitosan, Cellulose Acetate, and Polyethersulfone Nanofiber Scaffolds for Neural Differentiation

PubMed Central

Du, Jian; Tan, Elaine; Kim, Hyo Jun; Zhang, Allen; Bhattacharya, Rahul; Yarema, Kevin J

2013-01-01

Based on accumulating evidence that the 3D topography and the chemical features of a growth surface influence neuronal differentiation, we combined these two features by evaluating the cytotoxicity, proliferation, and differentiation of the rat PC12 line and human neural stem cells (hNSCs) on chitosan (CS), cellulose acetate (CA), and polyethersulfone (PES)-derived electrospun nanofibers that had similar diameters, centered in the 200 to 500 nm range. None of the nanofibrous materials were cytotoxic compared to 2D (e.g., flat surface) controls; however, proliferation generally was inhibited on the nanofibrous scaffolds although to a lesser extent on the polysaccharide-derived materials compared to PES. In an exception to the trend towards slower growth on the 3D substrates, hNSCs differentiated on the CS nanofibers proliferated faster than the 2D controls and both cell types showed enhanced indication of neuronal differentiation on the CS scaffolds. Together, these results demonstrate beneficial attributes of CS for neural tissue engineering when this polysaccharide is used in the context of the defined 3D topography found in electrospun nanofibers. PMID:24274534

Preparation and characterization of oriented poly(vinyl alcohol)/carbon nanotube composite nanofibers

NASA Astrophysics Data System (ADS)

Shimizu, Akikazu; Kato, Hayato; Sato, Taiga; Kushida, Masahito

2017-07-01

Oriented nanofiber mats blended with carbon nanotubes (CNTs) are expected to be applied as cell seeding scaffolds. Biomaterials that are often used for cell seeding scaffolds generally have low mechanical strength and low electrical conductivity; thus, it has been difficult to apply them to tissues such as heart and nerve. In this study, we prepared oriented poly(vinyl alcohol) (PVA) nanofiber mats blended with various CNT concentrations (up to 10 wt %) by electrospinning using the parallel plate electrodes as collectors with applied voltage. The morphology, mechanical properties, and electrical properties of the prepared oriented nanofiber mats were measured by using various techniques such as scanning electron microscopy (SEM). The tensile strength of the oriented nanofiber mats in the applied voltage direction increased from 2.5 to 9.7 MPa with CNT concentration. Furthermore, the electrical conductivity of the oriented nanofiber mats in the applied voltage direction increased from 0.67 × 10-7 to 4.3 × 10-7 S·m-1. Also, the mechanical strength and electrical conductivity of the oriented nanofiber mats in the applied voltage direction were 3-4 and 2-3 times higher than those in the perpendicular direction, respectively.

In vitro mineralization and bone osteogenesis in poly(ε-caprolactone)/gelatin nanofibers.

PubMed

Alvarez Perez, Marco A; Guarino, Vincenzo; Cirillo, Valentina; Ambrosio, Luigi

2012-11-01

The implementation of bio-inspired strategies in developing scaffolds for the reconstruction of oral, craniofacial and bone skeletal tissues after injury or resection remains a challenge. Currently, advanced scaffolds comprising nanofibers endowed with biochemical/biophysical signaling capability offer great advantages in bone regeneration, because of their faithful mimesis of the characteristic size scales encountered in the fibrous network of the native extracellular matrix (ECM). In this study, we investigate the biological potential of nanofibers made of polycaprolactone and gelatin on guiding the regenerative mechanisms of bone. Contact angle measurements and environmental SEM investigations indicate a weak linkage of gelatin molecules to PCL chains, facilitating an efficient adhesion signal to cells up to 3 days of culture. In vitro studies performed on human mesenchymal stem cells (hMSC) until 3 weeks in culture medium with osteogenic supplementation, clearly showing the effectiveness of PCL/Gelatin electrospun scaffolds in promoting bone osteogenesis and mineralization. The increase of alkaline phosphatase activity (ALP) and gene expression of bone-related molecules (bone sialoprotein, osteopontin and osteocalcin), indicated by immunodetection and upregulation level of mRNA, confirm that proposed nanofibers promote the osteogenic differentiation of hMSC, preferentially in osteogenic medium. Moreover, the evidence of newly formed collagen fibers synthesis by SIRCOL and their mineralization evaluated by Alizarin Red staining and EDS mapping of the elements Ca, P and Mg corroborate the idea that native osteoid matrix is ultimately deposited. All these data suggest that PCL and gelatin electrospun nanofibers have great potential as osteogenesis promoting scaffolds for successful application in bone surgery. Copyright © 2012 Wiley Periodicals, Inc.

Immobilization and Application of Electrospun Nanofiber Scaffold-based Growth Factor in Bone Tissue Engineering.

PubMed

Chen, Guobao; Lv, Yonggang

2015-01-01

Electrospun nanofibers have been extensively used in growth factor delivery and regenerative medicine due to many advantages including large surface area to volume ratio, high porosity, excellent loading capacity, ease of access and cost effectiveness. Their relatively large surface area is helpful for cell adhesion and growth factor loading, while storage and release of growth factor are essential to guide cellular behaviors and tissue formation and organization. In bone tissue engineering, growth factors are expected to transmit signals that stimulate cellular proliferation, migration, differentiation, metabolism, apoptosis and extracellular matrix (ECM) deposition. Bolus administration is not always an effective method for the delivery of growth factors because of their rapid diffusion from the target site and quick deactivation. Therefore, the integration of controlled release strategy within electrospun nanofibers can provide protection for growth factors against in vivo degradation, and can manipulate desired signal at an effective level with extended duration in local microenvironment to support tissue regeneration and repair which normally takes a much longer time. In this review, we provide an overview of growth factor delivery using biomimetic electrospun nanofiber scaffolds in bone tissue engineering. It begins with a brief introduction of different kinds of polymers that were used in electrospinning and their applications in bone tissue engineering. The review further focuses on the nanofiber-based growth factor delivery and summarizes the strategies of growth factors loading on the nanofiber scaffolds for bone tissue engineering applications. The perspectives on future challenges in this area are also pointed out.

Pectin-chitosan-PVA nanofibrous scaffold made by electrospinning and its potential use as a skin tissue scaffold.

PubMed

Lin, Hsin-Yi; Chen, Hsin-Hung; Chang, Shih-Hsin; Ni, Tsung-Sheng

2013-01-01

Scaffolds made of chitosan nanofibers are often too mechanically weak for their application and often their manufacturing processes involve the use of harmful and flammable organic solvents. In the attempt to improve the mechanical properties of nanofibrous scaffolds made of chitosan without the use of harmful chemicals, pectin, an anionic polymer was blended with chitosan, a cationic polymer, to form a polyelectrolyte complex and electrospun into nanofibers for the first time. The electrospun chitosan-pectin scaffolds, when compared to electrospun chitosan scaffolds, had a 58% larger diameter, a 21% higher Young's modulus, a 162% larger strain at break, and a 104% higher ultimate tensile strength. Compared to the chitosan scaffolds, the chitosan-pectin scaffolds' swelling ratios decreased by 55% after 60 min in a saline solution and more quickly released the preloaded tetracycline HCl. The L929 fibroblast cells proliferated slightly slower on the chitosan-pectin scaffolds than on the chitosan scaffolds. Nonetheless, cells on both materials deposited similar levels of extracellular type I collagen on a per DNA basis. In conclusion, a novel chitosan-pectin nanofibrous scaffold with superior mechanical properties than a chitosan nanofibrous scaffold was successfully made without the use of harmful solvents.

Mild process to design silk scaffolds with reduced β-sheet structure and various topographies at nanometer scale

PubMed Central

Pei, Yazhen; Liu, Xi; Liu, Shanshan; Lu, Qiang; Liu, Jing; Kaplan, David L; Zhu, Hesun

2014-01-01

Three-dimensional (3D) porous silk scaffolds with good biocompatibility and minimal immunogenicity, have promising applications in different tissue regenerations. However, a challenge remains to effectively fabricate their microstructures and mechanical properties to satisfy specific requirements of different tissues. In this study, silk scaffolds were fabricated to form extracellular matrix (ECM) mimetic nanofibrous architecture in a mild process. A slowly increasing concentration process was applied to regulate silk self-assembly into nanofibers in aqueous solution. Then glycerol was blended with the nanofiber solution and induced silk crystallization in lyophilization process, endowing freeze-dried scaffolds water-stability. The glycerol was leached from the scaffolds, leaving similar porous structure at a micrometer scale but different topographies at nanoscale. Compared to previous salt-leached and methanol annealed scaffolds, the present scaffolds showed lower β-sheet content, softer mechanical property, and improved cell growth and differentiation behaviors, implying their promising future as platforms for controlling stem cell fate and soft tissue regeneration. PMID:25463497

Living nanofiber yarn-based woven biotextiles for tendon tissue engineering using cell tri-culture and mechanical stimulation.

PubMed

Wu, Shaohua; Wang, Ying; Streubel, Philipp N; Duan, Bin

2017-10-15

Non-woven nanofibrous scaffolds have been developed for tendon graft application by using electrospinning strategies. However, electrospun nanofibrous scaffolds face some obstacles and limitations, including suboptimal scaffold structure, weak tensile and suture-retention strengths, and compact structure for cell infiltration. In this work, a novel nanofibrous, woven biotextile, fabricated based on electrospun nanofiber yarns, was implemented as a tissue engineered tendon scaffold. Based on our modified electrospinning setup, polycaprolactone (PCL) nanofiber yarns were fabricated with reproducible quality, and were further processed into plain-weaving fabrics interlaced with polylactic acid (PLA) multifilaments. Nonwoven nanofibrous PCL meshes with random or aligned fiber structures were generated using typical electrospinning as comparative counterparts. The woven fabrics contained 3D aligned microstructures with significantly larger pore size and obviously enhanced tensile mechanical properties than their nonwoven counterparts. The biological results revealed that cell proliferation and infiltration, along with the expression of tendon-specific genes by human adipose derived mesenchymal stem cells (HADMSC) and human tenocytes (HT), were significantly enhanced on the woven fabrics compared with those on randomly-oriented or aligned nanofiber meshes. Co-cultures of HADMSC with HT or human umbilical vein endothelial cells (HUVEC) on woven fabrics significantly upregulated the functional expression of most tenogenic markers. HADMSC/HT/HUVEC tri-culture on woven fabrics showed the highest upregulation of most tendon-associated markers than all the other mono- and co-culture groups. Furthermore, we conditioned the tri-cultured constructs with dynamic conditioning and demonstrated that dynamic stretch promoted total collagen secretion and tenogenic differentiation. Our nanofiber yarn-based biotextiles have significant potential to be used as engineered scaffolds to synergize the multiple cell interaction and mechanical stimulation for promoting tendon regeneration. Tendon grafts are essential for the treatment of various tendon-related conditions due to the inherently poor healing capacity of native tendon tissues. In this study, we combined electrospun nanofiber yarns with textile manufacturing strategies to fabricate nanofibrous woven biotextiles with hierarchical features, aligned fibrous topography, and sufficient mechanical properties as tendon tissue engineered scaffolds. Comparing to traditional electrospun random or aligned meshes, our novel nanofibrous woven fabrics possess strong tensile and suture-retention strengths and larger pore size. We also demonstrated that the incorporation of tendon cells and vascular cells promoted the tenogenic differentiation of the engineered tendon constructs, especially under dynamic stretch. This study not only presents a novel tissue engineered tendon scaffold fabrication technique but also provides a useful strategy to promote tendon differentiation and regeneration. Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Fabrication, biocompatibility, and tissue engineering substrate analysis of polyvinyl alcohol-gelatin core-shell electrospun nanofibers

NASA Astrophysics Data System (ADS)

Merkle, Valerie Marie

Cardiovascular disease is the leading cause of death in the United States with approximately 49% of the cardiovascular related deaths attributed to coronary heart disease (CHD). CHD is the accumulation of plaque resulting in the narrowing of the vessel lumen and a decrease in blood flow to the downstream heart muscle. In order to restore blood flow, arterial by-pass procedures can be undertaken. However, the patient's own arteries/veins may not be suitable for use as a vessel replacement, and synthetic grafts lack the compliancy and durability needed for these small diameter locations (< 5 mm). Therefore, the goal of this research is to develop a nanofibrous material that can be used in vascular applications such as this. In this study, we fabricate coaxial electrospun nanofibers with gelatin in the shell and polyvinyl alcohol (PVA) in the core using 1 Gelatin: 1 PVA and 3 Gelatin: 1 PVA mass ratios. Gelatin, derived from collagen, is highly bioactive while PVA, a synthetic polymer, has appealing mechanical properties. Therefore, by combining these materials in a core-shell structure, we hypothesize that the resulting nanofibers will have enhanced mechanical properties, cellular growth and migration, as well as minimal platelet deposition and activation compared to scaffolds composed solely of gelatin or PVA. First, the coaxial scaffolds exhibited an enhanced Young's modulus and ultimate strength compared to scaffolds composed of PVA or gelatin alone. Endothelial cells had high proliferation and migration on the coaxial electrospun scaffolds with higher migration seen on the stiffer, coaxial scaffolds. The smooth muscle cells had less proliferation and lower migration rates on the coaxial scaffolds than the endothelial cells. Using a modified prothrombinase assay, the coaxial scaffolds had minimal platelet activation. Lastly, when pre-seeding the coaxial scaffolds with endothelial cells or smooth muscle cells, the platelet deposition decreased in comparison to platelet deposition with no cell pre-seeding. Overall, the 1 Gel: 1 PVA coaxial scaffolds promoted endothelial cell growth and migration, minimized smooth muscle cell growth and migration, and had minimal platelet activation. Therefore, the 1 Gel: 1 PVA coaxial nanofibers are an intriguing material for use in vascular applications.

Modifying the mechanical properties of silk nanofiber scaffold by knitted orientation for regenerative medicine applications.

PubMed

Dodel, M; Hemmati Nejad, N; Bahrami, S H; Soleimani, M; Hanaee-Ahvaz, H

2016-08-31

Tissue reconstruction is among the increasing applications of polymer nanofibers. Fibrous scaffolds (mats) can be easily produced using the electrospinning method with structure and biomechanical properties similar to those of a cellular matrix. Electrospinning is widely used in the production of nanofibers and the GAP-method electrospinning is one of the means of producing fully aligned nanofibers. In this research, using the GAP-method, knitted fibrous scaffolds were made of silk fibroin, which is a biocompatible and biodegradable polymer. To extract fibroin from cocoons, the sodium chloride solution as well as dialysis and freeze-drying techniques were employed. The molecular weight of the extracted fibroin was measured with the SDS-Page electrophoresis technique. Moreover, the pure fibroin structure was examined using the ATR-FTIR method, and the viscosity of the solution used for electrospinning was measured with the Brookfield rotational viscometer. The scaffolds were prepared through electrospinning of the silk fibroin in pure formic acid solution. The following three structures were electrospun: 1) a random structure; 2) a knitted structure with an interstitial angle of 60 degrees; 3) a knitted structure with an interstitial angle of 90 degrees. Morphology of the resulting fibers was studied with a SEM (scanning electron microscope). Fibroin scaffolds are degradable in water. Therefore, they were fixated through immersion in methanol to be prepared for assays. The mechanical properties of the scaffolds were also studied using a tensile strength test device. The effect of methanol on the strength properties of the samples was also assessed. The hydrophilic potential of the samples was measured via a contact angle test. To increase the hydrophilicity of the scaffold surfaces, the cold oxygen plasma technique was employed. Finally, the biocompatibility and cell adhesion of the resulting scaffolds were examined through a HEK 293 cell culture, and the results were analyzed through the MTT, DAPI staining, and SEM imaging techniques. Results revealed that the oriented knitted structure contributed to the increase in Young's modulus and the maximum strength of scaffolds as compared to the random samples. Moreover, this structure can also be a suitable alternative to the typical chemical means of increasing strength.

Design of a nanocomposite substrate inducing adult stem cell assembly and progression toward an Epiblast-like or Primitive Endoderm-like phenotype via mechanotransduction.

PubMed

Morena, Francesco; Armentano, Ilaria; Montanucci, Pia; Argentati, Chiara; Fortunati, Elena; Montesano, Simona; Bicchi, Ilaria; Pescara, Teresa; Pennoni, Ilaria; Mattioli, Samantha; Torre, Luigi; Latterini, Loredana; Emiliani, Carla; Basta, Giuseppe; Calafiore, Riccardo; Kenny, Josè Maria; Martino, Sabata

2017-11-01

This work shows that the active interaction between human umbilical cord matrix stem cells and Poly (l-lactide)acid (PLLA) and PLLA/Multi Walled Carbon Nanotubes (MWCNTs) nanocomposite films results in the stem cell assembly as a spheroid conformation and affects the stem cell fate transition. We demonstrated that spheroids directly respond to a tunable surface and the bulk properties (electric, dielectric and thermal) of plain and nanocomposite PLLA films by triggering a mechanotransduction axis. This stepwise process starts from tethering of the cells' focal adhesion proteins to the surface, together with the adherens junctions between cells. Both complexes transmit traction forces to F-Actin stress fibres that link Filamin-A and Myosin-IIA proteins, generating a biological scaffold, with increased stiffening conformation from PLLA to PLLA/MWCNTs, and enable the nucleoskeleton proteins to boost chromatin reprogramming processes. Herein, the opposite expression of NANOG and GATA6 transcription factors, together with other lineage specification related proteins, steer spheroids toward an Epiblast-like or Primitive Endoderm-like lineage commitment, depending on the absence or presence of 1 wt% MWCNTs, respectively. This work represents a pioneering effort to create a stem cell/material interface that can model the stem cell fate transition under growth culture conditions. Copyright © 2017 Elsevier Ltd. All rights reserved.

Inflammation-sensitive in situ smart scaffolding for regenerative medicine.

PubMed

Patra, Hirak K; Sharma, Yashpal; Islam, Mohammad Mirazul; Jafari, Mohammad Javad; Murugan, N Arul; Kobayashi, Hisatoshi; Turner, Anthony P F; Tiwari, Ashutosh

2016-10-06

To cope with the rapid evolution of the tissue engineering field, it is now essential to incorporate the use of on-site responsive scaffolds. Therefore, it is of utmost importance to find new 'Intelligent' biomaterials that can respond to the physicochemical changes in the microenvironment. In this present report, we have developed biocompatible stimuli responsive polyaniline-multiwalled carbon nanotube/poly(N-isopropylacrylamide), (PANI-MWCNT/PNIPAm) composite nanofiber networks and demonstrated the physiological temperature coordinated cell grafting phenomenon on its surface. The composite nanofibers were prepared by a two-step process initiated with an assisted in situ polymerization followed by electrospinning. To obtain a smooth surface in individual nanofibers with the thinnest diameter, the component ratios and electrospinning conditions were optimized. The temperature-gated rearrangements of the molecular structure are characterized by FTIR spectroscopy with simultaneous macromolecular architecture changes reflected on the surface morphology, average diameter and pore size as determined by scanning electron microscopy. The stimuli responsiveness of the nanofibers has first been optimized with computational modeling of temperature sensitive components (coil-like and globular conformations) to tune the mechanism for temperature dependent interaction during in situ scaffolding with the cell membrane. The nanofiber networks show excellent biocompatibility, tested with fibroblasts and also show excellent sensitivity to inflammation to combat loco-regional acidosis that delay the wound healing process by an in vitro model that has been developed for testing the proposed responsiveness of the composite nanofiber networks. Cellular adhesion and detachment are regulated through physiological temperature and show normal proliferation of the grafted cells on the composite nanofibers. Thus, we report for the first time, the development of physiological temperature gated inflammation-sensitive smart biomaterials for advanced tissue regeneration and regenerative medicine.

Enhanced biocompatibility of PLGA nanofibers with gelatin/nano-hydroxyapatite bone biomimetics incorporation.

PubMed

Li, Daowei; Sun, Haizhu; Jiang, Liming; Zhang, Kai; Liu, Wendong; Zhu, Yang; Fangteng, Jiaozi; Shi, Ce; Zhao, Liang; Sun, Hongchen; Yang, Bai

2014-06-25

The biocompatibility of biomaterials is essentially for its application. The aim of current study was to evaluate the biocompatibility of poly(lactic-co-glycolic acid) (PLGA)/gelatin/nanohydroxyapatite (n-HA) (PGH) nanofibers systemically to provide further rationales for the application of the composite electrospun fibers as a favorable platform for bone tissue engineering. The PGH composite scaffold with diameter ranging from nano- to micrometers was fabricated by using electrospinning technique. Subsequently, we utilized confocal laser scanning microscopy (CLSM) and MTT assay to evaluate its cyto-compatibility in vitro. Besides, real-time quantitative polymerase chain reaction (qPCR) analysis and alizarin red staining (ARS) were performed to assess the osteoinductive activity. To further test in vivo, we implanted either PLGA or PGH composite scaffold in a rat subcutaneous model. The results demonstrated that PGH scaffold could better support osteoblasts adhesion, spreading, and proliferation and show better cyto-compatibility than pure PLGA scaffold. Besides, qPCR analysis and ARS showed that PGH composite scaffold exhibited higher osteoinductive activity owing to higher phenotypic expression of typical osteogenic genes and calcium deposition. The histology evaluation indicated that the incorporation of Gelatin/nanohydroxyapatite (GH) biomimetics could significantly reduce local inflammation. Our data indicated that PGH composite electrospun nanofibers possessed excellent cyto-compatibility, good osteogenic activity, as well as good performance of host tissue response, which could be versatile biocompatible scaffolds for bone tissue engineering.

Fabrication, characterization, and biocompatibility assessment of a novel elastomeric nanofibrous scaffold: A potential scaffold for soft tissue engineering.

PubMed

Shamirzaei Jeshvaghani, Elham; Ghasemi-Mobarakeh, Laleh; Mansurnezhad, Reza; Ajalloueian, Fatemeh; Kharaziha, Mahshid; Dinari, Mohammad; Sami Jokandan, Maryam; Chronakis, Ioannis S

2017-11-23

With regard to flexibility and strength properties requirements of soft biological tissue, elastomeric materials could be more beneficial in soft tissue engineering applications. The present work investigates the use of an elastic polymer, (polycaprolactone fumarate [PCLF]), for fabricating an electrospun scaffold. PCLF with number-average molecular weight of 13,284 g/mol was synthetized, electrospun PCLF:polycaprolactone (PCL) (70:30) nanofibrous scaffolds were fabricated and a novel strategy (in situ photo-crosslinking along with wet electrospinning) was applied for crosslinking of PCLF in the structure of PCLF:PCL nanofibers was presented. Sol fraction results, Fourier-transform infrared spectroscopy, and mechanical tests confirmed occurrence of crosslinking reaction. Strain at break and Young's modulus of crosslinked PCLF:PCL nanofibers fabricated was found to be 114.5 ± 3.9% and 0.6 ± 0.1 MPa, respectively, and dynamic mechanical analysis results revealed elasticity of nanofibers. MTS assay showed biocompatibility of PCLF:PCL (70:30) nanofibrous scaffolds. Our overall results showed that electrospun PCLF:PCL nanofibrous scaffold could be considered as a candidate for further in vitro and in vivo experiments and its application for engineering of soft tissues subjected to in vivo cyclic mechanical stresses. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2017. © 2017 Wiley Periodicals, Inc.

Biomacromolecule conjugated nanofiber scaffold for salivary gland tissue engineering

NASA Astrophysics Data System (ADS)

Jayarathanam, Kavitha

Xerostomia or dry mouth, resulting from loss of salivary gland secretion can be alleviated by tissue engineering approaches to restore glandular cell function. Engineering an artificial salivary gland structure requires closely mimicking the natural environment, both physically and functionally, to promote epithelial cell proliferation, monolayer formation and apico-basal polarization. While the physical structure of the salivary gland extracellular matrix (ECM) can be reconstructed using biocompatible nanofiber scaffolds, the chemical signals from ECM macromolecules are equally involved in the gland morphogenesis. In these glands, Hyaluronic acid (HA), a biomacromolecule that is a major component of the ECM, plays a crucial role in recruiting growth factors to improve cell viability and growth in these glands. Another molecule of interest that improved salivary epithelial cell viability and apico-basal differentiation is laminin, a major protein found in the basement membrane. We hypothesize that these biomacromolecules, when conjugated nanofiber scaffolds, will provide the essential chemical signals that promote cell viability, proliferation, polarity in the salivary cell line of interest. These morphological changes will in turn promote the secretory function (salivary production). The nanofiber scaffold consisting of poly(lactic-co-glycolic)acid is conjugated with HA using a polyethylene glycol (PEG) diamine crosslinker. This conjugation was confirmed using fluorescence spectrometry, water contact angle test and immunocytochemistry analysis using confocal microscopy. The effect of HA in promoting cell survival in-vitro was established with MTT assay using SIMS (mouse submandibular immortalized ductal SIMS cells) cells. The effect of HA in improving the apico - basal polarity of SIMS cells will be assessed. Chemical modification of synthetic nanopolymeric scaffolds with ECM molecules e.g., HA, laminin are the next step towards developing "smart scaffolds", that can be used to specifically induce proper salivary gland function. These scaffolds can potentially be used to provide a viable approach for creating future artificial tissue engineered glands.

Experimental and computational studies of poly-L-lactic acid for cardiovascular applications: recent progress

NASA Astrophysics Data System (ADS)

Naseem, Raasti; Zhao, Liguo; Liu, Yang; Silberschmidt, Vadim V.

2017-12-01

Stents are commonly used in medical procedures to alleviate the symptoms of coronary heart disease, a prevalent modern society disease. These structures are employed to maintain vessel patency and restore blood flow. Traditionally stents are made of metals such as stainless steel or cobalt chromium; however, these scaffolds have known disadvantages. An emergence of transient scaffolds is gaining popularity, with the structure engaged for a required period whilst healing of the diseased arterial wall occurs. Polymers dominate a medical device sector, with incorporation in sutures, scaffolds and screws. Thanks to their good mechanical and biological properties and their ability to degrade naturally. Polylactic acid is an extremely versatile polymer, with its properties easily tailored to applications. Its dominance in the stenting field increases continually, with the first polymer scaffold gaining FDA approval in 2016. Still some challenges with PLLA bioresorbable materials remain, especially with regard to understanding their mechanical response, assessment of its changes with degradation and comparison of their performance with that of metallic drug-eluting stent. Currently, there is still a lack of works on evaluating both the pre-degradation properties and degradation performance of these scaffolds. Additionally, there are no established material models incorporating non-linear viscoelastic behaviour of PLLA and its evolution with in-service degradation. Assessing these features through experimental analysis accompanied by analytical and numerical studies will provide powerful tools for design and optimisation of these structures endorsing their broader use in stenting. This overview assesses the recent studies investigating mechanical and computational performance of poly(l-lactic) acid and its use in stenting applications.

Pulp regeneration in a full-length human tooth root using a hierarchical nanofibrous microsphere system.

PubMed

Li, Xiangwei; Ma, Chi; Xie, Xiaohua; Sun, Hongchen; Liu, Xiaohua

2016-04-15

While pulp regeneration using tissue engineering strategy has been explored for over a decade, successful regeneration of pulp tissues in a full-length human root with a one-end seal that truly simulates clinical endodontic treatment has not been achieved. To address this challenge, we designed and synthesized a unique hierarchical growth factor-loaded nanofibrous microsphere scaffolding system. In this system, vascular endothelial growth factor (VEGF) binds with heparin and is encapsulated in heparin-conjugated gelatin nanospheres, which are further immobilized in the nanofibers of an injectable poly(l-lactic acid) (PLLA) microsphere. This hierarchical microsphere system not only protects the VEGF from denaturation and degradation, but also provides excellent control of its sustained release. In addition, the nanofibrous PLLA microsphere integrates the extracellular matrix-mimicking architecture with a highly porous injectable form, efficiently accommodating dental pulp stem cells (DPSCs) and supporting their proliferation and pulp tissue formation. Our in vivo study showed the successful regeneration of pulp-like tissues that fulfilled the entire apical and middle thirds and reached the coronal third of the full-length root canal. In addition, a large number of blood vessels were regenerated throughout the canal. For the first time, our work demonstrates the success of pulp tissue regeneration in a full-length root canal, making it a significant step toward regenerative endodontics. The regeneration of pulp tissues in a full-length tooth root canal has been one of the greatest challenges in the field of regenerative endodontics, and one of the biggest barriers for its clinical application. In this study, we developed a unique approach to tackle this challenge, and for the first time, we successfully regenerated living pulp tissues in a full-length root canal, making it a significant step toward regenerative endodontics. This study will make positive scientific impact and interest the broad and multidisciplinary readership in the dental biomaterials and craniofacial tissue engineering community. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Glycosaminoglycan-Mimetic Signals Direct the Osteo/Chondrogenic Differentiation of Mesenchymal Stem Cells in a Three-Dimensional Peptide Nanofiber Extracellular Matrix Mimetic Environment.

PubMed

Arslan, Elif; Guler, Mustafa O; Tekinay, Ayse B

2016-04-11

Recent efforts in bioactive scaffold development focus strongly on the elucidation of complex cellular responses through the use of synthetic systems. Designing synthetic extracellular matrix (ECM) materials must be based on understanding of cellular behaviors upon interaction with natural and artificial scaffolds. Hence, due to their ability to mimic both the biochemical and mechanical properties of the native tissue environment, supramolecular assemblies of bioactive peptide nanostructures are especially promising for development of bioactive ECM-mimetic scaffolds. In this study, we used glycosaminoglycan (GAG) mimetic peptide nanofiber gel as a three-dimensional (3D) platform to investigate how cell lineage commitment is altered by external factors. We observed that amount of fetal bovine serum (FBS) presented in the cell media had synergistic effects on the ability of GAG-mimetic nanofiber gel to mediate the differentiation of mesenchymal stem cells into osteogenic and chondrogenic lineages. In particular, lower FBS concentration in the culture medium was observed to enhance osteogenic differentiation while higher amount FBS promotes chondrogenic differentiation in tandem with the effects of the GAG-mimetic 3D peptide nanofiber network, even in the absence of externally administered growth factors. We therefore demonstrate that mesenchymal stem cell differentiation can be specifically controlled by the combined influence of growth medium components and a 3D peptide nanofiber environment.

Antibacterial performance and in vivo diabetic wound healing of curcumin loaded gum tragacanth/poly(ε-caprolactone) electrospun nanofibers.

PubMed

Ranjbar-Mohammadi, Marziyeh; Rabbani, Shahram; Bahrami, S Hajir; Joghataei, M T; Moayer, F

2016-12-01

In this study we describe the potential of electrospun curcumin-loaded poly(ε-caprolactone) (PCL)/gum tragacanth (GT) (PCL/GT/Cur) nanofibers for wound healing in diabetic rats. These scaffolds with antibacterial property against methicillin resistant Staphylococcus aureus as gram positive bacteria and extended spectrum β lactamase as gram negative bacteria were applied in two forms of acellular and cell-seeded for assessing their capability in healing full thickness wound on the dorsum of rats. After 15days, pathological study showed that the application of GT/PCL/Cur nanofibers caused markedly fast wound closure with well-formed granulation tissue dominated by fibroblast proliferation, collagen deposition, complete early regenerated epithelial layer and formation of sweat glands and hair follicles. No such appendage formation was observed in the untreated controls during this duration. Masson's trichrome staining confirmed the increased presence of collagen in the dermis of the nanofiber treated wounds on day 5 and 15, while the control wounds were largely devoid of collagen on day 5 and exhibited less collagen amount on day 15. Quantification analysis of scaffolds on day 5 confirmed that, tissue engineered scaffolds with increased amount of angiogenesis number, granulation tissue area (μ(2)), fibroblast number, and decreased epithelial gap (μ) can be more effective compared to GT/PCL/Cur nanofibers. Copyright © 2016 Elsevier B.V. All rights reserved.

Mitigating Scarring and Inflammation during Corneal Wound Healing using Nanofiber-Hydrogel Scaffolds

NASA Astrophysics Data System (ADS)

Fu, Amy

Due to the universal lack of donor tissue, there has been emerging interest in engineering materials to stimulate living cells to restore the features and functions of injured organs. We are particularly interested in developing materials for corneal use, where the necessity to maintain the tissue's transparency presents an additional challenge. Every year, there are 1.5 -- 2 million new cases of monocular blindness due to irregular healing of corneal injuries, dwarfing the approximately 150,000 corneal transplants performed. The large gap between the need and availability of cornea transplantation motivates us to develop a wound-healing scaffold that can prevent corneal blindness. To develop such a scaffold, it is necessary to regulate the cells responsible for repairing the damaged cornea, namely myofibroblasts, which are responsible for the disordered and non-refractive index matched scar that leads to corneal blindness. Using in vitro assays, we identified that protein nanofibers of certain orientation can promote cell migration and modulate the myofibroblast phenotype. The nanofibers are also transparent, easy to handle and non-cytotoxic. To adhere the nanofibers to a wound bed, we examined the use of two different in situ forming hydrogels: an artificial extracellular matrix protein (aECM)-based gel and a photo-crosslinkable heparin-based gel. Both hydrogels can be formed within minutes, are transparent upon gelation and are easily tunable. Using an in vivo mouse model for epithelial defects, we show that our corneal scaffolds (nanofibers together with hydrogel) are well-tolerated (no inflammatory response or turbidity) and support epithelium regrowth. We developed an ex vivo corneal tissue culture model where corneas that are wounded and treated with our scaffold can be cultured while retaining their ability to repair wounds for up to 21 days. Using this technique, we found that the aECM-based treatment induced a more favorable wound response than the heparin-based treatment, prompting us to further examine the efficacy of the aECM-based treatment in vivo using a rabbit model for stromal wounds. Results show that treated corneas have fewer myofibroblasts and immune cells than untreated ones, indicating that our corneal scaffold shows promise in promoting a calmer wound response and preventing corneal haze formation.

Spatiotemporal Oxygen Sensing Using Dual Emissive Boron Dye–Polylactide Nanofibers

PubMed Central

2015-01-01

Oxygenation in tissue scaffolds continues to be a limiting factor in regenerative medicine despite efforts to induce neovascularization or to use oxygen-generating materials. Unfortunately, many established methods to measure oxygen concentration, such as using electrodes, require mechanical disturbance of the tissue structure. To address the need for scaffold-based oxygen concentration monitoring, a single-component, self-referenced oxygen sensor was made into nanofibers. Electrospinning process parameters were tuned to produce a biomaterial scaffold with specific morphological features. The ratio of an oxygen sensitive phosphorescence signal to an oxygen insensitive fluorescence signal was calculated at each image pixel to determine an oxygenation value. A single component boron dye–polymer conjugate was chosen for additional investigation due to improved resistance to degradation in aqueous media compared to a boron dye polymer blend. Standardization curves show that in fully supplemented media, the fibers are responsive to dissolved oxygen concentrations less than 15 ppm. Spatial (millimeters) and temporal (minutes) ratiometric gradients were observed in vitro radiating outward from the center of a dense adherent cell grouping on scaffolds. Sensor activation in ischemia and cell transplant models in vivo show oxygenation decreases on the scale of minutes. The nanofiber construct offers a robust approach to biomaterial scaffold oxygen sensing. PMID:25426706

Bioactive nanofibers for fibroblastic differentiation of mesenchymal precursor cells for ligament/tendon tissue engineering applications.

PubMed

Sahoo, Sambit; Ang, Lay-Teng; Cho-Hong Goh, James; Toh, Siew-Lok

2010-02-01

Mesenchymal stem cells and precursor cells are ideal candidates for tendon and ligament tissue engineering; however, for the stem cell-based approach to succeed, these cells would be required to proliferate and differentiate into tendon/ligament fibroblasts on the tissue engineering scaffold. Among the various fiber-based scaffolds that have been used in tendon/ligament tissue engineering, hybrid fibrous scaffolds comprising both microfibers and nanofibers have been recently shown to be particularly promising. With the nanofibrous coating presenting a biomimetic surface, the scaffolds can also potentially mimic the natural extracellular matrix in function by acting as a depot for sustained release of growth factors. In this study, we demonstrate that basic fibroblast growth factor (bFGF) could be successfully incorporated, randomly dispersed within blend-electrospun nanofibers and released in a bioactive form over 1 week. The released bioactive bFGF activated tyrosine phosphorylation signaling within seeded BMSCs. The bFGF-releasing nanofibrous scaffolds facilitated BMSC proliferation, upregulated gene expression of tendon/ligament-specific ECM proteins, increased production and deposition of collagen and tenascin-C, reduced multipotency of the BMSCs and induced tendon/ligament-like fibroblastic differentiation, indicating their potential in tendon/ligament tissue engineering applications. 2009 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.

Electrospun Silk Biomaterial Scaffolds for Regenerative Medicine

PubMed Central

Zhang, Xiaohui; Reagan, Michaela R; Kaplan, David L.

2009-01-01

Electrospinning is a versatile technique that enables the development of nanofiber-based biomaterial scaffolds. Scaffolds can be generated that are useful for tissue engineering and regenerative medicine since they mimic the nanoscale properties of certain fibrous components of the native extracellular matrix in tissues. Silk is a natural protein with excellent biocompatibility, remarkable mechanical properties as well as tailorable degradability. Integrating these protein polymer advantages with electrospinning results in scaffolds with combined biochemical, topographical and mechanical cues with versatility for a range of biomaterial, cell and tissue studies and applications. This review covers research related to electrospinning of silk, including process parameters, post treatment of the spun fibers, functionalization of nanofibers, and the potential applications for these material systems in regenerative medicine. Research challenges and future trends are also discussed. PMID:19643154

Matrigel immobilization on the shish-kebab structured poly(ɛ-caprolactone) nanofibers for skin tissue engineering

NASA Astrophysics Data System (ADS)

Jing, Xin; Mi, Hao-Yang; Peng, Xiang-Fang; Turng, Lih-Sheng

2016-03-01

Surface properties of tissue engineering scaffolds such as topography, hydrophilicity, and functional groups play a vital role in cell adhesion, migration, proliferation, and apoptosis. First, poly(ɛ-caprolactone) (PCL) shish-kebab scaffolds (PCL-SK), which feature a three-dimensional structure comprised of electrospun PCL nanofibers covered by periodic, self-induced PCL crystal lamellae on the surface, was created to mimic the nanotopography of native collagen fibrils in the extracellular matrix (ECM). Second, matrigel was covalently immobilized on the surface of alkaline hydrolyzed PCL-SK scaffolds to enhance their hydrophilicity. This combined approach not only mimics the nanotopography of native collagen fibrils, but also simulates the surface features of collagen fibrils for cell growth. To investigate the viability of such scaffolds, HEF1 fibroblast cell assays were conducted and the results revealed that the nanotopography of the PCL-SK scaffolds facilitated cell adhesion and proliferation. The matrigel functionalization on PCL-SK scaffolds further enhanced cellular response, which suggested elevated biocompatibility and greater potential for skin tissue engineering applications.

Preparation and characterization of electrospun alginate/PLA nanofibers as tissue engineering material by emulsion eletrospinning.

PubMed

Xu, Weihong; Shen, Renzhe; Yan, Yurong; Gao, Jie

2017-01-01

Scaffolds made by biomaterials offer favorite environment for cell grow and show a wide potential application in tissue engineering. Novel biocompatibility materials polylatic acid (PLA) nanofiber membranes with favorable biocompatibility and good mechanical strength could serve as an innovative tissue engineering scaffold. Sodium alginate (SA) could be used in biomedical areas because of its anti-bacterial property, hydrophilicity and biocompatibility. In this article, we chose PLA as continuous phase and SA as dispersion phase to prepare a W/O emulsion and then electrospun it to get a SA/PLA composite nanofiber membranes. The CLSM images illustrated that the existence of SA was located on the surface of composite fibers and the FTIR results confirmed the result. A calcium ion replacement step was used as an after-treatment for SA/PLA nanofiber membranes in order to anchor the alginic ion in a form of gelated calcium alginate (CA). The single fiber tensile test shows a good mechanical property of CA/PLA nanofiber membranes, and the nanofiber membranes are beneficial for cell proliferation and differentiation owing to MTT array as well as Alizarin red S (ARS) staining test. Copyright © 2016 Elsevier Ltd. All rights reserved.

Engineering poly(hydroxy butyrate-co-hydroxy valerate) based vascular scaffolds to mimic native artery.

PubMed

Deepthi, S; Nivedhitha Sundaram, M; Vijayan, Ponni; Nair, Shantikumar V; Jayakumar, R

2018-04-01

Electrospun tri-layered fibrous scaffold incorporating VEGF and Platelet Factor Concentrate (PFC) in multiple layers having different layer architectures was designed to mimic native artery. The scaffold consisted of longitudinally aligned poly(hydroxy butyrate-co-hydroxy valerate) (PHBV) and poly(vinyl alcohol) (PVA) nanofibers (inner layer), radially aligned PHBV-elastin nanofibers (middle layer) to provide the bi-directional alignment and combination of longitudinally aligned PHBV-elastin and random PHBV/PVA multiscale fibers (peripheral layer). Tubular constructs of diameter <6 mm were developed. The developed electrospun fibers were characterised by Scanning Electron Microscope (SEM), Fourier Transform Infrared Spectroscopy and Tensile tests. Further the burst strength, compliance and stiffness index of tri-layered tubular scaffold was evaluated. SEM images of fibrous layers showed the typical longitudinal and radial alignment of fibers in the tubular construct. SEM images showed that the prepared PHBV nanofibers were in the range of 500-800 nm and PHBV microfibers were of 1-2 μm in diameter in the tri-layered electrospun membrane. PVA nanofibers were of size 200-250 nm. The tensile strength, percentage compliance and stiffness index of tri-layered membrane was in accordance with that of native small blood vessels. The developed tri-layered membrane was blood compatible, with hemolysis degree 0.85 ± 0.21% and did not activate platelets. Controlled release of VEGF and PFC was observed from the scaffold. The biocompatibility of the tri-layered scaffold was evaluated using HUVECs, SMCs and MSCs and SMCs infiltration from the outer layer was also evaluated. Specific protein expression for the HUVECs and SMCs was evaluated by flow cytometry and immunocytochemistry. HUVECs and SMCs exhibited good elongation and alignment along the direction of fibers and was found to maintain its CD31, VE-Cadherin and αSMA expression respectively. The results highlight the importance of bi-directional fiber alignment on the tri-layered electrospun scaffold as a suitable architectural prototype for vascular scaffolds to mimic the native arteries. Copyright © 2017 Elsevier B.V. All rights reserved.

A mild process to design silk scaffolds with reduced β-sheet structure and various topographies at the nanometer scale.

PubMed

Pei, Yazhen; Liu, Xi; Liu, Shanshan; Lu, Qiang; Liu, Jing; Kaplan, David L; Zhu, Hesun

2015-02-01

Three-dimensional (3-D) porous silk scaffolds with good biocompatibility and minimal immunogenicity show promise in a range of tissue regeneration applications. However, the challenge remains to effectively fabricate their microstructures and mechanical properties to satisfy the specific requirements of different tissues. In this study, silk scaffolds were fabricated to form an extracellular matrix (ECM) mimetic nanofibrous architecture using a mild process. A slowly increasing concentration process was applied to regulate silk self-assembly into nanofibers in aqueous solution. Then glycerol was blended with the nanofiber solution and induced silk crystallization in the lyophilization process, endowing freeze-dried scaffolds with water stability. The glycerol was leached from the scaffolds, leaving a similar porous structure at the micrometer scale but different topographies at the nanoscale. Compared to previous salt-leached and methanol-annealed scaffolds, the present scaffolds showed lower β-sheet content, softer mechanical property and improved cell growth and differentiation behaviors, suggesting their promising future as platforms for controlling stem cell fate and soft tissue regeneration. Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Silk scaffolds with tunable mechanical capability for cell differentiation

PubMed Central

Bai, Shumeng; Han, Hongyan; Huang, Xiaowei; Xu, Weian; Kaplan, David L.; Zhu, Hesun; Lu, Qiang

2015-01-01

Bombyx mori silk fibroin is a promising biomaterial for tissue regeneration and is usually considered an “inert” material with respect to actively regulating cell differentiation due to few specific cell signaling peptide domains in the primary sequence and the generally stiffer mechanical properties due to crystalline content formed in processing. In the present study, silk fibroin porous 3D scaffolds with nanostructures and tunable stiffness were generated via a silk fibroin nanofiber-assisted lyophilization process. The silk fibroin nanofibers with high β-sheet content were added into the silk fibroin solutions to modulate the self-assembly, and to directly induce water-insoluble scaffold formation after lyophilization. Unlike previously reported silk fibroin scaffold formation processes, these new scaffolds had lower overall β-sheet content and softer mechanical properties for improved cell compatibility. The scaffold stiffness could be further tuned to match soft tissue mechanical properties, which resulted in different differentiation outcomes with rat bone marrow-derived mesenchymal stem cells towards myogenic and endothelial cells, respectively. Therefore, these silk fibroin scaffolds regulate cell differentiation outcomes due to their mechanical features. PMID:25858557

Preparation of collagen/polyurethane/knitted silk as a composite scaffold for tendon tissue engineering.

PubMed

Sharifi-Aghdam, Maryam; Faridi-Majidi, Reza; Derakhshan, Mohammad Ali; Chegeni, Arash; Azami, Mahmoud

2017-07-01

The main objective of this study was to prepare a hybrid three-dimensional scaffold that mimics natural tendon tissues. It has been found that a knitted silk shows good mechanical strength; however, cell growth on the bare silk is not desirable. Hence, electrospun collagen/polyurethane combination was used to cover knitted silk. A series of collagen and polyurethane solutions (4%-7% w/v) in aqueous acetic acid were prepared and electrospun. According to obtained scanning electron microscopy images from pure collagen and polyurethane nanofibers, concentration was set constant at 5% (w/v) for blend solutions of collagen/polyurethane. Afterward, blend solutions with the weight ratios of 75/25, 50/50 and 25/75 were electrospun. Scanning electron microscopy images demonstrated the smooth and uniform morphology for the optimized nanofibers. The least fibers diameter among three weight ratios was found for collagen/polyurethane (25/75) which was 100.86 ± 40 nm and therefore was selected to be electrospun on the knitted silk. Attenuated total reflectance-Fourier transform infrared spectra confirmed the chemical composition of obtained electrospun nanofibers on the knitted silk. Tensile test of the specimens including blend nanofiber, knitted silk and commercial tendon substitute examined and indicated that collagen/polyurethane-coated knitted silk has appropriate mechanical properties as a scaffold for tendon tissue engineering. Then, Alamar Blue assay of the L929 fibroblast cell line seeded on the prepared scaffolds demonstrated appropriate viability of the cells with a significant proliferation on the scaffold containing more collagen content. The results illustrate that the designed structure would be promising for being used as a temporary substitute for tendon repair.

Towards an ideal polymer scaffold for tendon/ligament tissue engineering

NASA Astrophysics Data System (ADS)

Sahoo, Sambit; Ouyang, Hong Wei; Goh, James Cho-Hong; Tay, Tong-Earn; Toh, Siew Lok

2005-04-01

Tissue engineering holds promise in treating injured tendons and ligaments by replacing the injured tissues with "engineered tissues" with identical mechanical and functional characteristics. A biocompatible, biodegradable, porous scaffold with optimized architecture, sufficient surface area for cell attachment, growth and proliferation, faborable mechanical properties, and suitable degradation rate is a pre-requisite to achieve success with this aproach. Knitted poly(lactide-co-glycolide) (PLGA) scaffolds comprising of microfibers of 25 micron diameter were coated with PLGA nanofibers on their surfaces by electrospinning technique. A cell suspension of pig bone marrow stromal cells (BMSC) was seeded on the scaffolds by pipetting, and the cell-scaffold constructs were cultured in a CO2 incubator, at 37°C for 1-2 weeks. The "engineered tissues" were then assessed for cell attachment and proliferation, tissue formation, and mechanical properties. Nanofibers, of diameter 300-900 nm, were spread randomly over the knitted scaffold. The reduction in pore-size from about 1 mm (in the knitted scaffold) to a few micrometers (in the nano-microscaffold) allowed cell seeding by direct pipetting, and eliminated the need of a cell-delivery system like fibrin gel. BMSCs were seen to attach and proliferate well on the nano-microscaffold, producing abundant extracellular matrix. Mechanical testing revealed that the cell-seeded nano-microscaffolds possessed slightly higher values of failure load, elastic-region stiffness and toe-region stiffness, than the unseeded scaffolds. The combination of superior mechanical strength and integrity of knitted microfibers, with the large surface area and improved hydrophilicity of the electrospun nanofibers facilitated cell attachment and new tissue formation. This holds promise in tissue engineering of tendon/ligament.

Functional 3-D cardiac co-culture model using bioactive chitosan nanofiber scaffolds.

PubMed

Hussain, Ali; Collins, George; Yip, Derek; Cho, Cheul H

2013-02-01

The in vitro generation of a three-dimensional (3-D) myocardial tissue-like construct employing cells, biomaterials, and biomolecules is a promising strategy in cardiac tissue regeneration, drug testing, and tissue engineering applications. Despite significant progress in this field, current cardiac tissue models are not yet able to stably maintain functional characteristics of cardiomyocytes for long-term culture and therapeutic purposes. The objective of this study was to fabricate bioactive 3-D chitosan nanofiber scaffolds using an electrospinning technique and exploring its potential for long-term cardiac function in the 3-D co-culture model. Chitosan is a natural polysaccharide biomaterial that is biocompatible, biodegradable, non-toxic, and cost effective. Electrospun chitosan was utilized to provide structural scaffolding characterized by scale and architectural resemblance to the extracellular matrix (ECM) in vivo. The chitosan fibers were coated with fibronectin via adsorption in order to enhance cellular adhesion to the fibers and migration into the interfibrous milieu. Ventricular cardiomyocytes were harvested from neonatal rats and studied in various culture conditions (i.e., mono- and co-cultures) for their viability and function. Cellular morphology and functionality were examined using immunofluorescent staining for alpha-sarcomeric actin (SM-actin) and gap junction protein, Connexin-43 (Cx43). Scanning electron microscopy (SEM) and light microscopy were used to investigate cellular morphology, spatial organization, and contractions. Calcium indicator was used to monitor calcium ion flux of beating cardiomyocytes. The results demonstrate that the chitosan nanofibers retained their cylindrical morphology in long-term cell cultures and exhibited good cellular attachment and spreading in the presence of adhesion molecule, fibronectin. Cardiomyocyte mono-cultures resulted in loss of cardiomyocyte polarity and islands of non-coherent contractions. However, the cardiomyocyte-fibroblast co-cultures resulted in polarized cardiomyocyte morphology and retained their morphology and function for long-term culture. The Cx43 expression in the fibroblast co-culture was higher than the cardiomyocytes mono-culture and endothelial cells co-culture. In addition, fibroblast co-cultures demonstrated synchronized contractions involving large tissue-like cellular networks. To our knowledge, this is the first attempt to test chitosan nanofiber scaffolds as a 3-D cardiac co-culture model. Our results demonstrate that chitosan nanofibers can serve as a potential scaffold that can retain cardiac structure and function. These studies will provide useful information to develop a strategy that allows us to generate engineered 3-D cardiac tissue constructs using biocompatible and biodegradable chitosan nanofiber scaffolds for many tissue engineering applications. Copyright © 2012 Wiley Periodicals, Inc.

Epitope topography controls bioactivity in supramolecular nanofibers

PubMed Central

Sur, Shantanu; Tantakitti, Faifan; Matson, John B.; Stupp, Samuel I.

2015-01-01

Incorporating bioactivity into artificial scaffolds using peptide epitopes present in the extracellular matrix (ECM) is a well-known approach. A common strategy has involved epitopes that provide cells with attachment points and external cues through interaction with integrin receptors. Although a variety of bioactive sequences have been identified so far, less is known about their optimal display in a scaffold. We report here on the use of self-assembled peptide amphiphile (PA) nanofiber matrices to investigate the impact of spatial presentation of the fibronectin derived epitope RGDS on cell response. Using one, three, or five glycine residues, RGDS epitopes were systematically spaced out from the surface of the rigid nanofibers. We found that cell morphology was strongly affected by the separation of the epitope from the nanofiber surface, with the longest distance yielding the most cell-spreading, bundling of actin filaments, and a round-to-polygonal transformation of cell shape. Cell response to this type of epitope display was also accompanied with activated integrin-mediated signaling and formation of stronger adhesions between cells and substrate. Interestingly, unlike length, changing the molecular flexibility of the linker had minimal influence on cell behavior on the substrate for reasons that remain poorly understood. The use in this study of high persistence length nanofibers rather than common flexible polymers allows us to conclude that epitope topography at the nanoscale structure of a scaffold influences its bioactive properties independent of epitope density and mechanical properties. PMID:25745558

Fast degradable citrate-based bone scaffold promotes spinal fusion.

PubMed

Tang, Jiajun; Guo, Jinshan; Li, Zhen; Yang, Cheng; Xie, Denghui; Chen, Jian; Li, Shengfa; Li, Shaolin; Kim, Gloria B; Bai, Xiaochun; Zhang, Zhongmin; Yang, Jian

2015-07-21

It is well known that high rates of fusion failure and pseudoarthrosis development (5~35%) are concomitant in spinal fusion surgery, which was ascribed to the shortage of suitable materials for bone regeneration. Citrate was recently recognized to play an indispensable role in enhancing osteconductivity and osteoinductivity, and promoting bone formation. To address the material challenges in spinal fusion surgery, we have synthesized mechanically robust and fast degrading citrate-based polymers by incorporating N-methyldiethanolamine (MDEA) into clickable poly(1, 8-octanediol citrates) (POC-click), referred to as POC-M-click. The obtained POC-M-click were fabricated into POC-M-click-HA matchstick scaffolds by compositing with hydroxyapatite (HA) for interbody spinal fusion in a rabbit model. Spinal fusion was analyzed by radiography, manual palpation, biomechanical testing, and histological evaluation. At 4 and 8 weeks post surgery, POC-M-click-HA scaffolds presented optimal degradation rates that facilitated faster new bone formation and higher spinal fusion rates (11.2±3.7, 80±4.5 at week 4 and 8, respectively) than the poly(L-lactic acid)-HA (PLLA-HA) control group (9.3±2.4 and 71.1±4.4) (p<0.05). The POC-M-click-HA scaffold-fused vertebrates possessed a maximum load and stiffness of 880.8±14.5 N and 843.2±22.4 N/mm, respectively, which were also much higher than those of the PLLA-HA group (maximum: 712.0±37.5 N, stiffness: 622.5±28.4 N/mm, p<0.05). Overall, the results suggest that POC-M-click-HA scaffolds could potentially serve as promising bone grafts for spinal fusion applications.

Fast degradable citrate-based bone scaffold promotes spinal fusion

PubMed Central

Tang, Jiajun; Guo, Jinshan; Li, Zhen; Yang, Cheng; Xie, Denghui; Chen, Jian; Li, Shengfa; Li, Shaolin; Kim, Gloria B.; Bai, Xiaochun; Zhang, Zhongmin; Yang, Jian

2015-01-01

It is well known that high rates of fusion failure and pseudoarthrosis development (5~35%) are concomitant in spinal fusion surgery, which was ascribed to the shortage of suitable materials for bone regeneration. Citrate was recently recognized to play an indispensable role in enhancing osteconductivity and osteoinductivity, and promoting bone formation. To address the material challenges in spinal fusion surgery, we have synthesized mechanically robust and fast degrading citrate-based polymers by incorporating N-methyldiethanolamine (MDEA) into clickable poly(1, 8-octanediol citrates) (POC-click), referred to as POC-M-click. The obtained POC-M-click were fabricated into POC-M-click-HA matchstick scaffolds by compositing with hydroxyapatite (HA) for interbody spinal fusion in a rabbit model. Spinal fusion was analyzed by radiography, manual palpation, biomechanical testing, and histological evaluation. At 4 and 8 weeks post surgery, POC-M-click-HA scaffolds presented optimal degradation rates that facilitated faster new bone formation and higher spinal fusion rates (11.2±3.7, 80±4.5 at week 4 and 8, respectively) than the poly(L-lactic acid)-HA (PLLA-HA) control group (9.3±2.4 and 71.1±4.4) (p<0.05). The POC-M-click-HA scaffold-fused vertebrates possessed a maximum load and stiffness of 880.8±14.5 N and 843.2±22.4 N/mm, respectively, which were also much higher than those of the PLLA-HA group (maximum: 712.0±37.5 N, stiffness: 622.5±28.4 N/mm, p<0.05). Overall, the results suggest that POC-M-click-HA scaffolds could potentially serve as promising bone grafts for spinal fusion applications. PMID:26213625

Ternary Aligned Nanofibers of RGD Peptide-Displaying M13 Bacteriophage/PLGA/Graphene Oxide for Facilitated Myogenesis

PubMed Central

Shin, Yong Cheol; Kim, Chuntae; Song, Su-Jin; Jun, Seungwon; Kim, Chang-Seok; Hong, Suck Won; Hyon, Suong-Hyu; Han, Dong-Wook; Oh, Jin-Woo

2018-01-01

Recently, there have been tremendous efforts to develop the biofunctional scaffolds by incorporating various biochemical factors. In the present study, we fabricated poly(lactic-co-glycolic acid) (PLGA) nanofiber sheets decorated with graphene oxide (GO) and RGD peptide. The decoration of GO and RGD peptide was readily achieved by using RGD peptide-displaying M13 bacteriophage (RGD-M13 phage) and electrospinning. Furthermore, the aligned GO-decorated PLGA/RGD peptide (GO-PLGA/RGD) ternary nanofiber sheets were prepared by magnetic field-assisted electrospinning, and their potentials as bifunctional scaffolds for facilitating myogenesis were explored. We characterized the physicochemical and mechanical properties of the sheets by scanning electron microscopy, Raman spectroscopy, contact angle measurement, and tensile test. In addition, the C2C12 skeletal myoblasts were cultured on the aligned GO-PLGA/RGD nanofiber sheets, and their cellular behaviors, including initial attachment, proliferation and myogenic differentiation, were evaluated. Our results revealed that the GO-PLGA/RGD nanofiber sheets had suitable physicochemical and mechanical properties for supporting cell growth, and could significantly promote the spontaneous myogenic differentiation of C2C12 skeletal myoblasts. Moreover, it was revealed that the myogenic differentiation was further accelerated on the aligned GO-PLGA/RGD nanofiber sheets due to the synergistic effects of RGD peptide, GO and aligned nanofiber structure. Therefore, , it is suggested that the aligned GO-PLGA/RGD ternary nanofiber sheets are one of the most promising approaches for facilitating myogenesis and promoting skeletal tissue regeneration. PMID:29577018

Ternary Aligned Nanofibers of RGD Peptide-Displaying M13 Bacteriophage/PLGA/Graphene Oxide for Facilitated Myogenesis.

PubMed

Shin, Yong Cheol; Kim, Chuntae; Song, Su-Jin; Jun, Seungwon; Kim, Chang-Seok; Hong, Suck Won; Hyon, Suong-Hyu; Han, Dong-Wook; Oh, Jin-Woo

2018-01-01

Recently, there have been tremendous efforts to develop the biofunctional scaffolds by incorporating various biochemical factors. In the present study, we fabricated poly(lactic- co -glycolic acid) (PLGA) nanofiber sheets decorated with graphene oxide (GO) and RGD peptide. The decoration of GO and RGD peptide was readily achieved by using RGD peptide-displaying M13 bacteriophage (RGD-M13 phage) and electrospinning. Furthermore, the aligned GO-decorated PLGA/RGD peptide (GO-PLGA/RGD) ternary nanofiber sheets were prepared by magnetic field-assisted electrospinning, and their potentials as bifunctional scaffolds for facilitating myogenesis were explored. We characterized the physicochemical and mechanical properties of the sheets by scanning electron microscopy, Raman spectroscopy, contact angle measurement, and tensile test. In addition, the C2C12 skeletal myoblasts were cultured on the aligned GO-PLGA/RGD nanofiber sheets, and their cellular behaviors, including initial attachment, proliferation and myogenic differentiation, were evaluated. Our results revealed that the GO-PLGA/RGD nanofiber sheets had suitable physicochemical and mechanical properties for supporting cell growth, and could significantly promote the spontaneous myogenic differentiation of C2C12 skeletal myoblasts. Moreover, it was revealed that the myogenic differentiation was further accelerated on the aligned GO-PLGA/RGD nanofiber sheets due to the synergistic effects of RGD peptide, GO and aligned nanofiber structure. Therefore, , it is suggested that the aligned GO-PLGA/RGD ternary nanofiber sheets are one of the most promising approaches for facilitating myogenesis and promoting skeletal tissue regeneration.

Nanofiber Orientation and Surface Functionalization Modulate Human Mesenchymal Stem Cell Behavior In Vitro

PubMed Central

Kolambkar, Yash M.; Bajin, Mehmet; Wojtowicz, Abigail; Hutmacher, Dietmar W.; García, Andrés J.

2014-01-01

Electrospun nanofiber meshes have emerged as a new generation of scaffold membranes possessing a number of features suitable for tissue regeneration. One of these features is the flexibility to modify their structure and composition to orchestrate specific cellular responses. In this study, we investigated the effects of nanofiber orientation and surface functionalization on human mesenchymal stem cell (hMSC) migration and osteogenic differentiation. We used an in vitro model to examine hMSC migration into a cell-free zone on nanofiber meshes and mitomycin C treatment to assess the contribution of proliferation to the observed migration. Poly (ɛ-caprolactone) meshes with oriented topography were created by electrospinning aligned nanofibers on a rotating mandrel, while randomly oriented controls were collected on a stationary collector. Both aligned and random meshes were coated with a triple-helical, type I collagen-mimetic peptide, containing the glycine-phenylalanine-hydroxyproline-glycine-glutamate-arginine (GFOGER) motif. Our results indicate that nanofiber GFOGER peptide functionalization and orientation modulate cellular behavior, individually, and in combination. GFOGER significantly enhanced the migration, proliferation, and osteogenic differentiation of hMSCs on nanofiber meshes. Aligned nanofiber meshes displayed increased cell migration along the direction of fiber orientation compared to random meshes; however, fiber alignment did not influence osteogenic differentiation. Compared to each other, GFOGER coating resulted in a higher proliferation-driven cell migration, whereas fiber orientation appeared to generate a larger direct migratory effect. This study demonstrates that peptide surface modification and topographical cues associated with fiber alignment can be used to direct cellular behavior on nanofiber mesh scaffolds, which may be exploited for tissue regeneration. PMID:24020454

Antifungal nanofibers made by controlled release of sea animal derived peptide

NASA Astrophysics Data System (ADS)

Viana, Juliane F. C.; Carrijo, Jéssica; Freitas, Camila G.; Paul, Arghya; Alcaraz, Jarib; Lacorte, Cristiano C.; Migliolo, Ludovico; Andrade, César A.; Falcão, Rosana; Santos, Nuno C.; Gonçalves, Sónia; Otero-González, Anselmo J.; Khademhosseini, Ali; Dias, Simoni C.; Franco, Octávio L.

2015-03-01

Candida albicans is a common human-pathogenic fungal species with the ability to cause several diseases including surface infections. Despite the clear difficulties of Candida control, antimicrobial peptides (AMPs) have emerged as an alternative strategy for fungal control. In this report, different concentrations of antifungal Cm-p1 (Cencritchis muricatus peptide 1) were electrospun into nanofibers for drug delivery. The nanofibers were characterized by mass spectrometry confirming the presence of the peptide on the scaffold. Atomic force microscopy and scanning electronic microscopy were used to measure the diameters, showing that Cm-p1 affects fiber morphology as well as the diameter and scaffold thickness. The Cm-p1 release behavior from the nanofibers demonstrated peptide release from 30 min to three days, leading to effective yeast control in the first 24 hours. Moreover, the biocompatibility of the fibers were evaluated through a MTS assay as well as ROS production by using a HUVEC model, showing that the fibers do not affect cell viability and only nanofibers containing 10% Cm-p1-PVA improved ROS generation. In addition, the secretion of pro-inflammatory cytokines IL-6 and TNF-α by the HUVECs was also slightly modified by the 10% Cm-p1-PVA nanofibers. In conclusion, the electrospinning technique applied here allowed for the manufacture of biodegradable biomimetic nanofibrous extracellular membranes with the ability to control fungal infection.Candida albicans is a common human-pathogenic fungal species with the ability to cause several diseases including surface infections. Despite the clear difficulties of Candida control, antimicrobial peptides (AMPs) have emerged as an alternative strategy for fungal control. In this report, different concentrations of antifungal Cm-p1 (Cencritchis muricatus peptide 1) were electrospun into nanofibers for drug delivery. The nanofibers were characterized by mass spectrometry confirming the presence of the peptide on the scaffold. Atomic force microscopy and scanning electronic microscopy were used to measure the diameters, showing that Cm-p1 affects fiber morphology as well as the diameter and scaffold thickness. The Cm-p1 release behavior from the nanofibers demonstrated peptide release from 30 min to three days, leading to effective yeast control in the first 24 hours. Moreover, the biocompatibility of the fibers were evaluated through a MTS assay as well as ROS production by using a HUVEC model, showing that the fibers do not affect cell viability and only nanofibers containing 10% Cm-p1-PVA improved ROS generation. In addition, the secretion of pro-inflammatory cytokines IL-6 and TNF-α by the HUVECs was also slightly modified by the 10% Cm-p1-PVA nanofibers. In conclusion, the electrospinning technique applied here allowed for the manufacture of biodegradable biomimetic nanofibrous extracellular membranes with the ability to control fungal infection. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr00767d

Fabrication of mineralized electrospun PLGA and PLGA/gelatin nanofibers and their potential in bone tissue engineering.

PubMed

Meng, Z X; Li, H F; Sun, Z Z; Zheng, W; Zheng, Y F

2013-03-01

Surface mineralization is an effective method to produce calcium phosphate apatite coating on the surface of bone tissue scaffold which could create an osteophilic environment similar to the natural extracellular matrix for bone cells. In this study, we prepared mineralized poly(D,L-lactide-co-glycolide) (PLGA) and PLGA/gelatin electrospun nanofibers via depositing calcium phosphate apatite coating on the surface of these nanofibers to fabricate bone tissue engineering scaffolds by concentrated simulated body fluid method, supersaturated calcification solution method and alternate soaking method. The apatite products were characterized by the scanning electron microscopy (SEM), Fourier transform-infrared spectroscopy (FT-IR), and X-ray diffractometry (XRD) methods. A large amount of calcium phosphate apatite composed of dicalcium phosphate dihydrate (DCPD), hydroxyapatite (HA) and octacalcium phosphate (OCP) was deposited on the surface of resulting nanofibers in short times via three mineralizing methods. A larger amount of calcium phosphate was deposited on the surface of PLGA/gelatin nanofibers rather than PLGA nanofibers because gelatin acted as nucleation center for the formation of calcium phosphate. The cell culture experiments revealed that the difference of morphology and components of calcium phosphate apatite did not show much influence on the cell adhesion, proliferation and activity. Copyright © 2012 Elsevier B.V. All rights reserved.

Biodegradable electrospun nanofibers coated with platelet-rich plasma for cell adhesion and proliferation

PubMed Central

Díaz-Gómez, Luis; Alvarez-Lorenzo, Carmen; Concheiro, Angel; Silva, Maite; Dominguez, Fernando; Sheikh, Faheem A.; Cantu, Travis; Desai, Raj; Garcia, Vanessa L.; Macossay, Javier

2014-01-01

Biodegradable electrospun poly(ε-caprolactone) (PCL) scaffolds were coated with platelet-rich plasma (PRP) to improve cell adhesion and proliferation. PRP was obtained from human buffy coat, and tested on human adipose-derived mesenchymal stem cells (MSC) to confirm cell proliferation and cytocompatibility. Then, PRP was adsorbed on the PCL scaffolds via lyophilization, which resulted in uniform sponge-like coating of 2.85 (s.d. 0.14) mg/mg. The scaffolds were evaluated regarding mechanical properties (Young’s modulus, tensile stress and tensile strain), sustained release of total protein and growth factors (PDGF-BB, TGF-β1 and VEGF), and hemocompatibility. MSC seeded on the PRP-PCL nanofibers showed an increased adhesion and proliferation compared to pristine PCL fibers. Moreover, the adsorbed PRP enabled angiogenesis features observed as neovascularization in a chicken chorioallantoic membrane (CAM) model. Overall, these results suggest that PRP-PCL scaffolds hold promise for tissue regeneration applications. PMID:24857481

Matrigel immobilization on the shish-kebab structured poly(ε-caprolactone) nanofibers for skin tissue engineering

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jing, Xin, E-mail: jingxinscut@gmail.com; Mi, Hao-Yang; Wisconsin Institutes for Discovery, University of Wisconsin-Madison, 53715

Surface properties of tissue engineering scaffolds such as topography, hydrophilicity, and functional groups play a vital role in cell adhesion, migration, proliferation, and apoptosis. First, poly(ε-caprolactone) (PCL) shish-kebab scaffolds (PCL-SK), which feature a three-dimensional structure comprised of electrospun PCL nanofibers covered by periodic, self-induced PCL crystal lamellae on the surface, was created to mimic the nanotopography of native collagen fibrils in the extracellular matrix (ECM). Second, matrigel was covalently immobilized on the surface of alkaline hydrolyzed PCL-SK scaffolds to enhance their hydrophilicity. This combined approach not only mimics the nanotopography of native collagen fibrils, but also simulates the surface featuresmore » of collagen fibrils for cell growth. To investigate the viability of such scaffolds, HEF1 fibroblast cell assays were conducted and the results revealed that the nanotopography of the PCL-SK scaffolds facilitated cell adhesion and proliferation. The matrigel functionalization on PCL-SK scaffolds further enhanced cellular response, which suggested elevated biocompatibility and greater potential for skin tissue engineering applications.« less

Mechanical enhancement and in vitro biocompatibility of nanofibrous collagen-chitosan scaffolds for tissue engineering.

PubMed

Zou, Fengjuan; Li, Runrun; Jiang, Jianjun; Mo, Xiumei; Gu, Guofeng; Guo, Zhongwu; Chen, Zonggang

2017-12-01

The collagen-chitosan complex with a three-dimensional nanofiber structure was fabricated to mimic native ECM for tissue repair and biomedical applications. Though the three-dimensional hierarchical fibrous structures of collagen-chitosan composites could provide more adequate stimulus to facilitate cell adhesion, migrate and proliferation, and thus have the potential as tissue engineering scaffolding, there are still limitations in their applications due to the insufficient mechanical properties of natural materials. Because poly (vinyl alcohol) (PVA) and thermoplastic polyurethane (TPU) as biocompatible synthetic polymers can offer excellent mechanical properties, they were introduced into the collagen-chitosan composites to fabricate the mixed collagen/chitosan/PVA fibers and a sandwich structure (collagen/chitosan-TPU-collagen/chitosan) of nanofiber in order to enhance the mechanical properties of the nanofibrous collagen-chitosan scaffold. The results showed that the tensile behavior of materials was enhanced to different degrees with the difference of collagen content in the fibers. Besides the Young's modulus had no obvious changes, both the break strength and the break elongation of materials were heightened after reinforced by PVA. For the collagen-chitosan nanofiber reinforced by TPU, both the break strength and the Young's modulus of materials were heightened in different degrees with the variety of collagen content in the fibers despite the decrease of the break elongation of materials to some extent. In vitro cell test demonstrated that the materials could provide adequate environment for cell adhesion and proliferation. All these indicated that the reinforced collagen-chitosan nanofiber could be as potential scaffold for tissue engineering according to the different mechanical requirements in clinic.

Periodontal ligament cellular structures engineered with electrospun poly(DL-lactide-co-glycolide) nanofibrous membrane scaffolds.

PubMed

Inanç, Bülend; Arslan, Y Emre; Seker, Sükran; Elçin, A Eser; Elçin, Y Murat

2009-07-01

Periodontal tissue engineering is expected to overcome the limitations associated with the existing regenerative techniques for the treatment of periodontal defects involving alveolar bone, cementum, and periodontal ligament. Cell-based tissue engineering approaches involve the utilization of in vitro expanded cells with regenerative capacity and their delivery to the appropriate sites via biomaterial scaffolds. The aim of this study was to establish living periodontal ligament cell-containing structures on electrospun poly(DL-lactic-co-glycolic acid) (PLGA) nanofiber membrane scaffolds, assess their viability and characteristics, and engineer multilayered structures amenable to easy handling. Human periodontal ligament (hPDL) cells were expanded in explant culture and then characterized morphologically and immunohistochemically. PLGA nanofiber membranes were prepared by the electrospinning process; mechanical tensile properties were determined, surface topography, nanofiber size, and porosity status were investigated with SEM. Cells were seeded on the membranes at approximately 50,000 cell/cm(2) and cultured for 21 days either in expansion or in osteogenic induction medium. Cell adhesion and viability were demonstrated using SEM and MTT, respectively, and osteogenic differentiation was determined with IHC and immunohistomorphometric evaluation of osteopontin, osteocalcin, and bone sialoprotein marker expression. At days 3, 6, 9, and 12 additional cell/membrane layers were deposited on the existing ones and multilayered hybrid structures were established. Results indicate the feasibility of periodontal ligament cell-containing tissue-like structures engineering with PDL cells and electrospun nanofiber PLGA scaffolds supporting cell adhesion, viability and osteogenic differentiation properties of cells in hybrid structures amenable to macroscopic handling.

Nanofibrous scaffolds for the guidance of stem cell-derived neurons for auditory nerve regeneration.

PubMed

Hackelberg, Sandra; Tuck, Samuel J; He, Long; Rastogi, Arjun; White, Christina; Liu, Liqian; Prieskorn, Diane M; Miller, Ryan J; Chan, Che; Loomis, Benjamin R; Corey, Joseph M; Miller, Josef M; Duncan, R Keith

2017-01-01

Impairment of spiral ganglion neurons (SGNs) of the auditory nerve is a major cause for hearing loss occurring independently or in addition to sensory hair cell damage. Unfortunately, mammalian SGNs lack the potential for autonomous regeneration. Stem cell based therapy is a promising approach for auditory nerve regeneration, but proper integration of exogenous cells into the auditory circuit remains a fundamental challenge. Here, we present novel nanofibrous scaffolds designed to guide the integration of human stem cell-derived neurons in the internal auditory meatus (IAM), the foramen allowing passage of the spiral ganglion to the auditory brainstem. Human embryonic stem cells (hESC) were differentiated into neural precursor cells (NPCs) and seeded onto aligned nanofiber mats. The NPCs terminally differentiated into glutamatergic neurons with high efficiency, and neurite projections aligned with nanofibers in vitro. Scaffolds were assembled by seeding GFP-labeled NPCs on nanofibers integrated in a polymer sheath. Biocompatibility and functionality of the NPC-seeded scaffolds were evaluated in vivo in deafened guinea pigs (Cavia porcellus). To this end, we established an ouabain-based deafening procedure that depleted an average 72% of SGNs from apex to base of the cochleae and caused profound hearing loss. Further, we developed a surgical procedure to implant seeded scaffolds directly into the guinea pig IAM. No evidence of an inflammatory response was observed, but post-surgery tissue repair appeared to be facilitated by infiltrating Schwann cells. While NPC survival was found to be poor, both subjects implanted with NPC-seeded and cell-free control scaffolds showed partial recovery of electrically-evoked auditory brainstem thresholds. Thus, while future studies must address cell survival, nanofibrous scaffolds pose a promising strategy for auditory nerve regeneration.

Controlled surface morphology and hydrophilicity of polycaprolactone toward human retinal pigment epithelium cells.

PubMed

Shahmoradi, Saleheh; Yazdian, Fatemeh; Tabandeh, Fatemeh; Soheili, Zahra-Soheila; Hatamian Zarami, Ashraf Sadat; Navaei-Nigjeh, Mona

2017-04-01

Applying scaffolds as a bed to enhance cell proliferation and even differentiation is one of the treatment of retina diseases such as age-related macular degeneration (AMD) which deteriorating photoreceptors and finally happening blindness. In this study, aligned polycaprolactone (PCL) nanofibers were electrospun and at different conditions and their characteristics were measured by scanning electron microscope (SEM) and contact angle. Response surface methodology (RSM) was used to optimize the diameter of fabricated nanofibers. Two factors as solution concentration and voltage value were considered as independent variables and their effects on nanofibers' diameters were evaluated by central composite design and the optimum conditions were obtained as 0.12g/mL and 20kV, respectively. In order to decrease the hydrophobicity of PCL, the surface of the fabricated scaffolds was modified by alkaline hydrolysis method. Contact time of the scaffolds and alkaline solution and concentration of alkaline solution were optimized using Box Behnken design and (120min and 5M were the optimal, respectively). Contact angle measurement showed the high hydrophilicity of treated scaffolds (with contact angle 7.48°). Plasma surface treatment was applied to compare the effect of using two kinds of surface modification methods simultaneously on hydrolyzed scaffolds. The RPE cells grown on scaffolds were examined by immunocytochemistry (ICC), MTT and continuous inspection of cellular morphology. Interestingly, Human RPE cells revealed their characteristic morphology on hydrolyzed scaffold well. As a result, we introduced a culture substrate with low diameter (185.8nm), high porosity (82%) and suitable hydrophilicity (with contact angle 7.48 degree) which can be promising for hRPE cell transplantation. Copyright © 2016. Published by Elsevier B.V.

Enhanced chondrogenesis of human bone marrow mesenchymal Stem Cell (BMSC) on nanofiber-based polyethersulfone (PES) scaffold.

PubMed

Mahboudi, Hossein; Kazemi, Bahram; Soleimani, Masoud; Hanaee-Ahvaz, Hana; Ghanbarian, Hossein; Bandehpour, Mojgan; Enderami, Seyed Ehsan; Kehtari, Mousa; Barati, Ghasem

2018-02-15

Mesenchymal stem cells (MSC) from bone marrow hold great potential as a cell source for cartilage repair. The objective of our study was differentiation of MSC toward chondrocyte by using Nanofiber-based polyethersulfone (PES) scaffold and also enhanced chondrogenic differentiation of BMSC in vitro. MSCs were harvested from bone marrow of human and PES scaffold was fabricated via Electrospinning. The isolated cells were cultured on the PES scaffold and scaffold free method. After 21days, Real-time PCR was performed to evaluate the cartilage-specific genes in the mRNA levels. Also, in order to confirm our results, we have done immunocytochemistry and SEM imaging. Flowcytometry confirmed the nature of the isolated adherent cells. Immunocytochemistry and SEM imaging confirmed the differentiation of MSC toward chondrocyte. Also, real time PCR showed a significant increased gene expression of collagen type II and aggrecan on the PES scaffold method when compared to the mRNA levels measured in scaffold free method. Down regulation of Collagen type I was observed in PES scaffold compared to scaffold free at day 21. Also, both methods showed a similar pattern of expression of SOX9. Our results showed that PES scaffold maintains BMSC proliferation and differentiation, and can significantly enhance chondrogenic differentiation of BMSC. PES scaffold seeded BMSC showed the highest capacity for differentiation into chondrocyte-like cells. Copyright © 2017 Elsevier B.V. All rights reserved.

Porous SiO2 nanofiber grafted novel bioactive glass-ceramic coating: A structural scaffold for uniform apatite precipitation and oriented cell proliferation on inert implant.

PubMed

Das, Indranee; De, Goutam; Hupa, Leena; Vallittu, Pekka K

2016-05-01

A composite bioactive glass-ceramic coating grafted with porous silica nanofibers was fabricated on inert glass to provide a structural scaffold favoring uniform apatite precipitation and oriented cell proliferation. The coating surfaces were investigated thoroughly before and after immersion in simulated body fluid. In addition, the proliferation behavior of fibroblast cells on the surface was observed for several culture times. The nanofibrous exterior of this composite bioactive coating facilitated homogeneous growth of flake-like carbonated hydroxyapatite layer within a short period of immersion. Moreover, the embedded porous silica nanofibers enhanced hydrophilicity which is required for proper cell adhesion on the surface. The cells proliferated well following a particular orientation on the entire coating by the assistance of nanofibrous scaffold-like structural matrix. This newly engineered composite coating was effective in creating a biological structural matrix favorable for homogeneous precipitation of calcium phosphate, and organized cell growth on the inert glass surface. Copyright © 2016 Elsevier B.V. All rights reserved.

Emulsion Electrospinning as an Approach to Fabricate PLGA/Chitosan Nanofibers for Biomedical Applications

PubMed Central

Tavanai, Hossein; Hilborn, Jöns; Donzel-Gargand, Olivier; Leifer, Klaus; Arpanaei, Ayyoob

2014-01-01

Novel nanofibers from blends of polylactic-co-glycolic acid (PLGA) and chitosan have been produced through an emulsion electrospinning process. The spinning solution employed polyvinyl alcohol (PVA) as the emulsifier. PVA was extracted from the electrospun nanofibers, resulting in a final scaffold consisting of a blend of PLGA and chitosan. The fraction of chitosan in the final electrospun mat was adjusted from 0 to 33%. Analyses by scanning and transmission electron microscopy show uniform nanofibers with homogenous distribution of PLGA and chitosan in their cross section. Infrared spectroscopy verifies that electrospun mats contain both PLGA and chitosan. Moreover, contact angle measurements show that the electrospun PLGA/chitosan mats are more hydrophilic than electrospun mats of pure PLGA. Tensile strengths of 4.94 MPa and 4.21 MPa for PLGA/chitosan in dry and wet conditions, respectively, illustrate that the polyblend mats of PLGA/chitosan are strong enough for many biomedical applications. Cell culture studies suggest that PLGA/chitosan nanofibers promote fibroblast attachment and proliferation compared to PLGA membranes. It can be assumed that the nanofibrous composite scaffold of PLGA/chitosan could be potentially used for skin tissue reconstruction. PMID:24689041

Surface modification of endovascular stents with rosuvastatin and heparin-loaded biodegradable nanofibers by electrospinning.

PubMed

Janjic, Milka; Pappa, Foteini; Karagkiozaki, Varvara; Gitas, Christakis; Ktenidis, Kiriakos; Logothetidis, Stergios

2017-01-01

This study describes the development of drug-loaded nanofibrous scaffolds as a nanocoating for endovascular stents for the local and sustained delivery of rosuvastatin (Ros) and heparin (Hep) to injured artery walls after endovascular procedures via the electrospinning process. The proposed hybrid covered stents can promote re-endothelialization; improve endothelial function; reduce inflammatory reaction; inhibit neointimal hyperplasia of the injured artery wall, due to well-known pleiotropic actions of Ros; and prevent adverse events such as in-stent restenosis (ISR) and stent thrombosis (ST), through the antithrombotic action of Hep. Biodegradable nanofibers were prepared by dissolving cellulose acetate (AC) and Ros in N , N -dimethylacetamide (DMAc) and acetone-based solvents. The polymeric solution was electrospun (e-spun) into drug-loaded AC nanofibers onto three different commercially available stents (Co-Cr stent, Ni-Ti stent, and stainless steel stent), resulting in nonwoven matrices of submicron-sized fibers. Accordingly, Hep solution was further used for fibrous coating onto the engineered Ros-loaded stent. The functional encapsulation of Ros and Hep drugs into polymeric scaffolds further underwent physicochemical analysis. Morphological characterization took place via scanning electron microscopy (SEM) and atomic force microscopy (AFM) analyses, while scaffolds' wettability properties were obtained by contact angle (CA) measurements. The morphology of the drug-loaded AC nanofibers was smooth, with an average diameter of 200-800 nm, and after CA measurement, we concluded to the superhydrophobic nature of the engineered scaffolds. In vitro release rates of the pharmaceutical drugs were determined using a high-performance liquid chromatography assay, which showed that after the initial burst, drug release was controlled slowly by the degradation of the polymeric materials. These results imply that AC nanofibers encapsulated with Ros and Hep drugs have great potential in the development of endovascular grafts with anti-thrombogenic properties that can accelerate the re-endothelialization, reduce the neointimal hyperplasia and inflammatory reaction, and improve the endothelial function.

Incorporation of amoxicillin-loaded organic montmorillonite into poly(ester-urethane) urea nanofibers as a functional tissue engineering scaffold.

PubMed

Yu, Kui; Zhu, Tonghe; Wu, Yu; Zhou, Xiangxiang; Yang, Xingxing; Wang, Juan; Fang, Jun; El-Hamshary, Hany; Al-Deyab, Salem S; Mo, Xiumei

2017-03-01

A dual drug-loaded system is a promising alternative for the sustained drug release system and skin tissue engineering. In this study, a natural sodium montmorillonite (Na-MMT) modified by cetyl trimethyl ammonium bromide (CTAB) was prepared as a carrier to load a model drug - amoxicillin (AMX), the modified organic montmorillonite (CTAB-OMMT) loaded with AMX was marked as AMX@CTAB-OMMT and was subsequently incorporated into poly(ester-urethane) urea (PEUU) and gelatin hybrid nanofibers via electrospinning, resulting in a new drug-loaded nanofibrous scaffold (AMX@CTAB-OMMT-PU75). The scanning electron microscopy (SEM) result showed that the fiber morphology did not change after the embedding of AMX@CTAB-OMMT. Meanwhile, there was a significant increase of mechanical properties for PEUU/Gelatin hybrid nanofibers (PU75) after the incorporation of AMX@CTAB-OMMT and CTAB-OMMT. Importantly, AMX@CTAB-OMMT-PU75 nanofibers showed a kind of sustained drug release property which could be justified reasonably for the controlled release of AMX depending on the various application. The sustained release property could be identified roughly by the result of antibacterial test. The anaphylactic reaction test proved that there was no any anaphylactic reaction or inflammation on the back of rat for AMX@CTAB-OMMT-PU75 nanofibers. Consequently, the prepared drug-loaded AMX@CTAB-OMMT-PU75 nanofibrous scaffold is a promising candidate for application in the skin tissue engineering field and controlled drug release system. Copyright © 2016 Elsevier B.V. All rights reserved.

Enhancement of stem cell differentiation to osteogenic lineage on hydroxyapatite-coated hybrid PLGA/gelatin nanofiber scaffolds.

PubMed

Sanaei-Rad, Parisa; Jafarzadeh Kashi, Tahereh-Sadat; Seyedjafari, Ehsan; Soleimani, Masoud

2016-11-01

A combination of polymeric materials and bioceramics has recently received a great deal of attention for bone tissue engineering applications. In the present study, hybrid nanofibrous scaffolds were fabricated from PLGA and gelatin via electrospinning and then were coated with hydroxyapatite (HA). They were then characterized and used in stem cell culture studies for the evaluation of their biological behavior and osteogenic differentiation in vitro. This study showed that all PLGA, hybrid PLGA/gelatin and HA-PLGA/gelatin scaffolds were composed of ultrafine fibers with smooth morphology and interconnected pores. The MTT assay confirmed that the scaffolds can support the attachment and proliferation of stem cells. During osteogenic differentiation, bone-related gene expression, ALP activity and biomineralization on HA-PLGA/gelatin scaffolds were higher than those observed on other scaffolds and TCPS. PLGA/gelatin electrospun scaffolds also showed higher values of these markers than TCPS. Taking together, it was shown that nanofibrous structure enhanced osteogenic differentiation of adipose-tissue derived stem cells. Furthermore, surface-coated HA stimulated the effect of nanofibers on the commitment of stem cells toward osteolineage. In conclusion, HA-PLGA/gelatin electrospun scaffolds were demonstrated to have significant potential for bone tissue engineering applications. Copyright © 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

Optimization of electrospun TSF nanofiber alignment and diameter to promote growth and migration of mesenchymal stem cells

NASA Astrophysics Data System (ADS)

Qu, Jing; Zhou, Dandan; Xu, Xiaojing; Zhang, Feng; He, Lihong; Ye, Rong; Zhu, Ziyu; Zuo, Baoqi; Zhang, Huanxiang

2012-11-01

Silk fibroin scaffolds are a naturally derived biocompatible matrix with the potential for reconstructive surgical applications. In this study, tussah silk fibroin (TSF) nanofiber with different diameters (400 nm, 800 nm and 1200 nm) and alignment (random and aligned) were prepared by electrospinning, then the growth and migration of mesenchymal stem cells (MSCs) on these materials were further evaluated. CD90 immunofluorescence staining showed that fiber alignment exhibited a strong influence on the morphology of MSCs, indicating that the alignment of the scaffolds could determine the distribution of cells. Moreover, smaller diameter and aligned TSF scaffolds are more favorable to the growth of MSCs as compared with 800 nm and 1200 nm random TSF scaffolds. In addition, the increased migration speed and efficiency of MSCs induced by three-D TSF were verified, highlighting the guiding roles of TSF to the migrated MSCs. More importantly, 400 nm aligned TSF scaffolds dramatically improved cell migratory speed and further induced the most efficient migration of MSCs as compared with larger diameter TSF scaffolds. In conclusion, the data demonstrate that smaller diameter and aligned electrospun TSF represent valuable scaffolds for supporting and promoting MSCs growth and migration, thus raising the possibility of manipulating TSF scaffolds to enhance homing and therapeutic potential of MSCs in cellular therapy.

Stimulating effect of graphene oxide on myogenesis of C2C12 myoblasts on RGD peptide-decorated PLGA nanofiber matrices.

PubMed

Shin, Yong Cheol; Lee, Jong Ho; Kim, Min Jeong; Hong, Suck Won; Kim, Bongju; Hyun, Jung Keun; Choi, Yu Suk; Park, Jong-Chul; Han, Dong-Wook

2015-01-01

In the field of biomedical engineering, many studies have focused on the possible applications of graphene and related nanomaterials due to their potential for use as scaffolds, coating materials and delivery carriers. On the other hand, electrospun nanofiber matrices composed of diverse biocompatible polymers have attracted tremendous attention for tissue engineering and regenerative medicine. However, their combination is intriguing and still challenging. In the present study, we fabricated nanofiber matrices composed of M13 bacteriophage with RGD peptide displayed on its surface (RGD-M13 phage) and poly(lactic-co-glycolic acid, PLGA) and characterized their physicochemical properties. In addition, the effect of graphene oxide (GO) on the cellular behaviors of C2C12 myoblasts, which were cultured on PLGA decorated with RGD-M13 phage (RGD/PLGA) nanofiber matrices, was investigated. Our results revealed that the RGD/PLGA nanofiber matrices have suitable physicochemical properties as a tissue engineering scaffold and the growth of C2C12 myoblasts were significantly enhanced on the matrices. Moreover, the myogenic differentiation of C2C12 myoblasts was substantially stimulated when they were cultured on the RGD/PLGA matrices in the presence of GO. In conclusion, these findings propose that the combination of RGD/PLGA nanofiber matrices and GO can be used as a promising strategy for skeletal tissue engineering and regeneration.

Engineering PCL/lignin nanofibers as an antioxidant scaffold for the growth of neuron and Schwann cell.

PubMed

Wang, Jing; Tian, Lingling; Luo, Baiwen; Ramakrishna, Seeram; Kai, Dan; Loh, Xian Jun; Yang, In Hong; Deen, G Roshan; Mo, Xiumei

2018-05-12

Antioxidant is critical for the successful of nerve tissue regeneration, and biomaterials with antioxidant activity might be favorable for peripheral nerve repair. Lignin, a biopolymer from wood with excellent antioxidant properties, is still "unexplored" as biomaterials. To design an antioxidative bioscaffold for nerve regeneration, here we synthesized lignin-polycaprolactone (PCL) copolymers via solvent free ring-opening polymerization (ROP). Then such lignin-PCL copolymers were incorporated with PCL and engineered into nanofibrous scaffolds for supporting the growth of neuron and Schwann cell. Our results showed that the addition of lignin-PCL enhanced the mechanical properties of PCL nanofibers and endowed them with good antioxidant properties (up to 98.3 ± 1.9% free radical inhibition within 4 h). Cell proliferation assay showed that PCL/lignin-PCL nanofibers increased cell viability compared to PCL fibers, especially after an oxidative challenge. Moreover, Schwann cells and dorsal root ganglion (DRG) neurons cultured on the nanofibers to assess their potential for nerve regeneration. These results suggested that nanofibers with lignin copolymers promoted cell proliferation of both BMSCs and Schwann cells, enhanced myelin basic protein expressions of Schwann cells and stimulated neurite outgrowth of DRG neurons. In all, these sustainable, intrinsically antioxidant nanofibers may be a potential candidate for nerve TE applications. Copyright © 2018 Elsevier B.V. All rights reserved.

Microporous Poly(L-Lactic Acid) Membranes Fabricated by Polyethylene Glycol Solvent-Cast/Particulate Leaching Technique

PubMed Central

Selvam, Shivaram; Chang, Wenji V.; Nakamura, Tamako; Samant, Deedar M.; Thomas, Padmaja B.; Trousdale, Melvin D.; Mircheff, Austin K.; Schechter, Joel E.

2009-01-01

With the eventual goal of developing a tissue-engineered tear secretory system, we found that primary lacrimal gland acinar cells grown on solid poly(L-lactic acid) (PLLA) supports expressed the best histiotypic morphology. However, to be able to perform vectorial transport functions, epithelia must be supported by a permeable substratum. In the present study, we describe the use of a solvent-cast/particulate leaching technique to fabricate microporous PLLA membranes (mpPLLAm) from PLLA/polyethylene glycol blends. Scanning electron microscopy revealed pores on both the air-cured (∼4 μm) and glass-cured sides (<2 μm) of the mpPLLAm. Diffusion studies were performed with mpPLLAm fabricated from 57.1% PLLA/42.9% polyethylene glycol blends to confirm the presence of channelized pores. The data reveal that glucose, L-tryptophan, and dextran (a high molecular weight glucose polymer) readily permeate mpPLLAm. Diffusion of the immunoglobulin G through the mpPLLAm decreased with time, suggesting the possible adsorption and occlusion of the pores. Cells cultured on the mpPLLAm (57.1/42.9 wt%) grew to subconfluent monolayers but retained histiotypic morphological and physiological characteristics of lacrimal acinar cells in vivo. Our results suggest that mpPLLAm fabricated using this technique may be useful as a scaffold for a bioartificial lacrimal gland device. PMID:19260769

Comparative Study of Poly (ε-Caprolactone) and Poly(Lactic-co-Glycolic Acid) -Based Nanofiber Scaffolds for pH-Sensing.

PubMed

Di, Wenjun; Czarny, Ryan S; Fletcher, Nathan A; Krebs, Melissa D; Clark, Heather A

2016-10-01

This study aims to develop biodegradable and biocompatible polymer-based nanofibers that continuously monitor pH within microenvironments of cultured cells in real-time. In the future, these fibers will provide a scaffold for tissue growth while simultaneously monitoring the extracellular environment. Sensors to monitor pH were created by directly electrospinning the sensor components within a polymeric matrix. Specifically, the entire fiber structure is composed of the optical equivalent of an electrode, a pH-sensitive fluorophore, an ionic additive, a plasticizer, and a polymer to impart mechanical stability. The resulting poly(ε-caprolactone) (PCL) and poly(lactic-co-glycolic acid) (PLGA) based sensors were characterized by morphology, dynamic range, reversibility and stability. Since PCL-based nanofibers delivered the most desirable analytical response, this matrix was used for cellular studies. Electrospun nanofiber scaffolds (NFSs) were created directly out of optode material. The resulting NFS sensors respond to pH changes with a dynamic range centered at 7.8 ± 0.1 and 9.6 ± 0.2, for PCL and PLGA respectively. NFSs exhibited multiple cycles of reversibility with a lifetime of at least 15 days with preservation of response characteristics. By comparing the two NFSs, we found PCL-NFSs are more suitable for pH sensing due to their dynamic range and superior reversibility. The proposed sensing platform successfully exhibits a response to pH and compatibility with cultured cells. NSFs will be a useful tool for creating 3D cellular scaffolds that can monitor the cellular environment with applications in fields such as drug discovery and tissue engineering.

Evaluation of structural and mechanical properties of electrospun nano-micro hybrid of poly hydroxybutyrate-chitosan/silk scaffold for cartilage tissue engineering

PubMed Central

Karbasi, Saeed; Fekrat, Farnoosh; Semnani, Daryoush; Razavi, Shahnaz; Zargar, Elham Naghash

2016-01-01

Background: One of the new methods of scaffold fabrication is a nano-micro hybrid structure in which the properties of the scaffold are improved by introducing nanometer and micrometer structures. This method could be suitable for scaffold designing if some features improve. Materials and Methods: In this study, electrospun nanofibers of 9% weight solution of poly (3-hydroxybutyrate) (P3HB) and a 15% weight of chitosan by trifluoroacetic acid were coated on both the surface of a silk knitted substrate in the optimum condition to improve the mechanical properties of scaffolds for cartilage tissue engineering application. These hybrid nano-micro fibrous scaffolds were characterized by structural and mechanical evaluation methods. Results: Scanning electron microscopy values and porosity analysis showed that average diameter of nanofibers was 584.94 nm in electrospinning part and general porosity was more than 80%. Fourier transform infrared spectroscopy results indicated the presence of all elements without pollution. The tensile test also stated that by electrospinning, as well as adding chitosan, both maximum strength and maximum elongation increased to 187 N and 10 mm. It means that the microfibrous part of scaffold could affect mechanical properties of nano part of the hybrid scaffold, significantly. Conclusions: It could be concluded that P3HB-chitosan/silk hybrid scaffolds can be a good candidate for cartilage tissue engineering. PMID:28028520

Evaluation of structural and mechanical properties of electrospun nano-micro hybrid of poly hydroxybutyrate-chitosan/silk scaffold for cartilage tissue engineering.

PubMed

Karbasi, Saeed; Fekrat, Farnoosh; Semnani, Daryoush; Razavi, Shahnaz; Zargar, Elham Naghash

2016-01-01

One of the new methods of scaffold fabrication is a nano-micro hybrid structure in which the properties of the scaffold are improved by introducing nanometer and micrometer structures. This method could be suitable for scaffold designing if some features improve. In this study, electrospun nanofibers of 9% weight solution of poly (3-hydroxybutyrate) (P3HB) and a 15% weight of chitosan by trifluoroacetic acid were coated on both the surface of a silk knitted substrate in the optimum condition to improve the mechanical properties of scaffolds for cartilage tissue engineering application. These hybrid nano-micro fibrous scaffolds were characterized by structural and mechanical evaluation methods. Scanning electron microscopy values and porosity analysis showed that average diameter of nanofibers was 584.94 nm in electrospinning part and general porosity was more than 80%. Fourier transform infrared spectroscopy results indicated the presence of all elements without pollution. The tensile test also stated that by electrospinning, as well as adding chitosan, both maximum strength and maximum elongation increased to 187 N and 10 mm. It means that the microfibrous part of scaffold could affect mechanical properties of nano part of the hybrid scaffold, significantly. It could be concluded that P3HB-chitosan/silk hybrid scaffolds can be a good candidate for cartilage tissue engineering.

Functionalized hybrid nanofibers to mimic native ECM for tissue engineering applications

NASA Astrophysics Data System (ADS)

Karuppuswamy, Priyadharsini; Venugopal, Jayarama Reddy; Navaneethan, Balchandar; Laiva, Ashang Luwang; Sridhar, Sreepathy; Ramakrishna, Seeram

2014-12-01

Nanotechnology being one of the most promising technologies today shows an extremely huge potential in the field of tissue engineering to mimic the porous topography of natural extracellular matrix (ECM). Natural polymers are incorporated into the synthetic polymers to fabricate functionalized hybrid nanofibrous scaffolds, which improve cell and tissue compatibility. The present study identified the biopolymers - aloe vera, silk fibroin and curcumin incorporated into polycaprolactone (PCL) as suitable substrates for tissue engineering. Different combinations of PCL with natural polymers - PCL/aloe vera, PCL/silk fibroin, PCL/aloe vera/silk fibroin, PCL/aloe vera/silk fibroin/curcumin were electrospun into nanofibrous scaffolds. The fabricated two dimensional nanofibrous scaffolds showed high surface area, appropriate mechanical properties, hydrophilicity and porosity, required for the regeneration of diseased tissues. The nanofibrous scaffolds were characterized by Scanning electron microscope (SEM), porometry, Instron tensile tester, VCA optima contact angle measurement and FTIR to analyze the fiber diameter and morphology, porosity and pore size distribution, mechanical strength, wettability, chemical bonds and functional groups, respectively. The average fiber diameter of obtained fibers ranged from 250 nm to 350 nm and the tensile strength of PCL scaffolds at 4.49 MPa increased upto 8.3 MPa for PCL/silk fibroin scaffolds. Hydrophobicity of PCL decreased with the incorporation of natural polymers, especially for PCL/aloe vera scaffolds. The properties of as-spun nanofiber scaffolds showed their potential as promising scaffold materials in tissue engineering applications.

Novel biocompatible polymeric blends for bone regeneration: Material and matrix design and development

NASA Astrophysics Data System (ADS)

Deng, Meng

The first part of the work presented in this dissertation is focused on the design and development of novel miscible and biocompatible polyphosphazene-polyester blends as candidate materials for scaffold-based bone tissue engineering applications. Biodegradable polyesters such as poly(lactide-co-glycolide) (PLAGA) are among the most widely used polymeric materials for bone tissue engineering. However, acidic degradation products resulting from the bulk degradation mechanism often lead to catastrophic failure of the structure integrity, and adversely affect biocompatibility both in vitro and in vivo. One promising approach to circumvent these limitations is to blend PLAGA with other macromolecules that can buffer the acidic degradation products with a controlled degradation rate. Biodegradable polyphosphazenes (PPHOS), a new class of biomedical materials, have proved to be superior candidate materials to achieve this objective due to their unique buffering degradation products. A highly practical blending approach was adopted to develop novel biocompatible, miscible blends of these two polymers. In order to achieve this miscibility, a series of amino acid ester, alkoxy, aryloxy, and dipeptide substituted PPHOS were synthesized to promote hydrogen bonding interactions with PLAGA. Five mixed-substituent PPHOS compositions were designed and blended with PLAGA at different weight ratios producing candidate blends via a mutual solvent method. Preliminary characterization identified two specific side groups namely glycylglycine dipeptide and phenylphenoxy that resulted in improved blend miscibility and enhanced in vitro osteocompatibility. These findings led to the synthesis of a mixed-substituent polyphosphazene poly[(glycine ethyl glycinato)1(phenylphenoxy)1phosphazene] (PNGEGPhPh) for blending with PLAGA. Two dipeptide-based blends having weight ratios of PNGEGPhPh to PLAGA namely 25:75 (Matrix1) and 50:50 (Matrix2) were fabricated. Both of the blends were characterized for miscibility, mechanical properties, degradation kinetics, and in vitro osteocompatibility. Primary rat osteoblasts (PRO) isolated from rat calvaria were used to evaluate their in vitro osteocompatibility. The blends were also characterized for in vivo biodegradability and biocompatibility using a rat subcutaneous implantation model. Successful in vivo scaffold-based tissue regeneration greatly depends on the scaffold material biocompatibility, mechanical stability, and scaffold architecture to promote tissue in-growth. The other part of the work in the dissertation is focused on the development of mechanically competent bioresorbable nano-structured three-dimensional (3D) hiomimetic scaffolds for bone tissue engineering applications. Scaffold material selection was based on achieving improved mechanical stability, in vitro osteoblast performance, and in vivo biocompatibility. A miscible PNGEGPhPh-PLAGA blend system developed and characterized in the first part of the thesis work was chosen to fabricate a nanofiber-based mechanically competent biomimetic scaffold via electrospinning. Due to its versatility, controllability and reproducibility, the technique of electrospinning was adopted to produce blend nanofibers. The polymer solution concentration and electrospinning parameters were optimized to produce blend fibers in the range of 50-500 nm to mimic dimensions of collagen fibrils present in the natural extracellular matrix of native bone. These blend nanofiber matrices supported PRO adhesion, proliferation and showed an elevated phenotype expression compared to PLAGA nanofibers. Orienting electrospun nanofibers in a concentric manner with an open central cavity created a mechanically competent 3D scaffold mimicking the bone marrow cavity, as well as, the lamellar structure of bone. The 3D biomimetic scaffold exhibited a similar characteristic mechanical behavior to that of native bone. Compressive modulus of the scaffold was found to be within the range of human trabecular bone. To our knowledge this is the first mechanically competent 3D electrospun nanofiber scaffold with mechanical properties in the middle range of human trabecular bone. The potential of this scaffold for bone repair was further investigated by monitoring the cellular activity and mechanical performance over time using in vitro culture. This biomimetic scaffold supported the robust PRO growth throughout the scaffold architecture and maintained osteoblast phenotype expression in vitro, which resulted in a similar cell-matrix organization to that of native bone and maintenance of structure integrity. (Abstract shortened by UMI.)

Enhancing the Stiffness of Electrospun Nanofiber Scaffolds with Controlled Surface Coating and Mineralization

PubMed Central

Liu, Wenying; Yeh, Yi-Chun; Lipner, Justin; Xie, Jingwei; Sung, Hsing-Wen; Thomopoulos, Stavros; Xia, Younan

2011-01-01

A new method was developed to coat hydroxyapatite (HAp) onto electrospun poly(lactic-co-glycolic acid) (PLGA) nanofibers for tendon-to-bone insertion site repair applications. Prior to mineralization, chitosan and heparin were covalently immobilized onto the surface of the fibers to accelerate the nucleation of bone-like HAp crystals. Uniform coatings of HAp were obtained by immersing the nanofiber scaffolds into a modified 10 times concentrated simulated body fluid (m10SBF) for different periods of time. The new method resulted in thicker and denser coatings of mineral on the fibers compared to previously reported methods. Scanning electron microscopy measurements confirmed the formation of nanoscale HAp particles on the fibers. Mechanical property assessment demonstrated higher stiffness with respect to previous coating methods. A combination of the nanoscale fibrous structure and bone-like mineral coating could mimic the structure, composition, and function of mineralized tissues. PMID:21710996

Engineering and Modeling Carbon Nanofiller-Based Scaffolds for Tissue Regeneration

NASA Astrophysics Data System (ADS)

Al Habis, Nuha Hamad

Conductive biopolymers are starting to emerge as potential scaffolds of the future. These scaffolds exhibit some unique properties such as inherent conductivity, mechanical and surface properties. Traditionally, a conjugated polymer is used to constitute a conductive network. An alternative method currently being used is nanofillers as additives in the polymer. In this dissertation, we fabricated an intelligent scaffold for use in tissue engineering applications. The main idea was to enhance the mechanical, electrical properties and cell growth of scaffolds by using distinct types of nanofillers such as graphene, carbon nanofiber and carbon black. We identified the optimal concentrations of nano-additive in both fibrous and film scaffolds to obtain the highest mechanical and electrical properties without neglecting any of them. Lastly, we investigated the performance of these scaffold with cell biology. To accomplish these tasks, we first studied the mechanical properties of the scaffold as a function of morphology, concentration and variety of carbon nanofillers. Results showed that there was a gradual increase of the modulus and the fracture strength while using carbon black, carbon nanofiber and graphene, due to the small and strong carbon-to-carbon bonds and the length of the interlayer spacing. Moreover, regardless of the fabrication method, there was an increase in mechanical properties as the concentration of nanofillers increased until a threshold of 7 wt% was reached for the nanofiller film scaffold and 1%wt for the fibrous scaffold. Experimental results of carbon black exhibited a good agreement when compared with data obtained using numerical approaches and analytical models, especially in the case of lower carbon black fractions. Second, we examined the influence of electrical properties of nanofillers based on the concentration and the geometry of carbon nanofillers in the polymer matrix using experimental and numerical simulation approaches. The experimental results showed an increase in conductivity as the amount of nanofiller concentration increased. And regardless of nanofiller type, the trend remained the same. The percolation threshold was around 4-5wt% of nano-additive with PCL and PAN matrices, respectively. However, at the same concentrations, conductivity was higher in graphene-based nanocomposites than for CNF and carbon black-based nanocomposites. The numerical modeling highlighted the effect of nanofillers as constructing a conductive network due to the aggregation phenomenon. The conductivity trend for carbon black and carbon nanofiber-based composites by the numerical simulation approach was similar to the experimental approach. Lastly, we studied the effect of these carbon nanocomposite-based scaffolds on the behavior of cell growth. The results showed that regardless of the scaffold shape (film or fiber) and the additive's type, when the concentration of nano-additives was increased, electrical conductivity and cell density increased also. For a given nano-additive concentration and type, cell density increased in the scaffolds with fiber shape vs. the film. Importantly, as the conductivity of the scaffolds increased, so did the cell density. Consequently, this study has highlighted the close relationship between electrical conductivity, cell density and scaffold orientation. An increase in conductivity can be achieved in two ways: by molecular orientation of the nanofillers or by the appropriate selection of nano-additives such as graphene and carbon nanofiber.

Comparison between self-assembling peptide nanofiber scaffold (SAPNS) and fibrin sealant in neurosurgical hemostasis.

PubMed

Xu, Fei-Fan; Wang, Yue-Chun; Sun, Stella; Ho, Amy S W; Lee, Derek; Kiang, Karrie M Y; Zhang, Xiao-Qin; Lui, Wai-Man; Liu, Bai-Yun; Wu, Wu-Tian; Leung, Gilberto K K

2015-10-01

RADA16-I is a synthetic type I self-assembling peptide nanofiber scaffold (SAPNS) which may serve as a novel biocompatible hemostatic agent. Its application in neurosurgical hemostasis, however, has not been explored. Although RADA16-I is nontoxic and nonimmunogenic, its intrinsic acidity may potentially provoke inflammation in the surgically injured brain. We conducted an animal study to compare RADA16-I with fibrin sealant, a commonly used agent, with the hypothesis that the former would be a comparable alternative. Using a standardized surgical brain injury model, 30 Sprague-Dawley rats were randomized into three treatment groups: RADA16-I, fibrin sealant or gelatin sponge (control). Animals were sacrificed on day 3 and 42. Astrocytic and microglial infiltrations within the cerebral parenchyma adjacent to the operative site were significantly lower in the RADA16-I and fibrin sealant groups than control. RADA16-I did not cause more cellular inflammatory response despite its acidity when compared with fibrin sealant. Immunohistochemical studies showed infiltration by astrocytes and microglia into the fibrin sealant and RADA16-I grafts, suggesting their potential uses as tissue scaffolds. RADA16-I is a promising candidate for further translational and clinical studies that focus on its applications as a safe and effective hemostat, proregenerative nanofiber scaffold as well as drug and cell carrier. © 2015 Wiley Periodicals, Inc.

The surface grafting of graphene oxide with poly(ethylene glycol) as a reinforcement for poly(lactic acid) nanocomposite scaffolds for potential tissue engineering applications.

PubMed

Zhang, Chunmei; Wang, Liwei; Zhai, Tianliang; Wang, Xinchao; Dan, Yi; Turng, Lih-Sheng

2016-01-01

Graphene oxide (GO) was incorporated into poly(lactic acid) (PLA) as a reinforcing nanofiller to produce composite nanofibrous scaffolds using the electrospinning technique. To improve the dispersion of GO in PLA and the interfacial adhesion between the filler and matrix, GO was surface-grafted with poly(ethylene glycol) (PEG). Morphological, thermal, mechanical, and wettability properties, as well as preliminary cytocompatibility with Swiss mouse NIH 3T3 cells of PLA, PLA/GO, and PLA/GO-g-PEG electrospun nanofibers, were characterized. Results showed that the average diameter of PLA/GO-g-PEG electrospun nanofibers decreased with filler content. Both GO and GO-g-PEG improved the thermal stability of PLA, but GO-g-PEG was more effective. The water contact angle test of the nanofiber mats showed that the addition of GO in PLA did not change the surface wettability of the materials, but PLA/GO-g-PEG samples exhibited improved wettability with lower water contact angles. The tensile strength of the composite nanofiber mats was improved with the addition of GO, and it was further enhanced when GO was surface grafted with PEG. This suggested that improved interfacial adhesion between GO and PLA was achieved by grafting PEG onto the GO. The cell viability and proliferation results showed that the cytocompatibility of PLA was not compromised with the addition of GO and GO-g-PEG. With enhanced mechanical properties as well as good wettability and cytocompatibility, PLA/GO-g-PEG composite nanofibers have the potential to be used as scaffolds in tissue engineering. Copyright © 2015 Elsevier Ltd. All rights reserved.

Development of electrospun poly (vinyl alcohol)-based bionanocomposite scaffolds for bone tissue engineering.

PubMed

Enayati, Mohammad Saied; Behzad, Tayebeh; Sajkiewicz, Pawel; Rafienia, Mohammad; Bagheri, Rouhollah; Ghasemi-Mobarakeh, Laleh; Kolbuk, Dorota; Pahlevanneshan, Zari; Bonakdar, Shahin H

2018-04-01

The article is focused on the role of nanohydroxy apatite (nHAp) and cellulose nanofibers (CNFs) as fillers in the electrospun poly (vinyl alcohol) (ES-PVA) nanofibers for bone tissue engineering (TE). Fibrous scaffolds of PVA, PVA/nHAp (10 wt.%), and PVA/nHAp(10 wt.%)/CNF(3 wt.%) were successfully fabricated and characterized. Tensile test on electrospun PVA/nHAp10 and PVA/nHAp10/CNF3 revealed a three-fold and seven-fold increase in modulus compared with pure ES-PVA (45.45 ± 4.77). Although, nanofiller loading slightly reduced the porosity percentage, all scaffolds had porosity higher than 70%. In addition, contact angle test proved the great hydrophilicity of scaffolds. The presence of fillers reduced in vitro biodegradation rate in PBS while accelerates biomineralization in simulated body fluid (SBF). Furthermore, cell viability, cell attachment, and functional activity of osteoblast MG-63 cells were studied on scaffolds showing higher cellular activity for scaffolds with nanofillers. Generally, the obtained results confirm that the 3-componemnt fibrous scaffold of PVA/nHAp/CNF has promising potential in hard TE. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1111-1120, 2018. © 2018 Wiley Periodicals, Inc.

Fabrication of a biomimetic ZeinPDA nanofibrous scaffold impregnated with BMP-2 peptide conjugated TiO2 nanoparticle for bone tissue engineering.

PubMed

Babitha, S; Annamalai, Meenakshi; Dykas, Michal Marcin; Saha, Surajit; Poddar, Kingshuk; Venugopal, Jayarama Reddy; Ramakrishna, Seeram; Venkatesan, Thirumalai; Korrapati, Purna Sai

2018-04-01

A biomimetic Zein polydopamine based nanofiber scaffold was fabricated to deliver bone morphogenic protein-2 (BMP-2) peptide conjugated titanium dioxide nanoparticles in a sustained manner for investigating its osteogenic differentiation potential. To prolong its retention time at the target site, BMP-2 peptide has been conjugated to titanium dioxide nanoparticles owing to its high surface to volume ratio. The effect of biochemical cues from BMP-2 peptide and nanotopographical stimulation of electrospun Zein polydopamine nanofiber were examined for its enhanced osteogenic expression of human fetal osteoblast cells. The sustained delivery of bioactive signals, improved cell adhesion, mineralization, and differentiation could be attributed to its highly interconnected nanofibrous matrix with unique material composition. Further, the expression of osteogenic markers revealed that the fabricated nanofibrous scaffold possess better cell-biomaterial interactions. These promising results demonstrate the potential of the composite nanofibrous scaffold as an effective biomaterial substrate for bone regeneration. Copyright © 2017 John Wiley & Sons, Ltd.

Enhancement of Cell-Based Therapeutic Angiogenesis Using a Novel Type of Injectable Scaffolds of Hydroxyapatite-Polymer Nanocomposite Microspheres

PubMed Central

Koyama, Hidenori; Okada, Masahiro; Tanaka, Shinji; Shoji, Tetsuo; Emoto, Masanori; Furuzono, Tsutomu; Nishizawa, Yoshiki; Inaba, Masaaki

2012-01-01

Background Clinical trials demonstrate the effectiveness of cell-based therapeutic angiogenesis in patients with severe ischemic diseases; however, their success remains limited. Maintaining transplanted cells in place are expected to augment the cell-based therapeutic angiogenesis. We have reported that nano-hydroxyapatite (HAp) coating on medical devices shows marked cell adhesiveness. Using this nanotechnology, HAp-coated poly(l-lactic acid) (PLLA) microspheres, named nano-scaffold (NS), were generated as a non-biological, biodegradable and injectable cell scaffold. We investigate the effectiveness of NS on cell-based therapeutic angiogenesis. Methods and Results Bone marrow mononuclear cells (BMNC) and NS or control PLLA microspheres (LA) were intramuscularly co-implanted into mice ischemic hindlimbs. When BMNC derived from enhanced green fluorescent protein (EGFP)-transgenic mice were injected into ischemic muscle, the muscle GFP level in NS+BMNC group was approximate fivefold higher than that in BMNC or LA+BMNC groups seven days after operation. Kaplan-Meier analysis demonstrated that NS+BMNC markedly prevented hindlimb necrosis (P<0.05 vs. BMNC or LA+BMNC). NS+BMNC revealed much higher induction of angiogenesis in ischemic tissues and collateral blood flow confirmed by three-dimensional computed tomography angiography than those of BMNC or LA+BMNC groups. NS-enhanced therapeutic angiogenesis and arteriogenesis showed good correlations with increased intramuscular levels of vascular endothelial growth factor and fibroblast growth factor-2. NS co-implantation also prevented apoptotic cell death of transplanted cells, resulting in prolonged cell retention. Conclusion A novel and feasible injectable cell scaffold potentiates cell-based therapeutic angiogenesis, which could be extremely useful for the treatment of severe ischemic disorders. PMID:22529991

Genipin-crosslinked silk fibroin/hydroxybutyl chitosan nanofibrous scaffolds for tissue-engineering application.

PubMed

Zhang, Kuihua; Qian, Yongfang; Wang, Hongsheng; Fan, Linpeng; Huang, Chen; Yin, Anlin; Mo, Xiumei

2010-12-01

To improve water-resistant ability and mechanical properties of silk fibroin (SF)/hydroxybutyl chitosan (HBC) nanofibrous scaffolds for tissue-engineering applications, genipin, glutaraldehyde (GTA), and ethanol were used to crosslink electrospun nanofibers, respectively. The mechanical properties of nanofibrous scaffolds were obviously improved after 24 h of crosslinking with genipin and were superior to those crosslinked with GTA and ethanol for 24 h. SEM indicated that crosslinked nanofibers with genipin and GTA vapor had good water-resistant ability. Characterization of the microstructure (porosity and pore structure) demonstrated crosslinked nanofibrous scaffolds with genipin and GTA vapor had lager porosities and mean diameters than those with ethanol. Characterization of FTIR-ATR and (13)C NMR clarified both genipin and GTA acted as crosslinking agents for SF and HBC. Furthermore, genipin could induce SF conformation from random coil or α-helix to β-sheet. Although GTA could also successfully crosslink SF/HBC nanofibrous scaffolds, in long run, genipin maybe a better method due to lower cytotoxicity than GTA. Cell viability studies and wound-healing test in rats clarified that the genipin-crosslinked SF/HBC nanofibrous scaffolds had a good biocompatibility both in vitro and in vivo. These results suggested that genipin-crosslinked SF/HBC nanofibrous scaffolds might be potential candidates for wound dressing and tissue-engineering scaffolds. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2010.

A biodegradable antibiotic-eluting PLGA nanofiber-loaded deproteinized bone for treatment of infected rabbit bone defects.

PubMed

Gao, Jianting; Huang, Guofeng; Liu, Guojun; Liu, Yan; Chen, Qi; Ren, Lei; Chen, Changqing; Ding, Zhenqi

2016-08-01

We fabricated a biodegradable antibiotic-eluting poly(d,l)-lactide-co-glycolide nanofiber-loaded deproteinized bone (ANDB) scaffold that provided sustained delivery of vancomycin to repair methicillin-resistant Staphylococcus aureus bone defects. To fabricate the biodegradable ANDB, poly(d,l)-lactide-co-glycolide and vancomycin were first dissolved in 1,1,1,3,3,3-hexafluoro-2-propano. The solution was then electrospun to produce biodegradable antibiotic-eluting membranes that were deposited on the surface of bovine deproteinized cancellous bone. We used scanning electron microscopy to determine the properties of the scaffold. Both elution and high-performance liquid chromatography assays were used to evaluate the in vitro vancomycin release rate from the ANDB scaffold. Three types of scaffolds were co-cultured with bacteria to confirm the in vitro antibacterial activity. The infected bone defect rabbit model was induced by injecting 10(7) colony forming units of a methicillin-resistant Staphylococcus aureus strain into the radial defect of rabbits. Animals were then separated into treatment groups and implanted according to the following scheme: ANDB scaffold in group A, poly(d,l)-lactide-co-glycolide nanofiber-loaded deproteinized bone (NDB) scaffold with intravenous (i.v.) vancomycin in group B, and NDB scaffold alone in group C. Treatment efficacy was evaluated after eight weeks using radiological, microbiological, and histological examinations. In vitro results revealed that biodegradable ANDB scaffolds released concentrations of vancomycin that were greater than the minimum inhibitory concentration for more than four weeks. Bacterial inhibition tests also confirmed antibacterial efficacy lasted for approximately four weeks. Radiological and histological scores obtained in vivo revealed significant differences between groups A, B and C. Importantly, group A had significantly lower bacterial load and better bone regeneration when compared to either group B or C. Collectively, these results show that our fabricated ANDB scaffolds possess: (1) effective bactericidal activity against methicillin-resistant Staphylococcus aureus, (2) the ability to promote site-specific bone regeneration, and (3) the potential for use in the treatment of infected bone defects. © The Author(s) 2016.

Calcium phosphate deposition rate, structure and osteoconductivity on electrospun poly(l-lactic acid) matrix using electrodeposition or simulated body fluid incubation

PubMed Central

He, Chuanglong; Jin, Xiaobing; Ma, Peter X.

2013-01-01

Mineralized nanofibrous scaffolds have been proposed as promising scaffolds for bone regeneration due to their ability to mimic both nanoscale architecture and chemical composition of natural bone extracellular matrix (ECM). In this study, a novel electrodeposition method was compared with an extensively explored simulated body fluid (SBF) incubation method in terms of the deposition rate, chemical composition, and morphology of calcium phosphate formed on electrospun fibrous thin matrices with a fiber diameter in the range from about 200 nm to about 1400 nm prepared using 6, 8, 10 and 12 wt% poly(l-lactic acid) (PLLA) solutions in a mixture of dichloromethane and acetone (2:1 in volume). The effects of the surface modification using the two mineralization techniques on osteoblastic cell (MC3T3-E1) proliferation and differentiation were also examined. It was found that electrodeposition was two to three orders of magnitude faster than the SBF method in mineralizing the fibrous matrices, reducing the mineralization time from about two weeks to an hour to achieve the same amounts of mineralization. The mineralization rate also varied with the fiber diameter but in opposite directions between the two mineralization methods. As a general trend, the increase of fiber diameter resulted in a faster mineralization rate for the electrodeposition method but a slower mineralization rate for the SBF incubation method. Using the electrodeposition method, one can control the chemical composition and morphology of the calcium phosphate by varying the electric deposition potential and electrolyte temperature to tune the mixture of dicalcium phosphate dihydrate (DCPD) and hydroxy apatite (HAp). Using the SBF method, one can only obtain a low crystallinity HAp. The mineralized electrospun PLLA fibrous matrices from either method similarly facilitate the proliferation and osteogenic differentiation of preosteoblastic MC3T3-E1 cells as compared to neat PLLA matrices. Therefore, the electrodeposition method can be utilized as a fast and versatile technique to fabricate mineralized nanofibrous scaffolds for bone tissue engineering. PMID:24012605

Fabrication of Poly(ε-caprolactone) Scaffolds Reinforced with Cellulose Nanofibers, with and without the Addition of Hydroxyapatite Nanoparticles.

PubMed

Morouço, Pedro; Biscaia, Sara; Viana, Tânia; Franco, Margarida; Malça, Cândida; Mateus, Artur; Moura, Carla; Ferreira, Frederico C; Mitchell, Geoffrey; Alves, Nuno M

2016-01-01

Biomaterial properties and controlled architecture of scaffolds are essential features to provide an adequate biological and mechanical support for tissue regeneration, mimicking the ingrowth tissues. In this study, a bioextrusion system was used to produce 3D biodegradable scaffolds with controlled architecture, comprising three types of constructs: (i) poly( ε -caprolactone) (PCL) matrix as reference; (ii) PCL-based matrix reinforced with cellulose nanofibers (CNF); and (iii) PCL-based matrix reinforced with CNF and hydroxyapatite nanoparticles (HANP). The effect of the addition and/or combination of CNF and HANP into the polymeric matrix of PCL was investigated, with the effects of the biomaterial composition on the constructs (morphological, thermal, and mechanical performances) being analysed. Scaffolds were produced using a single lay-down pattern of 0/90°, with the same processing parameters among all constructs being assured. The performed morphological analyses showed a satisfactory distribution of CNF within the polymer matrix and high reliability was obtained among the produced scaffolds. Significant effects on surface wettability and thermal properties were observed, among scaffolds. Regarding the mechanical properties, higher scaffold stiffness in the reinforced scaffolds was obtained. Results from the cytotoxicity assay suggest that all the composite scaffolds presented good biocompatibility. The results of this first study on cellulose and hydroxyapatite reinforced constructs with controlled architecture clearly demonstrate the potential of these 3D composite constructs for cell cultivation with enhanced mechanical properties.

Electrodeposited nickel-cobalt sulfide nanosheet on polyacrylonitrile nanofibers: a binder-free electrode for flexible supercapacitors

NASA Astrophysics Data System (ADS)

Kamran Sami, Syed; Siddiqui, Saqib; Tajmeel Feroze, Muhammad; Chung, Chan-Hwa

2017-11-01

To pursue high-performance energy storage devices with both high energy density and power density, one-dimensional (1D) nanostructures play a key role in the development of functional devices including energy conversion, energy storage, and environmental devices. The polyacrylonitrile (PAN) nanofibers were obtained by the versatile electrospinning method. An ultra-thin nickel-cobalt sulfide (NiCoS) layer was conformably electrodeposited on a self-standing PAN nanofibers by cyclic voltammetry to fabricate the light-weighted porous electrodes for supercapacitors. The porous web of PAN nanofibers acts as a high-surface-area scaffold with significant electrochemical performance, while the electrodeposition of metal sulfide nanosheet further enhances the specific capacitance. The fabricated NiCoS on PAN (NiCoS/PAN) nanofibers exhibits a very high capacitance of 1513 F g-1 at 5 A g-1 in 1 M potassium chloride (KCl) aqueous electrolyte with superior rate capability and excellent electrochemical stability as a hybrid electrode. The high capacitance of the NiCoS is attributed to the large surface area of the electrospun PAN nanofibers scaffold, which has offered a large number of active sites for possible redox reaction of ultra-thin NiCoS layer. Benefiting from the compositional features and electrode architectures, the hybrid electrode of NiCoS/PAN nanofibers shows greatly improved electrochemical performance with an ultra-high capacitance (1124 F g-1 at 50 A g-1). Moreover, a binder-free asymmetric supercapacitor device is also fabricated by using NiCoS/PAN nanofibers as the positive electrode and activated carbon (MSP-20) on PAN nanofibers as the negative electrode; this demonstrates high energy density of 56.904 W h kg-1 at a power density of 1.445 kW kg-1, and it still delivers the energy density of 33.3923 W h kg-1 even at higher power density of 16.5013 kW kg-1.

γ-Fe2O3 nanoparticles filled polyvinyl alcohol as potential biomaterial for tissue engineering scaffold.

PubMed

Ngadiman, Nor Hasrul Akhmal; Idris, Ani; Irfan, Muhammad; Kurniawan, Denni; Yusof, Noordin Mohd; Nasiri, Rozita

2015-09-01

Maghemite (γ-Fe2O3) nanoparticle with its unique magnetic properties is recently known to enhance the cell growth rate. In this study, γ-Fe2O3 is mixed into polyvinyl alcohol (PVA) matrix and then electrospun to form nanofibers. Design of experiments was used to determine the optimum parameter settings for the electrospinning process so as to produce elctrospun mats with the preferred characteristics such as good morphology, Young's modulus and porosity. The input factors of the electrospinnning process were nanoparticles content (1-5%), voltage (25-35 kV), and flow rate (1-3 ml/h) while the responses considered were Young's modulus and porosity. Empirical models for both responses as a function of the input factors were developed and the optimum input factors setting were determined, and found to be at 5% nanoparticle content, 35 kV voltage, and 1 ml/h volume flow rate. The characteristics and performance of the optimum PVA/γ-Fe2O3 nanofiber mats were compared with those of neat PVA nanofiber mats in terms of morphology, thermal properties, and hydrophilicity. The PVA/γ-Fe2O3 nanofiber mats exhibited higher fiber diameter and surface roughness yet similar thermal properties and hydrophilicity compared to neat PVA PVA/γ-Fe2O3 nanofiber mats. Biocompatibility test by exposing the nanofiber mats with human blood cells was performed. In terms of clotting time, the PVA/γ-Fe2O3 nanofibers exhibited similar behavior with neat PVA. The PVA/γ-Fe2O3 nanofibers also showed higher cells proliferation rate when MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was done using human skin fibroblast cells. Thus, the PVA/γ-Fe2O3 electrospun nanofibers can be a promising biomaterial for tissue engineering scaffolds. Copyright © 2015 Elsevier Ltd. All rights reserved.

Synthesis of HAP nano rods and processing of nano-size ceramic reinforced poly(L)lactic acid composites

NASA Astrophysics Data System (ADS)

Flanigan, Kyle Yusef

2000-09-01

Bone is unique among the various connective tissues in that it is a composite of organic and inorganic components. Calcium phosphates occur principally in the form of hydroxyapatite crystals {Ca10(PO4) 6(OH)2}. Secreted apatite crystals are integral to the structural rigidity of the bone. When a bone breaks, there is often a need to implant an orthotic device to support the broken bone during remodeling. Current technologies use either metal pins and screws that need to be removed (by surgery) once the healing is complete or polymeric materials that either get resorbed or are porous enough to allow bone ingrowth. Poly(L)Lactic acid and copolymers of polyglycolic acid (PGA) are thermoplastics which show promise as the matrix material in biosorbable/load bearing implants. In service this material is hydrolyzed generating water and L-lactate. Orthoses composed of neat PLLA resins require greater than three years for complete resorbtion, however; 95% of strength is lost in 2 to 3 weeks in-vitro. This has limited the deployment of load bearing PLLA to screws, pins or short bracing spans. There exists a need for the development of an implantable and biosorbable orthotic device which will retain its structural integrity long enough for remodeling and healing process to generate new bone material, about 10 weeks. The scope of this dissertation is the development of HAP nano-whisker reinforcement and a HAP/PLLA thermoplastic composite. As proof of the feasibility of generating nano-reinforcement PLLA-composites, the surface of a galleried clay, montmorillonite, was modified and clay/PLLA composites processed and then characterized. Hydroxyapatite nano-whiskers were synthesized and functionalized using organosilanes and Menhaden fish-oil (common organic dispersant). The functionalized nano-fibers were used to process HAP/PLLA composites. Characterization techniques included thermal analysis, magnetic spectroscopy, XRD and ICP analysis and electron microscopy. The montmorillonite galleries were successfully intercalated and exfoliated. Gallery spacings increased as much as 30A with ODA. NMR results demonstrated that the ODA in the galleries of the highly loaded clay is in an extended trans-configuration at lower temperatures. Furthermore the alkyl chains are nano-constrained and have limited mobility. These configurations influence the gallery spacing. The appearance of two crystalline melts (DSC) may indicate that the desired physicochemical modification of the PLLA at the clay nano-reinforcement interface was achieved. For the growth of HAP nano-rods a hydrothermal synthesis route was developed. A kinetics study revealed several unique features of the method of growth. TEM analysis indicates that the synthesis procedure was successful in generating rod-like HAP structures of 100nm length and 10 nm in width. The effect of synthesis conditions on the phase purity and the morphology of the precipitates was investigated. The surface of the HAP rods was modified using OTS and OMS. The surface modified HAP was used to process HAP/PLLA composites. The properties of the composites depend strongly on the nature of the interface. Composites made with OMS or OTS demonstrated a higher elastic modulus. At 1% solids loading the OTS treated sample generated a 40% increase in modulus. Silane treated composites had DMA transitions shifted 10 to 20 degrees higher. "Well-ordered" SAMs improve the dispersion of the inorganic reinforcement in PLLA and promote the formation of mechanical entanglements at the HAP-PLLA interface. As a result load transfer is more complete resulting in higher modulus material.

Adhesion and proliferation of OCT-1 osteoblast-like cells on micro- and nano-scale topography structured poly(L-lactide).

PubMed

Wan, Yuqing; Wang, Yong; Liu, Zhimin; Qu, Xue; Han, Buxing; Bei, Jianzhong; Wang, Shenguo

2005-07-01

The impact of the surface topography of polylactone-type polymer on cell adhesion was to be concerned because the micro-scale texture of a surface can provide a significant effect on the adhesion behavior of cells on the surface. Especially for the application of tissue engineering scaffold, the pore size could have an influence on cell in-growth and subsequent proliferation. Micro-fabrication technology was used to generate specific topography to investigate the relationship between the cells and surface. In this study the pits-patterned surfaces of polystyrene (PS) film with diameters 2.2 and 0.45 microm were prepared by phase-separation, and the corresponding scale islands-patterned PLLA surface was prepared by a molding technique using the pits-patterned PS as a template. The adhesion and proliferation behavior of OCT-1 osteoblast-like cells morphology on the pits- and islands-patterned surface were characterized by SEM observation, cell attachment efficiency measurement and MTT assay. The results showed that the cell adhesion could be enhanced on PLLA and PS surface with nano-scale and micro-scale roughness compared to the smooth surfaces of the PLLA and PS. The OCT-1 osteoblast-like cells could grow along the surface with two different size islands of PLLA and grow inside the micro-scale pits of the PS. However, the proliferation of cells on the micro- and nano-scale patterned surface has not been enhanced compared with the controlled smooth surface.

Impedance Biosensors and Deep Crater Salivary Gland Scaffolds for Tissue Engineering

NASA Astrophysics Data System (ADS)

Schramm, Robert A.

The salivary gland is a complex, branching organ whose primary biological function is the production of the fluid critical to alimentary function and the lubrication and maintenance of the oral cavity, saliva. The most frequent disruption of the salivary organ system is one in which the rate of supply of saliva into the oral cavity is diminished, and this may vary from a minor reduction, to near cessation. Regenerative medicine is a field which seeks to find ways to overcome the symptoms of organ malfunction or damage by inducing regrowth, repair and replacement of partial or whole organ function. Historically, the only methods available to medical experts were certain chemical drugs and transplantation, each of which suffers from significant risks and drawbacks. Tissue Engineering arose in the past few decades thanks to the seminal work of Robert Langer with the charter mission of finding new biomaterials and techniques to achieve these ends. The original concept of tissue engineering was the cell or tissue scaffold, which is supports the regrowth of cells by making intimate contact with adherent cells, and induces improved regrowth in vitro or in vivo by providing mechanical or chemical signaling cues. Epithelial cell types such as salivary glands have structural functional polarity at the cellular level, an apical side which faces a void, and a basal side which faces the support substrate. While 3D scaffolds such as hydrogels maximize interaction area between cells and substrate, they struggle to develop cohesive tissues beyond the scale of small cellular clusters . 2D scaffolds enforce a defined polarity by allowing cell interaction at only one side of the cell. Langer pioneered the use of polymer nanofibers as the premier synthetic 2D scaffold biomaterial, due to their exceptionally high nano-scale surface area, and collagen-imitating structure. Prior work has established PLGA nanofibers, which allow salivary cells to attach, proliferate, and generate a thicker cobblestone-style cellular monolayer. In addition, providing shallow depressions in the nanofiber scaffold allows the salivary gland cells to experience a biomimetic substrate curvature, which further increases cell height, but not to the level of matching the height along the apico-basal vector of in vivo or 3D gels . This work endeavors to increase the depth of the depressions, in order to allow for an increase in substrate curvature and a maximization of cell height. It was also undertaken to develop an alternative method to grading the effectiveness of our scaffolds compared with one another. Analyzing protein structural localization with immunofluorescence and protein bulk concentration with western blot have some limitations. An electrochemical detection technique was developed to nondestructively assess the performance of scaffolds, specifically in inducing stronger resistance to fluid diffusion across the cell monolayer on a 2D pseudo-planar scaffold. This impedance spectroscopy technique, called trans-epithelial electrical resistance spectroscopy, requires the cells be suspended in media, with opposing electrodes above and below, generating an alternating current which drives free ions in the cell media across the scaffold membrane and cell layer, measuring the resistance that the membrane generates. Ions traverse the cell junctions preferentially, thus reporting on the junction barrier effectiveness. This method can be used to run large parallel experiments with multiple scaffold conditions, permitted that the scaffolds can be mounted within the apparatus. This research was able to eliminate once necessitated glass and polymer scaffold under layers, increasing scaffold perfusivity and allowing for a TEER analysis. Results show that salivary gland cells behave similarly on these thinned PLGA nanofiber scaffolds as on the control membrane.

Graphene-augmented nanofiber scaffolds demonstrate new features in cells behaviour

NASA Astrophysics Data System (ADS)

Kazantseva, Jekaterina; Ivanov, Roman; Gasik, Michael; Neuman, Toomas; Hussainova, Irina

2016-07-01

Three-dimensional (3D) customized scaffolds capable to mimic a native extracellular matrix open new frontiers in cells manipulation and advanced therapy. The major challenge is in a proper substrate for in vitro models on engineered scaffolds, capable to modulate cells differentiation. Here for the first time we demonstrate novel design and functionality of the 3D porous scaffolds of aligned, self-assembled ceramic nanofibers of ultra-high anisotropy ratio (~107), augmented into graphene shells. This unique hybrid nano-network allows an exceptional combination of selective guidance stimuli of stem cells differentiation, immune reactions variations, and local immobilization of cancer cells, which was not available before. The scaffolds were shown to be able to direct human mesenchymal stem cells (important for stimulation of neuronal and muscle cells) preferential orientation, to suppress major inflammatory factors, and to localize cancer cells; all without additions of specific culture media. The selective downregulation of specific cytokines is anticipated as a new tool for understanding of human immune system and ways of treatment of associated diseases. The effects observed are self-regulated by cells only, without side effects, usually arising from use of external factors. New scaffolds may open new horizons for stem cells fate control such as towards axons and neurites regeneration (Alzheimer’s disease) as well as cancer therapy development.

Controlled release of functional proteins through designer self-assembling peptide nanofiber hydrogel scaffold

PubMed Central

Koutsopoulos, Sotirios; Unsworth, Larry D.; Nagai, Yusuke; Zhang, Shuguang

2009-01-01

The release kinetics for a variety of proteins of a wide range of molecular mass, hydrodynamic radii, and isoelectric points through a nanofiber hydrogel scaffold consisting of designer self-assembling peptides were studied by using single-molecule fluorescence correlation spectroscopy (FCS). In contrast to classical diffusion experiments, the single-molecule approach allowed for the direct determination of diffusion coefficients for lysozyme, trypsin inhibitor, BSA, and IgG both inside the hydrogel and after being released into the solution. The results of the FCS analyses and the calculated pristine in-gel diffusion coefficients were compared with the values obtained from the Stokes–Einstein equation, Fickian diffusion models, and the literature. The release kinetics suggested that protein diffusion through nanofiber hydrogels depended primarily on the size of the protein. Protein diffusivities decreased, with increasing hydrogel nanofiber density providing a means of controlling the release kinetics. Secondary and tertiary structure analyses and biological assays of the released proteins showed that encapsulation and release did not affect the protein conformation and functionality. Our results show that this biocompatible and injectable designer self-assembling peptide hydrogel system may be useful as a carrier for therapeutic proteins for sustained release applications. PMID:19273853

An anisotropically and heterogeneously aligned patterned electrospun scaffold with tailored mechanical property and improved bioactivity for vascular tissue engineering.

PubMed

Xu, He; Li, Haiyan; Ke, Qinfei; Chang, Jiang

2015-04-29

The development of vascular scaffolds with controlled mechanical properties and stimulatory effects on biological activities of endothelial cells still remains a significant challenge to vascular tissue engineering. In this work, we reported an innovative approach to prepare a new type of vascular scaffolds with anisotropically and heterogeneously aligned patterns using electrospinning technique with unique wire spring templates, and further investigated the structural effects of the patterned electrospun scaffolds on mechanical properties and angiogenic differentiation of human umbilical vein endothelial cells (HUVECs). Results showed that anisotropically aligned patterned nanofibrous structure was obtained by depositing nanofibers on template in a structurally different manner, one part of nanofibers densely deposited on the embossments of wire spring and formed cylindrical-like structures in the transverse direction, while others loosely suspended and aligned along the longitudinal direction, forming a three-dimensional porous microstructure. We further found that such structures could efficiently control the mechanical properties of electrospun vascular scaffolds in both longitudinal and transverse directions by altering the interval distances between the embossments of patterned scaffolds. When HUVECs were cultured on scaffolds with different microstructures, the patterned scaffolds distinctively promoted adhesion of HUVECs at early stage and proliferation during the culture period. Most importantly, cells experienced a large shape change associated with cell cytoskeleton and nuclei remodeling, leading to a stimulatory effect on angiogenesis differentiation of HUVECs by the patterned microstructures of electrospun scaffolds, and the scaffolds with larger distances of intervals showed a higher stimulatory effect. These results suggest that electrospun scaffolds with the anisotropically and heterogeneously aligned patterns, which could efficiently control the mechanical properties and bioactivities of the scaffolds, might have great potential in vascular tissue engineering application.

Optimization of a biomimetic poly-(lactic acid) ligament scaffold

NASA Astrophysics Data System (ADS)

Uehlin, Andrew F.

The anterior cruciate ligament (ACL) is the most commonly injured ligament of the knee, often requiring orthopedic reconstruction using autograft or allograph tissue, both with significant disadvantages. As a result, tissue engineering an ACL replacement graft has been heavily investigated. The present study attempts to replicate the morphology and mechanical properties of the ACL using a nanomatrix composite of highly-aligned poly(lactic acid) (PLA) fibers with various surface and biochemical modifications. Additionally, this study attempts to recreate the natural mineralization gradient found at the ACL enthesis onto the scaffold, capable of inducing a favorable cellular response in vitro. Unidirectional electrospinning was used to create nanofibers of PLA, followed by an induced degradation of the nanofibers via 0.25M NaOH hydrolysis. The effects of the unidirectional electrospinning as well as the effects of NaOH hydrolysis on fiber alignment, fiber diameter, surface morphology, crystallinity, in vitro swelling, immobilization of fibrin, and mechanical properties were investigated, resulting in a modified morphology correlating to the microstructure of native ligament tissue with similar mechanical properties. Furthering the development of the PLA nanomatrix composite, a bioinkjet printer was used to immobilize nanoparticulate hydroxyapatite (HANP) on the surface of the scaffold. A series of 300pL droplets of HANP bioink were printed over a gradient pattern mimetic of (and spatially corresponding to) the mineralization gradient found over the microanatomy at the ACL enthesis. Proliferation and differentiation response of human mesenchymal stem cells (hMSCs) in vitro was assessed on a variety of conditions and combinations of the PLA nanofiber scaffold surface modifications (inclusive and exclusive of HANP, fibrin, and various time dependent NaOH treatments). It was found that a combinatory effect of the HANP gradient with fibrin on 20 minute NaOH treated PLA nanofibers enhanced the osteogenic differentiation of hMSCs, with an observable morphological change spatially corresponding to the compositional changes of the printed HANP gradient. Using the bioactive scaffold designed in this study as a template and expanding on the methods utilized, future studies can incorporate specific growth factors and other organic/inorganic biomolecules to further develop the engineered PLA nanomatrix into a functional ligament-replacement graft.

Electrospun photosensitive nanofibers: potential for photocurrent therapy in skin regeneration.

PubMed

Jin, Guorui; Prabhakaran, Molamma P; Kai, Dan; Kotaki, Masaya; Ramakrishna, Seeram

2013-01-01

Poly(3-hexylthiophene) (P3HT) is one of the most promising photovoltaic (PV) polymers in photocurrent therapy. A novel photosensitive scaffold for skin tissue engineering was fabricated by blending P3HT with polycaprolactone (PCL) and electrospun to obtain composite PCL/P3HT nanofibers with three different weight ratios of PCL : P3HT (w/w) of 150 : 2 [PCL/P3HT(2)], 150 : 10 [PCL/P3HT(10)] and 150 : 20 [PCL/P3HT(20)]. The photosensitive properties of the blend solutions and the composite nanofibers of PCL/P3HT were investigated. The incident photon-to-electron conversion efficiencies of the PCL/P3HT(2), PCL/P3HT(10), PCL/P3HT(20) were identified as 2.0 × 10(-6), 1.6 × 10(-5) and 2.9 × 10(-5), respectively, which confirm the photosensitive ability of the P3HT-containing scaffolds. The biocompatibility of the scaffold was evaluated by culturing human dermal fibroblasts and the results showed that the proliferation of HDFs under light stimulation on PCL/P3HT(10) was 12.8%, 11.9%, and 11.6% (p ≤ 0.05) higher than the cell growth on PCL, PCL/P3HT(2) and PCL/P3HT(20), respectively. Human dermal fibroblasts cultured under light stimulation on PCL/P3HT(10) not only showed better cell proliferation but also retained cell morphology similar to the phenotype observed on tissue culture plates (control). Our experimental results suggest novel and potential application of an optimized amount of P3HT-containing scaffold, especially PCL/P3HT(10) nanofibrous scaffold in photocurrent therapy for skin regeneration.

Myocardial tissue engineering using electrospun nanofiber composites

PubMed Central

Kim, Pyung-Hwan; Cho, Je-Yoel

2016-01-01

Emerging trends for cardiac tissue engineering are focused on increasing the biocompatibility and tissue regeneration ability of artificial heart tissue by incorporating various cell sources and bioactive molecules. Although primary cardiomyocytes can be successfully implanted, clinical applications are restricted due to their low survival rates and poor proliferation. To develop successful cardiovascular tissue regeneration systems, new technologies must be introduced to improve myocardial regeneration. Electrospinning is a simple, versatile technique for fabricating nanofibers. Here, we discuss various biodegradable polymers (natural, synthetic, and combinatorial polymers) that can be used for fiber fabrication. We also describe a series of fiber modification methods that can increase cell survival, proliferation, and migration and provide supporting mechanical properties by mimicking micro-environment structures, such as the extracellular matrix (ECM). In addition, the applications and types of nanofiber-based scaffolds for myocardial regeneration are described. Finally, fusion research methods combined with stem cells and scaffolds to improve biocompatibility are discussed. [BMB Reports 2016; 49(1): 26-36] PMID:26497579

Effect of negatively charged cellulose nanofibers on the dispersion of hydroxyapatite nanoparticles for scaffolds in bone tissue engineering.

PubMed

Park, Minsung; Lee, Dajung; Shin, Sungchul; Hyun, Jinho

2015-06-01

Nanofibrous 2,2,6,6-tetramethylpiperidine-1-oxyl(TEMPO)-oxidized bacterial cellulose (TOBC) was used as a dispersant of hydroxyapatite (HA) nanoparticles in aqueous solution. The surfaces of TOBC nanofibers were negatively charged after the reaction with the TEMPO/NaBr/NaClO system at pH 10 and room temperature. HA nanoparticles were simply adsorbed on the TOBC nanofibers (HA-TOBC) and dispersed well in DI water. The well-dispersed HA-TOBC colloidal solution formed a hydrogel after the addition of gelatin, followed by crosslinking with glutaraldehyde (HA-TOBC-Gel). The chemical modification of the fiber surfaces and the colloidal stability of the dispersion solution confirmed TOBC as a promising HA dispersant. Both the Young's modulus and maximum tensile stress increased as the amount of gelatin increased due to the increased crosslinking of gelatin. In addition, the well-dispersed HA produced a denser scaffold structure resulting in the increase of the Young's modulus and maximum tensile stress. The well-developed porous structures of the HA-TOBC-Gel composites were incubated with Calvarial osteoblasts. The HA-TOBC-Gel significantly improved cell proliferation as well as cell differentiation confirming the material as a potential candidate for use in bone tissue engineering scaffolds. Copyright © 2015 Elsevier B.V. All rights reserved.

Nanofiber Alignment Regulates NIH3T3 Cell Orientation and Cytoskeletal Gene Expression on Electrospun PCL+Gelatin Nanofibers.

PubMed

Fee, Timothy; Surianarayanan, Swetha; Downs, Crawford; Zhou, Yong; Berry, Joel

2016-01-01

To examine the influence of substrate topology on the behavior of fibroblasts, tissue engineering scaffolds were electrospun from polycaprolactone (PCL) and a blend of PCL and gelatin (PCL+Gel) to produce matrices with both random and aligned nanofibrous orientations. The addition of gelatin to the scaffold was shown to increase the hydrophilicity of the PCL matrix and to increase the proliferation of NIH3T3 cells compared to scaffolds of PCL alone. The orientation of nanofibers within the matrix did not have an effect on the proliferation of adherent cells, but cells on aligned substrates were shown to elongate and align parallel to the direction of substrate fiber alignment. A microarray of cyotoskeleton regulators was probed to examine differences in gene expression between cells grown on an aligned and randomly oriented substrates. It was found that transcriptional expression of eight genes was statistically different between the two conditions, with all of them being upregulated in the aligned condition. The proteins encoded by these genes are linked to production and polymerization of actin microfilaments, as well as focal adhesion assembly. Taken together, the data indicates NIH3T3 fibroblasts on aligned substrates align themselves parallel with their substrate and increase production of actin and focal adhesion related genes.

Development of VEGF-loaded PLGA matrices in association with mesenchymal stem cells for tissue engineering

PubMed Central

Rosa, A.R.; Steffens, D.; Santi, B.; Quintiliano, K.; Steffen, N.; Pilger, D.A.; Pranke, P.

2017-01-01

The association of bioactive molecules, such as vascular endothelial growth factor (VEGF), with nanofibers facilitates their controlled release, which could contribute to cellular migration and differentiation in tissue regeneration. In this research, the influence of their incorporation on a polylactic-co-glycolic acid (PLGA) scaffold produced by electrospinning on cell adhesion and viability and cytotoxicity was carried out in three groups: 1) PLGA/BSA/VEGF; 2) PLGA/BSA, and 3) PLGA. Morphology, fiber diameter, contact angle, loading efficiency and controlled release of VEGF of the biomaterials, among others, were measured. The nanofibers showed smooth surfaces without beads and with interconnected pores. PLGA/BSA/VEGF showed the smallest water contact angle and VEGF released for up to 160 h. An improvement in cell adhesion was observed for the PLGA/BSA/VEGF scaffolds compared to the other groups and the scaffolds were non-toxic for the cells. Therefore, the scaffolds were shown to be a good strategy for sustained delivery of VEGF and may be a useful tool for tissue engineering. PMID:28793048

Cytotoxicity assessment of polyhydroxybutyrate/chitosan/nano- bioglass nanofiber scaffolds by stem cells from human exfoliated deciduous teeth stem cells from dental pulp of exfoliated deciduous tooth

PubMed Central

Hashemi-Beni, Batool; Khoroushi, Maryam; Foroughi, Mohammad Reza; Karbasi, Saeed; Khademi, Abbas Ali

2018-01-01

Background: The aim of this study was to compare the cytotoxicity and the biocompatibility of three different nanofibers scaffolds after seeding of stem cells harvested from human deciduous dental pulp. Given the importance of scaffold and its features in tissue engineering, this study demonstrated the construction of polyhydroxybutyrate (PHB)/chitosan/nano-bioglass (nBG) nanocomposite scaffold using electrospinning method. Materials and Methods: This experimental study was conducted on normal exfoliated deciduous incisors obtained from 6-year-old to 11-year-old healthy children. The dental pulp was extracted from primary incisor teeth which are falling aseptically. After digesting the tissue with 4 mg/ml of type I collagenase, the cells were cultured in medium solution. Identification of stem cells from human exfoliated deciduous teeth was performed by flowcytometry using CD19, CD14, CD146, and CD90 markers. Then, 1 × 104 stem cells were seeded on the scaffold with a diameter of 10 mm × 0.3 mm. Cell viability was evaluated on days 3, 5, and 7 through methyl thiazol tetrazolium techniques (P < 0.05) on different groups that they are groups included (1) PHB scaffold (G1), (2) PHB/chitosan scaffold (G2), (3) the optimal PHB/chitosan/nBG scaffold (G3), (4) mineral trioxide aggregate (MTA), and (5) the G3 + MTA scaffold (G3 + MTA). Data were analyzed with two-way ANOVA at significance level of P < 0.05. Results: The results indicated that the PHB/chitosan/nBG scaffold and PHB/chitosan/nBG scaffold + MTA groups showed significant difference compared with the PHB/chitosan scaffold and PHB scaffold groups on the 7th day (P < 0.05). Conclusion: Thus, it can be concluded that the scaffold with nBG nanoparticles is more biocompatible than the other scaffolds and can be considered as a suitable scaffold for growth and proliferation of stem cells. PMID:29576778

Cytotoxicity assessment of polyhydroxybutyrate/chitosan/nano- bioglass nanofiber scaffolds by stem cells from human exfoliated deciduous teeth stem cells from dental pulp of exfoliated deciduous tooth.

PubMed

Hashemi-Beni, Batool; Khoroushi, Maryam; Foroughi, Mohammad Reza; Karbasi, Saeed; Khademi, Abbas Ali

2018-01-01

The aim of this study was to compare the cytotoxicity and the biocompatibility of three different nanofibers scaffolds after seeding of stem cells harvested from human deciduous dental pulp. Given the importance of scaffold and its features in tissue engineering, this study demonstrated the construction of polyhydroxybutyrate (PHB)/chitosan/nano-bioglass (nBG) nanocomposite scaffold using electrospinning method. This experimental study was conducted on normal exfoliated deciduous incisors obtained from 6-year-old to 11-year-old healthy children. The dental pulp was extracted from primary incisor teeth which are falling aseptically. After digesting the tissue with 4 mg/ml of type I collagenase, the cells were cultured in medium solution. Identification of stem cells from human exfoliated deciduous teeth was performed by flowcytometry using CD19, CD14, CD146, and CD90 markers. Then, 1 × 10 4 stem cells were seeded on the scaffold with a diameter of 10 mm × 0.3 mm. Cell viability was evaluated on days 3, 5, and 7 through methyl thiazol tetrazolium techniques ( P < 0.05) on different groups that they are groups included (1) PHB scaffold (G1), (2) PHB/chitosan scaffold (G2), (3) the optimal PHB/chitosan/nBG scaffold (G3), (4) mineral trioxide aggregate (MTA), and (5) the G3 + MTA scaffold (G3 + MTA). Data were analyzed with two-way ANOVA at significance level of P < 0.05. The results indicated that the PHB/chitosan/nBG scaffold and PHB/chitosan/nBG scaffold + MTA groups showed significant difference compared with the PHB/chitosan scaffold and PHB scaffold groups on the 7 th day ( P < 0.05). Thus, it can be concluded that the scaffold with nBG nanoparticles is more biocompatible than the other scaffolds and can be considered as a suitable scaffold for growth and proliferation of stem cells.

Synthesis of Three-dimensional Polymer Nanostructures via Chemical Vapor Deposition

NASA Astrophysics Data System (ADS)

Cheng, Kenneth

Chemical vapor deposition (CVD) is a widely practiced methodology for preparing thin film polymer coatings, and the coatings can be applied to a broad range of materials, including three-dimensional solid structures and low-vapor pressure liquids. Reactive poly(p-xylylene) (PPX) coatings prepared by CVD can be used as a powerful tool for surface functionalization and bio-conjugation. The first portion of this dissertation serves to extend the use of CVD-based reactive PPX coatings as a surface functionalization strategy for the conjugation of biomolecules. Micro-structured PPX coatings having multiple surface reactive groups were fabricated. Multiple orthogonal click reactions were then employed to selectively immobilize galactose and mannobiose to the micro-structured polymer coatings. The presence of different types of carbohydrate enables lectins binding for examining ligands/cell receptor interactions. This dissertation also demonstrates the use of CVD-based reactive PPX coatings as intermediate layers to immobilize adenoviral vectors onto tissue scaffolds. The ability to tether adenoviral vectors on tissue scaffolds localizes the transduction near the scaffold surface and reduces acute toxicity and hepatic pathology cause by direct administration of the viral vector, providing a safe and efficient gene therapy delivery strategy. In the second portion of this dissertation, we explore the CVD of PPX onto surfaces coated with a thin layer of liquid crystal (LC). Instead of forming a conformal PPX coating encapsulating the LC layer, PPX assembled into an array of high-aspect ratio nanofibers inside the LC layer. The LC layer was demonstrated to act as a template where the anisotropic internal ordering of the LC facilitated the formation of nanofibers. The diameter of the nanofibers was in the range of 100 nm and could be tuned by type of LC template used, and the length of the nanofibers could be precisely controlled by varying the thickness of the LC film. The overall shape of the nanofibers could be controlled by the internal ordering of the LC template, as exemplified by the assembly of helical nanofibers using cholesteric LC as the template. PPX nanofibers could be applied to a broad range of materials, such as curved surface, metal meshes and microparticles. We successfully created nanofibers with different surface functionalities and utilized them to capture molecules of interest. We also demonstrated the synthesis of twisted nanofibers using chiral-substituted precursors. The direction and the degree of twisting of nanofibers could be controlled by the handedness and the enantiomeric excess of the chiral precursor. Finally, we showed that the LC-templated CVD method could be extended to fabricating nanofibers made of other CVD-based polymer systems, such as poly(lutidine) and poly(p-phenylene vinylene). Our work opens a new platform for designing functional polymer nanostructures with programmable geometry, alignment and chemistry. The polymer nanostructures can be attractive for applications ranging from sensors, affinity filtration, and catalytic supports.

Suspended, Shrinkage-Free, Electrospun PLGA Nanofibrous Scaffold for Skin Tissue Engineering.

PubMed

Ru, Changhai; Wang, Feilong; Pang, Ming; Sun, Lining; Chen, Ruihua; Sun, Yu

2015-05-27

Electrospinning is a technique for creating continuous nanofibrous networks that can architecturally be similar to the structure of extracellular matrix (ECM). However, the shrinkage of electrospun mats is unfavorable for the triggering of cell adhesion and further growth. In this work, electrospun PLGA nanofiber assemblies are utilized to create a scaffold. Aided by a polypropylene auxiliary supporter, the scaffold is able to maintain long-term integrity without dimensional shrinkage. This scaffold is also able to suspend in cell culture medium; hence, keratinocyte cells seeded on the scaffold are exposed to air as required in skin tissue engineering. Experiments also show that human skin keratinocytes can proliferate on the scaffold and infiltrate into the scaffold.

Putting Electrospun Nanofibers to Work for Biomedical Research

PubMed Central

Xie, Jingwei; Li, Xiaoran; Xia, Younan

2009-01-01

Electrospinning has been exploited for almost one century to process polymers and related materials into nanofibers with controllable compositions, diameters, porosities, and porous structures for a variety of applications. Owing to its high porosity and large surface area, a non-woven mat of electrospun nanofibers can serve as an ideal scaffold to mimic the extracellular matrix for cell attachment and nutrient transportation. The nanofiber itself can also be functionalized through encapsulation or attachment of bioactive species such as extracellular matrix proteins, enzymes, and growth factors. In addition, the nanofibers can be further assembled into a variety of arrays or architectures by manipulating their alignment, stacking, or folding. All these attributes make electrospinning a powerful tool for generating nanostructured materials for a range of biomedical applications that include controlled release, drug delivery, and tissue engineering. PMID:20011452

Influence of ciprofloxacin-based additives on the hydrolysis of nanofiber polyurethane membranes.

PubMed

Wright, Meghan E E; Wong, Andrew T; Levitt, Daniel; Parrag, Ian C; Yang, Meilin; Santerre, J Paul

2018-05-01

A degradable polycarbonate urethane (PCNU) and an antimicrobial oligomer (AO) were used to generate anti-infective nanofiber scaffolds through blend electrospinning. The AO consists of two molecules of ciprofloxacin (CF) bound through hydrolysable linkages to triethylene glycol. The membranes were conceived for use as tissue engineering scaffolds for the regeneration of soft tissues for the periodontium, where there would be a need for a local dose of antibiotic to the periodontal space as the scaffold degrades in order to prevent biomaterial-associated infection. Scaffolds were made using AO at 7 and 15% w/w equivalent CF, and compared to scaffolds with 15% w/w CF (with HCl counterion). AO was hydrolyzed and released CF continuously over 28 days, while the 15% w/w CF HCl scaffolds showed a burst release within hours, with no subsequent release in the subsequent 28 day period. Released CF from both the AO and CF HCl scaffolds had a similar minimum inhibitory concentration to that of off-the-shelf CF. Interestingly, the introduction of drug in either form (AO or CF HCl) was found to increase the hydrolytic stability of the electrospun degradable PCNU scaffold matrix itself. The alteration of hydrolysis kinetics was attributed to changes in the hydrogen bonding character and microstructure within the scaffolds, introduced by the presence of CF. This study has revealed that in generating in situ drug release systems, the secondary effects of the added drug on the degradation properties of the polymeric carriers must be considered, particularly for systems that act dually as tissue engineering scaffolds. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1211-1222, 2018. © 2018 Wiley Periodicals, Inc.

Interwoven Aligned Conductive Nanofiber Yarn/Hydrogel Composite Scaffolds for Engineered 3D Cardiac Anisotropy.

PubMed

Wu, Yaobin; Wang, Ling; Guo, Baolin; Ma, Peter X

2017-06-27

Mimicking the anisotropic cardiac structure and guiding 3D cellular orientation play a critical role in designing scaffolds for cardiac tissue regeneration. Significant advances have been achieved to control cellular alignment and elongation, but it remains an ongoing challenge for engineering 3D cardiac anisotropy using these approaches. Here, we present a 3D hybrid scaffold based on aligned conductive nanofiber yarns network (NFYs-NET, composition: polycaprolactone, silk fibroin, and carbon nanotubes) within a hydrogel shell for mimicking the native cardiac tissue structure, and further demonstrate their great potential for engineering 3D cardiac anisotropy for cardiac tissue engineering. The NFYs-NET structures are shown to control cellular orientation and enhance cardiomyocytes (CMs) maturation. 3D hybrid scaffolds were then fabricated by encapsulating NFYs-NET layers within hydrogel shell, and these 3D scaffolds performed the ability to promote aligned and elongated CMs maturation on each layer and individually control cellular orientation on different layers in a 3D environment. Furthermore, endothelialized myocardium was constructed by using this hybrid strategy via the coculture of CMs on NFYs-NET layer and endothelial cells within hydrogel shell. Therefore, these 3D hybrid scaffolds, containing NFYs-NET layer inducing cellular orientation, maturation, and anisotropy and hydrogel shell providing a suitable 3D environment for endothelialization, has great potential in engineering 3D cardiac anisotropy.

Preparation, structural characterization, and in vitro cell studies of three-dimensional SiO2-CaO binary glass scaffolds built ofultra-small nanofibers.

PubMed

Luo, Honglin; Li, Wei; Ao, Haiyong; Li, Gen; Tu, Junpin; Xiong, Guangyao; Zhu, Yong; Wan, Yizao

2017-07-01

Three-dimensional (3D) nanofibrous scaffolds hold great promises in tissue engineering and regenerative medicine. In this work, for the first time, 3D SiO 2 -CaO binary glass nanofibrous scaffolds have been fabricated via a combined method of template-assisted sol-gel and calcination by using bacterial cellulose as the template. SEM with EDS, TEM, and AFM confirm that the molar ratio of Ca to Si and fiber diameter of the resultant SiO 2 -CaO nanofibers can be controlled by immersion time in the solution of tetraethyl orthosilicate and ethanol. The optimal immersion time was 6h which produced the SiO 2 -CaO binary glass containing 60at.% Si and 40at.% Ca (named 60S40C). The fiber diameter of 60S40C scaffold is as small as 29nm. In addition, the scaffold has highly porous 3D nanostructure with dominant mesopores at 10.6nm and macropores at 20μm as well as a large BET surface area (240.9m 2 g -1 ), which endow the 60S40C scaffold excellent biocompatibility and high ALP activity as revealed by cell studies using osteoblast cells. These results suggest that the 60S40C scaffold has great potential in bone tissue regeneration. Copyright © 2017 Elsevier B.V. All rights reserved.

Characterization of Electrospun Nanofibrous Scaffolds for Nanobiomedical Applications

NASA Astrophysics Data System (ADS)

Emul, E.; Saglam, S.; Ates, H.; Korkusuz, F.; Saglam, N.

2016-08-01

The electrospinning method is employed in the production of porous fiber scaffolds, and the usage of electrospun scaffolds especially as drug carrier and bone reconstructive material such as implants is promising for future applications in tissue engineering. The number of publications has grown very rapidly in this field through the fabrication of complex scaffolds, novel approaches in nanotechnology, and improvements of imaging methods. Hence, characterization of these materials has also grown significantly important for getting satisfied and accurate results. This advantageous and versatile method is ideal for mimicking bone extracellular matrix, and many biodegradable and biocompatible polymers are preferred in the field of bone reconstruction. In this study, gelatin, gelatin/nanohydroxyapatite (nHAp) and gelatin/PLLA/nHAp scaffolds were fabricated by the electrospinning process. These composite fibers showed clear and continuous morphology according to observation through a scanning electron microscope and their component analyses were also determined by Fourier transform infrared spectrometer analyses. These characterization experiments revealed the great effects of the electrospinning method for biomedical applications and have an especially important role in bone reconstruction and production of implant coating material.

Dynamic Mechanical and Nanofibrous Topological Combinatory Cues Designed for Periodontal Ligament Engineering.

PubMed

Kim, Joong-Hyun; Kang, Min Sil; Eltohamy, Mohamed; Kim, Tae-Hyun; Kim, Hae-Won

2016-01-01

Complete reconstruction of damaged periodontal pockets, particularly regeneration of periodontal ligament (PDL) has been a significant challenge in dentistry. Tissue engineering approach utilizing PDL stem cells and scaffolding matrices offers great opportunity to this, and applying physical and mechanical cues mimicking native tissue conditions are of special importance. Here we approach to regenerate periodontal tissues by engineering PDL cells supported on a nanofibrous scaffold under a mechanical-stressed condition. PDL stem cells isolated from rats were seeded on an electrospun polycaprolactone/gelatin directionally-oriented nanofiber membrane and dynamic mechanical stress was applied to the cell/nanofiber construct, providing nanotopological and mechanical combined cues. Cells recognized the nanofiber orientation, aligning in parallel, and the mechanical stress increased the cell alignment. Importantly, the cells cultured on the oriented nanofiber combined with the mechanical stress produced significantly stimulated PDL specific markers, including periostin and tenascin with simultaneous down-regulation of osteogenesis, demonstrating the roles of topological and mechanical cues in altering phenotypic change in PDL cells. Tissue compatibility of the tissue-engineered constructs was confirmed in rat subcutaneous sites. Furthermore, in vivo regeneration of PDL and alveolar bone tissues was examined under the rat premaxillary periodontal defect models. The cell/nanofiber constructs engineered under mechanical stress showed sound integration into tissue defects and the regenerated bone volume and area were significantly improved. This study provides an effective tissue engineering approach for periodontal regeneration-culturing PDL stem cells with combinatory cues of oriented nanotopology and dynamic mechanical stretch.

Dynamic Mechanical and Nanofibrous Topological Combinatory Cues Designed for Periodontal Ligament Engineering

PubMed Central

Kim, Joong-Hyun; Kang, Min Sil; Eltohamy, Mohamed; Kim, Tae-Hyun; Kim, Hae-Won

2016-01-01

Complete reconstruction of damaged periodontal pockets, particularly regeneration of periodontal ligament (PDL) has been a significant challenge in dentistry. Tissue engineering approach utilizing PDL stem cells and scaffolding matrices offers great opportunity to this, and applying physical and mechanical cues mimicking native tissue conditions are of special importance. Here we approach to regenerate periodontal tissues by engineering PDL cells supported on a nanofibrous scaffold under a mechanical-stressed condition. PDL stem cells isolated from rats were seeded on an electrospun polycaprolactone/gelatin directionally-oriented nanofiber membrane and dynamic mechanical stress was applied to the cell/nanofiber construct, providing nanotopological and mechanical combined cues. Cells recognized the nanofiber orientation, aligning in parallel, and the mechanical stress increased the cell alignment. Importantly, the cells cultured on the oriented nanofiber combined with the mechanical stress produced significantly stimulated PDL specific markers, including periostin and tenascin with simultaneous down-regulation of osteogenesis, demonstrating the roles of topological and mechanical cues in altering phenotypic change in PDL cells. Tissue compatibility of the tissue-engineered constructs was confirmed in rat subcutaneous sites. Furthermore, in vivo regeneration of PDL and alveolar bone tissues was examined under the rat premaxillary periodontal defect models. The cell/nanofiber constructs engineered under mechanical stress showed sound integration into tissue defects and the regenerated bone volume and area were significantly improved. This study provides an effective tissue engineering approach for periodontal regeneration—culturing PDL stem cells with combinatory cues of oriented nanotopology and dynamic mechanical stretch. PMID:26989897

Fabrication and Biocompatibility of Electrospun Silk Biocomposites

PubMed Central

Wei, Kai; Kim, Byoung-Suhk; Kim, Ick-Soo

2011-01-01

Silk fibroin has attracted great interest in tissue engineering because of its outstanding biocompatibility, biodegradability and minimal inflammatory reaction. In this study, two kinds of biocomposites based on regenerated silk fibroin are fabricated by electrospinning and post-treatment processes, respectively. Firstly, regenerated silk fibroin/tetramethoxysilane (TMOS) hybrid nanofibers with high hydrophilicity are prepared, which is superior for fibroblast attachment. The electrospinning process causes adjacent fibers to ‘weld’ at contact points, which can be proved by scanning electron microscope (SEM). The water contact angle of silk/tetramethoxysilane (TMOS) composites shows a sharper decrease than pure regenerated silk fibroin nanofiber, which has a great effect on the early stage of cell attachment behavior. Secondly, a novel tissue engineering scaffold material based on electrospun silk fibroin/nano-hydroxyapatite (nHA) biocomposites is prepared by means of an effective calcium and phosphate (Ca–P) alternate soaking method. nHA is successfully produced on regenerated silk fibroin nanofiber within several min without any pre-treatments. The osteoblastic activities of this novel nanofibrous biocomposites are also investigated by employing osteoblastic-like MC3T3-E1 cell line. The cell functionality such as alkaline phosphatase (ALP) activity is ameliorated on mineralized silk nanofibers. All these results indicate that this silk/nHA biocomposite scaffold material may be a promising biomaterial for bone tissue engineering. PMID:24957869

Potential of Electrospun Nanofibers for Biomedical and Dental Applications

PubMed Central

Zafar, Muhammad; Najeeb, Shariq; Khurshid, Zohaib; Vazirzadeh, Masoud; Zohaib, Sana; Najeeb, Bilal; Sefat, Farshid

2016-01-01

Electrospinning is a versatile technique that has gained popularity for various biomedical applications in recent years. Electrospinning is being used for fabricating nanofibers for various biomedical and dental applications such as tooth regeneration, wound healing and prevention of dental caries. Electrospun materials have the benefits of unique properties for instance, high surface area to volume ratio, enhanced cellular interactions, protein absorption to facilitate binding sites for cell receptors. Extensive research has been conducted to explore the potential of electrospun nanofibers for repair and regeneration of various dental and oral tissues including dental pulp, dentin, periodontal tissues, oral mucosa and skeletal tissues. However, there are a few limitations of electrospinning hindering the progress of these materials to practical or clinical applications. In terms of biomaterials aspects, the better understanding of controlled fabrication, properties and functioning of electrospun materials is required to overcome the limitations. More in vivo studies are definitely required to evaluate the biocompatibility of electrospun scaffolds. Furthermore, mechanical properties of such scaffolds should be enhanced so that they resist mechanical stresses during tissue regeneration applications. The objective of this article is to review the current progress of electrospun nanofibers for biomedical and dental applications. In addition, various aspects of electrospun materials in relation to potential dental applications have been discussed. PMID:28787871

Manufacture of porous biodegradable polymer conduits by an extrusion process for guided tissue regeneration

NASA Technical Reports Server (NTRS)

Widmer, M. S.; Gupta, P. K.; Lu, L.; Meszlenyi, R. K.; Evans, G. R.; Brandt, K.; Savel, T.; Gurlek, A.; Patrick, C. W. Jr; Mikos, A. G.;

Mechanical properties and cellular response of novel electrospun nanofibers for ligament tissue engineering: Effects of orientation and geometry.

PubMed

Pauly, Hannah M; Kelly, Daniel J; Popat, Ketul C; Trujillo, Nathan A; Dunne, Nicholas J; McCarthy, Helen O; Haut Donahue, Tammy L

2016-08-01

Electrospun nanofibers are a promising material for ligamentous tissue engineering, however weak mechanical properties of fibers to date have limited their clinical usage. The goal of this work was to modify electrospun nanofibers to create a robust structure that mimics the complex hierarchy of native tendons and ligaments. The scaffolds that were fabricated in this study consisted of either random or aligned nanofibers in flat sheets or rolled nanofiber bundles that mimic the size scale of fascicle units in primarily tensile load bearing soft musculoskeletal tissues. Altering nanofiber orientation and geometry significantly affected mechanical properties; most notably aligned nanofiber sheets had the greatest modulus; 125% higher than that of random nanofiber sheets; and 45% higher than aligned nanofiber bundles. Modifying aligned nanofiber sheets to form aligned nanofiber bundles also resulted in approximately 107% higher yield stresses and 140% higher yield strains. The mechanical properties of aligned nanofiber bundles were in the range of the mechanical properties of the native ACL: modulus=158±32MPa, yield stress=57±23MPa and yield strain=0.38±0.08. Adipose derived stem cells cultured on all surfaces remained viable and proliferated extensively over a 7 day culture period and cells elongated on nanofiber bundles. The results of the study suggest that aligned nanofiber bundles may be useful for ligament and tendon tissue engineering based on their mechanical properties and ability to support cell adhesion, proliferation, and elongation. Copyright © 2016 Elsevier Ltd. All rights reserved.

"Clickable" Polymeric Nanofibers through Hydrophilic-Hydrophobic Balance: Fabrication of Robust Biomolecular Immobilization Platforms.

PubMed

Kalaoglu-Altan, Ozlem I; Sanyal, Rana; Sanyal, Amitav

2015-05-11

Fabrication of hydrophilic polymeric nanofibers that undergo facile and selective functionalization through metal catalyst-free Diels-Alder "click" reaction in aqueous environment is outlined. Electrospinning of copolymers containing an electron-rich furan moiety, hydrophobic methyl methacrylate units and hydrophilic poly(ethylene glycol)s as side chains provide specifically functionalizable yet antibiofouling fibers that remain stable in aqueous media due to appropriate hydrophobic hydrophilic balance. Efficient functionalization of these nanofibers is accomplished through the Diels-Alder reaction by exposing them to maleimide-containing molecules and ligands. Diels-Alder conjugation based functionalization is demonstrated through attachment of fluorescein-maleimide and a maleimide tethered biotin ligand. Biotinylated nanofibers were utilized to mediate immobilization of the protein streptavidin, as well as streptavidin coated quantum dots. Facile fabrication from readily available polymers and their effective functionalization under mild and reagent-free conditions in aqueous media make these "clickable" nanofibers attractive candidates as functionalizable scaffolds for various biomedical applications.

Towards optimization of odonto/osteogenic bioengineering: in vitro comparison of simvastatin, sodium fluoride, melanocyte-stimulating hormone.

PubMed

Zijah, Vahid; Salehi, Roya; Aghazadeh, Marziyeh; Samiei, Mohammad; Alizadeh, Effat; Davaran, Soodabeh

2017-06-01

Tissue engineering has emerged as a potential therapeutic option for dental problems in recent years. One of the policies in tissue engineering is to use both scaffolds and additive factors for enhancing cell responses. This study aims to evaluate and compare the effect of three types of biofactors on poly-caprolactone-poly-ethylene glycol-poly caprolactone (PCL-PEG-PCL) nanofibrous scaffold on human dental pulp stem cell (hDPSCs) engineering. The PCL-PEG-PCL copolymer was synthesized with ring opening polymerization method, and its nanofiber scaffold was prepared by electrospinning method. Nanofibrous scaffold-seeded hDPSCs were treated with sodium fluoride (NaF), melanocyte-stimulating hormone (MSH), or simvastatin (SIM). Non-treated nanofiber seeded cells were utilized as control. The viability, biocompatibility, adhesion, proliferation rate, morphology, osteo/odontogenic potential, and the expression of tissue-specific genes were studied. The results showed that significant higher results demonstrated significant higher adhesive behavior, viability, alizarin red activity, and dentin specific gene expression in MSH- and SIM-treated cells (p < 0.05). This study is unique; in that, it compares the effects of different treatments for optimization of dental tissue engineering.

Nanomaterials for Craniofacial and Dental Tissue Engineering.

PubMed

Li, G; Zhou, T; Lin, S; Shi, S; Lin, Y

2017-07-01

Tissue engineering shows great potential as a future treatment for the craniofacial and dental defects caused by trauma, tumor, and other diseases. Due to the biomimetic features and excellent physiochemical properties, nanomaterials are of vital importance in promoting cell growth and stimulating tissue regeneration in tissue engineering. For craniofacial and dental tissue engineering, the frequently used nanomaterials include nanoparticles, nanofibers, nanotubes, and nanosheets. Nanofibers are attractive for cell invasion and proliferation because of their resemblance to extracellular matrix and the presence of large pores, and they have been used as scaffolds in bone, cartilage, and tooth regeneration. Nanotubes and nanoparticles improve the mechanical and chemical properties of scaffold, increase cell attachment and migration, and facilitate tissue regeneration. In addition, nanofibers and nanoparticles are also used as a delivery system to carry the bioactive agent in bone and tooth regeneration, have better control of the release speed of agent upon degradation of the matrix, and promote tissue regeneration. Although applications of nanomaterials in tissue engineering remain in their infancy with numerous challenges to face, the current results indicate that nanomaterials have massive potential in craniofacial and dental tissue engineering.

Structure and properties of slow-resorbing nanofibers obtained by (co-axial) electrospinning as tissue scaffolds in regenerative medicine

PubMed Central

Gola, Joanna; Ghavami, Saeid; Skonieczna, Magdalena; Markowski, Jarosław; Likus, Wirginia; Lewandowska, Magdalena; Maziarz, Wojciech

2017-01-01

With the rapid advancement of regenerative medicine technologies, there is an urgent need for the development of new, cell-friendly techniques for obtaining nanofibers—the raw material for an artificial extracellular matrix production. We investigated the structure and properties of PCL10 nanofibers, PCL5/PCL10 core-shell type nanofibers, as well as PCL5/PCLAg nanofibres prepared by electrospinning. For the production of the fiber variants, a 5–10% solution of polycaprolactone (PCL) (Mw = 70,000–90,000), dissolved in a mixture of formic acid and acetic acid at a ratio of 70:30 m/m was used. In order to obtain fibers containing PCLAg 1% of silver nanoparticles was added. The electrospin was conducted using the above-described solutions at the electrostatic field. The subsequent bio-analysis shows that synthesis of core-shell nanofibers PCL5/PCL10, and the silver-doped variant nanofiber core shell PCL5/PCLAg, by using organic acids as solvents, is a robust technique. Furthermore, the incorporation of silver nanoparticles into PCL5/PCLAg makes such nanofibers toxic to model microbes without compromising its biocompatibility. Nanofibers obtained such way may then be used in regenerative medicine, for the preparation of extracellular scaffolds: (i) for controlled bone regeneration due to the long decay time of the PCL, (ii) as bioscaffolds for generation of other types of artificial tissues, (iii) and as carriers of nanocapsules for local drug delivery. Furthermore, the used solvents are significantly less toxic than the solvents for polycaprolactone currently commonly used in electrospin, like for example chloroform (CHCl3), methanol (CH3OH), dimethylformamide (C3H7NO) or tetrahydrofuran (C4H8O), hence the presented here electrospin technique may allow for the production of multilayer nanofibres more suitable for the use in medical field. PMID:29302386

Composites structures for bone tissue reconstruction

NASA Astrophysics Data System (ADS)

Neto, W.; Santos, João.; Avérous, L.; Schlatter, G.; Bretas, Rosario.

2015-05-01

The search for new biomaterials in the bone reconstitution field is growing continuously as humane life expectation and bone fractures increase. For this purpose, composite materials with biodegradable polymers and hydroxyapatite (HA) have been used. A composite material formed by a film, nanofibers and HA has been made. Both, the films and the non-woven mats of nanofibers were formed by nanocomposites made of butylene adipate-co-terephthalate (PBAT) and HA. The techniques used to produce the films and nanofibers were spin coating and electrospinning, respectively. The composite production and morphology were evaluated. The composite showed an adequate morphology and fibers size to be used as scaffold for cell growth.

Nano/micro hybrid scaffold of PCL or P3HB nanofibers combined with silk fibroin for tendon and ligament tissue engineering.

PubMed

Naghashzargar, Elham; Farè, Silvia; Catto, Valentina; Bertoldi, Serena; Semnani, Dariush; Karbasi, Saeed; Tanzi, Maria Cristina

2015-07-04

A novel biodegradable nano/micro hybrid structure was obtained by electrospinning P3HB or PCL nanofibers onto a twisted silk fibroin (SF) structure, with the aim of fabricating a suitable scaffold for tendon and ligament tissue engineering. The electrospinning (ES) processing parameters for P3HB and PCL were optimized on 2D samples, and applied to produce two different nano/micro hybrid constructs (SF/ES-PCL and SF/ES-P3HB).Morphological, chemico-physical and mechanical properties of the novel hybrid scaffolds were evaluated by SEM, ATR FT-IR, DSC, tensile and thermodynamic mechanical tests. The results demonstrated that the nanofibers were tightly wrapped around the silk filaments, and the crystallinity of the SF twisted yarns was not influenced by the presence of the electrospun polymers. The slightly higher mechanical properties of the hybrid constructs confirmed an increase of internal forces due to the interaction between nano and micro components. Cell culture tests with L929 fibroblasts, in the presence of the sample eluates or in direct contact with the hybrid structures, showed no cytotoxic effects and a good level of cytocompatibility of the nano/micro hybrid structures in term of cell viability, particularly at day 1. Cell viability onto the nano/micro hybrid structures decreased from the first to the third day of culture when compared with the control culture plastic, but appeared to be higher when compared with the uncoated SF yarns. Although additional in vitro and in vivo tests are needed, the original fabrication method here described appears promising for scaffolds suitable for tendon and ligament tissue engineering.

Preparation of polypyrrole-embedded electrospun poly(lactic acid) nanofibrous scaffolds for nerve tissue engineering

PubMed Central

Zhou, Jun-feng; Wang, Yi-guo; Cheng, Liang; Wu, Zhao; Sun, Xiao-dan; Peng, Jiang

2016-01-01

Polypyrrole (PPy) is a biocompatible polymer with good conductivity. Studies combining PPy with electrospinning have been reported; however, the associated decrease in PPy conductivity has not yet been resolved. We embedded PPy into poly(lactic acid) (PLA) nanofibers via electrospinning and fabricated a PLA/PPy nanofibrous scaffold containing 15% PPy with sustained conductivity and aligned topography. There was good biocompatibility between the scaffold and human umbilical cord mesenchymal stem cells as well as Schwann cells. Additionally, the direction of cell elongation on the scaffold was parallel to the direction of fibers. Our findings suggest that the aligned PLA/PPy nanofibrous scaffold is a promising biomaterial for peripheral nerve regeneration. PMID:27904497

Assembling new technologies at the interface of materials science and biology

NASA Astrophysics Data System (ADS)

Stendahl, John C.

Molecular self-assembly can be used to construct advanced materials by taking cues from nature and harnessing noncovalent interactions. This bottom-up approach affords molecular level precision that can cultivate pathways to improved materials function. The graduate research presented in this thesis integrates molecular self-assembly with traditional concepts in chemistry and materials science, with the ultimate goal of developing innovative solutions in technology and medicine. In the field of polymer engineering, self-assembly was used to create supramolecular nanoribbons that, when incorporated into polystyrene, modify its microstructure and significantly enhance its toughness and ductility. In medicine, self-assembly was used to create ordered, chemically functional materials to improve interactions with cells and other constituents of the biological environment. One system that was investigated is based on a triblock molecule in which cholesterol is connected to a lysine dendron by a flexible oligo-(L-lactic acid) spacer. These molecules self-assemble into polar surface coatings on fibrous poly(L-lactic acid) scaffolds that improve the scaffold's wettability and increase its retention of cells during seeding. Another self-assembling system that was investigated for biomedical applications is a family of molecules referred to as peptide amphiphiles (PA's). PA's consist of hydrophobic alkyl tails connected to short, hydrophilic peptides that incorporate biological signaling epitopes. These molecules spontaneously assemble into networks of well-defined nanofibers in aqueous environments, with the signaling epitopes presented in high density on the nanofiber exteriors. Nanofiber assembly is triggered by charge screening on the peptides and is able to produce self-supporting gels in concentrations of less than 1.0 wt.-%. The assembly process and mechanical properties of PA gels was investigated in detail with vibrational spectroscopy and oscillatory rheology. PA nanofibers were used in conjunction with fibrous poly(L-lactic acid] fabrics to create chemically functional scaffolds to facilitate islet cell transplantation. In transplant studies in diabetic mice, the use of scaffolds for islet delivery was shown to significantly improve transplant outcomes over free islet injections. Together, these studies illustrate that molecular self-assembly can be used to create functional materials for a variety of applications. These materials utilize noncovalent interactions to produce supramolecular structures that have important impacts on properties.

Poly(hydroxybutyrate)/chitosan Aligned Electrospun Scaffold as a Novel Substrate for Nerve Tissue Engineering.

PubMed

Karimi, Afarin; Karbasi, Saeed; Razavi, Shahnaz; Zargar, Elham Naghash

2018-01-01

Reconstruction of nervous system is a great challenge in the therapeutic medical field. Nerve tissue engineering is a novel method to regenerate nervous system in human health care. Tissue engineering has introduced novel approaches to promote and guide peripheral nerve regeneration using submicron and nanoscale fibrous scaffolds. In this study, 9 wt% poly(3-hydroxybutyrate) (PHB) solutions with two different ratios of chitosan (CTS) (15%, and 20%) were mixed in trifluoroacetic acid as a cosolvent. Thereafter, random and aligned PHB/CTS scaffolds were fabricated by electrospinning method in an appropriate condition. Average diameters for aligned PHB, PHB/CTS 85:15 and PHB/CTS 80:20 were obtained as 675 nm, 740.3 nm, and 870.74 nm, which was lesser than random fibers. The solution components entity authenticity was approved by Fourier transform infrared. The addition of CTS decreased both water droplet contact angle from 124.79° to 43.14° in random and 110.87° to 33.49° in aligned PHB/CTS fibrous scaffold. Moreover, alignment of fibers causes tremendous increase in hydrophilicity of fibrous PHB/CTS substrate. Tensile strength increased from 6.41 MPa for random to 8.73 MPa for aligned PHB/CTS 85:15. Our results indicated that aligned PHB/CTS 85:15 nanofibers are the desired scaffold than the random PHB/CTS nanofibers for application in nerve tissue regeneration.

Effects of Structural Properties of Electrospun TiO2 Nano-fiber Meshes on their Osteogenic Potential

PubMed Central

Wang, Xiaokun; Gittens, Rolando A.; Song, Rosemary; Tannenbaum, Rina; Olivares-Navarrete, Rene; Schwartz, Zvi; Chen, Haifeng; Boyan, Barbara D.

2011-01-01

Ideal outcomes in the field of tissue engineering and regenerative medicine involve biomaterials that can enhance cell differentiation and production of local factors for natural tissue regeneration without the use of systemic drugs. Biomaterials typically used in tissue engineering applications include polymeric scaffolds that mimic the 3-D structural environment of the native tissue, but these are often functionalized with proteins or small peptides to improve their biological performance. For bone applications, titanium (Ti) implants, or more appropriately the titania (TiO2) passive oxide layer formed on their surface, have been shown to enhance osteoblast differentiation in vitro and to promote osseointegration in vivo. In this study we evaluated the effect on osteoblast differentiation of pure TiO2 nano-fiber meshes with different surface micro-roughness and nano-fiber diameters, prepared by the electrospinning method. MG63 cells were seeded on TiO2 meshes, and cell number, differentiation markers and local factor production were analyzed. The results showed that cells grew throughout the entire surfaces and with similar morphology in all groups. Cell number was sensitive to surface micro-roughness, whereas cell differentiation and local factor production was regulated by both surface roughness and nano-fiber diameter. These results indicate that scaffold structural cues alone can be used to drive cell differentiation and create an osteogenic environment without the use of exogenous factors. PMID:22075122

G-CSF loaded nanofiber/nanoparticle composite coated with collagen promotes wound healing in vivo.

PubMed

Tanha, Shima; Rafiee-Tehrani, Morteza; Abdollahi, Mohamad; Vakilian, Saeid; Esmaili, Zahra; Naraghi, Zahra Safaei; Seyedjafari, Ehsan; Javar, Hamid Akbari

2017-10-01

Sustained release of functional growth factors can be considered as a beneficial methodology for wound healing. In this study, recombinant human granulocyte colony-stimulating factor (G-CSF)-loaded chitosan nanoparticles were incorporated in Poly(ε-caprolactone) (PCL) nanofibers, followed by surface coating with collagen type I. Physical and mechanical properties of the PCL nanofibers containing G-CSF loaded chitosan nanoparticles PCL/NP(G-CSF) and in vivo performance for wound healing were investigated. G-CSF structural stability was evaluated through SDS_PAGE, reversed phase (RP) HPLC and size-exclusion chromatography, as well as circular dichroism. Nanofiber/nanoparticle composite scaffold was demonstrated to have appropriate mechanical properties as a wound dresser and a sustained release of functional G-CSF. The PCL/NP(G-CSF) scaffold showed a suitable proliferation and well-adherent morphology of stem cells. In vivo study and histopathological evaluation outcome revealed that skin regeneration was dramatically accelerated under PCL/NP(G-CSF) as compared with control groups. Superior fibroblast maturation, enhanced collagen deposition and minimum inflammatory cells were also the beneficial properties of PCL/NP(G-CSF) over the commercial dressing. The synergistic effect of extracellular matrix-mimicking nanofibrous membrane and G-CSF could develop a suitable supportive substrate in order to extensive utilization for the healing of skin wounds. © 2017 Wiley Periodicals Inc. J Biomed Mater Res Part A: 105A: 2830-2842, 2017. © 2017 Wiley Periodicals, Inc.

Surface changes of biopolymers PHB and PLLA induced by Ar+ plasma treatment and wet etching

NASA Astrophysics Data System (ADS)

Slepičková Kasálková, N.; Slepička, P.; Sajdl, P.; Švorčík, V.

2014-08-01

Polymers, especially group of biopolymers find potential application in a wide range of disciplines due to their biodegradability. In biomedical applications these materials can be used as a scaffold or matrix. In this work, the influence of the Ar+ plasma treatment and subsequent wet etching (acetone/water) on the surface properties of polymers were studied. Two biopolymers - polyhydroxybutyrate with 8% polyhydroxyvalerate (PHB) and poly-L-lactic acid (PLLA) were used in these experiments. Modified surface layers were analyzed by different methods. Surface wettability was characterized by determination of water contact angle. Changes in elemental composition of modified surfaces were performed by X-ray Photoelectron Spectroscopy (XPS). Surface morphology and roughness was examined using Atomic Force Microscopy (AFM). Gravimetry method was used to study the mass loss. It was found that the modification from both with plasma and wet etching leads to dramatic changes of surface properties (surface chemistry, morphology and roughness). Rate of changes of these features strongly depends on the modification parameters.

Mechanical behavior of polymer-based vs. metallic-based bioresorbable stents

PubMed Central

Ang, Hui Ying; Huang, Ying Ying; Lim, Soo Teik; Wong, Philip; Joner, Michael

2017-01-01

Bioresorbable scaffolds (BRS) were developed to overcome the drawbacks of current metallic drug-eluting stents (DES), such as late in-stent restenosis and caging of the vessel permanently. The concept of the BRS is to provide transient support to the vessel during healing before being degraded and resorbed by the body, freeing the vessel and restoring vasomotion. The mechanical properties of the BRS are influenced by the choice of the material and processing methods. Due to insufficient radial strength of the bioresorbable material, BRS often required large strut profile as compared to conventional metallic DES. Having thick struts will in turn affect the deliverability of the device and may cause flow disturbance, thereby increasing the incidence of acute thrombotic events. Currently, the bioresorbable poly-l-lactic acid (PLLA) polymer and magnesium (Mg) alloys are being investigated as materials in BRS technologies. The bioresorption process, mechanical properties, in vitro observations and clinical outcomes of PLLA-based and Mg-based BRS will be examined in this review. PMID:28894598

Nanostructured thick 3D nanofibrous scaffold can induce bone.

PubMed

Eap, Sandy; Morand, David; Clauss, François; Huck, Olivier; Stoltz, Jean-François; Lutz, Jean-Christophe; Gottenberg, Jacques-Eric; Benkirane-Jessel, Nadia; Keller, Laetitia; Fioretti, Florence

2015-01-01

Designing unique nanostructured biomimetic materials is a new challenge in modern regenerative medicine. In order to develop functional substitutes for damaged organs or tissues, several methods have been used to create implants able to regenerate robust and durable bone. Electrospinning produces nonwoven scaffolds based on polymer nanofibers mimicking the fibrillar organization of bone extracellular matrix. Here, we describe a biomimetic 3D thick nanofibrous scaffold obtained by electrospinning of the biodegradable, bioresorbable and FDA-approved polymer, poly(ε-caprolactone). Such scaffold presents a thickness reaching one centimeter. We report here the demonstration that the designed nanostructured implant is able to induce in vivo bone regeneration.

Gelatine/PLLA sponge-like scaffolds: morphological and biological characterization.

PubMed

Lazzeri, Luigi; Cascone, Maria Grazia; Danti, Serena; Serino, Lorenzo Pio; Moscato, Stefania; Bernardini, Nunzia

2006-12-01

Biodegradable synthetic polymers such as poly(lactic acid) are widely used to prepare scaffolds for cell transplantation and tissue growth, using different techniques set up for the purpose. However the poor hydrophilicity of these polymers represents the main limitation to their use as scaffolds because it causes a low affinity for the cells. An effective way to solve this problem could be represented by the addition of biopolymers that are in general highly hydrophilic. The present work concerns porous biodegradable sponge-like systems based on poly(L-lactic acid) and gelatine. Morphology and porosity characteristics of the sponges were studied by scanning electron microscopy and mercury intrusion porosimetry respectively. Blood compatibility was investigated by bovine plasma fibrinogen adsorption test and platelet adhesion test. The cell culture method was used in order to evaluate the ability of the matrices to work as scaffolds for tissue regeneration. The obtained results indicate that the sponges have interesting porous characteristics, good blood compatibility and above all good ability to support cell adhesion and growth. In fact viable and metabolically active animal cells were found inside the sponges after 8 weeks in culture. On this basis the systems produced seem to be good candidates as scaffolds for tissue regeneration.

Electrospun nanofibrous scaffolds of segmented polyurethanes based on PEG, PLLA and PTMC blocks: Physico-chemical properties and morphology.

PubMed

Trinca, Rafael Bergamo; Abraham, Gustavo A; Felisberti, Maria Isabel

2015-11-01

Biocompatible polymeric scaffolds are crucial for successful tissue engineering. Biomedical segmented polyurethanes (SPUs) are an important and versatile class of polymers characterized by a broad spectrum of compositions, molecular architectures, properties and applications. Although SPUs are versatile materials that can be designed by different routes to cover a wide range of properties, they have been infrequently used for the preparation of electrospun nanofibrous scaffolds. This study reports the preparation of new electrospun polyurethane scaffolds. The segmented polyurethanes were synthesized using low molar masses macrodyols (poly(ethylene glycol), poly(l-lactide) and poly(trimethylene carbonate)) and 1,6-hexane diisocyanate and 1,4-butanodiol as isocyanate and chain extensor, respectively. Different electrospinning parameters such as solution properties and processing conditions were evaluated to achieve smooth, uniform bead-free fibers. Electrospun micro/nanofibrous structures with mean fiber diameters ranging from 600nm to 770nm were obtained by varying the processing conditions. They were characterized in terms of thermal and dynamical mechanical properties, swelling degree and morphology. The elastomeric polyurethane scaffolds exhibit interesting properties that could be appropriate as biomimetic matrices for soft tissue engineering applications. Copyright © 2015 Elsevier B.V. All rights reserved.

Sustained delivery of siRNA/mesoporous silica nanoparticle (siRNA/MSN) complexes from nanofiber scaffolds for long-term gene silencing.

PubMed

Pinese, Coline; Lin, Junquan; Milbreta, Ulla; Li, Mingqiang; Wang, Yucai; Leong, Kam W; Chew, Sing Yian

2018-06-08

A low toxicity and efficient delivery system is needed to deliver small interfering RNAs (siRNA) in vitro and in vivo. The use of mesoporous silica nanoparticles (MSN) is becoming increasingly common due to its biocompatibility, tunable pore size and customizable properties. However, bolus delivery of siRNA/MSN complexes remains suboptimal, especially when a sustained and long-term administration is required. Here, we utilized electrospun scaffolds for sustained delivery of siRNA/MSN-PEI through surface adsorption and nanofiber encapsulation. As a proof-of-concept, we targeted collagen type I expression to modulate fibrous capsule formation. Surface adsorption of siRNA/MSN-PEI provided sustained availability of siRNA for at least 30 days in vitro. As compared to conventional bolus delivery, such scaffold-mediated transfection provided more effective gene silencing (p < 0.05). On the contrary, a longer sustained release was attained (at least 5 months) when siRNA/MSN-PEI complexes were encapsulated within the electrospun fibers. In vivo subcutaneous implantation and biodistribution analysis of these scaffolds revealed that siRNA remained localized up to ∼290 μm from the implants. Finally, a fibrous capsule reduction of ∼45.8 % was observed after 4 weeks in vivo as compared to negative scrambled siRNA treatment. Taken together, these results demonstrate the efficacy of scaffold-mediated sustained delivery of siRNA/MSN-PEI for long-term non-viral gene silencing applications. The bolus delivery of siRNA/ Mesoporous Silica Nanoparticles (MSN) complexes shows high efficiency to silence protein agonists of tumoral processes as cancer treatments. However, in tissue engineering area, scaffold mediated delivery is desired to achieve a local and sustained release of therapeutics. We showed the feasibility and the efficacy of siRNA/MSN delivered from electrospun scaffolds through surface adsorption and nanofiber encapsulation. We showed that this method enhances siRNA transfection efficiency and sustained targeted proteins silencing in vitro and in vivo. As a proof of concept, in this study, we targeted collagen type I expression to modulate fibrous capsule formation. However this platform can be applied to the release and transfection of siRNA or miRNA in cancer and tissue engineering applications. Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

An in vivo evaluation of PLLA/PLLA-gHA nano-composite for internal fixation of mandibular bone fractures.

PubMed

Peng, Weihai; Zheng, Wei; Shi, Kai; Wang, Wangshu; Shao, Ying; Zhang, Duo

2015-11-09

Internal fixation of bone fractures using biodegradable poly(L-lactic-acid) (PLLA)-based materials has attracted the attention of many researchers. In the present study, 36 male beagle dogs were randomly assigned to two groups: PLLA/PLLA-gHA (PLLA-grafted hydroxyapatite) group and PLLA group. PLLA/PLLA-gHA and PLLA plates were embedded in the muscular bags of the erector spinae and also implanted to fix mandibular bone fractures in respective groups. At 1, 2, 3, 6, 9, and 12 months postoperatively, the PLLA/PLLA-gHA and PLLA plates were evaluated by adsorption and degradation tests, and the mandibles were examined through radiographic analysis, biomechanical testing, and histological analysis. The PLLA/PLLA-gHA plates were non-transparent and showed a creamy white color, and the PLLA plates were transparent and faint yellow in color. At all time points following surgery, adsorption and degradation of the PLLA/PLLA-gHA plates were significantly less than those of the PLLA plates, and the lateral and longitudinal bending strengths of the surgically treated mandibles of the beagle dogs in the PLLA/PLLA-gHA group were significantly greater than those of the PLLA group and reached almost the value of intact mandibles at 12 months postoperatively. Additionally, relatively rapid bone healing was observed in the PLLA/PLLA-gHA group with the formation of new lamellar bone tissues at 12 months after the surgery. The PLLA/PLLA-gHA nano-composite can be employed as a biodegradable material for internal fixation of mandibular bone fractures.

Mechanical, Biological and Electrochemical Investigations of Advanced Micro/Nano Materials for Tissue Engineering and Energy Storage

NASA Astrophysics Data System (ADS)

Pu, Juan

Various micro/nano materials have been extensively studied for applications in tissue engineering and energy storage. Tissue engineering seeks to repair or replace damaged tissue by integrating approaches from cellular/molecular biology and material chemistry/engineering. A major challenge is the consistent design of three-dimensional (3D) scaffolds that mimic the structure and biological functions of extracellular matrix (ECM), guide cell migration, provide mechanical support, and regulate cell activity. Electrospun micro/nanofibers have been investigated as promising tissue engineering scaffolds because they resemble native ECM and possess tunable surface morphologies. Supercapacitors, one of the energy storage devices, bridge the performance gap between rechargeable batteries and conventional capacitors. Active electrode materials of supercapacitors must possess high specific surface area, high conductivity, and good electrochemical properties. Carbon-based micro/nano-particles, such as graphene, activated carbon (AC), and carbon nanotubes, are commonly used as active electrode materials for storing charge in supercapacitors by the electrical double layer mechanism due to their high specific surface area and excellent conductivity. In this thesis, the mechanical properties of electrospun bilayer microfibrous membranes were investigated for potential applications in tissue engineering. Bilayer microfibrous membranes of poly(l-lactic acid) (PLLA) were fabricated by electrospinning using a parallel-disk mandrel configuration, which resulted in the sequential deposition of a layer with aligned fibers (AFL) across the two parallel disks and a layer with random fibers (RFL), both deposited by a single process step. The membrane structure and fiber alignment were characterized by scanning electron microscopy and two-dimensional fast Fourier transform. Because of the intricacies of the generated electric field, the bilayer membranes exhibited higher porosity than the membranes fabricated with a single drum collector. Furthermore, the bilayer PLLA scaffolds showed gradual variation in through-thickness porosity and fiber alignment and an average porosity much higher than that of conventionally electrospun scaffolds (controls) with randomly distributed fibers. The biocompatibility and biological performance of the bilayer fibrous scaffolds was evaluated by in vivo experiments involving subcutaneous scaffold implantation in Sprague-Dawley rats, followed by histology and immunohistochemistry studies. The results illustrate the potential of bilayer scaffolds to overcome major limitations of conventionally electrospun scaffolds associated with intrinsically small pores, low porosity and, consequently, poor cell infiltration. The significantly higher porosity and larger pore size of the RFL enhanced cell motility through the scaffold thickness, whereas the relatively dense structure of the AFL provided adequate mechanical strength. The bilayer scaffolds showed more than two times higher cell infiltration than controls during implantation in vivo. Moreover, the unique structure of bilayer scaffolds promoted collagen fiber deposition, cell proliferation, and ingrowth of smooth muscle cells and endothelial cells in vivo.. Novel all-solid-state microsupercapacitors (MSCs) with 3D electrodes consisting of active materials and a polymer electrolyte (PE) designed for high-energy-density storage applications were fabricated and tested. The incorporation of a PE in the electrode material enhanced the accessibility of the surface of active materials by electrolyte ions and decreased the ion diffusion path during electrochemical charging/discharging. For a scan rate of 5 mV s -1, the MSCs with graphene/PE and AC/PE composite electrodes demonstrated a very high areal capacitance of 95 and 134 mF cm-2 , respectively, comparable with that of 3D MSCs having a liquid electrolyte. In addition, the graphene/PE MSCs showed 70% increase in specific capacitance after 10,000 charge/discharge cycles, attributed to an electro-activation process resulting from ion intercalation between the graphene nanosheets. The AC/PE MSCs also demonstrated excellent stability. Single-walled carbon nanotube (SWCNT) networks were deposited on an ultrathin polyimide substrate using the spray-deposition technique and patterned into interdigital electrodes to construct ultrahigh-power, extremely flexible, and foldable MSCs capable of operating at an ultrahigh scan rate and delivering a stack capacitance of 18 F cm-3 and an energy density of 1.6 mWh cm-3, which is comparable with that of lithium thin-film batteries. An ultrahigh power density of 1125 W cm-3 and extremely small time constant of 1 ms were obtained with SWCNT MSCs, comparable with aluminum electrolytic capacitors. A honeycomb polydimethylsiloxane substrate was introduced for stretchable MSC arrays based on SWNCT interdigital electrodes, which enables facile integration in flexible or wearable electronics. The honeycomb structure accommodates large deformation without generating large strains in the MSCs and interconnects. The results show that such stretchable MSC arrays with SWCNT electrodes demonstrate excellent rate capability and power performance as well as electrochemical stability up to 150% or 275% stretching and under excessive bending or twisting. The present stretchable MSC arrays with honeycomb structures show high potential for integration with other electronics, such as energy harvesters, power management circuits, wireless charging circuits, and various sensors, encompassing a wide range of wearable, bio-implantable electronic systems. (Abstract shortened by ProQuest.).

Fabrication and characterization of electrospun cellulose/nano-hydroxyapatite nanofibers for bone tissue engineering.

PubMed

Ao, Chenghong; Niu, Yan; Zhang, Ximu; He, Xu; Zhang, Wei; Lu, Canhui

2017-04-01

Nanofibrous scaffolds from cotton cellulose and nano-hydroxyapatite (nano-HA) were electrospun for bone tissue engineering. The solution properties of cellulose/nano-HA spinning dopes and their associated electrospinnability were characterized. Morphological, thermal and mechanical properties of the electrospun cellulose/nano-HA nanocomposite nanofibers (ECHNN) were measured and the biocompatibility of ECHNN with human dental follicle cells (HDFCs) was evaluated. Scanning electron microscope (SEM) images indicated that the average diameter of ECHNN increased with a higher nano-HA loading and the fiber diameter distributions were well within the range of natural ECM (extra cellular matrix) fibers (50-500nm). The ECHNN exhibited extraordinary mechanical properties with a tensile strength and a Young's modulus up to 70.6MPa and 3.12GPa respectively. Moreover, it was discovered that the thermostability of the ECHNN could be enhanced with the incorporation of nano-HA. Cell culture experiments demonstrated that the ECHNN scaffolds were quite biocompatible for HDFCs attachment and proliferation, suggesting their great potentials as scaffold materials in bone tissue engineering. Copyright © 2017 Elsevier B.V. All rights reserved.

Electrospinning of Biocompatible Nanofibers

NASA Astrophysics Data System (ADS)

Coughlin, Andrew J.; Queen, Hailey A.; McCullen, Seth D.; Krause, Wendy E.

2006-03-01

Artificial scaffolds for growing cells can have a wide range of applications including wound coverings, supports in tissue cultures, drug delivery, and organ and tissue transplantation. Tissue engineering is a promising field which may resolve current problems with transplantation, such as rejection by the immune system and scarcity of donors. One approach to tissue engineering utilizes a biodegradable scaffold onto which cells are seeded and cultured, and ideally develop into functional tissue. The scaffold acts as an artificial extracellular matrix (ECM). Because a typical ECM contains collagen fibers with diameters of 50-500 nm, electrostatic spinning (electrospinning) was used to mimic the size and structure of these fibers. Electrospinning is a novel way of spinning a nonwoven web of fibers on the order of 100 nm, much like the web of collagen in an ECM. We are investigating the ability of several biocompatible polymers (e.g., chitosan and polyvinyl alcohol) to form defect-free nanofiber webs and are studying the influence of the zero shear rate viscosity, molecular weight, entanglement concentration, relaxation time, and solvent on the resulting fiber size and morphology.

Laminin- and basement membrane-polycaprolactone blend nanofibers as a scaffold for regenerative medicine.

PubMed

Neal, Rebekah A; Lenz, Steven M; Wang, Tiffany; Abebayehu, Daniel; Brooks, Benjamin P C; Ogle, Roy C; Botchwey, Edward A

2014-09-01

Mimicking one or more components of the basement membrane (BM) holds great promise for overcoming insufficiencies in tissue engineering therapies. We have electrospun laminin nanofibers (NFs) isolated from the murine Engelbreth-Holm Swarm (EHS) tumor and evaluated them as a scaffold for embryonic stem cell culture. Seeded human embryonic stem cells were found to better maintain their undifferentiated, colony environment when cultured on laminin NFs compared to laminin mats, with 75% remaining undifferentiated on NFs. Mouse embryonic stem cells cultured on 10% laminin-polycaprolactone (PCL) NFs maintained their colony formation for twice as long without passage compared to those on PCL or gelatin substrates. In addition, we have established a protocol for electrospinning reconstituted basement membrane aligned (RBM)-PCL NFs within 10° of angular deviation. Neuron-like PC12 cells show significantly greater attachment (p < 0.001) and percentage of neurite-extending cells in vitro on 10% RBM-PCL NFs when compared to 1% and 0% RBM-PCL NFs (p < 0.015 and p < 0.001, respectively). Together, these results implicate laminin- and RBM-PCL scaffolds as a promising biomimetic substrate for regenerative medicine applications.

Cell Attachment and Proliferation of Human Adipose-Derived Stem Cells on PLGA/Chitosan Electrospun Nano-Biocomposite.

PubMed

Razavi, Shahnaz; Karbasi, Saeed; Morshed, Mohammad; Zarkesh Esfahani, Hamid; Golozar, Mohammad; Vaezifar, Sedigheh

2015-01-01

In this study, nano-biocomposite composed of poly (lactide-co-glycolide) (PLGA) and chitosan (CS) were electrospun through a single nozzle by dispersing the CS nano-powders in PLGA solution. The cellular behavior of human adipose derived stem cells (h-ADSCs) on random and aligned scaffolds was then evaluated. In this experimental study, the PLGA/CS scaffolds were prepared at the different ratios of 90/10, 80/20, and 70/30 (w/w) %. Morphology, cell adhesion and prolif- eration rate of h-ADSCs on the scaffolds were assessed using scanning electron microscope (SEM), 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay and trypan blue staining respectively. H-ADSCs seeded on the matrices indicated that the PLGA/CS composite matrix with aligned nanofibres and higher content of CS nano-powders gave significantly better performance than others in terms of cell adhesion and proliferation rate (P<0.05). We found that CS enhanced cell adhesion and proliferation rate, and aligned nanofibers guided cell growth along the longitudinal axis of the nanofibers, which would provide a beneficial approach for tissue engineering.

Composites structures for bone tissue reconstruction

DOE Office of Scientific and Technical Information (OSTI.GOV)

Neto, W.; Santos, João; Avérous, L.

2015-05-22

The search for new biomaterials in the bone reconstitution field is growing continuously as humane life expectation and bone fractures increase. For this purpose, composite materials with biodegradable polymers and hydroxyapatite (HA) have been used. A composite material formed by a film, nanofibers and HA has been made. Both, the films and the non-woven mats of nanofibers were formed by nanocomposites made of butylene adipate-co-terephthalate (PBAT) and HA. The techniques used to produce the films and nanofibers were spin coating and electrospinning, respectively. The composite production and morphology were evaluated. The composite showed an adequate morphology and fibers size tomore » be used as scaffold for cell growth.« less

Recent Advances in Electrospun Nanofiber Interfaces for Biosensing Devices

PubMed Central

Sapountzi, Eleni; Braiek, Mohamed; Chateaux, Jean-François; Lagarde, Florence

2017-01-01

Electrospinning has emerged as a very powerful method combining efficiency, versatility and low cost to elaborate scalable ordered and complex nanofibrous assemblies from a rich variety of polymers. Electrospun nanofibers have demonstrated high potential for a wide spectrum of applications, including drug delivery, tissue engineering, energy conversion and storage, or physical and chemical sensors. The number of works related to biosensing devices integrating electrospun nanofibers has also increased substantially over the last decade. This review provides an overview of the current research activities and new trends in the field. Retaining the bioreceptor functionality is one of the main challenges associated with the production of nanofiber-based biosensing interfaces. The bioreceptors can be immobilized using various strategies, depending on the physical and chemical characteristics of both bioreceptors and nanofiber scaffolds, and on their interfacial interactions. The production of nanobiocomposites constituted by carbon, metal oxide or polymer electrospun nanofibers integrating bioreceptors and conductive nanomaterials (e.g., carbon nanotubes, metal nanoparticles) has been one of the major trends in the last few years. The use of electrospun nanofibers in ELISA-type bioassays, lab-on-a-chip and paper-based point-of-care devices is also highly promising. After a short and general description of electrospinning process, the different strategies to produce electrospun nanofiber biosensing interfaces are discussed. PMID:28813013

Synthesis and characterization of curcumin loaded PLA-Hyperbranched polyglycerol electrospun blend for wound dressing applications.

PubMed

Perumal, Govindaraj; Pappuru, Sreenath; Chakraborty, Debashis; Maya Nandkumar, A; Chand, Dillip Kumar; Doble, Mukesh

2017-07-01

This study is aimed to develop curcumin (Cur) incorporated electrospun nanofibers of a blend of poly (lactic acid) (PLA) and hyperbranched polyglycerol (HPG) for wound healing applications. Both the polymers are synthesized and fabricated by electrospinning technique. The produced nanofibers were characterized by Scanning Electron Microscopy (SEM), X-Ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FT-IR), Differential Scanning Colorimetry (DSC) and Thermogravimetric Analysis (TGA). Electrospun scaffolds (PLA/HPG/Cur) exhibits very high hydrophilicity, high swelling and drug uptake and promotes better cell viability, adhesion and proliferation when compared to PLA/Cur electrospun nanofibers. Biodegradation study revealed that the morphology of the nanofibers were unaffected even after 14days immersion in Phosphate Buffered Saline. In vitro scratch assay indicates that migration of the cells in the scratch treated with PLA/HPG/Cur is complete within 36h. These results suggest that PLA/HPG/Cur nanofibers can be a potential wound patch dressing for acute and chronic wound applications. Copyright © 2017 Elsevier B.V. All rights reserved.

Chondroitin sulfate immobilization at the surface of electrospun nanofiber meshes for cartilage tissue regeneration approaches

NASA Astrophysics Data System (ADS)

Piai, Juliana Francis; da Silva, Marta Alves; Martins, Albino; Torres, Ana Bela; Faria, Susana; Reis, Rui L.; Muniz, Edvani Curti; Neves, Nuno M.

2017-05-01

Aiming at improving the biocompatibility of biomaterial scaffolds, surface modification presents a way to preserve their mechanical properties and to improve the surface bioactivity. In this work, chondroitin sulfate (CS) was immobilized at the surface of electrospun poly(caprolactone) nanofiber meshes (PCL NFMs), previously functionalized by UV/O3 exposure and aminolysis. Contact angle, SEM, optical profilometry, FTIR, X-ray photoelectron spectroscopy techniques confirmed the success of CS-immobilization in PCL NFMs. Furthermore, CS-immobilized PCL NFMs showed lower roughness and higher hydrophilicity than the samples without CS. Human articular chondrocytes (hACs) were cultured on electrospun PCL NFMs with or without CS immobilization. It was observed that hACs proliferated through the entire time course of the experiment in both types of nanofibrous scaffolds, as well as for the production of glycosaminoglycans. Quantitative-PCR results demonstrated over-expression of cartilage-related genes such as Aggrecan, Collagen type II, COMP and Sox9 on both types of nanofibrous scaffolds. Morphological observations from SEM and LSCM revealed that hACs maintained their characteristic round shape and cellular agglomeration exclusively on PCL NFMs with CS immobilization. In conclusion, CS immobilization at the surface of PCL NFMs was achieved successfully and provides a valid platform enabling further surface functionalization methods in scaffolds to be developed for cartilage tissue engineering.

Effect of Polyhydroxybutyrate/Chitosan/Bioglass nanofiber scaffold on proliferation and differentiation of stem cells from human exfoliated deciduous teeth into odontoblast-like cells.

PubMed

Khoroushi, Maryam; Foroughi, Mohammad Reza; Karbasi, Saeed; Hashemibeni, Batool; Khademi, Abbas Ali

2018-08-01

Scaffolds and their characteristics play a central role in tissue engineering. The purpose of this study was to determine the effects of Polyhydroxybutyrate (PHB)/Chitosan/nano-bioglass (nBG) nanofiber scaffold made using the electrospinning method, on the proliferation and differentiation of stem cells obtained from human exfoliated deciduous teeth into odontoblast-like cells. In this experimental study, the pulps of the molten deciduous teeth were isolated, thereafter, the stem cells from human exfoliated deciduous teeth (SHED) were extracted and then the 3-(4,5-dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the cell viability percentage. The expression of some stem cell genes was studied by flowcytometry. These cells were then subjected to odontoblast by using the bone morphogenetic proteins-2 (BMP2) growth factor in the differentiation medium and for the expression of their specific genes. Primers of collagen type-I, dentin sialophosphoprotein (DSPP) and alkaline phosphatase (ALP) were used and the percentage of differentiation to odontoblast cells in induction scaffolds was investigated using real-time PCR and immunohistochemistry methods. The results revealed a 6-fold increase in the expression of DSPP genes and collagen type-I, and a 2-fold increase in the expression of ALP in scaffold with BMP2 group compared to the scaffold as control group which according to the immunohistochemical test results, showed the extracted SHED to have been differentiated into dentin odontoblast-like cells. As a result, this scaffold can be used as a suitable substrate to apply in dentin tissue engineering. Copyright © 2018. Published by Elsevier B.V.

Surface modification of electrospun PVA/chitosan nanofibers by dielectric barrier discharge plasma at atmospheric pressure and studies of their mechanical properties and biocompatibility.

PubMed

Das, Punamshree; Ojah, Namita; Kandimalla, Raghuram; Mohan, Kiranjyoti; Gogoi, Dolly; Dolui, Swapan Kumar; Choudhury, Arup Jyoti

2018-03-22

In this paper, surface of electrospun PVA/Cs nanofibers is modified using dielectric barrier discharge (DBD) plasma and the relationship between the observed mechanical properties and biocompatibility of the nanofibers and plasma-induced surface properties is discussed. Plasma treatment of electrospun PVA/Cs nanofibers is carried out with both inert (argon, Ar) and reactive (oxygen, O 2 ) gases at atmospheric pressure. Incorporation of oxygen-containing polar functional groups on the surface of Ar-plasma treated (PVA/Cs/Ar) and O 2 -plasma treated (PVA/Cs/O 2 ) nanofibers and increase in surface roughness contribute to the improvement of surface wettability and the decrease of contact angle with water of the nanofibers. Both PVA/Cs/Ar and PVA/Cs/O 2 nanofibers show high tensile strength (11.6-15.6%) and Young's modulus (33.8-37.3%) as compared to the untreated one. Experimental results show that in terms of haemolytic activity the PVA/Cs/Ar and PVA/Cs/O 2 nanofibers do not cause structural changes of blood cells and meet the biocompatibility requirements for blood-contacting polymeric materials. MTT cell viability results further reveals improvement in biocompatibility of PVA/Cs nanofibers after Ar and O 2 plasma treatment. The results suggest that DBD plasma treated electrospun PVA/Cs nanofibers have the potential to be used as wound dressing and scaffolds for tissue engineering. Copyright © 2018 Elsevier B.V. All rights reserved.

Mechanical properties of electrospun bilayer fibrous membranes as potential scaffolds for tissue engineering.

PubMed

Pu, Juan; Komvopoulos, Kyriakos

2014-06-01

Bilayer fibrous membranes of poly(l-lactic acid) (PLLA) were fabricated by electrospinning, using a parallel-disk mandrel configuration that resulted in the sequential deposition of a layer with fibers aligned across the two parallel disks and a layer with randomly oriented fibers, both layers deposited in a single process step. Membrane structure and fiber alignment were characterized by scanning electron microscopy and two-dimensional fast Fourier transform. Because of the intricacies of the generated electric field, bilayer membranes exhibited higher porosity than single-layer membranes consisting of randomly oriented fibers fabricated with a solid-drum collector. However, despite their higher porosity, bilayer membranes demonstrated generally higher elastic modulus, yield strength and toughness than single-layer membranes with random fibers. Bilayer membrane deformation at relatively high strain rates comprised multiple abrupt microfracture events characterized by discontinuous fiber breakage. Bilayer membrane elongation yielded excessive necking of the layer with random fibers and remarkable fiber stretching (on the order of 400%) in the layer with fibers aligned in the stress direction. In addition, fibers in both layers exhibited multiple localized necking, attributed to the nonuniform distribution of crystalline phases in the fibrillar structure. The high membrane porosity, good mechanical properties, and good biocompatibility and biodegradability of PLLA (demonstrated in previous studies) make the present bilayer membranes good scaffold candidates for a wide range of tissue engineering applications. Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Electrospinning: An enabling nanotechnology platform for drug delivery and regenerative medicine.

PubMed

Chen, Shixuan; Li, Ruiquan; Li, Xiaoran; Xie, Jingwei

2018-05-02

Electrospinning provides an enabling nanotechnology platform for generating a rich variety of novel structured materials in many biomedical applications including drug delivery, biosensing, tissue engineering, and regenerative medicine. In this review article, we begin with a thorough discussion on the method of producing 1D, 2D, and 3D electrospun nanofiber materials. In particular, we emphasize on how the 3D printing technology can contribute to the improvement of traditional electrospinning technology for the fabrication of 3D electrospun nanofiber materials as drug delivery devices/implants, scaffolds or living tissue constructs. We then highlight several notable examples of electrospun nanofiber materials in specific biomedical applications including cancer therapy, guiding cellular responses, engineering in vitro 3D tissue models, and tissue regeneration. Finally, we finish with conclusions and future perspectives of electrospun nanofiber materials for drug delivery and regenerative medicine. Copyright © 2018 Elsevier B.V. All rights reserved.

Chitosan based nanofibers in bone tissue engineering.

PubMed

Balagangadharan, K; Dhivya, S; Selvamurugan, N

2017-11-01

Bone tissue engineering involves biomaterials, cells and regulatory factors to make biosynthetic bone grafts with efficient mineralization for regeneration of fractured or damaged bones. Out of all the techniques available for scaffold preparation, electrospinning is given priority as it can fabricate nanostructures. Also, electrospun nanofibers possess unique properties such as the high surface area to volume ratio, porosity, stability, permeability and morphological similarity to that of extra cellular matrix. Chitosan (CS) has a significant edge over other materials and as a graft material, CS can be used alone or in combination with other materials in the form of nanofibers to provide the structural and biochemical cues for acceleration of bone regeneration. Hence, this review was aimed to provide a detailed study available on CS and its composites prepared as nanofibers, and their associated properties found suitable for bone tissue engineering. Copyright © 2016 Elsevier B.V. All rights reserved.

Recent Applications of Coaxial and Emulsion Electrospinning Methods in the Field of Tissue Engineering

PubMed Central

McClellan, Phillip; Landis, William J.

2016-01-01

Abstract Electrospinning has emerged as an effective method of producing nanoscale fibers for use in multiple fields of study. One area of significant interest is nanofiber utilization for tissue engineering because the nanofibrous mats can mimic the native extracellular matrix of biological tissues. A logical next step is the inclusion of certain molecules and compounds to accelerate or increase the efficacy of tissue regeneration. Two methods are under scrutiny for their capability to encapsulate therapeutic compounds within electrospun nanofibers: emulsion and coaxial electrospinning. Both have advantages and disadvantages, which need to be taken into careful consideration when deciding to use them in a specific application. Several examples are provided here to highlight the vast potential of multilayered nanofibers as well as the emergence of new techniques to produce three-dimensional scaffolds of nanofibers for use in the field of tissue engineering. PMID:27610268

Poly(hydroxybutyrate)/chitosan Aligned Electrospun Scaffold as a Novel Substrate for Nerve Tissue Engineering

PubMed Central

Karimi, Afarin; Karbasi, Saeed; Razavi, Shahnaz; Zargar, Elham Naghash

2018-01-01

Background: Reconstruction of nervous system is a great challenge in the therapeutic medical field. Nerve tissue engineering is a novel method to regenerate nervous system in human health care. Tissue engineering has introduced novel approaches to promote and guide peripheral nerve regeneration using submicron and nanoscale fibrous scaffolds. Materials and Methods: In this study, 9 wt% poly(3-hydroxybutyrate) (PHB) solutions with two different ratios of chitosan (CTS) (15%, and 20%) were mixed in trifluoroacetic acid as a cosolvent. Thereafter, random and aligned PHB/CTS scaffolds were fabricated by electrospinning method in an appropriate condition. Results: Average diameters for aligned PHB, PHB/CTS 85:15 and PHB/CTS 80:20 were obtained as 675 nm, 740.3 nm, and 870.74 nm, which was lesser than random fibers. The solution components entity authenticity was approved by Fourier transform infrared. The addition of CTS decreased both water droplet contact angle from 124.79° to 43.14° in random and 110.87° to 33.49° in aligned PHB/CTS fibrous scaffold. Moreover, alignment of fibers causes tremendous increase in hydrophilicity of fibrous PHB/CTS substrate. Tensile strength increased from 6.41 MPa for random to 8.73 MPa for aligned PHB/CTS 85:15. Conclusions: Our results indicated that aligned PHB/CTS 85:15 nanofibers are the desired scaffold than the random PHB/CTS nanofibers for application in nerve tissue regeneration. PMID:29657929

Braided nanofibrous scaffold for tendon and ligament tissue engineering.

PubMed

Barber, John G; Handorf, Andrew M; Allee, Tyler J; Li, Wan-Ju

2013-06-01

Tendon and ligament (T/L) injuries present an important clinical challenge due to their intrinsically poor healing capacity. Natural healing typically leads to the formation of scar-like tissue possessing inferior mechanical properties. Therefore, tissue engineering has gained considerable attention as a promising alternative for T/L repair. In this study, we fabricated braided nanofibrous scaffolds (BNFSs) as a potential construct for T/L tissue engineering. Scaffolds were fabricated by braiding 3, 4, or 5 aligned bundles of electrospun poly(L-lactic acid) nanofibers, thus introducing an additional degree of flexibility to alter the mechanical properties of individual scaffolds. We observed that the Young's modulus, yield stress, and ultimate stress were all increased in the 3-bundle compared to the 4- and 5-bundle BNFSs. Interestingly, acellular BNFSs mimicked the normal tri-phasic mechanical behavior of native tendon and ligament (T/L) during loading. When cultured on the BNFSs, human mesenchymal stem cells (hMSCs) adhered, aligned parallel to the length of the nanofibers, and displayed a concomitant realignment of the actin cytoskeleton. In addition, the BNFSs supported hMSC proliferation and induced an upregulation in the expression of key pluripotency genes. When cultured on BNFSs in the presence of tenogenic growth factors and stimulated with cyclic tensile strain, hMSCs differentiated into the tenogenic lineage, evidenced most notably by the significant upregulation of Scleraxis gene expression. These results demonstrate that BNFSs provide a versatile scaffold capable of supporting both stem cell expansion and differentiation for T/L tissue engineering applications.

Superelastic, superabsorbent and 3D nanofiber-assembled scaffold for tissue engineering.

PubMed

Chen, Weiming; Ma, Jun; Zhu, Lei; Morsi, Yosry; -Ei-Hamshary, Hany; Al-Deyab, Salem S; Mo, Xiumei

2016-06-01

Fabrication of 3D scaffold to mimic the nanofibrous structure of the nature extracellular matrix (ECM) with appropriate mechanical properties and excellent biocompatibility, remain an important technical challenge in tissue engineering. The present study reports the strategy to fabricate a 3D nanofibrous scaffold with similar structure to collagen in ECM by combining electrospinning and freeze-drying technique. With the technique reported here, a nanofibrous structure scaffold with hydrophilic and superabsorbent properties can be readily prepared by Gelatin and Polylactic acid (PLA). In wet state the scaffold also shows a super-elastic property, which could bear a compressive strain as high as 80% and recovers its original shape afterwards. Moreover, after 6 days of culture, L-929 cells grow, proliferate and infiltrated into the scaffold. The results suggest that this 3D nanofibrous scaffold would be promising for varied field of tissue engineering application. Copyright © 2016 Elsevier B.V. All rights reserved.

Electrospun nanofibrous 3D scaffold for bone tissue engineering.

PubMed

Eap, Sandy; Ferrand, Alice; Palomares, Carlos Mendoza; Hébraud, Anne; Stoltz, Jean-François; Mainard, Didier; Schlatter, Guy; Benkirane-Jessel, Nadia

2012-01-01

Tissue engineering aims at developing functional substitutes for damaged tissues by mimicking natural tissues. In particular, tissue engineering for bone regeneration enables healing of some bone diseases. Thus, several methods have been developed in order to produce implantable biomaterial structures that imitate the constitution of bone. Electrospinning is one of these methods. This technique produces nonwoven scaffolds made of nanofibers which size and organization match those of the extracellular matrix. Until now, seldom electrospun scaffolds were produced with thickness exceeding one millimeter. This article introduces a new kind of electrospun membrane called 3D scaffold of thickness easily exceeding one centimeter. The manufacturing involves a solution of poly(ε-caprolactone) in DMF/DCM system. The aim is to establish parameters for electrospinning in order to characterize these 3D scaffolds and, establish whether such scaffolds are potentially interesting for bone regeneration.

Physical and Biological Modification of Polycaprolactone Electrospun Nanofiber by Panax Ginseng Extract for Bone Tissue Engineering Application.

PubMed

Pajoumshariati, Seyedramin; Yavari, Seyedeh Kimia; Shokrgozar, Mohammad Ali

2016-05-01

Medicinal plants as a therapeutic agent with osteogenic properties can enhance fracture-healing process. In this study, the osteo-inductive potential of Asian Panax Ginseng root extract within electrospun polycaprolactone (PCL) based nanofibers has been investigated. Scanning electron microscopy images revealed that all nanofibers were highly porous and beadles with average diameter ranging from 250 to 650 nm. The incorporation of ginseng extract improved the physical characteristics (i.e., hydrophilicity) of PCL nanofibers, as well as the mechanical properties. Although ginseng extract increased the degradation rate of pure PCL nanofibers, the porous structure and morphology of fibers did not change significantly after 42 days. It was found that nanofibrous scaffolds containing ginseng extract had higher proliferation (up to ~1.5 fold) compared to the pristine PCL. The qRT-PCR analysis demonstrated the addition of ginseng extract into PCL nanofibers induced significant expression of osteogenic genes (Osteocalcin, Runx-2 and Col-1) in MSCs in a concentration dependent manner. Moreover, higher calcium content, alkaline phosphatase activity and higher mineralization of MSCs were observed compared to the pristine PCL fibers. Our results indicated the promising potential of ginseng extract as an additive to enhance osteo-inductivity, mechanical and physical properties of PCL nanofibers for bone tissue engineering application.

Local myocardial insulin-like growth factor 1 (IGF-1) delivery with biotinylated peptide nanofibers improves cell therapy for myocardial infarction

NASA Astrophysics Data System (ADS)

Davis, Michael E.; Hsieh, Patrick C. H.; Takahashi, Tomosaburo; Song, Qing; Zhang, Shuguang; Kamm, Roger D.; Grodzinsky, Alan J.; Anversa, Piero; Lee, Richard T.

2006-05-01

Strategies for cardiac repair include injection of cells, but these approaches have been hampered by poor cell engraftment, survival, and differentiation. To address these shortcomings for the purpose of improving cardiac function after injury, we designed self-assembling peptide nanofibers for prolonged delivery of insulin-like growth factor 1 (IGF-1), a cardiomyocyte growth and differentiation factor, to the myocardium, using a "biotin sandwich" approach. Biotinylated IGF-1 was complexed with tetravalent streptavidin and then bound to biotinylated self-assembling peptides. This biotin sandwich strategy allowed binding of IGF-1 but did not prevent self-assembly of the peptides into nanofibers within the myocardium. IGF-1 that was bound to peptide nanofibers activated Akt, decreased activation of caspase-3, and increased expression of cardiac troponin I in cardiomyocytes. After injection into rat myocardium, biotinylated nanofibers provided sustained IGF-1 delivery for 28 days, and targeted delivery of IGF-1 in vivo increased activation of Akt in the myocardium. When combined with transplanted cardiomyocytes, IGF-1 delivery by biotinylated nanofibers decreased caspase-3 cleavage by 28% and increased the myocyte cross-sectional area by 25% compared with cells embedded within nanofibers alone or with untethered IGF-1. Finally, cell therapy with IGF-1 delivery by biotinylated nanofibers improved systolic function after experimental myocardial infarction, demonstrating how engineering the local cellular microenvironment can improve cell therapy. engineering | maturation | scaffold

Controlled Bioactive Molecules Delivery Strategies for Tendon and Ligament Tissue Engineering using Polymeric Nanofibers.

PubMed

Hiong Teh, Thomas Kok; Hong Goh, James Cho; Toh, Siew Lok

2015-01-01

The interest in polymeric nanofibers has escalated over the past decade given its promise as tissue engineering scaffolds that can mimic the nanoscale structure of the native extracellular matrix. With functionalization of the polymeric nanofibers using bioactive molecules, localized signaling moieties can be established for the attached cells, to stimulate desired biological effects and direct cellular or tissue response. The inherently high surface area per unit mass of polymeric nanofibers can enhance cell adhesion, bioactive molecules loading and release efficiencies, and mass transfer properties. In this review article, the application of polymeric nanofibers for controlled bioactive molecules delivery will be discussed, with a focus on tendon and ligament tissue engineering. Various polymeric materials of different mechanical and degradation properties will be presented along with the nanofiber fabrication techniques explored. The bioactive molecules of interest for tendon and ligament tissue engineering, including growth factors and small molecules, will also be reviewed and compared in terms of their nanofiber incorporation strategies and release profiles. This article will also highlight and compare various innovative strategies to control the release of bioactive molecules spatiotemporally and explore an emerging tissue engineering strategy involving controlled multiple bioactive molecules sequential release. Finally, the review article concludes with challenges and future trends in the innovation and development of bioactive molecules delivery using polymeric nanofibers for tendon and ligament tissue engineering.

In vitro and in vivo biological characterization of poly(lactic acid) fiber scaffolds synthesized by air jet spinning.

PubMed

Granados-Hernández, Marco Vladimir; Serrano-Bello, Janeth; Montesinos, Juan José; Alvarez-Gayosso, Carlos; Medina-Velázquez, Luis Alberto; Alvarez-Fregoso, Octavio; Alvarez-Perez, Marco Antonio

2017-11-30

Poly(lactic acid) (PLA) is one of the most promising renewable and biodegradable polymers for mimic extracellular matrix for tissue engineering applications. In this work, PLA spun membrane scaffold were successfully prepared by air jet spinning technology. Morphology, mechanical properties, in vitro biocompatibility, and in vitro and in vivo degradation of PLA fibrous scaffold were characterized by X-ray diffraction, Fourier Transform Infrared, and scanning electron microscope (SEM). Morphological results assessed by SEM analyses indicated that PLA scaffolds possessed an average fiber diameter of approximately 0.558 ± 0.141 µm for 7% w/v of PLA and approximately 0.647 ± 0.137 µm for 10% w/v. Interestingly, our results showed that the nanofiber size of PLA scaffold allow structural stability after 100 days of in vitro degradation in Ringer solution where the average fiber diameter were of approximately 0.633 ± 0.147 µm for 7% w/v and approximately 0.645 ± 0.140 µm for 10% w/v of PLA. Mechanical properties of PLA fibers scaffold after in vitro degradation showed decrease in terms of flexibility elongation, and less energy was needed to achieve maximal elastic deformation. The fiber size exerts an influence on the biological response of human Bone Marrow Mesenchymal Stromal Cells as confirmed by MTT assay after 9 days of cell culture and the in vivo degradation assay of 7% w/v and 10% w/v of PLA scaffold, did not demonstrate evidence of toxicity with a mild inflammatory respond. In conclusion, airbrushing technology promises to be a viable and attractive alternative technique for producing a biocompatible PLA nanofiber scaffold that could be considered for tissue engineering regeneration. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2017. © 2017 Wiley Periodicals, Inc.

Cryogenic 3D printing for producing hierarchical porous and rhBMP-2-loaded Ca-P/PLLA nanocomposite scaffolds for bone tissue engineering.

PubMed

Wang, Chong; Zhao, Qilong; Wang, Min

2017-06-07

The performance of bone tissue engineering scaffolds can be assessed through cell responses to scaffolds, including cell attachment, infiltration, morphogenesis, proliferation, differentiation, etc, which are determined or heavily influenced by the composition, structure, mechanical properties, and biological properties (e.g. osteoconductivity and osteoinductivity) of scaffolds. Although some promising 3D printing techniques such as fused deposition modeling and selective laser sintering could be employed to produce biodegradable bone tissue engineering scaffolds with customized shapes and tailored interconnected pores, effective methods for fabricating scaffolds with well-designed hierarchical porous structure (both interconnected macropores and surface micropores) and tunable osteoconductivity/osteoinductivity still need to be developed. In this investigation, a novel cryogenic 3D printing technique was investigated and developed for producing hierarchical porous and recombinant human bone morphogenetic protein-2 (rhBMP-2)-loaded calcium phosphate (Ca-P) nanoparticle/poly(L-lactic acid) nanocomposite scaffolds, in which the Ca-P nanoparticle-incorporated scaffold layer and rhBMP-2-encapsulated scaffold layer were deposited alternatingly using different types of emulsions as printing inks. The mechanical properties of the as-printed scaffolds were comparable to those of human cancellous bone. Sustained releases of Ca 2+ ions and rhBMP-2 were achieved and the biological activity of rhBMP-2 was well-preserved. Scaffolds with a desirable hierarchical porous structure and dual delivery of Ca 2+ ions and rhBMP-2 exhibited superior performance in directing the behaviors of human bone marrow-derived mesenchymal stem cells and caused improved cell viability, attachment, proliferation, and osteogenic differentiation, which has suggested their great potential for bone tissue engineering.

Cell Attachment and Proliferation of Human Adipose-Derived Stem Cells on PLGA/Chitosan Electrospun Nano-Biocomposite

PubMed Central

Razavi, Shahnaz; Karbasi, Saeed; Morshed, Mohammad; Zarkesh Esfahani, Hamid; Golozar, Mohammad; Vaezifar, Sedigheh

2015-01-01

Objective In this study, nano-biocomposite composed of poly (lactide-co-glycolide) (PLGA) and chitosan (CS) were electrospun through a single nozzle by dispersing the CS nano-powders in PLGA solution. The cellular behavior of human adipose derived stem cells (h-ADSCs) on random and aligned scaffolds was then evaluated. Materials and Methods In this experimental study, the PLGA/CS scaffolds were prepared at the different ratios of 90/10, 80/20, and 70/30 (w/w) %. Morphology, cell adhesion and prolif- eration rate of h-ADSCs on the scaffolds were assessed using scanning electron microscope (SEM), 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay and trypan blue staining respectively. Results H-ADSCs seeded on the matrices indicated that the PLGA/CS composite matrix with aligned nanofibres and higher content of CS nano-powders gave significantly better performance than others in terms of cell adhesion and proliferation rate (P<0.05). Conclusion We found that CS enhanced cell adhesion and proliferation rate, and aligned nanofibers guided cell growth along the longitudinal axis of the nanofibers, which would provide a beneficial approach for tissue engineering. PMID:26464814

Novel electrospun nanofibers of modified gelatin-tyrosine in cartilage tissue engineering.

PubMed

Agheb, Maria; Dinari, Mohammad; Rafienia, Mohammad; Salehi, Hossein

2017-02-01

In natural cartilage tissues, chondrocytes are linked to extracellular matrix (ECM) through cell-surface binding proteins. Surface modification of gelatin can provide a new generation of biopolymers and fibrous scaffolds with chemical, mechanical, and biological properties. In this study tyrosine protein and 1,2,3-triazole ring were utilized to functionalize gelatin without Cu catalyst. Their molecular structure was characterized by Fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy ( 1 HNMR). Chemical cross-linkers such as glutaraldehyde (GA) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysulfosuccinimide (NHS) were used to electrospin the modified gelatin. The modification of gelatin and cross-linking effects were confirmed by scanning electron microscopy (SEM), contact angle measurement, and mechanical tests. MTT assay using chondrocyte cells showed cell viability of electrospun modified gelatin scaffolds. In vitro cell culture studies showed that electrospun engineered protein scaffolds would support the attachment and growth of cells. The results also showed that cross-linked nanofibers with EDC/NHS could be considered excellent matrices in cell adhesion and proliferation before electrospinning process and their potential substrate in tissue engineering applications, especially in the field of cartilage engineering. Copyright © 2016. Published by Elsevier B.V.

In-situ synthesis of AgNPs in the natural/synthetic hybrid nanofibrous scaffolds: Fabrication, characterization and antimicrobial activities.

PubMed

Maharjan, Bikendra; Joshi, Mahesh Kumar; Tiwari, Arjun Prasad; Park, Chan Hee; Kim, Cheol Sang

2017-01-01

Silver nanoparticles embedded within a nanofibrous polymer matrix have significant attention in recent years as an antimicrobial wound dressing materials. Herein, we have fabricated a novel Ag-polyurethane-zein hybrid nanofibrous scaffold for wound dressing applications. AgNPs were synthesized in-situ via reduction of silver nitrate in electrospinning solution. Varying mass composition of the components showed the pronounced effect on the morphology and physicochemical properties of the composite fibers. Field-Emission Scanning Electron Microscopy (FESEM) images revealed that PU and zein with mass ratio 2:1 produced the bead-free continuous and uniformly distributed nanofibers. Fourier-transform Infrared Spectroscopy (FTIR), X-ray diffraction (XRD) and Thermogravimetric Analysis (TGA) confirmed the well interaction between component polymers. Compared to the pristine PU nanofibers, composite fibers showed enhanced tensile strength, young׳s modulus and surface wettability. The antibacterial capacity of the nanofibrous membrane was evaluated against gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacterial strains via a zone of inhibition test, and the results showed high antibacterial performance for Ag incorporated composite mat. Experimental results of cell viability assay and microscopic imaging revealed that as-fabricated scaffolds have an excellent ability for fibroblast cell adhesion, proliferation and growth. Overall, as-fabricated antibacterial natural/synthetic composite scaffold can be a promising substrate for repairing skin defects. Copyright © 2016. Published by Elsevier Ltd.

Peptide modified nanofibrous scaffold promotes human mesenchymal stem cell proliferation and long-term passaging.

PubMed

Mobasseri, Rezvan; Tian, Lingling; Soleimani, Masoud; Ramakrishna, Seeram; Naderi-Manesh, Hossein

2018-03-01

Long-term culture, passage and proliferation of human mesenchymal stem cells (hMSCs) cause loss of their stemness properties including self-renewal and multipotency. By optimizing the MSCs environment in vitro, maintaining the stemness state and better controlling the cell fate might be possible. We have recently reported the significant effects of bioactive Tat protein-derived peptide named R-peptide on hMSC adhesion, morphology and proliferation, which has demonstrated R-peptide enhanced MSC early adhesion and proliferation in comparison to other bioactive molecules including RGD peptide, fibronectin and collagen. In this study, R-peptide was used to evaluate stemness properties of MSCs after long-term passaging. R-peptide conjugated poly caprolactone (PCL) nanofibrous scaffold and unmodified nanofibrous scaffold were used to study the impact of R-peptide modified PCL nanofibers and PCL nanofibers on cell behavior. The results showed early formation of focal adhesion (FA) complex on R-peptide modified scaffolds at 30min after cell seeding. The rate of cell proliferation was significantly increased due to presence of R-peptide, and the MSCs marker analyses using flow cytometry and immunocytochemistry staining proved the ability of R-peptide to maintain mesenchymal stem cell properties (high proliferation, expression of multipotent markers and differentiation capacity) even after long-term passage culturing. Accordingly, our (The) results concluded that bioactive R-peptide in combination with nanofibrous scaffold can mimic the native ECM comprising micro/nano architecture and biochemical molecules in a best way. The designed scaffold can link extracellular matrix (ECM) to nucleus via formation of FA and organization of cytoskeleton, causing fast and strong attachment of MSCs and allowing integrin-mediated signaling to start. Copyright © 2017 Elsevier B.V. All rights reserved.

Biomimetic Silk Scaffolds with an Amorphous Structure for Soft Tissue Engineering.

PubMed

Sang, Yonghuan; Li, Meirong; Liu, Jiejie; Yao, Yuling; Ding, Zhaozhao; Wang, Lili; Xiao, Liying; Lu, Qiang; Fu, Xiaobing; Kaplan, David L

2018-03-21

Fine tuning physical cues of silk fibroin (SF) biomaterials to match specific requirements for different soft tissues would be advantageous. Here, amorphous SF nanofibers were used to fabricate scaffolds with better hierarchical extracellular matrix (ECM) mimetic microstructures than previous silk scaffolds. Kinetic control was introduced into the scaffold forming process, resulting in the direct production of water-stable scaffolds with tunable secondary structures and thus mechanical properties. These biomaterials remained with amorphous structures, offering softer properties than prior scaffolds. The fine mechanical tunability of these systems provides a feasible way to optimize physical cues for improved cell proliferation and enhanced neovascularization in vivo. Multiple physical cues, such as partly ECM mimetic structures and optimized stiffness, provided suitable microenvironments for tissue ingrowth, suggesting the possibility of actively designing bioactive SF biomaterials. These systems suggest a promising strategy to develop novel SF biomaterials for soft tissue repair and regenerative medicine.

Potential of inherent RGD containing silk fibroin-poly (Є-caprolactone) nanofibrous matrix for bone tissue engineering.

PubMed

Bhattacharjee, Promita; Kundu, Banani; Naskar, Deboki; Kim, Hae-Won; Bhattacharya, Debasis; Maiti, T K; Kundu, S C

2016-02-01

The current study deals with the fabrication and characterization of blended nanofibrous scaffolds of tropical tasar silk fibroin of Antheraea mylitta and poly (Є-caprolactone) to act as an ideal scaffold for bone regeneration. The use of poly (Є-caprolactone) in osteogenesis is well-recognized. At the same time, the osteoconductive nature of the non-mulberry tasar fibroin is also established due to its internal integrin binding peptide RGD (Arg-Gly-Asp) sequences, which enhance cellular interaction and proliferation. Considering that the materials have the required and favorable properties, the blends are formed using an equal volume ratio of fibroin (2 and 4 wt%) and poly (Є-caprolactone) solution (10 wt%) to fabricate nanofibers. The nanofibers possess an average diameter of 152 ± 18 nm (2 % fibroin/PCL) and 175 ± 15 nm (4% fibroin/PCL). The results of Fourier transform infrared spectroscopy substantiates the preservation of the secondary structure of the fibroin in the blends indicating the structural stability of the neo-matrix. With an increase in the fibroin percentage, the hydrophobicity and thermal stability of the matrices as measured from melting temperature Tm (using DSC) decrease, while the mechanical strength is improved. The blended nanofibrous scaffolds are biodegradable, and support the viability and proliferation of human osteoblast-like cells as observed through scanning electron and confocal microscopes. Alkaline phosphatase assay indicates the cell proliferation and the generation of the neo-bone matrix. Taken together, these findings illustrate that the silk-poly (Є-caprolactone) blended nanofibrous scaffolds have an excellent prospect as scaffolding material in bone tissue engineering.

Cell-adhesive RGD peptide-displaying M13 bacteriophage/PLGA nanofiber matrices for growth of fibroblasts.

PubMed

Shin, Yong Cheol; Lee, Jong Ho; Jin, Linhua; Kim, Min Jeong; Oh, Jin-Woo; Kim, Tai Wan; Han, Dong-Wook

2014-01-01

M13 bacteriophages can be readily fabricated as nanofibers due to non-toxic bacterial virus with a nanofiber-like shape. In the present study, we prepared hybrid nanofiber matrices composed of poly(lactic-co-glycolic acid, PLGA) and M13 bacteriophages which were genetically modified to display the RGD peptide on their surface (RGD-M13 phage). The surface morphology and chemical composition of hybrid nanofiber matrices were characterized by scanning electron microscopy (SEM) and Raman spectroscopy, respectively. Immunofluorescence staining was conducted to investigate the existence of M13 bacteriophages in RGD-M13 phage/PLGA hybrid nanofibers. In addition, the attachment and proliferation of three different types of fibroblasts on RGD-M13 phage/PLGA nanofiber matrices were evaluated to explore how fibroblasts interact with these matrices. SEM images showed that RGD-M13 phage/PLGA hybrid matrices had the non-woven porous structure, quite similar to that of natural extracellular matrices, having an average fiber diameter of about 190 nm. Immunofluorescence images and Raman spectra revealed that RGD-M13 phages were homogeneously distributed in entire matrices. Moreover, the attachment and proliferation of fibroblasts cultured on RGD-M13 phage/PLGA matrices were significantly enhanced due to enriched RGD moieties on hybrid matrices. These results suggest that RGD-M13 phage/PLGA matrices can be efficiently used as biomimetic scaffolds for tissue engineering applications.

Fabrication of a novel scaffold of clotrimazole-microemulsion-containing nanofibers using an electrospinning process for oral candidiasis applications.

PubMed

Tonglairoum, Prasopchai; Ngawhirunpat, Tanasait; Rojanarata, Theerasak; Kaomongkolgit, Ruchadaporn; Opanasopit, Praneet

2015-02-01

Clotrimazole (CZ)-loaded microemulsion-containing nanofiber mats were developed as an alternative for oral candidiasis applications. The microemulsion was composed of oleic acid (O), Tween 80 (T80), and a co-surfactant such as benzyl alcohol (BzOH), ethyl alcohol (EtOH) or isopropyl alcohol (IPA). The nanofiber mats were obtained by electrospinning a blended solution of a CZ-loaded microemulsion and a mixed polymer solution of 2% (w/v) chitosan (CS) and 10% (w/v) polyvinyl alcohol (PVA) at a weight ratio of 30:70. The nanofiber mats were characterized using various analytical techniques. The entrapment efficiency, drug release, antifungal activity and cytotoxicity were investigated. The average diameter of the nanofiber mats was in the range of 105.91-125.56 nm. The differential scanning calorimetry (DSC) and powder X-ray diffractometry (PXRD) results revealed the amorphous state of the CZ-loaded microemulsions incorporated into the nanofiber mats. The entrapment efficiency of CZ in the mats was approximately 72.58-98.10%, depended on the microemulsion formulation. The release experiment demonstrated different CZ release characteristics from nanofiber mats prepared using different CZ-loaded microemulsions. The extent of drug release from the fiber mats at 4h was approximately 64.81-74.15%. The release kinetics appeared to follow Higuchi's model. In comparison with CZ lozenges (10mg), the nanofiber mats exhibited more rapid killing activity. Moreover, the nanofiber mats demonstrated desirable mucoadhesive properties and were safe for 2h. Therefore, the nanofiber mats have the potential to be promising candidates for oral candidiasis applications. Copyright © 2014 Elsevier B.V. All rights reserved.

Regeneration of Bombyx mori silk nanofibers and nanocomposite fibrils by the electrospinning process

NASA Astrophysics Data System (ADS)

Ayutsede, Jonathan Eyitouyo

In recent years, there has been significant interest in the utilization of natural materials for novel nanoproducts such as tissue engineered scaffolds. Silkworm silk fibers represent one of the strongest natural fibers known. Silkworm silk, a protein-based natural biopolymer, has received renewed interest in recent years due to its unique properties (strength, toughness) and potential applications such as smart textiles, protective clothing and tissue engineering. The traditional 10--20 mum diameter, triangular-shaped Bombyx mori fibers have remained unchanged over the years. However, in our study, we examine the scientific implication and potential applications of reducing the diameter to the nanoscale, changing the triangular shape of the fiber and adding nanofillers in the form of single wall carbon nanotubes (SWNT) by the electrospinning process. The electrospinning process preserves the natural conformation of the silk (random and beta-sheet). The feasibility of changing the properties of the electrospun nanofibers by post processing treatments (annealing and chemical treatment) was investigated. B. mori silk fibroin solution (formic acid) was successfully electrospun to produce uniform nanofibers (as small as 12 nm). Response Surface Methodology (RSM) was applied for the first time to experimental results of electrospinning, to develop a processing window that can reproduce regenerated silk nanofibers of a predictable size (d < 100nm). SWNT-silk multifunctional nanocomposite fibers were fabricated for the first time with anticipated properties (mechanical, thermal and electrically conductive) that may have scientific applications (nerve regeneration, stimulation of cell-scaffold interaction). In order to realize these applications, the following areas need to be addressed: a systematic investigation of the dispersion of the nanotubes in the silk matrix, a determination of new methodologies for characterizing the nanofiber properties and establishing the nature of the silk-SWNT interactions. A new visualization system was developed to characterize the transport properties of the nanofibrous assemblies. The morphological, chemical, structural and mechanical properties of the nanofibers were determined by field emission environmental scanning microscopy, Fourier transform infrared and Raman spectroscopy, wide angle x-ray diffraction and microtensile tester respectively.

Protective therapeutic effects of peptide nanofiber and hyaluronic acid hybrid membrane in in vivo osteoarthritis model.

PubMed

Arslan, Elif; Sardan Ekiz, Melis; Eren Cimenci, Cagla; Can, Nuray; Gemci, M Hanifi; Ozkan, Huseyin; Guler, Mustafa O; Tekinay, Ayse B

2018-06-01

Osteoarthritis (OA) is a condition where tissue function is lost through a combination of secondary inflammation and deterioration in articular cartilage. One of the most common causes of OA is age-related tissue impairment because of wear and tear due to mechanical erosion. Hyaluronic acid-based viscoelastic supplements have been widely used for the treatment of knee injuries. However, the current formulations of hyaluronic acid are unable to provide efficient healing and recovery. Here, a nanofiber-hyaluronic acid membrane system that was prepared by using a quarter of the concentration of commercially available hyaluronic acid supplement, Hyalgan®, was used for the treatment of an osteoarthritis model, and Synvisc®, which is another commercially available hyaluronic acid containing viscoelastic supplement, was used as a control. The results show that this system provides efficient protection of arthritic cartilage tissue through the preservation of cartilage morphology with reduced osteophyte formation, protection of the subchondral region from deterioration, and maintenance of cartilage specific matrix proteins in vivo. In addition, the hybrid nanofiber membrane enabled chondrocyte encapsulation and provided a suitable culturing environment for stem cell growth in vitro. Overall, our results suggest that this hybrid nanofibrous scaffold provides a potential platform the treatment of OA. Osteoarthritis is a debilitating joint disease affecting millions of people worldwide. It occurs especially in knees due to aging, sport injuries or obesity. Although hyaluronic acid-based viscoelastic supplements are widely used, there is still no effective treatment method for osteoarthritis, which necessitates surgical operation as an only choice for severe cases. Therefore, there is an urgent need for efficient therapeutics. In this study, a nanofiber-HA membrane system was developed for the efficient protection of arthritic cartilage tissue from degeneration. This hybrid nanofiber system provided superior therapeutic activity at a relatively lower concentration of hyaluronic acid than Hyalgan® and Synvisc® gels, which are currently used in clinics. This work demonstrates for the first time that this hybrid nanofiber membrane scaffold can be utilized as a potential candidate for osteoarthritis treatment. Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Types of neural guides and using nanotechnology for peripheral nerve reconstruction

PubMed Central

Biazar, Esmaeil; Khorasani, MT; Montazeri, Naser; Pourshamsian, Khalil; Daliri, Morteza; T, Mostafa Rezaei; B, Mahmoud Jabarvand; Khoshzaban, Ahad; K, Saeed Heidari; Jafarpour, Mostafa; Roviemiab, Ziba

2010-01-01

Peripheral nerve injuries can lead to lifetime loss of function and permanent disfigurement. Different methods, such as conventional allograft procedures and use of biologic tubes present problems when used for damaged peripheral nerve reconstruction. Designed scaffolds comprised of natural and synthetic materials are now widely used in the reconstruction of damaged tissues. Utilization of absorbable and nonabsorbable synthetic and natural polymers with unique characteristics can be an appropriate solution to repair damaged nerve tissues. Polymeric nanofibrous scaffolds with properties similar to neural structures can be more effective in the reconstruction process. Better cell adhesion and migration, more guiding of axons, and structural features, such as porosity, provide a clearer role for nanofibers in the restoration of neural tissues. In this paper, basic concepts of peripheral nerve injury, types of artificial and natural guides, and methods to improve the performance of tubes, such as orientation, nanotechnology applications for nerve reconstruction, fibers and nanofibers, electrospinning methods, and their application in peripheral nerve reconstruction are reviewed. PMID:21042546

Physical properties and morphology of electrospun composite fiber mats of polyhydroxyalkanoate containing nanoclay and tricalcium phosphate additives

NASA Astrophysics Data System (ADS)

Tanadchangsaeng, N.; Boonyagul, S.

2018-05-01

Recently, nanofiber research has gained substantial attention from scientists. In this study, the main component of the nanofiber sheet is polyhydroxyalkanoate (PHA) polymer, which is strong, ductile, flexible and adhesive to human skin. Two major additives of nanofiber sheet that we applied are nanoclay and tricalcium phosphate. The additives are generally synthetic substances that can be chemically synthesized and compatible with tissues body. Nanoclay has a low density, strong, durable to compressive strength and humidity. While, tricalcium phosphate is a calcium phosphate ceramic that is biocompatible to human tissue. From the reasons above, we proposed to choose both nanoclay and tricalcium phosphate for adding into PHA nanofibers for film formation. Thus, this study aims to investigate the morphological and mechanical properties of the fiber mat by using PHA added with various amount of nanoclay and tricalcium phosphate at 0.1%, 1% and 10% by weight, and fabricate nanofiber samples by electrospinning technique. The tested results of scanning electron microscope (SEM) morphology show that the fibers have a uniformed pattern. The PHA containing nanoclay of all additive contents exhibited micrometer diameter distributions, while PHA loaded with 1% tricalcium phosphate still had the nano-scale diameter range, and might be the optimum additive load for further nanometer medical applications. A tensile test was performed to determine the effect of nanoclay and tricalcium phosphate contents on the mechanical properties of the electrospun PHA films, and reflect the level of modularity. With nanoclay components being integrated into the polymer matrix, subsequent reduction in fiber crystallinity was occurred after addition of nanoclay with an increase of modulus value. The results confirmed that PHA fiber mat containing 1% nanoclay may have a potential for using as a rigid scaffold which bearing force loading in human organ system. Whereas, it can be indicated that PHA fiber mat containing 1% tricalcium phosphate might be employed as a flexible scaffold for biomedical materials application due to a high elongation at break value.

Comparison of cell behavior on pva/pva-gelatin electrospun nanofibers with random and aligned configuration

NASA Astrophysics Data System (ADS)

Huang, Chen-Yu; Hu, Keng-Hsiang; Wei, Zung-Hang

2016-12-01

Electrospinning technique is able to create nanofibers with specific orientation. Poly(vinyl alcohol) (PVA) have good mechanical stability but poor cell adhesion property due to the low affinity of protein. In this paper, extracellular matrix, gelatin is incorporated into PVA solution to form electrospun PVA-gelatin nanofibers membrane. Both randomly oriented and aligned nanofibers are used to investigate the topography-induced behavior of fibroblasts. Surface morphology of the fibers is studied by optical microscopy and scanning electron microscopy (SEM) coupled with image analysis. Functional group composition in PVA or PVA-gelatin is investigated by Fourier Transform Infrared (FTIR). The morphological changes, surface coverage, viability and proliferation of fibroblasts influenced by PVA and PVA-gelatin nanofibers with randomly orientated or aligned configuration are systematically compared. Fibroblasts growing on PVA-gelatin fibers show significantly larger projected areas as compared with those cultivated on PVA fibers which p-value is smaller than 0.005. Cells on PVA-gelatin aligned fibers stretch out extensively and their intracellular stress fiber pull nucleus to deform. Results suggest that instead of the anisotropic topology within the scaffold trigger the preferential orientation of cells, the adhesion of cell membrane to gelatin have substantial influence on cellular behavior.

Development of Omniphobic Desalination Membranes Using a Charged Electrospun Nanofiber Scaffold.

PubMed

Lee, Jongho; Boo, Chanhee; Ryu, Won-Hee; Taylor, André D; Elimelech, Menachem

2016-05-04

In this study, we present a facile and scalable approach to fabricate omniphobic nanofiber membranes by constructing multilevel re-entrant structures with low surface energy. We first prepared positively charged nanofiber mats by electrospinning a blend polymer-surfactant solution of poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP) and cationic surfactant (benzyltriethylammonium). Negatively charged silica nanoparticles (SiNPs) were grafted on the positively charged electrospun nanofibers via dip-coating to achieve multilevel re-entrant structures. Grafted SiNPs were then coated with fluoroalkylsilane to lower the surface energy of the membrane. The fabricated membrane showed excellent omniphobicity, as demonstrated by its wetting resistance to various low surface tension liquids, including ethanol with a surface tension of 22.1 mN/m. As a promising application, the prepared omniphobic membrane was tested in direct contact membrane distillation to extract water from highly saline feed solutions containing low surface tension substances, mimicking emerging industrial wastewaters (e.g., from shale gas production). While a control hydrophobic PVDF-HFP nanofiber membrane failed in the desalination/separation process due to low wetting resistance, our fabricated omniphobic membrane exhibited a stable desalination performance for 8 h of operation, successfully demonstrating clean water production from the low surface tension feedwater.

Improved human endometrial stem cells differentiation into functional hepatocyte-like cells on a glycosaminoglycan/collagen-grafted polyethersulfone nanofibrous scaffold.

PubMed

Khademi, Farzaneh; Ai, Jafar; Soleimani, Masoud; Verdi, Javad; Mohammad Tavangar, Seyed; Sadroddiny, Esmaeil; Massumi, Mohammad; Mahmoud Hashemi, Seyed

2017-11-01

Liver tissue engineering (TE) is rapidly emerging as an effective technique which combines engineering and biological processes to compensate for the shortage of damaged or destroyed liver tissues. We examined the viability, differentiation, and integration of hepatocyte-like cells on an electrospun polyethersulfone (PES) scaffold, derived from human endometrial stem cells (hEnSCs). Natural polymers were separately grafted on plasma-treated PES nanofibers, that is, collagen, heparan sulfate (HS) and collagen-HS. Galactosilated PES (PES-Gal) nanofibrous were created. The engineering and cell growth parameters were considered and compared with each sample. The cellular studies revealed increased cell survival, attachment, and normal morphology on the bioactive natural polymer-grafted scaffolds after 30 days of hepatic differentiation. The chemical and molecular assays displayed hepatocyte differentiation. These cells were also functional, showing glycogen storage, α-fetoprotein, and albumin secretion. The HS nanoparticle-grafted PES nanofibers demonstrated a high rate of cell proliferation, differentiation, and integration. Based on the observations mentioned above, engineered tissue is a good option in the future, for the commercial production of three-dimensional liver tissues for clinical purposes. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2516-2529, 2017. © 2016 Wiley Periodicals, Inc.

Silk fibroin/poly (vinyl alcohol) blend scaffolds for controlled delivery of curcumin

PubMed Central

Li, Xiaomeng; Qin, Jinli; Ma, Jun

2015-01-01

A silk fibroin/poly (vinyl alcohol) porous scaffold with a water vapor transmission rate of 2125 ± 464 g/m2/day has been developed via thermally induced phase separation (gelation) and freeze-drying process. A hierarchical architecture of micropores and nanofibers was observed inside the scaffolds, and the related structures were analyzed. The viability and proliferation of 3T3 fibroblasts were examined, which indicated that the scaffolds exerted low cytotoxicity. After loading curcumin, the scaffolds can suppress the growth of 3T3 fibroblasts. The release behavior of curcumin from the scaffolds was investigated. At pH = 7.2, the release profiles showed no significant difference for the loading amounts of 0.5 mg and 0.25 mg per sample. Meanwhile, the cumulative amount of released drug at pH = 5.7 was significantly more than that in neutral solution due to more degradation of the scaffolds. It was suggested that the silk fibroin/poly (vinyl alcohol) blend scaffolds could be potentially used as wound dressing materials. PMID:26816634

Doxorubicin-loaded PLA/pearl electrospun nanofibrous scaffold for drug delivery and tumor cell treatment

NASA Astrophysics Data System (ADS)

Dai, Jiamu; Jin, Junhong; Yang, Shenglin; Li, Guang

2017-07-01

A drug-loaded implantable scaffold is a promising substitute for the treatment of tissue defects after a tumor resection operation. In this work, natural pearl powder with good biocompatibility and osteoconductivity was incorporated into polylactic (PLA) nanofibers via electrospinning, and doxorubicin hydrochloride (DOX) was also loaded in the PLA/pearl scaffold, resulting in a drug-loaded composite nanofibrous scaffold (DOX@PLA/pearl). In vitro drug delivery of DOX from a PLA/pearl composite scaffold was measured and in vitro anti-tumor efficacy was also examined, in particular the effect of the pearl content on both key properties were studied. The results showed that DOX was successfully loaded into PLA/pearl composite nanofibrous scaffolds with different pearl content. More importantly, the delivery rate of DOX kept rising as the pearl content increased, and the anti-tumor efficacy of the drug-loaded scaffold on HeLa cells was improved at an appropriate pearl powder concentration. Thus, we expect that the prepared DOX@PLA/pearl powder nanofibrous mat is a highly promising implantable scaffold that has great potential in postoperative cancer treatment.

Effect of tissue scaffold topography on protein structure monitored by fluorescence spectroscopy.

PubMed

Portugal, Carla A M; Truckenmüller, Roman; Stamatialis, Dimitrios; Crespo, João G

2014-11-10

The impact of surface topography on the structure of proteins upon adhesion was assessed through non-invasive fluorescence monitoring. This study aimed at obtaining a better understanding about the role of protein structural status on cell-scaffold interactions. The changes induced upon adsorption of two model proteins with different geometries, trypsin (globular conformation) and fibrinogen (rod-shaped conformation) on poly-l-lactic acid (PLLA) scaffolds with different surface topographies, flat, fibrous and surfaces with aligned nanogrooves, were assessed by fluorescence spectroscopy monitoring, using tryptophan as structural probe. Hence, the maximum emission blue shift and the increase of fluorescence anisotropy observed after adsorption of globular and rod-like shaped proteins on surfaces with parallel nanogrooves were ascribed to more intense protein-surface interactions. Furthermore, the decrease of fluorescence anisotropy observed upon adsorption of proteins to scaffolds with fibrous morphology was more significant for rod-shaped proteins. This effect was associated to the ability of these proteins to adjust to curved surfaces. The additional unfolding of proteins induced upon adsorption on scaffolds with a fibrous morphology may be the reason for better cell attachment there, promoting an easier access of cell receptors to initially hidden protein regions (e.g. RGDS sequence), which are known to have a determinant role in cell attaching processes. Copyright © 2014 Elsevier B.V. All rights reserved.

Bilayered nanofibrous 3D hierarchy as skin rudiment by emulsion electrospinning for burn wound management.

PubMed

Pal, Pallabi; Dadhich, Prabhash; Srivas, Pavan Kumar; Das, Bodhisatwa; Maulik, Dhrubajyoti; Dhara, Santanu

2017-08-22

Mimicking skin extracellular matrix hierarchy, the present work aims to develop a bilayer skin graft comprising a porous cotton-wool-like 3D layer with membranous structure of PCL-chitosan nanofibers. Emulsion electrospinning with differential stirring periods of PCL-chitosan emulsion results in development of a bilayer 3D structure with varied morphology. The electrospun membrane has fiber diameter ∼274 nm and pore size ∼1.16 μm while fluffy 3D layer has fiber diameter ∼1.62 μm and pore size ∼62 μm. The 3D layer was further coated with collagen I isolated from Cirrhinus cirrhosus fish scales to improve biofunctionality. Surface coating with collagen I resulted in bundling the fibers together, thereby increasing their average diameter to 2.80 μm and decreasing pore size to ∼45 μm. The architecture and composition of the scaffold promotes efficient cellular activity where interconnected porosity with ECM resembling collagen I coating assists cellular adhesion, infiltration, and proliferation from initial days of fibroblast seeding, while keratinocytes migrate on the surface only without infiltrating in the membranous nanofiber layer. Anatomy of the scaffold arising due to variation in pore size distribution at different layers thereby facilitates compartmentalization and prevents initial cellular transmigration. The scaffold also assists in extracellular matrix protein synthesis and keratinocyte stratification in vitro. Further, the scaffold effectively integrates and attaches with third-degree burn wound margins created in rat models and accelerates healing in comparison to standard Tegaderm dressing™. The bilayer scaffold is thus a promising, readily available, cost-effective, off-the-shelf matrix as a skin substitute.

Outlines on nanotechnologies applied to bladder tissue engineering.

PubMed

Alberti, C

2012-01-01

Tissue engineering technologies are more and more expanding as consequence of recent developments in the field of biomaterial science and nanotechnology research. An important issue in designing scaffold materials is that of recreating the ECM (extra-cellular matrix) functional features - particularly ECM-derived complex molecule signalling - to mimic its capability of directing cell-growth and neotissue morphogenesis. In this way the nanotechnology may offer intriguing chances, biomaterial nanoscale-based scaffold geometry behaving as nanomechanotransducer complex interacting with different cell nanosize proteins, especially with those of cell surface mechanoreceptors. To fabricate 3D-scaffold complex architectures, endowed with controlled geometry and functional properties, bottom-up approaches, based on molecular self-assembling of small building polymer units, are used, sometimes functionalizing them by incorporation of bioactive peptide sequences such as RDG (arginine - glycine - aspartic acid, a cell-integrin binding domain of fibronectin), whereas the top-down approaches are useful to fabricate micro/nanoscale structures, such as a microvasculature within an existing complex bioarchitecture. Synthetic polymer-based nanofibers, produced by electrospinning process, may be used to create fibrous scaffolds that can facilitate, given their nanostructured geometry and surface roughness, cell adhesion and growth. Also bladder tissue engineering may benefit by nanotechnology advances to achieve a better reliability of the bladder engineered tissue. Particularly, bladder smooth muscle cell adhesion to nanostructured polymeric surfaces is significantly enhanced in comparison with that to conventional biomaterials. Moreover nanostructured surfaces of bladder engineered tissue show a decreased calcium stone production. In a bladder tumor animal model, the dispersion of carbon nanofibers in a polymeric scaffold-based tissue engineered replacement neobladder, appears to inhibit a carcinogenic relapse in bladder prosthetic material. Facing the future, a full success of bladder tissue engineering will mainly depend on the progress of both biomaterial nanotechnologies and stem cell biology research.

Sustained delivery of VEGF from designer self-assembling peptides improves cardiac function after myocardial infarction

DOE Office of Scientific and Technical Information (OSTI.GOV)

Guo, Hai-dong; Cui, Guo-hong; Yang, Jia-jun

Highlights: Black-Right-Pointing-Pointer The designer peptide LRKKLGKA could self-assemble into nanofibers. Black-Right-Pointing-Pointer Injection of LRKKLGKA peptides could promote the sustained delivery of VEGF. Black-Right-Pointing-Pointer Injection of VEGF with LRKKLGKA peptides lead to sufficient angiogenesis. Black-Right-Pointing-Pointer Injection of VEGF with LRKKLGKA peptides improves heart function. -- Abstract: Poor vascularization and insufficient oxygen supply are detrimental to the survival of residual cardiomyocytes or transplanted stem cells after myocardial infarction. To prolong and slow the release of angiogenic factors, which stimulate both angiogenesis and vasculogenesis, we constructed a novel self-assembling peptide by attaching the heparin-binding domain sequence LRKKLGKA to the self-assembling peptide RADA16. Thismore » designer self-assembling peptide self-assembled into nanofiber scaffolds under physiological conditions, as observed by atomic force microscopy. The injection of designer self-assembling peptides can efficiently provide the sustained delivery of VEGF for at least 1 month. At 4 weeks after transplantation, cardiac function was improved, and scar size and collagen deposition were markedly reduced in the group receiving VEGF with the LRKKLGKA scaffolds compared with groups receiving VEGF alone, LRKKLGKA scaffolds alone or VEGF with RADA16 scaffolds. The microvessel density in the VEGF with LRKKLGKA group was higher than that in the VEGF with RADA16 group. TUNEL and cleaved caspase-3 expression assays showed that the transplantation of VEGF with LRKKLGKA enhanced cell survival in the infarcted heart. These results present the tailor-made peptide scaffolds as a new generation of sustained-release biomimetic biomaterials and suggest that the use of angiogenic factors along with designer self-assembling peptides can lead to myocardial protection, sufficient angiogenesis, and improvement in cardiac function.« less

Nitrate removal properties of solid-phase denitrification processes using acid-blended poly(L-lactic acid) as the sole substrate

NASA Astrophysics Data System (ADS)

Yamada, T.; Matsuoka, H.; Sun, J.; Yoshikawa, S.; Tsuji, H.; Hiraishi, A.

2013-04-01

The large amount of waste that is discharged along with the diffusion of poly(L-lactic acid) (PLLA) articles in use is persistent concern. Previously, we studied solid-phase denitrification (SPD) processes using PLLA to establish an effective re-use of PLLA waste. We found that PLLA with a weight-average molecular weight (Mw) of approximately 10,000 was suitable for SPD processes; however, the recycling of PLLA waste consumes a high energy. A new PLLA plastic including 5% poly(ethylene oxalate) (PEOxPLLA) as a blend material has attracted attention because recycling of PEOxPLLA consumes less electricity than that of PLLA. In this study, our main objectives were to evaluate whether PEOxPLLA can be used for SPD processes by changing its Mw and to investigate the bioavailability for denitrification of hydrolysates released from PEOxPLLA. The predicted hydrolysates, including oxalic acid, ethylene glycol, and lactate, are abiotically released, leading to different biological nitrate removal rates. Consequently, the nitrate removal rate of PEOxPLLA ranged from 0.9-4.1 mg-NO3--N·g-MLSS·h-1 by changing the Mw in the range of 8,500-238,000. In culture-dependent approaches, denitrifying bacteria using each substrate as an electron donor are found in activated sludge, suggesting that all hydrolysates functioned in the SPD processes using PEOxPLLA.

The Favorable Effect of Mesenchymal Stem Cell Treatment on the Antioxidant Protective Mechanism in the Corneal Epithelium and Renewal of Corneal Optical Properties Changed after Alkali Burns.

PubMed

Cejka, Cestmir; Holan, Vladimir; Trosan, Peter; Zajicova, Alena; Javorkova, Eliska; Cejkova, Jitka

2016-01-01

The aim of this study was to examine whether mesenchymal stem cells (MSCs) and/or corneal limbal epithelial stem cells (LSCs) influence restoration of an antioxidant protective mechanism in the corneal epithelium and renewal of corneal optical properties changed after alkali burns. The injured rabbit corneas (with 0.25 N NaOH) were untreated or treated with nanofiber scaffolds free of stem cells, with nanofiber scaffolds seeded with bone marrow MSCs (BM-MSCs), with adipose tissue MSCs (Ad-MSCs), or with LSCs. On day 15 following the injury, after BM-MSCs or LSCs nanofiber treatment (less after Ad-MSCs treatment) the expression of antioxidant enzymes was restored in the regenerated corneal epithelium and the expressions of matrix metalloproteinase 9 (MMP9), inducible nitric oxide synthase (iNOS), α-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), and vascular endothelial factor (VEGF) were low. The central corneal thickness (taken as an index of corneal hydration) increased after the injury and returned to levels before the injury. In injured untreated corneas the epithelium was absent and numerous cells revealed the expressions of iNOS, MMP9, α-SMA, TGF-β1, and VEGF. In conclusion, stem cell treatment accelerated regeneration of the corneal epithelium, restored the antioxidant protective mechanism, and renewed corneal optical properties.

Collagen-PVA aligned nanofiber on collagen sponge as bi-layered scaffold for surface cartilage repair.

PubMed

Lin, Hsin-Yi; Tsai, Wen-Chi; Chang, Shih-Hsing

2017-05-01

Researchers have made bi-layered scaffolds but mostly for osteochondral repairs. The anatomic structure of human cartilage has different zones and that each has varying matrix morphology and mechanical properties is often overlooked. Two bi-layered collagen-based composites were made to replicate the superficial and transitional zones of an articular cartilage. Aligned and random collagen-PVA nanofibers were electrospun onto a freeze-dried collagen sponge to make the aligned and random composites, respectively. The morphology, swelling ratio, degradation and tensile properties of the two composites were examined. Primary porcine chondrocytes were cultured on the composites for three weeks and their proliferation and secretion of glycosaminoglycan (GAG) and type II collagen were measured. The influences of the cell culture on the tensile properties of the composites were studied. The nanofiber layer remained adhered to the sponge after three weeks of cell culture. Both composites lost 30-35% of their total weight in a saline buffer after three weeks. The tensile strength and Young's modulus of both composites increased after three weeks of chondrocyte culture (p < 0.05). The aligned composite with extracellular matrix deposition had a Young's modulus (0.35 MPa) similar to that of articular cartilage reported in literature (0.36-0.8 MPa). The chondrocytes on both aligned and random composites proliferated and secreted similar amounts of GAG and type II collagen. They were seen embedded in lacunae after three weeks. The aligned composite may be more suitable for articular cartilage repair because of the higher tensile strength from the aligned nanofibers on the surface that can better resist wear.

Vancomycin containing PLLA/β-TCP controls experimental osteomyelitis in vivo.

PubMed

Kankilic, Berna; Bilgic, Elif; Korkusuz, Petek; Korkusuz, Feza

2014-11-19

Implant-related osteomyelitis (IRO) is recently controlled with local antibiotic delivery systems to overcome conventional therapy disadvantages. In vivo evaluation of such systems is however too little. We asked whether vancomycin (V)-containing poly-l-lactic acid/β-tricalcium phosphate (PLLA/β-TCP) composites control experimental IRO and promote bone healing in vivo. Fifty-six rats were distributed to five groups in this longitudinal controlled study. Experimental IRO was established at tibiae by injecting methicillin-resistant Staphylococcus aureus (MRSA) suspensions with titanium particles in 32 rats. Vancomycin-free PLLA/β-TCP composites were implanted into the normal and infected tibiae, whereas V-PLLA/β-TCP composites and coated (C)-V-PLLA/β-TCP composites were implanted into IRO sites. Sham-operated tibiae established the control group. Radiological and histological scores were quantified with microbiological findings on weeks 1 and 6. IRO is resolved in the CV- and the V-PLLA/β-TCP groups but not in the PLLA/β-TCP group. MRSA was not isolated in the CV- and the V-PLLA/β-TCP groups at all times whereas the bacteria were present in the PLLA/β-TCP group. Radiological signs secondary to infection are improved from 10.9 ± 0.9 to 3.0 ± 0.3 in the V-PLLA/β-TCP group but remained constant in the PLLA/β-TCP group. Histology scores are improved from 24.7 ± 6.5 to 17.6 ± 4.8 and from 27.6 ± 7.9 to 32.4 ± 8.9 in the CV-PLLA/β-TCP and the V-PLLA/β-TCP groups, respectively. New bone was formed in all the PLLA/β-TCP group at weeks 1 and 6. CV- and V-PLLA/β-TCP composites controlled experimental IRO and promoted bone healing. CV- and V-PLLA/β-TCP composites have the potential of controlling experimental IRO and promoting bone healing.

Tough and flexible CNT-polymeric hybrid scaffolds for engineering cardiac constructs.

PubMed

Kharaziha, Mahshid; Shin, Su Ryon; Nikkhah, Mehdi; Topkaya, Seda Nur; Masoumi, Nafiseh; Annabi, Nasim; Dokmeci, Mehmet R; Khademhosseini, Ali

2014-08-01

In the past few years, a considerable amount of effort has been devoted toward the development of biomimetic scaffolds for cardiac tissue engineering. However, most of the previous scaffolds have been electrically insulating or lacked the structural and mechanical robustness to engineer cardiac tissue constructs with suitable electrophysiological functions. Here, we developed tough and flexible hybrid scaffolds with enhanced electrical properties composed of carbon nanotubes (CNTs) embedded aligned poly(glycerol sebacate):gelatin (PG) electrospun nanofibers. Incorporation of varying concentrations of CNTs from 0 to 1.5% within the PG nanofibrous scaffolds (CNT-PG scaffolds) notably enhanced fiber alignment and improved the electrical conductivity and toughness of the scaffolds while maintaining the viability, retention, alignment, and contractile activities of cardiomyocytes (CMs) seeded on the scaffolds. The resulting CNT-PG scaffolds resulted in stronger spontaneous and synchronous beating behavior (3.5-fold lower excitation threshold and 2.8-fold higher maximum capture rate) compared to those cultured on PG scaffold. Overall, our findings demonstrated that aligned CNT-PG scaffold exhibited superior mechanical properties with enhanced CM beating properties. It is envisioned that the proposed hybrid scaffolds can be useful for generating cardiac tissue constructs with improved organization and maturation. Copyright © 2014 Elsevier Ltd. All rights reserved.

Differentiation of Wharton's Jelly-Derived Mesenchymal Stem Cells into Motor Neuron-Like Cells on Three-Dimensional Collagen-Grafted Nanofibers.

PubMed

Bagher, Zohreh; Azami, Mahmoud; Ebrahimi-Barough, Somayeh; Mirzadeh, Hamid; Solouk, Atefeh; Soleimani, Mansooreh; Ai, Jafar; Nourani, Mohammad Reza; Joghataei, Mohammad Taghi

2016-05-01

Cell transplantation strategies have provided potential therapeutic approaches for treatment of neurodegenerative diseases. Mesenchymal stem cells from Wharton's jelly (WJMSCs) are abundant and available adult stem cells with low immunological incompatibility, which could be considered for cell replacement therapy in the future. However, MSC transplantation without any induction or support material causes poor control of cell viability and differentiation. In this study, we investigated the effect of the nanoscaffolds on WJMSCs differentiation into motor neuronal lineages in the presence of retinoic acid (RA) and sonic hedgehog (Shh). Surface properties of scaffolds have been shown to significantly influence cell behaviors such as adhesion, proliferation, and differentiation. Therefore, polycaprolactone (PCL) nanofibers were constructed via electrospinning, surface modified by plasma treatment, and grafted by collagen. Characterization of the scaffolds by means of ATR-FTIR, contact angel, and Bradford proved grafting of the collagen on the surface of the scaffolds. WJMSCs were seeded on nanofibrous and tissue culture plate (TCP) and viability of WJMSCs were measured by MTT assay and then induced to differentiate into motor neuron-like cells for 15 days. Differentiated cells were evaluated morphologically, and real-time PCR and immunocytochemistry methods were done to evaluate expression of motor neuron-like cell markers in mRNA and protein levels. Our results showed that obtained cells could express motor neuron biomarkers at both RNA and protein levels, but the survival and differentiation of WJMSCs into motor neuron-like cells on the PCL/collagen scaffold were higher than cultured cells in the TCP and PCL groups. Taken together, WJMSCs are an attractive stem cell source for inducing into motor neurons in vitro especially when grown on nanostructural scaffolds and PCL/collagen scaffolds can provide a suitable, three-dimensional situation for neuronal survival and differentiation that suggest their potential application towards nerve regeneration.

Phase Structure and Properties of a Biodegradable Block Copolymer Coalesced from It's Crystalline Inclusion Compound Formed with alpha-Cyclodextrin

NASA Astrophysics Data System (ADS)

Shuai, Xintao; Wei, Min; Probeni, Francis; Bullions, Todd A.; Shin, I. Daniel; Tonelli, Alan E.

2002-03-01

A well-defined biodegradable block copolymer of poly(epsilon caprolactone) (PCL) and poly(L-lactic acid) (PLLA) was synthesized and characterized and then included as a guest in an inclusion compound (IC) formed with the host alpha-cyclodextrin (CD). The PCL-b-PLLA block copolymer was subsequently coalesced from it's CD-IC crystals by either treatment with hot water (50 C) or an aqueous amylase solution at 25 C. The coalesced PCL-b-PLLA was examined by FTIR, DSC, TGA, and WAXD and was found to be much more homogeneosly organized, with much less segregation and crystallinity of the PCL and PLLA microphases. The morpholgy, crystallization kinetics, thermal behavior, and biodegradability of the coalesced PCL-b-PLLA block copolymer was studied by comparison to similar observations made on as-synthesized PCL-b-PLLA, PCL and PLLA homopolymers, and their solution-cast blend. The PCL and PLLA blocks are found to be more intimately mixed, with less phase segregation, in the coalesced diblock copolymer, and this leads to homogeneous bulk crystallization, which is not observed for the as-synthesized diblock copolymer. The coalesced PCL-b-PLLA was also found to be more quickly biodegraded (lipase from Rhizopus arrhizus)than the as-synthesized PCL-b-PLLA or the physical blend of PCL and PLLA homopolymers. Overall, the coalescence of the inherently phase segregated diblock copolymer PCL-b-PLLA results in a small amount of compact, chain-extended PCL and PLLA crystals embedded in an amorphous phase, largely consisting of well-mixed PCL and PLLA blocks. Thus, we have demonstrated that it is possible to control the morpholgy of a biodegradable diblock copolymer, thereby significantly modifying it's properties, by coalescence from it's CD-IC crystals.

A three-dimensional hierarchical collagen scaffold fabricated by a combined solid freeform fabrication (SFF) and electrospinning process to enhance mesenchymal stem cell (MSC) proliferation

NASA Astrophysics Data System (ADS)

Ahn, SeungHyun; Koh, Young Ho; Kim, GeunHyung

2010-06-01

Collagen has the advantage of being very similar to macromolecular substances that can be recognized and metabolized in the biological environment. Although the natural material has superior property for this purpose, its use to fabricate reproducible and pore-structure-controlled 3D structures, which are designed to allow the entry of sufficient cells and the easy diffusion of nutrients, has been limited due to its low processability. Here, we propose a hybrid technology that combines a cryogenic plotting system with an electrospinning process. Using this technique, an easily pore-size-controllable hierarchical 3D scaffold consisting of micro-sized highly porous collagen strands and micro/nano-sized collagen fibers was fabricated. The pore structure of the collagen scaffold was controlled by the collagen micro/nanofibers, which were layered in the scaffold. The hierarchical scaffolds were characterized with respect to initial cell attachment and proliferation of bone marrow-derived mesenchymal stem cells within the scaffolds. The hierarchical scaffold exhibited incredibly enhanced initial cell attachment and cell compactness between pores of the plotted scaffold relative to the normally designed 3D collagen scaffold.

Formation of methotrexate-PLLA-PEG-PLLA composite microspheres by microencapsulation through a process of suspension-enhanced dispersion by supercritical CO2

PubMed Central

Chen, Ai-Zheng; Wang, Guang-Ya; Wang, Shi-Bin; Li, Li; Liu, Yuan-Gang; Zhao, Chen

2012-01-01

Background The aim of this study was to improve the drug loading, encapsulation efficiency, and sustained-release properties of supercritical CO2-based drug-loaded polymer carriers via a process of suspension-enhanced dispersion by supercritical CO2 (SpEDS), which is an advanced version of solution-enhanced dispersion by supercritical CO2 (SEDS). Methods Methotrexate nanoparticles were successfully microencapsulated into poly (L-lactide)-poly(ethylene glycol)-poly(L-lactide) (PLLA-PEG-PLLA) by SpEDS. Methotrexate nanoparticles were first prepared by SEDS, then suspended in PLLA-PEG-PLLA solution, and finally microencapsulated into PLLA-PEG-PLLA via SpEDS, where an “injector” was utilized in the suspension delivery system. Results After microencapsulation, the composite methotrexate (MTX)-PLLA-PEG-PLLA microspheres obtained had a mean particle size of 545 nm, drug loading of 13.7%, and an encapsulation efficiency of 39.2%. After an initial burst release, with around 65% of the total methotrexate being released in the first 3 hours, the MTX-PLLA-PEG-PLLA microspheres released methotrexate in a sustained manner, with 85% of the total methotrexate dose released within 23 hours and nearly 100% within 144 hours. Conclusion Compared with a parallel study of the coprecipitation process, microencapsulation using SpEDS offered greater potential to manufacture drug-loaded polymer microspheres for a drug delivery system. PMID:22787397

Growth of multiwalled-carbon nanotubes using vertically aligned carbon nanofibers as templates/scaffolds and improved field-emission properties

NASA Astrophysics Data System (ADS)

Cui, H.; Yang, X.; Baylor, L. R.; Lowndes, D. H.

2005-01-01

Multiwalled-carbon nanotubes (MWCNTs) are grown on top of vertically aligned carbon nanofibers (VACNFs) via microwave plasma-enhanced chemical vapor deposition (MPECVD). The VACNFs are first grown in a direct-current plasma-enhanced chemical vapor deposition reactor using nickel catalyst. A layer of carbon-silicon materials is then deposited on the VACNFs and the nickel catalyst particle is broken down into smaller nanoparticles during an intermediate reactive-ion-plasma deposition step. These nickel nanoparticles nucleate and grow MWCNTs in the following MPECVD process. Movable-probe measurements show that the MWCNTs have greatly improved field-emission properties relative to the VACNFs.

Covalent-supramolecular hybrid polymers as muscle-inspired anisotropic actuators.

PubMed

Chin, Stacey M; Synatschke, Christopher V; Liu, Shuangping; Nap, Rikkert J; Sather, Nicholas A; Wang, Qifeng; Álvarez, Zaida; Edelbrock, Alexandra N; Fyrner, Timmy; Palmer, Liam C; Szleifer, Igal; Olvera de la Cruz, Monica; Stupp, Samuel I

2018-06-19

Skeletal muscle provides inspiration on how to achieve reversible, macroscopic, anisotropic motion in soft materials. Here we report on the bottom-up design of macroscopic tubes that exhibit anisotropic actuation driven by a thermal stimulus. The tube is built from a hydrogel in which extremely long supramolecular nanofibers are aligned using weak shear forces, followed by radial growth of thermoresponsive polymers from their surfaces. The hierarchically ordered tube exhibits reversible anisotropic actuation with changes in temperature, with much greater contraction perpendicular to the direction of nanofiber alignment. We identify two critical factors for the anisotropic actuation, macroscopic alignment of the supramolecular scaffold and its covalent bonding to polymer chains. Using finite element analysis and molecular calculations, we conclude polymer chain confinement and mechanical reinforcement by rigid supramolecular nanofibers are responsible for the anisotropic actuation. The work reported suggests strategies to create soft active matter with molecularly encoded capacity to perform complex tasks.

A culture system to study oligodendrocyte myelination-processes using engineered nanofibers

PubMed Central

Lee, Seonok; Leach, Michelle K.; Redmond, Stephanie A.; Chong, S.Y. Christin; Mellon, Synthia H.; Tuck, Samuel J.; Feng, Zhang-Qi; Corey, Joseph M.; Chan, Jonah R.

2012-01-01

Current methods for studying central nervous system myelination necessitate permissive axonal substrates conducive for myelin wrapping by oligodendrocytes. We have developed a neuron-free culture system in which electron-spun nanofibers of varying sizes substitute for axons as a substrate for oligodendrocyte myelination, thereby allowing manipulation of the biophysical elements of axonal-oligodendroglial interactions. To investigate axonal regulation of myelination, this system effectively uncouples the role of molecular (inductive) cues from that of biophysical properties of the axon. We use this method to uncover the causation and sufficiency of fiber diameter in the initiation of concentric wrapping by rat oligodendrocytes. We also show that oligodendrocyte precursor cells display sensitivity to the biophysical properties of fiber diameter and initiate membrane ensheathment prior to differentiation. The use of nanofiber scaffolds will enable screening for potential therapeutic agents that promote oligodendrocyte differentiation and myelination as well as provide valuable insight into the processes involved in remyelination. PMID:22796663

Polymer based nanocomposites with nanofibers and exfoliated clay

NASA Technical Reports Server (NTRS)

Meador, Michael A.; Reneker, Darrell H.

2005-01-01

Polymer solutions, containing clay sheets, were electrospun into nanofibers and microfibers that contained clay sheets inside. Controllable removal of polymer by plasma etching from the surface of fibers revealed the arrangement of clay. The shape, flexibility, size distribution and arrangement of clay sheets were observed by transmission and scanning electron microscopy. The clay sheets were partially aligned in big fibers with normal direction of clay sheets perpendicular to fiber axis. Crumpling of clay sheets inside fibers was observed when the fiber diameter was comparable to the lateral size of clay sheets. Single sheets of clay were observed both by catching clay sheets dispersed in water with electrospun nanofiber mats and by the deliberate removal of most of the polymer in the fibers. Thin, flexible gas barrier films, that are reasonably strong, were assembled from clay sheets and polymer nanofibers. Structure of composite films was characterized with scanning electron microscopy. Continuous film of clay sheets were physically attached to the surface of fiber mats. Spincoating film of polymer and clay sheets was reinforced by electrospun fiber scaffold. Certain alignment of clay sheets was observed in the vicinity of fibers.

Dual-source dual-power electrospinning and characteristics of multifunctional scaffolds for bone tissue engineering.

PubMed

Wang, Chong; Wang, Min

2012-10-01

Electrospun tissue engineering scaffolds are attractive due to their distinctive advantages over other types of scaffolds. As both osteoinductivity and osteoconductivity play crucial roles in bone tissue engineering, scaffolds possessing both properties are desirable. In this investigation, novel bicomponent scaffolds were constructed via dual-source dual-power electrospinning (DSDPES). One scaffold component was emulsion electrospun poly(D,L-lactic acid) (PDLLA) nanofibers containing recombinant human bone morphogenetic protein (rhBMP-2), and the other scaffold component was electrospun calcium phosphate (Ca-P) particle/poly(lactic-co-glycolic acid) (PLGA) nanocomposite fibers. The mass ratio of rhBMP-2/PDLLA fibers to Ca-P/PLGA fibers in bicomponent scaffolds could be controlled in the DSDPES process by adjusting the number of syringes used to supply solutions for electrospinning. Through process optimization, both types of fibers could be evenly distributed in bicomponent scaffolds. The structure and properties of each type of fibers in the scaffolds were studied. The morphological and structural properties and wettability of scaffolds were assessed. The effects of emulsion composition for rhBMP-2/PDLLA fibers and mass ratio of fibrous components in bicomponent scaffolds on in vitro release of rhBMP-2 from scaffolds were investigated. In vitro degradation of scaffolds was also studied by monitoring their morphological changes, weight losses and decreases in average molecular weight of fiber matrix polymers.

Spatially selective modification of PLLA surface: From hydrophobic to hydrophilic or to repellent

NASA Astrophysics Data System (ADS)

Bastekova, Kristina; Guselnikova, Olga; Postnikov, Pavel; Elashnikov, Roman; Kunes, Martin; Kolska, Zdenka; Švorčík, Vaclav; Lyutakov, Oleksiy

2017-03-01

A universal approach to controlled surface modification of polylactic acid (PLLA) films using diazonium chemistry was proposed. The multistep procedure includes surface activation of PLLA by argon plasma treatment and chemical activation of arenediazonium tosylates by NaBH4. The surface of PLLA film was grafted with different functional organic groups (OFGs), changing the PLLA surface properties (wettability, morphology, zeta potential, chemical composition, and mechanical response). Three approaches of OFG grafting were examined: (i) plasma treatment following by PLLA immersion into diazonium salt aqueous solution; (ii) grafting of PLLA surface through the reaction with chemically created aryl radicals; (iii) mutual combination of both methods The best results were achieved in the last case, where the previous plasma treatment was combined with further reaction of PLLA surface with generated aryl radicals. Using this method PLLA surface was successfully grafted with amino, carboxyl, aliphatic and fluorinated OFGs. Further investigation of surface properties from potential biological and medical points of view was performed using zeta potential, biodegradation and biofouling tests. It was shown that proposed technique allows preparation of biorepellent or bioabsorptive surfaces, tuning of PLLA biodegradation rate and nanomechanical properties, as well as the introduction of inverse properties (such as hydrophilic and hydrophobic) on both sides of PLLA films.

Conjugation Magnetic PAEEP-PLLA Nanoparticles with Lactoferrin as a Specific Targeting MRI Contrast Agent for Detection of Brain Glioma in Rats

NASA Astrophysics Data System (ADS)

Luo, Binhua; Wang, Siqi; Rao, Rong; Liu, Xuhan; Xu, Haibo; Wu, Yun; Yang, Xiangliang; Liu, Wei

2016-04-01

The diagnosis of malignant brain gliomas is largely based on magnetic resonance imaging (MRI) with contrast agents. In recent years, nano-sized contrast agents have been developed for improved MRI diagnosis. In this study, oleylamine-coated Fe3O4 magnetic nanoparticles (OAM-MNPs) were synthesized with thermal decomposition method and encapsulated in novel amphiphilic poly(aminoethyl ethylene phosphate)/poly(L-lactide) (PAEEP-PLLA) copolymer nanoparticles. The OAM-MNP-loaded PAEEP-PLLA nanoparticles (M-PAEEP-PLLA-NPs) were further conjugated with lactoferrin (Lf) for glioma tumor targeting. The Lf-conjugated M-PAEEP-PLLA-NPs (Lf-M-PAEEP-PLLA-NPs) were characterized by photon correlation spectroscopy (PCS), transmission electron microscopy (TEM), Fourier transform infrared (FTIR), thermo-gravimetric analysis (TGA), X-ray diffraction (XRD), and vibrating sample magnetometer (VSM). The average size of OAM-MNPs, M-PAEEP-PLLA-NPs, and Lf-M-PAEEP-PLLA-NPs were 8.6 ± 0.3, 165.7 ± 0.6, and 218.2 ± 0.4 nm, with polydispersity index (PDI) of 0.185 ± 0.023, 0.192 ± 0.021, and 0.224 ± 0.036, respectively. TEM imaging showed that OAM-MNPs were monodisperse and encapsulated in Lf-M-PAEEP-PLLA-NPs. TGA analysis showed that the content of iron oxide nanoparticles was 92.8 % in OAM-MNPs and 45.2 % in Lf-M-PAEEP-PLLA-NPs. VSM results indicated that both OAM-MNPs and Lf-M-PAEEP-PLLA-NPs were superparamagnetic, and the saturated magnetic intensity were 77.1 and 74.8 emu/g Fe. Lf-M-PAEEP-PLLA-NPs exhibited good biocompatibility in cytotoxicity assay. The high cellular uptake of Lf-M-PAEEP-PLLA-NPs in C6 cells indicated that Lf provided effective targeting for the brain tumor cells. The T 2 relaxation rate ( r 2) of M-PAEEP-PLLA-NPs and Lf-M-PAEEP-PLLA-NPs were calculated to be 167.2 and 151.3 mM-1 s-1. In MRI on Wistar rat-bearing glioma tumor, significant contrast enhancement could clearly appear at 4 h after injection and last 48 h. Prussian blue staining of the section clearly showed the retention of Lf-M-PAEEP-PLLA-NPs in tumor tissues. The results from the in vitro and in vivo MRI indicated that Lf-M-PAEEP-PLLA-NPs possessed strong, long-lasting, tumor targeting, and enhanced tumor MRI contrast ability. Lf-M-PAEEP-PLLA-NPs represent a promising nano-sized MRI contrast agent for brain glioma targeting MRI.

Large animal in vivo evaluation of a binary blend polymer scaffold for skeletal tissue-engineering strategies; translational issues.

PubMed

Smith, James O; Tayton, Edward R; Khan, Ferdous; Aarvold, Alexander; Cook, Richard B; Goodship, Allen; Bradley, Mark; Oreffo, Richard O C

2017-04-01

Binary blend polymers offer the opportunity to combine different desirable properties into a single scaffold, to enhance function within the field of tissue engineering. Previous in vitro and murine in vivo analysis identified a polymer blend of poly(l-lactic acid)-poly(ε-caprolactone) (PLLA:PCL 20:80) to have characteristics desirable for bone regeneration. Polymer scaffolds in combination with marrow-derived skeletal stem cells (SSCs) were implanted into mid-shaft ovine 3.5 cm tibial defects, and indices of bone regeneration were compared to groups implanted with scaffolds alone and with empty defects after 12 weeks, including micro-CT, mechanical testing and histological analysis. The critical nature of the defect was confirmed via all modalities. Both the scaffold and scaffold/SSC groups showed enhanced quantitative bone regeneration; however, this was only found to be significant in the scaffold/SSCs group (p = 0.04) and complete defect bridging was not achieved in any group. The mechanical strength was significantly less than that of contralateral control tibiae (p < 0.01) and would not be appropriate for full functional loading in a clinical setting. This study explored the hypothesis that cell therapy would enhance bone formation in a critical-sized defect compared to scaffold alone, using an external fixation construct, to bridge the scale-up gap between small animal studies and potential clinical translation. The model has proved a successful critical defect and analytical techniques have been found to be both valid and reproducible. Further work is required with both scaffold production techniques and cellular protocols in order to successfully scale-up this stem cell/binary blend polymer scaffold. © 2015 The Authors. Journal of Tissue Engineering and Regenerative Medicine published by John Wiley & Sons, Ltd. © 2015 The Authors. Journal of Tissue Engineering and Regenerative Medicine published by John Wiley & Sons, Ltd.

Electrospinning of PLGA/gum tragacanth nanofibers containing tetracycline hydrochloride for periodontal regeneration.

PubMed

Ranjbar-Mohammadi, Marziyeh; Zamani, M; Prabhakaran, M P; Bahrami, S Hajir; Ramakrishna, S

2016-01-01

Controlled drug release is a process in which a predetermined amount of drug is released for longer period of time, ranging from days to months, in a controlled manner. In this study, novel drug delivery devices were fabricated via blend electrospinning and coaxial electrospinning using poly lactic glycolic acid (PLGA), gum tragacanth (GT) and tetracycline hydrochloride (TCH) as a hydrophilic model drug in different compositions and their performance as a drug carrier scaffold was evaluated. Scanning electron microscopy (SEM) results showed that fabricated PLGA, blend PLGA/GT and core shell PLGA/GT nanofibers had a smooth and bead-less morphology with the diameter ranging from 180 to 460 nm. Drug release studies showed that both the fraction of GT within blend nanofibers and the core-shell structure can effectively control TCH release rate from the nanofibrous membranes. By incorporation of TCH into core-shell nanofibers, drug release was sustained for 75 days with only 19% of burst release within the first 2h. The prolonged drug release, together with proven biocompatibility, antibacterial and mechanical properties of drug loaded core shell nanofibers make them a promising candidate to be used as drug delivery system for periodontal diseases. Copyright © 2015 Elsevier B.V. All rights reserved.

An in vitro evaluation of various biomaterials for the development of a tissue-engineered lacrimal gland

NASA Astrophysics Data System (ADS)

Selvam, Shivaram

The most common cause of ocular morbidity in developed countries is dry eye, many cases of which are due to lacrimal insufficiency. It has been established that lacrimal insufficiency results from processes caused by both immune-related and non-immune related events such as Sjogren's syndrome, Stevens-Johnson syndrome, chemical and thermal injuries and ocular cicatricial pemphigoid. Patients with these conditions would benefit from repair of their damaged lacrimal tissue by the creation of a replacement for the lacrimal gland. The new field of tissue engineering built on the interface between principles and methods of the life sciences with those of engineering to develop biocompatible materials has created the possibility for repairing or replacing damaged tissues. This thesis explores the use of tissue engineering principles for the development of a tissue-engineered lacrimal gland. This thesis also contributes to the development of a novel model for addressing lacrimal gland physiology and epithelial fluid transport. The first part of the research work focused on the evaluation of morphological and physiological properties of purified lacrimal gland acinar cells (pLGACs) cultured on various biopolymers: silicone, collagen I, poly-D,L-lactide-co-glycolide (PLGA; 85:15 and 50:50), and poly-L-lactic acid (PLLA) in the presence and absence of an extracellular matrix, MatrigelRTM. Results indicated that PLLA demonstrated the best support expression of acinar cell-like morphology. The second part demonstrated the ex vivo reconstitution of an electrophysiologically functional lacrimal gland tissue on porous polyester membrane scaffolds. Results showed that pLGACs were capable of establishing continuous epithelial monolayers that generate active ionic fluxes consistent with current models for Na +-dependent Cl-- secretion. The third part outlined the fabrication of porous PLLA membranes, the optimal biomaterial for culturing lacrimal epithelial cells. Microporous PLLA-Polyethylene glycol (PEG) blend membranes (mpPLLAbm) with interconnected pores were prepared by the water extraction of PEG from solution cast blend membranes using the solvent-cast/particulate leaching technique. Diffusion experiments on mpPLLAbm (57.1/42.9 wt%) were performed to demonstrate that the membrane was permeable to glucose, L-tryptophan, and dextran.

A chemistry/physics pathway with nanofibrous scaffolds for gene delivery.

PubMed

Wan, Fen; Tang, Zhaohui; He, Weidong; Chu, Benjamin

2010-10-21

This perspective is to introduce a new pathway for non-viral gene delivery by taking advantage of nanofibrous scaffolds as gene storage devices, gene carriers and homing devices. During gene delivery to the target, the DNA has to be protected in order to pass through a set of barriers before reaching the nucleus. The DNA can form a complex with polycations, and numerous publications exist on how to stabilize the DNA fragments by natural and synthetic materials. Electrospun nanofibrous scaffolds can be used to store the DNA, especially in the form of a more stabilized polyplex, and then to deliver the DNA (polyplex) to cells that are attached to the scaffold. While each essential step has been tested experimentally, the overall yet untested process, especially for in vivo experiments, may lead to a promising specific approach for gene/drug storage and delivery. The pathway described herein is based mainly on our understanding of the physics and chemistry of gene storage and delivery processes, in contrast to using pure biological concepts. Novel biodegradable, biocompatible nanofibrous materials with imbedded DNA (e.g., in the polyplex form) can then be designed to fabricate an intelligent scaffold for gene delivery. To achieve the above goal, the first step is to stabilize the DNA so that it can be incorporated into nanofibrous scaffolds. In this respect, we shall discuss the different methods of DNA/gene condensation and complex formation, and then explain the strategy used to incorporate DNA into electrospun nanofibers. Solvent-induced DNA condensation and then encapsulation were achieved. However, the released naked DNA was not sufficiently protected for gene transfection in cells. The objective of the current perspective is to suggest that, instead of the solvent-induced DNA condensation, one can combine the recently developed polyplex formation by using branched polyethyleneimine (bPEI). More importantly, free bPEI can be incorporated into the nanofibers separately so that during the gene delivery step, the presence of a predesigned amount of free bPEI can greatly increase the gene transfection efficiency, as has been reported recently by Chi Wu and his coworkers. Thus, a physics/chemistry-based pathway that utilizes nanofibrous scaffolds for gene delivery is within reach.

Fabrication and characterization of curcumin-loaded silk fibroin/P(LLA-CL) nanofibrous scaffold

NASA Astrophysics Data System (ADS)

Lian, Yuan; Zhan, Jian-Chao; Zhang, Kui-Hua; Mo, Xiu-Mei

2014-12-01

Curcumin exhibited excellent properties including antioxidant, antiinflammatory, antiviral, antibacterial, antifungal, anticancer, and anticoagulant activities. In this study, curcumin was incorporated into silk fibroin (SF)/poly(L-lactic acid- co-e-caprolactone) (P(LLA-CL)) nanofibrous scaffolds via electrospinning, and changes brought about by raising the curcumin content were observed: SEM images showed that the average nanofibrous diameter decreased at the beginning and then increased, and the nanofibers became uniform; FTIR showed that the conformation of SF transforming from random coil form to β-sheet structure had not been induced, while SF conformation converted to β-sheet after being treated with 75% ethanol vapor; XRD results confirmed that the crystal structure of (P(LLA-CL)) had been destroyed; The mechanical test illustrated that nanofibrous scaffolds still maintained good mechanical properties. Further, curcumin-loaded nanofibrous scaffolds were evaluated for drug release, antioxidant and antimicrobial activities in vitro. The results showed that curcumin presented a sustained release behavior from nanofibrous scaffolds and maintained its free radical scavenging ability, and such scaffolds could effectively inhibit S. aureus growth (> 95%). Thus, curcumin-loaded SF/P(LLA-CL) nanofibrous scaffolds might be potential candidates for wound dressing and tissue engineering scaffolds.

Mechanical and biodegradable properties of porous titanium filled with poly-L-lactic acid by modified in situ polymerization technique.

PubMed

Nakai, Masaaki; Niinomi, Mitsuo; Ishii, Daisuke

2011-10-01

Porous titanium (pTi) can possess a low Young's modulus equal to that of human bone, depending on its porosity. However, the mechanical strength of pTi deteriorates greatly with increasing porosity. On the other hand, certain medical polymers exhibit biofunctionalities, which are not possessed intrinsically by metallic materials. Therefore, a biodegradable medical polymer, poly-L-lactic acid (PLLA), was used to fill in the pTi pores using a modified in-situ polymerization technique. The mechanical and biodegradable properties of pTi filled with PLLA (pTi/PLLA) as fabricated by this technique and the effects of the PLLA filling were evaluated in this study. The pTi pores are almost completely filled with PLLA by the developed process (i.e., technique). The tensile strength and tensile Young's modulus of pTi barely changes with the PLLA filling. However, the PLLA filling improves the compressive 0.2% proof stress of pTi having any porosity and increases the compressive Young's modulus of pTi having relatively high porosity. This difference between the tensile and compressive properties of pTi/PLLA is considered to be caused by the differing resistances of PLLA in the pores to tensile and compressive deformations. The PLLA filled into the pTi pores degrades during immersion in Hanks' solution at 310 K. The weight loss due to PLLA degradation increases with increasing immersion time. However, the rate of weight loss of pTi/PLLA during immersion decreases with increasing immersion time. Hydroxyapatite formation is observed on the surface of pTi/PLLA after immersion for ≥8 weeks. The decrease in the weight-loss rate may be caused by weight gain due to hydroxyapatite formation and/or the decrease in contact area with Hanks' solution caused by its formation on the surface of pTi/PLLA. Copyright © 2011 Elsevier Ltd. All rights reserved.

Hydrogel Delivery of Mesenchymal Stem Cell–Expressing Bone Morphogenetic Protein-2 Enhances Bone Defect Repair

PubMed Central

Hsiao, Hui-Yi; Yang, Shu-Rui; Brey, Eric M.; Chu, I-Ming

2016-01-01

Background: The application of bone tissue engineering for repairing bone defects has gradually shown some satisfactory progress. One of the concerns raising scientific attention is the poor supply of growth factors. A number of growth factor delivery approaches have been developed for promoting bone formation. However, there is no systematic comparison of those approaches on efficiency of neobone formation. In this study, the approaches using periosteum, direct supply of growth factors, or gene transfection of growth factors were evaluated to determine the osteogenic capacity on the repair of bone defect. Methods: In total, 42 male 21-week-old Sprague-Dawley rats weighing 250 to 400 g were used as the bone defect model to evaluate the bone repair efficiency. Various tissue engineered constructs of poly(ethylene glycol)-poly(l-lactic acid) (PEG-PLLA) copolymer hydrogel with periosteum, with external supply of bone morphogenetic protein-2 (BMP2), or with BMP2-transfected bone marrow–derived mesenchymal stem cells (BMMSCs) were filled in a 7-mm bone defect region. Animals were euthanized at 3 months, and the hydrogel constructs were harvested. The evaluation with histological staining and radiography analysis were performed for the volume of new bone formation. Results: The PEG-PLLA scaffold with BMMSCs promotes bone regeneration with the addition of periosteum. The group with BMP2-transfected BMMSCs demonstrated the largest volume of new bone among all the testing groups. Conclusions: Altogether, the results of this study provide the evidence that the combination of PEG-PLLA hydrogels with BMMSCs and sustained delivery of BMP2 resulted in the maximal bone regeneration. PMID:27622106

Bioinspired Design of Polycaprolactone Composite Nanofibers as Artificial Bone Extracellular Matrix for Bone Regeneration Application.

PubMed

Gao, Xiang; Song, Jinlin; Zhang, Yancong; Xu, Xiao; Zhang, Siqi; Ji, Ping; Wei, Shicheng

2016-10-07

The design and development of functional biomimetic systems for programmed stem cell response is a field of topical interest. To mimic bone extracellular matrix, we present an innovative strategy for constructing drug-loaded composite nanofibrous scaffolds in this study, which could integrate multiple cues from calcium phosphate mineral, bioactive molecule, and highly ordered fiber topography for the control of stem cell fate. Briefly, inspired by mussel adhesion mechanism, a polydopamine (pDA)-templated nanohydroxyapatite (tHA) was synthesized and then surface-functionalized with bone morphogenetic protein-7-derived peptides via catechol chemistry. Afterward, the resulting peptide-loaded tHA (tHA/pep) particles were blended with polycaprolactone (PCL) solution to fabricate electrospun hybrid nanofibers with random and aligned orientation. Our research demonstrated that the bioactivity of grafted peptides was retained in composite nanofibers. Compared to controls, PCL-tHA/pep composite nanofibers showed improved cytocompatibility. Moreover, the incorporated tHA/pep particles in nanofibers could further facilitate osteogenic differentiation potential of human mesenchymal stem cells (hMSCs). More importantly, the aligned PCL-tHA/pep composite nanofibers showed more osteogenic activity than did randomly oriented counterparts, even under nonosteoinductive conditions, indicating excellent performance of biomimetic design in cell fate decision. After in vivo implantation, the PCL-tHA/pep composite nanofibers with highly ordered structure could significantly promote the regeneration of lamellar-like bones in a rat calvarial critical-sized defect. Accordingly, the presented strategy in our work could be applied for a wide range of potential applications in not only bone regeneration application but also pharmaceutical science.

Aligned poly(ε-caprolactone)/graphene oxide and reduced graphene oxide nanocomposite nanofibers: Morphological, mechanical and structural properties.

PubMed

Ramazani, Soghra; Karimi, Mohammad

2015-11-01

A number of studies have demonstrated that the mechanical properties of electrospun polymeric nanofibrous scaffolds are enhanced with the incorporation of graphene and its derivatives, thus developing their applications in hard tissue engineering. However, our understanding of the relationship between the microstructure and properties of these fibrous scaffolds and how they are influenced by graphene oxide (GO) and reduced graphene oxide (RGO) loading is much more limited. Thus, in this paper, poly(ε-caprolactone) (PCL)/GO and RGO nanocomposite nanofibers containing 0, 0.1, 0.5 and 1wt.% GO and RGO were prepared using an electrospinning technique. With the addition of 0.1wt.% of GO and RGO nanosheets in PCL, the tensile strength of PCL scaffolds increased over ~160 and 304% respectively and elastic modulus increased over 103 and 163% due to the good dispersion of the nanosheets and their interaction with the molecular chains of PCL. These were supported by the parallel increase in relaxation time and molecular orientation of PCL chains at the presence of nanosheets with a loading of 0.1wt.%. The enhancement effect of the nanosheets was weakened with an increase in GO and RGO loading up to 1wt.% in which it is connected to a partial exfoliation of the nanosheets. Copyright © 2015 Elsevier B.V. All rights reserved.

The Preparation, Characterization, Mechanical and Antibacterial Properties of GO-ZnO Nanocomposites with a Poly(l-lactide)-Modified Surface

PubMed Central

Yuan, Mingwei; Xiong, Chengdong; Jiang, Lin; Li, Hongli

2018-01-01

Graphene oxide (GO) was employed for the preparation of GO-zinc oxide (ZnO). The hydroxyl group on the surface was exploited to trigger the l-lactide ring-opening polymerization. A composite material with poly(l-lactide) (PLLA) chains grafted to the GO-ZnO surface, GO-ZnO-PLLA, was prepared. The results demonstrated that the employed method allowed one-step, rapid grafting of PLLA to the GO-ZnO surface. The chemical structure of the GO surface was altered by improved dispersion of GO-ZnO in organic solvents, thus enhancing the GO-ZnO dispersion in the PLLA matrix and the interface bonding with PLLA. Subsequently, composite films, GO-ZnO-PLLA and GO-ZnO-PLLA/PLLA, were prepared. The changes in interface properties and mechanical properties were studied. Furthermore, the antibacterial performance of nano-ZnO was investigated. PMID:29473891

Long-term drug release from electrospun fibers for in vivo inflammation prevention in the prevention of peritendinous adhesions.

PubMed

Hu, Changmin; Liu, Shen; Zhang, Yang; Li, Bin; Yang, Huilin; Fan, Cunyi; Cui, Wenguo

2013-07-01

Physical barriers such as electrospun fibrous membranes are potentially useful in preventing peritendinous adhesions after surgery. However, inflammatory responses caused by degradation of barrier materials remain a major challenge. This study aimed to fabricate electrospun composite fibrous membranes based on drug-loaded modified mesoporous silica (MMS) and poly (l-lactic acid) (PLLA). Using a co-solvent-based electrospinning method ibuprofen (IBU)-loaded MMS was successfully and uniformly encapsulated in the PLLA fibers. The electrospun PLLA-MMS-IBU composite fibrous membranes showed significantly lower initial burst release (6% release in the first 12h) compared with that of electrospun PLLA-IBU fibrous membranes (46% release in the first 12h) in in vitro release tests. Moreover, the release from PLLA-MMS-IBU was also for significantly longer than that from PLLA-IBU (100 vs. 20days). In animal studies both PLLA-IBU and PLLA-MMS-IBU showed improved anti-adhesion properties and anti-inflammatory effects compared with PLLA fibrous membrane alone 4weeks after implantation. Further, animals implanted with PLLA-MMS-IBU for 8weeks showed the lowest inflammation and best recovery compared with those implanted with PLLA-IBU and PLLA, most likely as a result of its long-term IBU release profile. Therefore, this study provides a platform technique for fabricating fibrous membranes with long-term sustained drug release characteristics which may function as a novel carrier for long-term anti-inflammation and anti-adhesion to prevent peritendinous adhesions. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Mechanical Sensing with Flexible Metallic Nanowires

NASA Astrophysics Data System (ADS)

Dobrokhotov, Vladimir; Yazdanpanah, Mehdi; Pabba, Santosh; Safir, Abdelilah; Cohn, Robert

2008-03-01

A calibrated method of force sensing is demonstrated in which the buckled shape of a long flexible metallic nanowire is interpreted to determine the applied force. Using a nanomanipulator the nanowire is buckled in the chamber of a scanning electron microscope (SEM) and the buckled shapes are recorded in SEM images. Force is determined as a function of deflection for an assumed elastic modulus by fitting the shapes using the generalized elastica model. In this calibration the elastic modulus was determined using an auxiliary AFM measurement, with the needle in the same orientation as in the SEM. Following this calibration the needle was used as a sensor in a different orientation than the AFM coordinates to deflect a suspended PLLA polymer fiber from which the elastic modulus (2.96 GPa) was determined. In this study the same needle remained rigidly secured to the AFM cantilever throughout the entire SEM/AFM calibration procedure and the characterization of the nanofiber.

Nanofibrous nonmulberry silk/PVA scaffold for osteoinduction and osseointegration.

PubMed

Bhattacharjee, Promita; Kundu, Banani; Naskar, Deboki; Maiti, Tapas K; Bhattacharya, Debasis; Kundu, Subhas C

2015-05-01

Poly-vinyl alcohol and nonmulberry tasar silk fibroin of Antheraea mylitta are blended to fabricate nanofibrous scaffolds for bone regeneration. Nanofibrous matrices are prepared by electrospinning the equal volume ratio blends of silk fibroin (2 and 4 wt%) with poly-vinyl alcohol solution (10 wt%) and designated as 2SF/PVA and 4SF/PVA, respectively with average nanofiber diameters of 177 ± 13 nm (2SF/PVA) and 193 ± 17 nm (4SF/PVA). Fourier transform infrared spectroscopy confirms retention of the secondary structure of fibroin in blends indicating the structural stability of neo-matrix. Both thermal stability and contact angle of the blends decrease with increasing fibroin percentage. Conversely, fibroin imparts mechanical stability to the blends; greater tensile strength is observed with increasing fibroin concentration. Blended scaffolds are biodegradable and support well the neo-bone matrix synthesis by human osteoblast like cells. The findings indicate the potentiality of nanofibrous scaffolds of nonmulberry fibroin as bone scaffolding material. © 2014 Wiley Periodicals, Inc.

Potential applications of three-dimensional structure of silk fibroin/poly(ester-urethane) urea nanofibrous scaffold in heart valve tissue engineering

NASA Astrophysics Data System (ADS)

Du, Juan; Zhu, Tonghe; Yu, Haiyan; Zhu, Jingjing; Sun, Changbing; Wang, Jincheng; Chen, Sihao; Wang, Jihu; Guo, Xuran

2018-07-01

Tissue engineering heart valves (TEHV) are thought to have many advantages in low immunogenicity, good histocompatibility, excellent mechanical properties. In this paper, we reported the fabrication and characterization of a novel composite nanofibrous scaffold consisting of silk fibroin (SF) and poly(ester-urethane) urea (LDI-PEUU) by using electrospinning. Chemical and physical properties of scaffolds were evaluated using scanning electron microscopy, attenuated total reflectance Fourier transform infrared, X-ray diffraction, contact angle measurement, thermogravimetric analysis, biodegradation test and tensile strength analysis. We determined that the composite scaffolds supported the growth of human umbilical vein endothelial cell (HUVEC). The results of cell proliferation and cell morphology indicate that SF/LDI-PEUU nanofibers promoted cell viability, which supporting the application in tissue engineering. All results clarified that SF/LDI-PEUU (40:60) nanofibrous scaffolds meet the required specifications for tissue engineering and could be used as a promising construct for heart valve tissue engineering.

PLLA-PHB fiber membranes obtained by solvent-free electrospinning for short-time drug delivery.

PubMed

Cao, K; Liu, Y; Olkhov, A A; Siracusa, V; Iordanskii, A L

2018-02-01

Fibers of poly(L-lactic acid) (PLLA)/polyhydroxybutyrate (PHB) with different concentrations of the drug dipyridamole (DPD) were prepared using solvent-free melt electrospinning to obtain a polymeric drug delivery system. The electrospun fibers were morphologically, structurally, thermally, and dynamically characterized. Crazes that resemble lotus root crevices were interestingly observed in the 7:3 PLLA/PHB fibers with 1% DPD. The crystallinity of PLLA slightly decreased as PHB was incorporated, and the addition of DPD significantly reduced the melting temperature of the composite. The interactions between PLLA and PHB mainly occurred at a proportion of 7:3, and drug encapsulation in the fibers was verified. The kinetic profiles of drug release demonstrated the predominant multiple patterns involving a diffusional stage in the short-term mode of release and kinetic process related to the hydrolysis of the biopolymers. Furthermore, the dynamic behavior of the polymer molecules was evaluated based on the segmental mobility using probe electron spin resonance spectroscopy. The segmental mobility in the amorphous fraction of PLLA decreased with increasing PLLA content. The 9:1 PLLA/PHB system was more resistant to polymer hydrolysis than to the 7:3 system and the rate of diffusion transport was approximately two times higher for the 7:3 PLLA/PHB fibers than for the 9:1 PLLA/PHB fibers.

Surface modification of poly(L-lactic acid) to improve its cytocompatibility via assembly of polyelectrolytes and gelatin.

PubMed

Lin, Yuan; Wang, Luling; Zhang, Peibiao; Wang, Xin; Chen, Xuesi; Jing, Xiabin; Su, Zhaohui

2006-03-01

Poly(L-lactide) (PLLA) surface was modified via aminolysis by poly(allylamine hydrochloride) (PAH) at high pH and subsequent electrostatic self-assembly of poly(sodium styrenesulfonate) (PSS) and PAH, and the process was monitored by X-ray photoelectron spectroscopy (XPS) and contact angle measurement. These modified PLLAs were then used as charged substrates for further incorporation of gelatin to improve their cytocompatibility. The amphoteric nature of the gelatin was exploited and the gelatin was adsorbed to the negatively charged PLLA/PSS and positively charged PLLA/PAH at pH=3.4 and 7.4, respectively. XPS and water contact angle data indicated that the gelatin adsorption at pH=3.4 resulted in much higher surface coverage by gelatin than at pH=7.4. All the modified PLLA surfaces became more hydrophilic than the virgin PLLA. Chondrocyte culture was used to test the cell attachment, cell morphology and cell viability on the modified PLLA substrates. The results showed that the PAH and PSS modified PLLA exhibited better cytocompatibility than virgin PLLA, and the incorporation of the gelatin on these modified PLLA substrates further improved their cytocompatibility, with the PLLA/PSS substrate treated with the gelatin at pH=3.4 being the best, exceeding the chondrocyte compatibility of the tissue culture polystyrene.

A poly(lactide) stereocomplex structure with modified magnesium oxide and its effects in enhancing the mechanical properties and suppressing inflammation.

PubMed

Kum, Chang Hun; Cho, Youngjin; Seo, Seong Ho; Joung, Yoon Ki; Ahn, Dong June; Han, Dong Keun

2014-09-24

Biodegradable polymers such as poly(L-lactide) (PLLA) have been widely utilized as materials for biomedical applications. However, the relatively poor mechanical properties of PLLA and its acid-induced cell inflammation brought about by the acidic byproducts during biodegradation pose severe problems. In this study, these drawbacks of PLLA are addressed using a stereocomplex structure, where oligo-D-lactide-grafted magnesium hydroxide (MgO-ODLA) is synthesized by grafting d-lactide onto the surface of magnesium hydroxide, which is then blended with a PLLA film. The structure, morphology, pH change, thermal and mechanical properties, in-vitro cytotoxicity, and inflammation effect of the MgO-ODLAs and their PLLA composites are evaluated through various analyses. The PLLA/MgO70-ODLA30 (0-20 wt%) composite with a stereocomplex structure shows a 20% increase in its tensile strength and an improvement in the modulus compared to its oligo-L-lactide (PLLA/MgO70-OLLA30) counterpart. The interfacial interaction parameter of PLLA/MgO70-ODLA30 (5.459) has superior properties to those of PLLA/MgO70-OLLA30 (4.013) and PLLA/Mg(OH)2 (1.774). The cell cytotoxicity and acid-induced inflammatory response are suppressed by the neutralizing effect of the MgO-ODLAs. In addition, the inflammatory problem caused by the rapid acidification of the stereocomplex structure is also addressed. As a result, the stereocomplex structure of the MgO-ODLA/PLLA composite can be used to overcome the problems associated with the biomedical applications of PLLA films. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Design and manufacture of neural tissue engineering scaffolds using hyaluronic acid and polycaprolactone nanofibers with controlled porosity.

PubMed

Entekhabi, Elahe; Haghbin Nazarpak, Masoumeh; Moztarzadeh, Fathollah; Sadeghi, Ali

2016-12-01

Given the large differences in nervous tissue and other tissues of the human body and its unique features, such as poor and/or lack of repair, there are many challenges in the repair process of this tissue. Tissue engineering is one of the most effective approaches to repair neural damages. Scaffolds made from electrospun fibers have special potential in cell adhesion, function and cell proliferation. This research attempted to design a high porous nanofibrous scaffold using hyaluronic acid and polycaprolactone to provide ideal conditions for nerve regeneration by applying proper physicochemical and mechanical signals. Chemical and mechanical properties of pure PCL and PCL/HA nanofibrous scaffolds were measured by FTIR and tensile test. Morphology, swelling behavior, and biodegradability of the scaffolds were evaluated too. Porosity of various layers of scaffolds was measured by image analysis method. To assess the cell-scaffold interaction, SH-SY5Y human neuroblastoma cell line were cultured on the electrospun scaffolds. Taken together, these results suggest that the blended nanofibrous scaffolds PCL/HA 95:5 exhibit the most balanced properties to meet all of the required specifications for neural cells and have potential application in neural tissue engineering. Copyright © 2016 Elsevier B.V. All rights reserved.

Crystallization and Microphase Separation in Chiral Block Copolymers

NASA Astrophysics Data System (ADS)

Ho, Rong-Ming

2012-02-01

Block copolymers composed of chiral entities, denoted as chiral block copolymers (BCP*s), were designed to fabricate helical architectures from self-assembly. A helical phase (denoted H*) was discovered in the self-assembly of poly(styrene)-b-poly(L-lactide) (PS-PLLA) BCPs*. To examine the phase behavior of the PS-PLLA, self-assembled superstructures resulting from the competition between crystallization and microphase separation of the PS-PLLA in solution were examined. A kinetically controlled process by changing non-solvent addition rate was utilized to control the BCP* self-assembly. Single-crystal lozenge lamellae were obtained by the slow self-assembly (i.e., slow non-solvent addition rate) of PS-PLLA whereas amorphous helical ribbon superstructures were obtained from the fast self-assembly (i.e., fast non-solvent addition rate). As a result, the formation of helical architectures from the self-assembly of the PS-PLLA reflects the impact of chirality on microphase separation, but the chiral effect might be overwhelmed by crystallization. Consequently, various crystalline PS-PLLA nanostructures in bulk were obtained by controlling the crystallization temperature of PLLA (Tc,PLLA) at which crystalline helices and crystalline cylinders occur while Tc,PLLA=x Tg,PS, respectively. Anisotropic arrangement of the PLLA crystallites grown within the microdomains was identified. The formation of this exclusive crystalline growth is attributed to the spatial confinement effect for crystallization. While Tc,PLLA=x Tg,PS, the preferential growth may modulate the curvature of microdomains by shifting the molecular chains to access the fast path for crystalline growth due to the increase in chain mobility. As a result, a spring-like behavior of the helical nanostructure can be driven by crystallization so as to dictate the transformation of helices and to result in crystalline cylinders.

In-situ polymerized cellulose nanocrystals (CNC)-poly(l-lactide) (PLLA) nanomaterials and applications in nanocomposite processing.

PubMed

Miao, Chuanwei; Hamad, Wadood Y

2016-11-20

CNC-PLLA nanomaterials were synthesized via in-situ ring-opening polymerization of l-lactide in the presence of CNC, resulting in hydrophobic, homogeneous mixture of PLLA-grafted-CNC and free PLLA homopolymer. The free PLLA serves two useful functions: as barrier to further prevent PLLA-g-CNC from forming aggregates, and in creating improved interfacial properties when these nanomaterials are blended with other polymers, hence enhancing their performance. CNC-PLLA nanomaterials can be used for medical or engineering applications as-they-are or by compounding with suitable biopolymers using versatile techniques, such as solution casting, co-extrusion or injection molding, to form hybrid nanocomposites of tunable mechanical properties. When compounded with commercial-grade PLA, the resulting CNC-PLA nanocomposites appear transparent and have tailored (dynamic and static) mechanical and barrier properties, approaching those of poly(ethylene terephthalate), PET. The effect of reaction conditions on the properties of CNC-PLLA nanomaterials have been carefully studied and detailed throughout the paper. Copyright © 2016 Elsevier Ltd. All rights reserved.

Poly(ɛ-caprolactone)/gelatin composite electrospun scaffolds with porous crater-like structures for tissue engineering.

PubMed

Hwang, Patrick T J; Murdock, Kyle; Alexander, Grant C; Salaam, Amanee D; Ng, Joshua I; Lim, Dong-Jin; Dean, Derrick; Jun, Ho-Wook

2016-04-01

Electrospinning has been widely used to fabricate scaffolds imitating the structure of natural extracellular matrix (ECM). However, conventional electrospinning produces tightly compacted nanofiber layers with only small superficial pores and a lack of bioactivity, which limit the usefulness of electrospinning in biomedical applications. Thus, a porous poly(ε-caprolactone) (PCL)/gelatin composite electrospun scaffold with crater-like structures was developed. Porous crater-like structures were created on the scaffold by a gas foaming/salt leaching process; this unique fiber structure had more large pore areas and higher porosity than the conventional electrospun fiber network. Various ratios of PCL/gelatin (concentration ratios: 100/0, 75/25, and 50/50) composite electrospun scaffolds with and without crater-like structures were characterized by their microstructures, surface chemistry, degradation, mechanical properties, and ability to facilitate cell growth and infiltration. The combination of PCL and gelatin endowed the scaffold with both structural stability of PCL and bioactivity of gelatin. All ratios of scaffolds with crater-like structures showed fairly similar surface chemistry, degradation rates, and mechanical properties to equivalent scaffolds without crater-like structures; however, craterized scaffolds displayed higher human mesenchymal stem cell (hMSC) proliferation and infiltration throughout the scaffolds after 7-day culture. Therefore, these results demonstrated that PCL/gelatin composite electrospun scaffolds with crater-like structures can provide a structurally and biochemically improved three-dimensional ECM-mimicking microenvironment. © 2016 Wiley Periodicals, Inc.

Poly(ε-caprolactone)/gelatin composite electrospun scaffolds with porous crater-like structures for tissue engineering

PubMed Central

Hwang, Patrick T.J.; Murdock, Kyle; Alexander, Grant C.; Salaam, Amanee D.; Ng, Joshua I.; Lim, Dong-Jin; Dean, Derrick; Jun, Ho-Wook

2016-01-01

Electrospinning has been widely used to fabricate scaffolds imitating the structure of natural extracellular matrix (ECM). However, conventional electrospinning produces tightly compacted nanofiber layers with only small superficial pores and a lack of bioactivity, which limit the usefulness of electrospinning in biomedical applications. Thus, a porous poly(ε-caprolactone) (PCL)/gelatin composite electrospun scaffold with crater-like structures was developed. Porous crater-like structures were created on the scaffold by a gas foaming/salt leaching process; this unique fiber structure had more large pore areas and higher porosity than the conventional electrospun fiber network. Various ratios of PCL/gelatin (concentration ratios: 100/0, 75/25, and 50/50) composite electrospun scaffolds with and without crater-like structures were characterized by their microstructures, surface chemistry, degradation, mechanical properties, and ability to facilitate cell growth and infiltration. The combination of PCL and gelatin endowed the scaffold with both structural stability of PCL and bioactivity of gelatin. All ratios of scaffolds with crater-like structures showed fairly similar surface chemistry, degradation rates, and mechanical properties to equivalent scaffolds without crater-like structures; however, craterized scaffolds displayed higher human mesenchymal stem cell (hMSC) proliferation and infiltration throughout the scaffolds after 7-day culture. Therefore, these results demonstrated that PCL/gelatin composite electrospun scaffolds with crater-like structures can provide a structurally and biochemically improved three-dimensional ECM-mimicking microenvironment. PMID:26567028

Optimization of protein cross-linking in bicomponent electrospun scaffolds for therapeutic use

NASA Astrophysics Data System (ADS)

Papa, Antonio; Guarino, Vincenzo; Cirillo, Valentina; Oliviero, Olimpia; Ambrosio, Luigi

2015-12-01

Bio-instructive electrospun scaffolds based on the combination of synthetic polymers, such as PCL or PLLA, and natural polymers (e.g., collagen) have been extensively investigated as temporary extracellular matrix (ECM) analogues able to support cell proliferation and stem cell differentiation for the regeneration of several tissues. The growing use of natural polymers as carrier of bioactive molecules is introducing new ideas for the design of polymeric drug delivery systems based on electrospun fibers with improved bioavailability, therapeutic efficacy and programmed drug release. In particular, the release mechanism is driven by the use of water soluble proteins (i.e., collagen, gelatin) which fully degrade in in vitro microenvironment, thus delivering the active principles. However, these protein are generally rapidly digested by enzymes (i.e., collagenase) produced by many different cell types, both in vivo and in vitro with significant drawbacks in tissue engineering and controlled drug delivery. Here, we aim at investigating different chemical strategies to improve the in vitro stability and mechanical strength of scaffolds against enzymatic degradation, by modifying the biodegradation rates of proteins embedded in bicomponent fibers. By comparing scaffolds treated by different cross-linking agents (i.e., GC, EDC, BDDGE), we have provided an extensive morphological/chemical/physical characterization via SEM and TGA to identify the best conditions to control drug release via protein degradation from bicomponent fibers without compromising in vitro cell response.

Optimization of protein cross-linking in bicomponent electrospun scaffolds for therapeutic use

DOE Office of Scientific and Technical Information (OSTI.GOV)

Papa, Antonio; IMAST SCaRL, Piazza Bovio 22, 80133 Naples; Guarino, Vincenzo, E-mail: vincenzo.guarino@cnr.it

Bio-instructive electrospun scaffolds based on the combination of synthetic polymers, such as PCL or PLLA, and natural polymers (e.g., collagen) have been extensively investigated as temporary extracellular matrix (ECM) analogues able to support cell proliferation and stem cell differentiation for the regeneration of several tissues. The growing use of natural polymers as carrier of bioactive molecules is introducing new ideas for the design of polymeric drug delivery systems based on electrospun fibers with improved bioavailability, therapeutic efficacy and programmed drug release. In particular, the release mechanism is driven by the use of water soluble proteins (i.e., collagen, gelatin) which fullymore » degrade in in vitro microenvironment, thus delivering the active principles. However, these protein are generally rapidly digested by enzymes (i.e., collagenase) produced by many different cell types, both in vivo and in vitro with significant drawbacks in tissue engineering and controlled drug delivery. Here, we aim at investigating different chemical strategies to improve the in vitro stability and mechanical strength of scaffolds against enzymatic degradation, by modifying the biodegradation rates of proteins embedded in bicomponent fibers. By comparing scaffolds treated by different cross-linking agents (i.e., GC, EDC, BDDGE), we have provided an extensive morphological/chemical/physical characterization via SEM and TGA to identify the best conditions to control drug release via protein degradation from bicomponent fibers without compromising in vitro cell response.« less

Bioactive Nano-Fibrous Scaffolds for Bone and Cartilage Tissue Engineering

NASA Astrophysics Data System (ADS)

Feng, Kai

Scaffolds that can mimic the structural features of natural extracellular matrix and can deliver biomolecules in a controlled fashion may provide cells with a favorable microenvironment to facilitate tissue regeneration. Biodegradable nanofibrous scaffolds with interconnected pore network have previously been developed in our laboratory to mimic collagen matrix and advantageously support both bone and cartilage regeneration. This dissertation project aims to expand both the structural complexity and the biomolecule delivery capacity of such biomimetic scaffolds for tissue engineering. We first developed a nanofibrous scaffold that can release an antibiotic (doxycycline) with a tunable release rate and a tunable dosage, which was demonstrated to be able to inhibit bacterial growth over a prolonged time period. We then developed a nanofibrous tissue-engineciing scaffold that can release basic fibroblast growth factor (bFGF) in a spatially and temporally controlled fashion. In a mouse subcutaneous implantation model, the bFGF-releasing scaffold was shown to enhance cell penetration, tissue ingrowth and angiogenesis. It was also found that both the dose and the release rate of bFGF play roles in the biologic function of the scaffold. After that, we developed a nanofibrous PLLA scaffold that can release both bone morphogenetic protein 7 (BMP-7) and platelet-derived growth factor (PDGF) with distinct dosages and release kinetics. It was demonstrated that BMP-7 and PDGF could synergistically enhance bone regeneration using a mouse ectopic bone formation model and a rat periodontal fenestration defect regeneration model. The regeneration outcome was dependent on the dosage, the ratio and the release kinetics of the two growth factors. Last, we developed an anisotropic composite scaffold with an upper layer mimicking the superficial zone of cartilage and a lower layer mimicking the middle zone of cartilage. The thin superficial layer was fabricated using an electrospinning technique to support a more parallel ECM orientation to the cartilage surface. The lower layer was fabricated using a phase-separation technique to support a more isotropic ECM distribution. Human bone marrow-derived mesenchymal stem cells (hMSCs) were seeded on this complex scaffold and cultured under chondrogenic conditions. The results showed that the composite scaffold was indeed able to support anisotropic cartilage tissue structure formation.

Integrating Microtissues in Nanofiber Scaffolds for Regenerative Nanomedicine

PubMed Central

Keller, Laetitia; Wagner, Quentin; Offner, Damien; Eap, Sandy; Musset, Anne-Marie; Arruebo, Manuel; Kelm, Jens M.; Schwinté, Pascale; Benkirane-Jessel, Nadia

2015-01-01

A new generation of biomaterials focus on smart materials incorporating cells. Here, we describe a novel generation of synthetic nanofibrous implant functionalized with living microtissues for regenerative nanomedicine. The strategy designed here enhances the effectiveness of therapeutic implants compared to current approaches used in the clinic today based on single cells added to the implant. PMID:28793604

Development of a nanostructured DNA delivery scaffold via electrospinning of PLGA and PLA-PEG block copolymers

NASA Technical Reports Server (NTRS)

Luu, Y. K.; Kim, K.; Hsiao, B. S.; Chu, B.; Hadjiargyrou, M.; Hadjiargyou, M. (Principal Investigator)

2003-01-01

The present work utilizes electrospinning to fabricate synthetic polymer/DNA composite scaffolds for therapeutic application in gene delivery for tissue engineering. The scaffolds are non-woven, nano-fibered, membranous structures composed predominantly of poly(lactide-co-glycolide) (PLGA) random copolymer and a poly(D,L-lactide)-poly(ethylene glycol) (PLA-PEG) block copolymer. Release of plasmid DNA from the scaffolds was sustained over a 20-day study period, with maximum release occurring at approximately 2 h. Cumulative release profiles indicated amounts released were approximately 68-80% of the initially loaded DNA. Variations in the PLGA to PLA-PEG block copolymer ratio vastly affected the overall structural morphology, as well as both the rate and efficiency of DNA release. Results indicated that DNA released directly from these electrospun scaffolds was indeed intact, capable of cellular transfection, and successfully encoded the protein beta-galactosidase. When tested under tensile loads, the electrospun polymer/DNA composite scaffolds exhibited tensile moduli of approximately 35 MPa, with approximately 45% strain initially. These values approximate those of skin and cartilage. Taken together, this work represents the first successful demonstration of plasmid DNA incorporation into a polymer scaffold using electrospinning.

Synthesis of PDLLA/PLLA-bentonite nanocomposite through sonication

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sitompul, Johnner, E-mail: sitompul@che.itb.ac.id; Setyawan, Daru, E-mail: daru.setyawan@gmail.com; Kim, Daniel Young Joon, E-mail: daniel.kim12321@gmail.com

2016-04-19

This paper concerns the synthesis of poly(D,L-lactic acid)/poly(L-lactic acid) bentonite nanocomposites. Poly (D,L-lactic acid) (PDLLA) was synthesized using lactic acid through the ZnO-catalyzed direct polycondensation method at vacuum pressure and poly(L-lactic acid) (PLLA) was synthesized with L-lactide by ring-opening polymerization method. The PDLLA/PLLA-bentonite nanocomposite films were synthesized using the solvent casting method. The nanoclay, bentonite, was prepared using the solution-intercalation method by dissolving the nanoparticles into chloroform before sonication. In this study, PDLLA/PLLA-bentonite nanocomposite films were produced using variable amounts of nanoclay and sonication times during the mixing of PDLLA/PLLA and bentonite. The properties of the PDLLA/PLLA nanocomposites were thenmore » characterized using the X-ray Diffraction (XRD), Universal Testing Machine (UTM), Water Vapor Permeability (WVP) tests, and the enzymatic biodegradability test. The XRD test was used to measure the intercalation of nanoclay layers in the PDLLA/PLLA matrix and the PDLLA/PLLA-bentonite intercalated nanocomposite films. It was found through these various tests that adding bentonite to the PDLLA/PLLA increases tensile strength to 56.76 MP. Furthermore, the biodegradability increases as well as the barrier properties of the polymers The different sonication time used during the mixing of the polymer solution with bentonite also affected the properties of the PDLLA/PLLA-bentonite nanocomposite films.« less

AFM feature definition for neural cells on nanofibrillar tissue scaffolds.

PubMed

Tiryaki, Volkan M; Khan, Adeel A; Ayres, Virginia M

2012-01-01

A diagnostic approach is developed and implemented that provides clear feature definition in atomic force microscopy (AFM) images of neural cells on nanofibrillar tissue scaffolds. Because the cellular edges and processes are on the same order as the background nanofibers, this imaging situation presents a feature definition problem. The diagnostic approach is based on analysis of discrete Fourier transforms of standard AFM section measurements. The diagnostic conclusion that the combination of dynamic range enhancement with low-frequency component suppression enhances feature definition is shown to be correct and to lead to clear-featured images that could change previously held assumptions about the cell-cell interactions present. Clear feature definition of cells on scaffolds extends the usefulness of AFM imaging for use in regenerative medicine. © Wiley Periodicals, Inc.

[Anti-tumor effects of DDP-PLLA-CNTs on human cholangiocarcinoma cell line in vitro].

PubMed

Li, Maolan; Lu, Wei; Zhang, Fei; Ding, Qichen; Wu, Xiangsong; Tan, Zhujun; Wu, Wenguang; Weng, Hao; Wang, Xuefeng; Shi, Weibin; Dong, Ping; Gu, Jun; Liu, Yingbin

2014-11-04

To explore the antitumor effects of DDP-PLLA-CNTs on human cholangiocarcinoma cell line. DDP-PLLA-CNTs were prepared with the method of ultrasound emulsification. The morphology of DDP-PLLA-CNTs was determined by scanning electron microscope (SEM). And its drug loading and drug release curve in vitro was detected by UV-Vis-NIR spectrophotometer. CCK8 was used to test the cytotoxic effects of DDP-PLLA-CNTs at different concentrations on QBC939 cell proliferation.Flow cytometry was employed to measure the changes of apoptotic rate. With excellent controlled-release characteristic of in vitro drug release, DDP-PLLA-CNTs inhibited the proliferation and significantly increased the apoptotic rate of QBC939 cell line. DDP-PLLA-CNTs have drug sustained-release characteristics and can significantly inhibit the proliferation of QBC939 cell line.

Fibrous scaffolds fabricated by emulsion electrospinning: from hosting capacity to in vivo biocompatibility

NASA Astrophysics Data System (ADS)

Spano, F.; Quarta, A.; Martelli, C.; Ottobrini, L.; Rossi, R. M.; Gigli, G.; Blasi, L.

2016-04-01

Electrospinning is a versatile method for preparing functional three-dimensional scaffolds. Synthetic and natural polymers have been used to produce micro- and nanofibers that mimic extracellular matrices. Here, we describe the use of emulsion electrospinning to prepare blended fibers capable of hosting aqueous species and releasing them in solution. The existence of an aqueous and a non-aqueous phase allows water-soluble molecules to be introduced without altering the structure and the degradation of the fibers, and means that their release properties under physiological conditions can be controlled. To demonstrate the loading capability and flexibility of the blend, various species were introduced, from magnetic nanoparticles and quantum rods to biological molecules. Cellular studies showed the spontaneous adhesion and alignment of cells along the fibers. Finally, in vivo experiments demonstrated the high biocompatibility and safety of the scaffolds up to 21 days post-implantation.Electrospinning is a versatile method for preparing functional three-dimensional scaffolds. Synthetic and natural polymers have been used to produce micro- and nanofibers that mimic extracellular matrices. Here, we describe the use of emulsion electrospinning to prepare blended fibers capable of hosting aqueous species and releasing them in solution. The existence of an aqueous and a non-aqueous phase allows water-soluble molecules to be introduced without altering the structure and the degradation of the fibers, and means that their release properties under physiological conditions can be controlled. To demonstrate the loading capability and flexibility of the blend, various species were introduced, from magnetic nanoparticles and quantum rods to biological molecules. Cellular studies showed the spontaneous adhesion and alignment of cells along the fibers. Finally, in vivo experiments demonstrated the high biocompatibility and safety of the scaffolds up to 21 days post-implantation. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr00782a

Electrospun Fe3O4/TiO2 hybrid nanofibers and their in vitro biocompatibility: prospective matrix for satellite cell adhesion and cultivation.

PubMed

Amna, Touseef; Hassan, M Shamshi; Van Ba, Hoa; Khil, Myung-Seob; Lee, Hak-Kyo; Hwang, I H

2013-03-01

We report the fabrication of novel Fe3O4/TiO2 hybrid nanofibers with the improved cellular response for potential tissue engineering applications. In this study, Fe3O4/TiO2 hybrid nanofibers were prepared by facile sol-gel electrospinning using titanium isopropoxide and iron(III) nitrate nonahydrate as precursors. The obtained electrospun nanofibers were vacuum dried at 80 °C and then calcined at 500 °C. The physicochemical characterization of the synthesized composite nanofibers was carried out by scanning electron microscopy, energy dispersive X-ray spectroscopy, transmission electron microscopy and X-ray diffraction pattern. To examine the in vitro cytotoxicity, satellite cells were treated with as-prepared Fe3O4/TiO2 and the viability of cells was analyzed by Cell Counting Kit-8 assay at regular time intervals. The morphological features of unexposed satellite cells and exposed to Fe3O4/TiO2 composite were examined with a phase contrast microscope whereas the quantification of cell viability was carried out via confocal laser scanning microscopy. The morphology of the cells attached to hybrid matrix was observed by Bio-SEM. Cytotoxicity experiments indicated that the satellite cells could attach to the Fe3O4/TiO2 composite nanofibers after being cultured. We observed that Fe3O4-TiO2 composite nanofibers could support cell adhesion and growth. Results from this study therefore suggest that Fe3O4/TiO2 composite scaffold with small diameters (approximately 200 nm) can mimic the natural extracellular matrix well and provide possibilities for diverse applications in the field of tissue engineering and regenerative medicine. Copyright © 2012 Elsevier B.V. All rights reserved.

Modeling and process optimization of electrospinning of chitosan-collagen nanofiber by response surface methodology

NASA Astrophysics Data System (ADS)

Amiri, Nafise; Moradi, Ali; Abolghasem Sajjadi Tabasi, Sayyed; Movaffagh, Jebrail

2018-04-01

Chitosan-collagen composite nanofiber is of a great interest to researchers in biomedical fields. Since the electrospinning is the most popular method for nanofiber production, having a comprehensive knowledge of the electrospinning process is beneficial. Modeling techniques are precious tools for managing variables in the electrospinning process, prior to the more time- consuming and expensive experimental techniques. In this study, a central composite design of response surface methodology (RSM) was employed to develop a statistical model as well as to define the optimum condition for fabrication of chitosan-collagen nanofiber with minimum diameter. The individual and the interaction effects of applied voltage (10–25 kV), flow rate (0.5–1.5 mL h‑1), and needle to collector distance (15–25 cm) on the fiber diameter were investigated. ATR- FTIR and cell study were done to evaluate the optimized nanofibers. According to the RSM, a two-factor interaction (2FI) model was the most suitable model. The high regression coefficient value (R 2 ≥ 0.9666) of the fitted regression model and insignificant lack of fit (P = 0.0715) indicated that the model was highly adequate in predicting chitosan-collagen nanofiber diameter. The optimization process showed that the chitosan-collagen nanofiber diameter of 156.05 nm could be obtained in 9 kV, 0.2 ml h‑1, and 25 cm which was confirmed by experiment (155.92 ± 18.95 nm). The ATR-FTIR and cell study confirmed the structure and biocompatibility of the optimized membrane. The represented model could assist researchers in fabricating chitosan-collagen electrospun scaffolds with a predictable fiber diameter, and optimized chitosan-collagen nanofibrous mat could be a potential candidate for wound healing and tissue engineering.

Tuning Molecular Weights of Bombyx mori (B. mori) Silk Sericin to Modify Its Assembly Structures and Materials Formation

PubMed Central

2015-01-01

Bombyx mori (B. mori) silk sericin is a protein with features desirable as a biomaterial, such as increased hydrophilicity and biodegradation, as well as resistance to oxidation, bacteria, and ultraviolet light. In contrast to other widely studied B. mori silk proteins such as fibroin, sericin is still unexplored as a building block for fabricating biomaterial, and thus a facile technique of processing it into a material is needed. Here, electrospinning technology was used to fabricate it into biomaterials from two forms of B. mori silk sericin with different molecular weights, one is a low (12.0 kDa) molecular sericin (LS) form and another is a high (66.0 kDa) molecular weight sericin (HS) form. Circular dichroism (CD) spectra showed that LS in hexafluoroacetone (HFA) solvent adopted a predominantly random coil conformation, whereas HS tended to form a β-sheet structure along with a large content of random coils. In addition, LS and HS in HFA solvent were found to form cylinder-like smaller nanoparticles and larger irregular aggregates before electrospinning, respectively. As a result, biomaterials based on microparticles and nanofibers were successfully fabricated by electrospinning of LS and HS dissolved in HFA, respectively. The cell viability and differentiation assay indicated that nanofibers and microparticles improved cell adhesion, growth, and differentiation, proving that the scaffolds electrospun from sericin are biocompatible regardless of its molecular weight. The microparticles, not common in electrospinning of silk proteins reported previously, were found to promote the osteogenic differentiation of mesenchymal stem cells in comparison to the nanofibers. This study suggested that molecular weight of sericin mediates its secondary structure and assembly structure, which in turn leads to a control of final morphology of the electrospun materials. The microparticles and nanofibers of sericin can be potentially used as building blocks for fabricating the scaffolds for tissue engineering. PMID:25050697

Tuning molecular weights of Bombyx mori (B. mori) silk sericin to modify its assembly structures and materials formation.

PubMed

Yang, Mingying; Shuai, Yajun; Zhou, Guanshan; Mandal, Namita; Zhu, Liangjun; Mao, Chuanbin

2014-08-27

Bombyx mori (B. mori) silk sericin is a protein with features desirable as a biomaterial, such as increased hydrophilicity and biodegradation, as well as resistance to oxidation, bacteria, and ultraviolet light. In contrast to other widely studied B. mori silk proteins such as fibroin, sericin is still unexplored as a building block for fabricating biomaterial, and thus a facile technique of processing it into a material is needed. Here, electrospinning technology was used to fabricate it into biomaterials from two forms of B. mori silk sericin with different molecular weights, one is a low (12.0 kDa) molecular sericin (LS) form and another is a high (66.0 kDa) molecular weight sericin (HS) form. Circular dichroism (CD) spectra showed that LS in hexafluoroacetone (HFA) solvent adopted a predominantly random coil conformation, whereas HS tended to form a β-sheet structure along with a large content of random coils. In addition, LS and HS in HFA solvent were found to form cylinder-like smaller nanoparticles and larger irregular aggregates before electrospinning, respectively. As a result, biomaterials based on microparticles and nanofibers were successfully fabricated by electrospinning of LS and HS dissolved in HFA, respectively. The cell viability and differentiation assay indicated that nanofibers and microparticles improved cell adhesion, growth, and differentiation, proving that the scaffolds electrospun from sericin are biocompatible regardless of its molecular weight. The microparticles, not common in electrospinning of silk proteins reported previously, were found to promote the osteogenic differentiation of mesenchymal stem cells in comparison to the nanofibers. This study suggested that molecular weight of sericin mediates its secondary structure and assembly structure, which in turn leads to a control of final morphology of the electrospun materials. The microparticles and nanofibers of sericin can be potentially used as building blocks for fabricating the scaffolds for tissue engineering.

Fabrication and Evaluation of Electrospun, 3D-Bioplotted, and Combination of Electrospun/3D-Bioplotted Scaffolds for Tissue Engineering Applications

PubMed Central

Mellor, Liliana F.; Huebner, Pedro; Cai, Shaobo; Taylor, Michael A.; Spang, Jeffrey

2017-01-01

Electrospun scaffolds provide a dense framework of nanofibers with pore sizes and fiber diameters that closely resemble the architecture of native extracellular matrix. However, it generates limited three-dimensional structures of relevant physiological thicknesses. 3D printing allows digitally controlled fabrication of three-dimensional single/multimaterial constructs with precisely ordered fiber and pore architecture in a single build. However, this approach generally lacks the ability to achieve submicron resolution features to mimic native tissue. The goal of this study was to fabricate and evaluate 3D printed, electrospun, and combination of 3D printed/electrospun scaffolds to mimic the native architecture of heterogeneous tissue. We assessed their ability to support viability and proliferation of human adipose derived stem cells (hASC). Cells had increased proliferation and high viability over 21 days on all scaffolds. We further tested implantation of stacked-electrospun scaffold versus combined electrospun/3D scaffold on a cadaveric pig knee model and found that stacked-electrospun scaffold easily delaminated during implantation while the combined scaffold was easier to implant. Our approach combining these two commonly used scaffold fabrication technologies allows for the creation of a scaffold with more close resemblance to heterogeneous tissue architecture, holding great potential for tissue engineering and regenerative medicine applications of osteochondral tissue and other heterogeneous tissues. PMID:28536700

Fabrication of bioinspired composite nanofiber membranes with robust superhydrophobicity for direct contact membrane distillation.

PubMed

Liao, Yuan; Wang, Rong; Fane, Anthony G

2014-06-03

The practical application of membrane distillation (MD) for water purification is hindered by the absence of desirable membranes that can fulfill the special requirements of the MD process. Compared to the membranes fabricated by other methods, nanofiber membranes produced by electrospinning are of great interest due to their high porosity, low tortuosity, large surface pore size, and high surface hydrophobicity. However, the stable performance of the nanofiber membranes in the MD process is still unsatisfactory. Inspired by the unique structure of the lotus leaf, this study aimed to develop a strategy to construct superhydrophobic composite nanofiber membranes with robust superhydrophobicity and high porosity suitable for use in MD. The newly developed membrane consists of a superhydrophobic silica-PVDF composite selective skin formed on a polyvinylidene fluoride (PVDF) porous nanofiber scaffold via electrospinning. This fabrication method could be easily scaled up due to its simple preparation procedures. The effects of silica diameter and concentration on membrane contact angle, sliding angle, and MD performance were investigated thoroughly. For the first time, the direct contact membrane distillation (DCMD) tests demonstrate that the newly developed membranes are able to present stable high performance over 50 h of testing time, and the superhydrophobic selective layer exhibits excellent durability in ultrasonic treatment and a continuous DCMD test. It is believed that this novel design strategy has great potential for MD membrane fabrication.

Fabrication and characterization of poly(L-lactic acid) gels induced by fibrous complex crystallization with solvents

DOE Office of Scientific and Technical Information (OSTI.GOV)

Matsuda, Yasuhiro; Fukatsu, Akinobu; Wang, Yangyang

2014-01-01

Complex crystal induced gelation of poly(L-lactic acid) (PLLA) solutions was studied for a series of solvents, including N,N-dimethylformamide (DMF). By cooling the solutions prepared at elevated temperatures, PLLA gels were produced in solvents that induced complex crystals ( -crystals) with PLLA. Fibrous structure of PLLA in the gel with DMF was observed by polarizing optical microscopy, field emission electron microscopy, and atomic force microscopy. Upon heating, the crystal form of PLLA in the DMF gel changed from -crystal to a-crystal, the major crystal form in common untreated PLLA films, but the morphology and high elastic modulus of the gel remainedmore » until the a-crystal dissolved at higher temperature. In addition, a solvent exchanging method was developed, which allowed PLLA gels to be prepared in other useful solvents that do not induce -crystals without losing the morphology and mechanical properties.« less

Chitin butyrate coated electrospun nylon-6 fibers for biomedical applications

NASA Astrophysics Data System (ADS)

Pant, Hem Raj; Kim, Han Joo; Bhatt, Lok Ranjan; Joshi, Mahesh Kumar; Kim, Eun Kyo; Kim, Jeong In; Abdal-hay, Abdalla; Hui, K. S.; Kim, Cheol Sang

2013-11-01

In this study, we describe the preparation and characterizations of chitin butyrate (CB) coated nylon-6 nanofibers using single-spinneret electrospinning of blends solution. The physicochemical properties of nylon-6 composite fibers with different proportions of CB to nylon-6 were determined using FE-SEM, TEM, FT-IR spectroscopy, and water contact angle measurement. FE-SEM and TEM images revealed that the nylon-6 and CB were immiscible in the as-spun nanofibers, and phase separated nanofiber morphology becomes more pronounced with increasing amounts of CB. The bone formation ability of composite fibers was evaluated by incubating in biomimetic simulated body fluid. In order to assay the cytocompatibility and cell behavior on the composite scaffolds, osteoblast cells were seeded on the matrix. Results suggest that the deposition of CB layer on the surface of nylon-6 could increase its cell compatibility and bone formation ability. Therefore, as-synthesized nanocomposite fibrous mat has great potentiality in hard tissue engineering.

Design of electrospun nanofibrous mats for osteogenic differentiation of mesenchymal stem cells.

PubMed

Wang, Shige; Hu, Fei; Li, Jingchao; Zhang, Shuping; Shen, Mingwu; Huang, Mingxian; Shi, Xiangyang

2017-05-26

The clinical translation potential of mesenchymal stem cells (MSCs) in regenerative medicine has been greatly exploited. With the merits of high surface area to volume ratio, facile control of components, well retained topography, and the capacity to mimic the native extracellular matrix (ECM), nanofibers have received a great deal of attention as bone tissue engineering scaffolds. Electrospinning has been considered as an efficient approach for scale-up fabrication of nanofibrous materials. Electrospun nanofibers are capable of stimulating cell-matrix interaction to form a cell niche, directing cellular behavior, and promoting the MSCs adhesion and proliferation. In this review, we give a comprehensive literature survey on the mechanisms of electrospun nanofibers in supporting the MSCs differentiation. Specifically, the influences of biological and physical osteogenic inductive cues on the MSCs osteogenic differentiation are reviewed. Along with the significant advances in the field, current research challenges and future perspectives are also discussed. Copyright © 2017 Elsevier Inc. All rights reserved.

Fabrication of Microfibrous and Nano-/Microfibrous Scaffolds: Melt and Hybrid Electrospinning and Surface Modification of Poly(L-lactic acid) with Plasticizer

PubMed Central

Yoon, Young Il; Park, Ko Eun; Lee, Seung Jin; Park, Won Ho

2013-01-01

Biodegradable poly(L-lactic acid) (PLA) fibrous scaffolds were prepared by electrospinning from a PLA melt containing poly(ethylene glycol) (PEG) as a plasticizer to obtain thinner fibers. The effects of PEG on the melt electrospinning of PLA were examined in terms of the melt viscosity and fiber diameter. Among the parameters, the content of PEG had a more significant effect on the average fiber diameter and its distribution than those of the spinning temperature. Furthermore, nano-/microfibrous silk fibroin (SF)/PLA and PLA/PLA composite scaffolds were fabricated by hybrid electrospinning, which involved a combination of solution electrospinning and melt electrospinning. The SF/PLA (20/80) scaffolds consisted of a randomly oriented structure of PLA microfibers (average fiber diameter = 8.9 µm) and SF nanofibers (average fiber diameter = 820 nm). The PLA nano-/microfiber (20/80) scaffolds were found to have similar pore parameters to the PLA microfiber scaffolds. The PLA scaffolds were treated with plasma in the presence of either oxygen or ammonia gas to modify the surface of the fibers. This approach of controlling the surface properties and diameter of fibers could be useful in the design and tailoring of novel scaffolds for tissue engineering. PMID:24381937

[Anti-tumor effect of 5-FU-PLLA-CNTs on human gastric carcinoma cell lines in vitro].

PubMed

Gu, Jun; Li, Maolan; Wu, Xiangsong; Wu, Wenguang; Zhang, Lin; Ding, Qichen; Yang, Jiahua; Weng, Hao; Ding, Qian; Bao, Runfa; Shu, Yijun; Liu, Yingbin

2014-04-01

To prepare cisPLLAtin-loaded polylactic acid/cnts, and to study the anti-tumor effect of 5-FU-PLLA-CNTs on human gastric carcinoma cell lines(MGC803 and MNK45). 5-FU-PLLA-CNTs were prepared with ultrasound emulsification. The morphology of 5-FU-PLLA-CNTs was determined by scanning electron microscope(SEM), and its drug loading and drug release curve in vitro were detected by UV-Vis-NIR spectrophotometer. Cells were divided into experiment, positive control and negative control groups. CCK8 method was used to test the cytotoxic effect of 5-FU-PLLA-CNTs in different concentrations on MGC803 and MNK45 cell proliferation. Flow cytometry was employed to measure the apoptotic rate of MGC803 and MNK45 cells before and after the intervention of 5-FU-PLLA-CNTs. Deep layer film of 5-FU-PLLA-CNTs was successfully established, whose drug-load rate was(4.54±0.43)%, entrapment rate was(21.56±2.36)%. In vitro release test showed release rate within 24 h of 5-FU-PLLA-CNTs was 23.9% in a as lowly increasing manner, and accumulating release rate was 85.3% at day 31. CCk8 experiment revealed, as compared to control group, 5-FU-PLLA-CNTs significantly inhibited the proliferation of two cell lines in dose-dependent and time-dependent manner. The best 5-FU-PLLA-CNTs concentration of inhibition for human gastric cancer cell lines was 1 mg/well. Flow cytometry indicated the apoptotic rate of MGC803 and MNK45 cells in experiment group treated by 1 mg/well 5-FU-PLLA-CNTs significantly increased as compared to negative control group (P<0.05), while the difference was not significant as compared to positive control group (P>0.05). The 5-FU-PLLA-CNTs has good drug sustained-release capacity, and can significantly kill and inhibit the proliferation of MGC803 and MNK45 cell lines.

Co-precipitation of asiatic acid and poly( l-lactide) using rapid expansion of subcritical solutions into liquid solvents

NASA Astrophysics Data System (ADS)

Sane, Amporn; Limtrakul, Jumras

2011-09-01

Poly( l-lactide) (PLLA) nanoparticles loaded with asiatic acid (AA) were successfully produced by rapid expansion of a subcritical solution into an aqueous receiving solution containing a dispersing agent. A mixture of carbon dioxide (CO2) and ethanol (EtOH) with a weight ratio of 1:1 was used as the solvent for AA and PLLA. Two surfactants, Pluronic F127 and sodium dodecyl sulfate were employed. The former was found to be more effective for stabilizing AA-loaded PLLA nanoparticles, as a rapid expansion into a 0.1 wt% Pluronic F127 solution produced a stable nanosuspension consisting mainly of well-dispersed, individual nanoparticles. The effects of rapid expansion-processing conditions—AA to PLLA weight ratio and pre-expansion temperature (Tpre)—on the size and morphology of composite nanoparticles, and the loading capacity and entrapment efficiency of AA in PLLA nanoparticles, were systematically investigated. It was found that AA-loaded PLLA nanoparticles with a size range of 30-100 nm were consistently fabricated by rapid expansion at Tpre of 70-100 °C and AA to PLLA weight ratios of 1:2 and 1:4, and with a constant pre-expansion pressure of 330 bar. The Tpre and AA to PLLA weight ratio had no significant effects on the size of the nanoparticles. The AA to PLLA weight ratio is a controlling parameter for both the loading capacity and the entrapment efficiency of AA in PLLA nanoparticles. The loading capacity and entrapment efficiency increased from 8-11 to 16-21 wt%, and 38-57 to 50-62 wt%, respectively, when the AA to PLLA weight ratio changed from 1:4 to 1:2. However, increasing the Tpre from 70 to 100 °C decreased both the loading capacity and entrapment efficiency of AA in PLLA nanoparticles by 20-30%.

Preparation and characterization of multi-layer biodegradable nanofibers by coaxial electrospinning and their potential for tissue engineering

NASA Astrophysics Data System (ADS)

Liu, Wenwen

As an evolution of conventional electrospinning, coaxial electrospinning became popular soon after its debut as a novel way to develop nanofibers with special structures, such as core-shell and hollow interior. In recent years, there has been an increasing interest in a modified coaxial electrospinning, tri-layer coaxial electrospinning, to develop more complex structures, such as multi-layer and nanowire-in-microtube. Previous studies have primarily concentrated on the fabrication of tri-layered inorganic fibers while studies on tri-layered coaxial polymeric fibers has not been reported until very recently. Our research focuses on the fabrication of core-shell and tri-layer structured biodegradable polymeric nanofibers with coaxial electrospinning. Different characterization methods have been applied to observe the internal structure in single nanofibers and the potential application of tri-layer coaxial electrospinning has been discussed. The material system consists of biodegradable natural polymer gelatin, synthetic polymers poly (epsilon-caprolactone) (PCL) and poly (lactic-co-glycolic acid) (PLGA). A uniquely designed three-needle concentric spinneret is developed to perform tri-layer coaxial electrospinning. Different kinds of core-shell structured nanofibers, including gelatin/PCL, PCL/gelatin, gelatin/PLGA and PCL/PLGA, have been fabricated with a customized coaxial electrospinning apparatus. Two kinds of tri-layer coaxial nanofibers, two-component ABA structured gelatin/PCL/gelatin biodegradable nanofibers and tri-component ABC structured gelatin/PCL/PLGA biodegradable nanofibers, have been developed with the customized three needle coaxial electrospinning setup. The core-shell and tri-layered structures of electrospun nanofibers have been characterized by several commonly used techniques, such as laser scanning confocal microscopy (LSCM) and transmission electron microscopy (TEM). Besides the conventional methods, other newer techniques, including focused ion beam-scanning electron microscopy (FIB-SEM), super-resolution structured illumination microscopy (SR-SIM) and nanoscale-infrared spectroscopy (nano-IR), have been explored to investigate the internal structure in singles fibers. Additionally, the potential application of coaxial electrospinning in the fabrication of bioactive scaffolds for tissue engineering has been studied. Different kinds of coaxial nanofibers were fabricated and studied to determine the potential for BSA and growth factor release and some preliminary results were obtained.

In situ tissue engineering with synthetic self-assembling peptide nanofiber scaffolds, PuraMatrix, for mucosal regeneration in the rat middle-ear

PubMed Central

Akiyama, Naotaro; Yamamoto-Fukuda, Tomomi; Takahashi, Haruo; Koji, Takehiko

2013-01-01

Middle-ear mucosa maintains middle-ear pressure. However, the majority of surgical cases exhibit inadequate middle-ear mucosal regeneration, and mucosal transplantation is necessary in such cases. The aim of the present study was to assess the feasibility of transplantation of isolated mucosal cells encapsulated within synthetic self-assembling peptide nanofiber scaffolds using PuraMatrix, which has been successfully used as scaffolding in tissue engineering, for the repair of damaged middle-ear. Middle-ear bullae with mucosa were removed from Sprague Dawley (SD) transgenic rats, transfected with enhanced green fluorescent protein (EGFP) transgene and excised into small pieces, then cultured up to the third passage. After surgical elimination of middle-ear mucosa in SD recipient rats, donor cells were encapsulated within PuraMatrix and transplanted into these immunosuppressed rats. Primary cultured cells were positive for pancytokeratin but not for vimentin, and retained the character of middle-ear epithelial cells. A high proportion of EGFP-expressing cells were found in the recipient middle-ear after transplantation with PuraMatrix, but not without PuraMatrix. These cells retained normal morphology and function, as confirmed by histological examination, immunohistochemistry, and electron microscopy, and multiplied to form new epithelial and subepithelial layers together with basement membrane. The present study demonstrated the feasibility of transplantation of cultured middle-ear mucosal epithelial cells encapsulated within PuraMatrix for regeneration of surgically eliminated mucosa of the middle-ear in SD rats. PMID:23926427

In situ tissue engineering with synthetic self-assembling peptide nanofiber scaffolds, PuraMatrix, for mucosal regeneration in the rat middle-ear.

PubMed

Akiyama, Naotaro; Yamamoto-Fukuda, Tomomi; Takahashi, Haruo; Koji, Takehiko

2013-01-01

Middle-ear mucosa maintains middle-ear pressure. However, the majority of surgical cases exhibit inadequate middle-ear mucosal regeneration, and mucosal transplantation is necessary in such cases. The aim of the present study was to assess the feasibility of transplantation of isolated mucosal cells encapsulated within synthetic self-assembling peptide nanofiber scaffolds using PuraMatrix, which has been successfully used as scaffolding in tissue engineering, for the repair of damaged middle-ear. Middle-ear bullae with mucosa were removed from Sprague Dawley (SD) transgenic rats, transfected with enhanced green fluorescent protein (EGFP) transgene and excised into small pieces, then cultured up to the third passage. After surgical elimination of middle-ear mucosa in SD recipient rats, donor cells were encapsulated within PuraMatrix and transplanted into these immunosuppressed rats. Primary cultured cells were positive for pancytokeratin but not for vimentin, and retained the character of middle-ear epithelial cells. A high proportion of EGFP-expressing cells were found in the recipient middle-ear after transplantation with PuraMatrix, but not without PuraMatrix. These cells retained normal morphology and function, as confirmed by histological examination, immunohistochemistry, and electron microscopy, and multiplied to form new epithelial and subepithelial layers together with basement membrane. The present study demonstrated the feasibility of transplantation of cultured middle-ear mucosal epithelial cells encapsulated within PuraMatrix for regeneration of surgically eliminated mucosa of the middle-ear in SD rats.

Development of PVA/gelatin nanofibrous scaffolds for Tissue Engineering via electrospinning

NASA Astrophysics Data System (ADS)

Perez-Puyana, V.; Jiménez-Rosado, M.; Romero, A.; Guerrero, A.

2018-03-01

The electrospinning process is an emerging and relatively easy technique to prepare three-dimensional matrices with micro- and nanofibers. To achieve it, aqueous polymer solutions from synthetic or natural polymers are used. PVA was selected as polymer and gelatin because of its biocompatibility and biodegradability. A complete characterization of the polymeric solutions (density, surface tension, etc) was previously performed. Subsequently, a standard electrospinning process (15 kV, 0.4 ml h-1 and 10 cm) was carried out to obtain scaffolds. The influence of the polymer concentration and the protein addition was observed by performing FTIR analyses and studied by analyzing the water contact angle and SEM images.

Emerging chitin and chitosan nanofibrous materials for biomedical applications

NASA Astrophysics Data System (ADS)

Ding, Fuyuan; Deng, Hongbing; Du, Yumin; Shi, Xiaowen; Wang, Qun

2014-07-01

Over the past several decades, we have witnessed significant progress in chitosan and chitin based nanostructured materials. The nanofibers from chitin and chitosan with appealing physical and biological features have attracted intense attention due to their excellent biological properties related to biodegradability, biocompatibility, antibacterial activity, low immunogenicity and wound healing capacity. Various methods, such as electrospinning, self-assembly, phase separation, mechanical treatment, printing, ultrasonication and chemical treatment were employed to prepare chitin and chitosan nanofibers. These nanofibrous materials have tremendous potential to be used as drug delivery systems, tissue engineering scaffolds, wound dressing materials, antimicrobial agents, and biosensors. This review article discusses the most recent progress in the preparation and application of chitin and chitosan based nanofibrous materials in biomedical fields.

Fabrication and characterization of Antheraea pernyi silk fibroin-blended P(LLA-CL) nanofibrous scaffolds for peripheral nerve tissue engineering

NASA Astrophysics Data System (ADS)

Wang, Juan; Sun, Binbin; Bhutto, Muhammad Aqeel; Zhu, Tonghe; Yu, Kui; Bao, Jiayu; Morsi, Yosry; El-Hamshary, Hany; El-Newehy, Mohamed; Mo, Xiumei

2017-03-01

Electrospun nanofibers have gained widespreading interest for tissue engineering application. In the present study, ApF/P(LLA-CL) nanofibrous scaffolds were fabricated via electrospinning. The feasibility of the material as tissue engineering nerve scaffold was investigated in vitro. The average diameter increased with decreasing the blend ratio of ApF to P(LLA-CL). Characterization of 13C NMR and FTIR clarified that there is no obvious chemical bond reaction between ApF and P(LLA-CL). The tensile strength and elongation at break increased with the content increase of P(LLA-CL). The surface hydrophilic property of nanofibrous scaffolds enhanced with the increased content of ApF. Cell viability studies with Schwann cells demonstrated that ApF/P(LLA-CL) blended nanofibrous scaffolds significantly promoted cell growth as compare to P(LLA-CL), especially when the weight ratio of ApF to P(LLA-CL) was 25:75. The present work provides a basis for further studies of this novel nanofibrous material (ApF/P(LLA-CL)) in peripheral nerve tissue repair or regeneration.

Electrospun nanofibers: Formation, characterization, and evaluation for nerve tissue engineering applications

NASA Astrophysics Data System (ADS)

Zander, Nicole E.

The effects of fiber alignment and surface chemistry, including the covalent attachment and physical adsorption of the extracellular matrix (ECM) proteins laminin and collagen, on the neurite outgrowth of neuron-like PC12 cells were examined. Neuron-like PC12 cells responded to fiber orientation, and were successfully contact-guided by aligned electrospun nanofibers. In addition, fibers with attached protein, either physically adsorbed or covalently attached, improved neurite outgrowth lengths. Furthermore, aligning the fibers and attaching the ECM protein laminin, in particular, significantly improved neurite outgrowth over randomly oriented fibers with laminin. Since this research suggested that protein concentration on the fibers was the dominant driving force for improved neurite outgrowth, the effect of protein concentration, incorporated onto the surface of the nanofibers, on neurite outgrowth was examined. Two ways to control protein concentration on the fibers were explored—the variation of the fiber-protein reaction time and the variation of the protein soaking solution concentration. In addition, analytical methods to quantify the concentration of protein, as well as the protein coverage, on the surface of the fibers were developed. Although most of the fiber mats had multilayer protein coverage, and hence physically adsorbed proteins which could potentially mean a loss in bioactivity, the neuron-like PC12 cell neurites responded in a dose-dependent manner with increased neurite lengths on scaffolds with higher protein concentrations. The work was extended further by forming protein gradients on the fiber mats in hopes of locally directing neurite outgrowth and orientation. Fiber mats with both linear gradients (continuous change in protein concentration) and step gradients (six regions of uniform protein coverage, with protein concentration increasing from region to region) were fabricated and analyzed. The step gradients formed in the aligned fiber direction showed the most promise for use in cell culture assays. While surface chemistry and topography are important, porosity of the scaffold is also critical to control cellular infiltration and tissue formation. To enhance the porosity of our electrospun nanofiber scaffolds and improve the infiltration of cells, two methods were explored to control porosity. In the first method, the scaffold polymer polycaprolactone was co-electrospun with a sacrificial polymer polyethylene oxide, which was removed after the bi-component fiber mat was formed. In doing so, the void space was increased. In the second method, the spinning solution concentration of polycaprolactone was varied to control fiber diameter and porosity. The second method proved to be more effective at improving the cellular infiltration of PC12 cells. Two orders of magnitude range of fiber diameters were achieved, and nearly full infiltration of PC12 cells was observed for the mats with the highest porosity. The pore sizes of these mats were on the order of the size of the cell bodies (approximately 6-10 µm). Although the majority of this work focuses on using conventional electrospinning to generate solid-core fibers, core-shell fibers, have many applications in tissue engineering, among other fields. We explored an efficient way to generate these fibers from an emulsion solution using a conventional electrospinning apparatus. We characterized the fibers using an atomic force microscope (AFM) elastic modulus mapping technique, along with AFM phase imaging, angle-resolved x-ray photoelectron spectroscopy and thermal gravimetric analysis, to determine the chemical and molar composition of the core and shell layers. This work presents novel analytical techniques for the characterization of core-shell nanofibers in order to better predict and understand their material properties. (Abstract shortened by UMI.).

An aligned porous electrospun fibrous membrane with controlled drug delivery - An efficient strategy to accelerate diabetic wound healing with improved angiogenesis.

PubMed

Ren, Xiaozhi; Han, Yiming; Wang, Jie; Jiang, Yuqi; Yi, Zhengfang; Xu, He; Ke, Qinfei

2018-04-01

A chronic wound in diabetic patients is usually characterized by poor angiogenesis and delayed wound closure. The exploration of efficient strategy to significantly improve angiogenesis in the diabetic wound bed and thereby accelerate wound healing is still a significant challenge. Herein, we reported a kind of aligned porous poly (l-lactic acid) (PlLA) electrospun fibrous membranes containing dimethyloxalylglycine (DMOG)-loaded mesoporous silica nanoparticles (DS) for diabetic wound healing. The PlLA electrospun fibers aligned in a single direction and there were ellipse-shaped nano-pores in situ generated onto the surface of fibers, while the DS were well distributed in the fibers and the DMOG as well as Si ion could be controlled released from the nanopores on the fibers. The in vitro results revealed that the aligned porous composite membranes (DS-PL) could stimulate the proliferation, migration and angiogenesis-related gene expression of human umbilical vein endothelial cells (HUVECs) compared with the pure PlLA membranes. The in vivo study further demonstrated that the prepared DS-PL membranes significantly improved neo-vascularization, re-epithelialization and collagen formation as well as inhibited inflammatory reaction in the diabetic wound bed, which eventually stimulated the healing of the diabetic wound. Collectively, these results suggest that the combination of hierarchical structures (nanopores on the aligned fibers) with the controllable released DMOG drugs as well as Si ions from the membranes, which could create a synergetic effect on the rapid stimulation of angiogenesis in the diabetic wound bed, is a potential novel therapeutic strategy for highly efficient diabetic wound healing. A chronic wound in diabetic patients is usually characterized by the poor angiogenesis and the delayed wound closure. The main innovation of this study is to design a new kind of skin tissue engineered scaffold, aligned porous poly (l-lactic acid) (PlLA) electrospun membranes containing dimethyloxalylglycine (DMOG)-loaded mesoporous silica nanoparticles (DS), which could significantly improve angiogenesis in the diabetic wound bed and thereby accelerate diabetic wound healing. The results revealed that the electrospun fibers with ellipse-shaped nano-pores on the surface were aligned in a single direction, while there were DS particles distributed in the fibers and the DMOG as well as Si ions could be controllably released from the nanopores on the fibers. The in vitro studies demonstrated that the hierarchical nanostructures (nanopores on the aligned fibers) and the controllable released chemical active agents (DMOG drugs and Si ions) from the DS-PL membranes could exert a synergistic effect on inducing the endothelial cell proliferation, migration and differentiation. Above all, the scaffolds distinctly induced the angiogenesis, collagen deposition and re-epithelialization as well as inhibited inflammation reaction in the wound sites, which eventually stimulated the healing of diabetic wounds in vivo. The significance of the current study is that the combination of the hierarchical aligned porous nanofibrous structure with DMOG-loaded MSNs incorporated in electrospun fibers may suggest a high-efficiency strategy for chronic wound healing. Copyright © 2018 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Cytocompatibility of Siloxane-Containing Vaterite/Poly(l-lactic acid) Composite Coatings on Metallic Magnesium.

PubMed

Yamada, Shinya; Maeda, Hirotaka; Obata, Akiko; Lohbauer, Ulrich; Yamamoto, Akiko; Kasuga, Toshihiro

2013-12-12

Poly(l-lactic acid)-based films which include 60 wt % of vaterite (V) or siloxane-containing vaterite (SiV) were coated on a pure magnesium substrate, denoted by PLLA/V or PLLA/SiV, respectively, to suppress early corrosion and improve its cytocompatibility. Both coating films adhered to the Mg substrate with 2.3-2.8 MPa of tensile bonding strength. Soaking test for 7 days in α-modified minimum essential medium revealed that the morphological instability of the PLLA/V film caused a higher amount of Mg 2+ ion to be released from the coating sample. On the other hand, in the case of the coating with the PLLA/SiV film, no morphological change even after the soaking test was observed, owing to the suppression of the degradation rate. In cell culture tests, the proliferation of mouse osteoblast-like cell (MC3T3-E1) was significantly enhanced by both coatings, in comparison with the uncoated magnesium substrate. The cell morphology revealed that a few less-spread cells were observed on the PLLA/V film, while more elongated cells were done on the PLLA/SiV film. The cells on the PLLA/SiV film exhibited an extremely higher alkaline phosphatase activity after 21 days of incubation than that on the PLLA/V one. The PLLA/SiV film suppressed the early corrosion and enhanced cytocompatibility on metallic magnesium.

Competing Stereocomplexation and Homocrystallization of Poly(l-lactic acid)/Poly(d-lactic acid) Racemic Mixture: Effects of Miscible Blending with Other Polymers.

PubMed

Bao, Jianna; Xue, Xiaojia; Li, Kai; Chang, Xiaohua; Xie, Qing; Yu, Chengtao; Pan, Pengju

2017-07-20

Promoting the stereocomplexation ability of high-molecular-weight poly(l-lactic acid) (PLLA) and poly(d-lactic acid) (PDLA) is an efficient way to improve the thermal resistance of the resulting materials. Herein, we studied the competing crystallization kinetics, polymorphic crystalline structure, and lamellae structure of the PLLA/PDLA component in its miscible blends with poly(vinyl acetate) (PVAc) and proposed a method to improve the stereocomplexation ability of PLLA and PDLA through miscible blending with the other polymer. Crystallization of the PLLA/PDLA component is suppressed after the addition of PVAc, due to the dilution effect. The stereocomplexation ability of PLLA and PDLA is enhanced by blending with PVAc; this becomes more obvious at a high PVAc content (≥50 wt %) but less significant with the further increase of PLLA, PDLA molecular weights. Almost exclusive formation of SCs is achieved for PLLA and PDLA after blending with a large proportion of PVAc (e.g., 75 wt %). Incorporation of PVAc also facilitates the HC-to-SC structural reorganization upon heating. The increased chain mobility, decreased equilibrium melting point, and enhanced intermolecular interactions may account for the preferential stereocomplexation in PLLA/PDLA/PVAc blends.

Combination of Bioactive Polymeric Membranes and Stem Cells for Periodontal Regeneration: In Vitro and In Vivo Analyses.

PubMed

Gonçalves, Flávia; de Moraes, Míriam Santos; Ferreira, Lorraine Braga; Carreira, Ana Cláudia Oliveira; Kossugue, Patrícia Mayumi; Boaro, Letícia Cristina Cidreira; Bentini, Ricardo; Garcia, Célia Regina da Silva; Sogayar, Mari Cleide; Arana-Chavez, Victor Elias; Catalani, Luiz Henrique

2016-01-01

Regeneration of periodontal tissues requires a concerted effort to obtain consistent and predictable results in vivo. The aim of the present study was to test a new family of bioactive polymeric membranes in combination with stem cell therapy for periodontal regeneration. In particular, the novel polyester poly(isosorbide succinate-co-L-lactide) (PisPLLA) was compared with poly(L-lactide) (PLLA). Both polymers were combined with collagen (COL), hydroxyapatite (HA) and the growth factor bone morphogenetic protein-7 (BMP7), and their osteoinductive capacity was evaluated via in vitro and in vivo experiments. Membranes composed of PLLA/COL/HA or PisPLLA/COL/HA were able to promote periodontal regeneration and new bone formation in fenestration defects in rat jaws. According to quantitative real-time polymerase chain reaction (qRT-PCR) and Alizarin Red assays, better osteoconductive capacity and increased extracellular mineralization were observed for PLLA/COL/HA, whereas better osteoinductive properties were associated with PisPLLA/COL/HA. We concluded that membranes composed of either PisPLLA/COL/HA or PLLA/COL/HA present promising results in vitro as well as in vivo and that these materials could be potentially applied in periodontal regeneration.

Electrospinning of gelatin and SMPU with carbon nanotubes for tissue engineering scaffolds.

PubMed

Mejia, Monica A; Hoyos, Lina M; Zapata, Jenniffer; Restrepo, Luz M; Moneada, Maria E

2016-08-01

The nanofibres created by electrospinning technique are currently used for a variety of applications in tissue engineering; and Gelatin and Polyurethane Shape-Memory (SMPU) have important results in biomedicine. Similarly, carbon nanotubes combined with other biomaterials change important properties, opening new opportunities for biomedical applications. In this work, we constructed scaffold using electrospinning technique based in bovine-hide gelatin, SMPU and both materials hybrid with carbon nanotube. Morphology and cytotoxicity were evaluated and mechanical properties for two materials were obtained in scaffold building. Morphological, mechanical and citotoxic properties of the electrospun fibers were found to be dependent of alteration in materials concentration, electrospinning conditions and MWCNT concentration. According to morphological, cytotoxic and mechanical analysis, SMPU more MWCNT were the best material, with nanofibers of 451 nm, tensile strength of 1.912 MPa, and a high ratio surface volume.

Effect of initiators on synthesis of poly(L-lactide) by ring opening polymerization

NASA Astrophysics Data System (ADS)

Pholharn, D.; Srithep, Y.; Morris, J.

2017-06-01

We studied the effect of several aliphatic alcohols, including1-dodecanol, 1-octanol and methanol, as initiators on synthesis of poly(L-lactide) (PLLA) by ring opening polymerization. The reaction starts with L-lactide monomer and uses stannous octoate as catalyst. Fourier transform infrared spectroscopy and X-ray diffraction analysis verified that PLLAs were produced successfully. Weight, number average molecular weight and polydispersity index of PLLAs were measured by gel permeation chromatography. The PLLA initiated by methanol (PLLA-Meth) presented the highest molecular weight and yield percent. From differential scanning calorimetry, PLLA-Meth showed the highest melting temperature at ∼167°C, crystallization temperature at 110°C and degree of crystallinity 80%. The thermal stability was assessed by thermogravimetric analysis: this confirmed that PLLA-Meth was superior with the highest degradation temperature compared to PLLA initiated by other initiators. We concluded that methanol was the most appropriate initiator for PLLA synthesis by ring opening polymerization.

Mechanoactive Scaffold Induces Tendon Remodeling and Expression of Fibrocartilage Markers

PubMed Central

Spalazzi, Jeffrey P.; Vyner, Moira C.; Jacobs, Matthew T.; Moffat, Kristen L.

2008-01-01

Biological fixation of soft tissue-based grafts for anterior cruciate ligament (ACL) reconstruction poses a major clinical challenge. The ACL integrates with subchondral bone through a fibrocartilage enthesis, which serves to minimize stress concentrations and enables load transfer between two distinct tissue types. Functional integration thus requires the reestablishment of this fibrocartilage interface on reconstructed ACL grafts. We designed and characterized a novel mechanoactive scaffold based on a composite of poly-α-hydroxyester nanofibers and sintered microspheres; we then used the scaffold to test the hypothesis that scaffold-induced compression of tendon grafts would result in matrix remodeling and the expression of fibrocartilage interface-related markers. Histology coupled with confocal microscopy and biochemical assays were used to evaluate the effects of scaffold-induced compression on tendon matrix collagen distribution, cellularity, proteoglycan content, and gene expression over a 2-week period. Scaffold contraction resulted in over 15% compression of the patellar tendon graft and upregulated the expression of fibrocartilage-related markers such as Type II collagen, aggrecan, and transforming growth factor-β3 (TGF-β3). Additionally, proteoglycan content was higher in the compressed tendon group after 1 day. The data suggest the potential of a mechanoactive scaffold to promote the formation of an anatomic fibrocartilage enthesis on tendon-based ACL reconstruction grafts. PMID:18512112

Mechanoactive scaffold induces tendon remodeling and expression of fibrocartilage markers.

PubMed

Spalazzi, Jeffrey P; Vyner, Moira C; Jacobs, Matthew T; Moffat, Kristen L; Lu, Helen H

2008-08-01

Biological fixation of soft tissue-based grafts for anterior cruciate ligament (ACL) reconstruction poses a major clinical challenge. The ACL integrates with subchondral bone through a fibrocartilage enthesis, which serves to minimize stress concentrations and enables load transfer between two distinct tissue types. Functional integration thus requires the reestablishment of this fibrocartilage interface on reconstructed ACL grafts. We designed and characterized a novel mechanoactive scaffold based on a composite of poly-alpha-hydroxyester nanofibers and sintered microspheres; we then used the scaffold to test the hypothesis that scaffold-induced compression of tendon grafts would result in matrix remodeling and the expression of fibrocartilage interface-related markers. Histology coupled with confocal microscopy and biochemical assays were used to evaluate the effects of scaffold-induced compression on tendon matrix collagen distribution, cellularity, proteoglycan content, and gene expression over a 2-week period. Scaffold contraction resulted in over 15% compression of the patellar tendon graft and upregulated the expression of fibrocartilage-related markers such as Type II collagen, aggrecan, and transforming growth factor-beta3 (TGF-beta3). Additionally, proteoglycan content was higher in the compressed tendon group after 1 day. The data suggest the potential of a mechanoactive scaffold to promote the formation of an anatomic fibrocartilage enthesis on tendon-based ACL reconstruction grafts.

Nucleated Poly(L-lactic acid) with N, N‧-oxalyl bis(benzoic acid) dihydrazide

NASA Astrophysics Data System (ADS)

Tian, Liang-Liang; Cai, Yan-Hua

2018-04-01

One of the major challenges in the field of Poly(L-lactic acid) (PLLA) is the enhancement of crystallization. In the present work, the evaluation of the influence of N, N‧-oxalyl bis(benzoic acid) dihydrazide (TBOD), as a novel organic nucleating agent, on the non-isothermal crystallization, melting behavior, and thermal stability of PLLA was performed using differential scanning calorimeter and thermogravimetric analysis. Non-isothermal crystallization measurement revealed that TBOD had an excellent accelerating effect for the crystallization of PLLA in cooling, and upon the addition of 3 wt% TBOD, PLLA exhibited the highest onset crystallization temperature and the crystallization peak temperature, as well as the largest non-isothermal crystallization enthalpy. In particular, when the TBOD concentration was 1 wt% ∼ 3 wt%, the onset crystallization temperatures were higher than the theoretical ceiling temperature of crystallization, thoroughly demonstrating the powerful crystallization promoting ability of TBOD. Additionally, the non-isothermal crystallization behavior of PLLA/TBOD depended on the TBOD concentration, cooling rate as well as the final melting temperature. The melting behavior of PLLA/TBOD after non-isothermal crystallization further confirmed the effect of TBOD on the crystallization process and crystal structure of PLLA, and the appearance of the double melting peaks during melting stages was attribute to the melting-recrystallization. For melting behavior after isothermal crystallization, the crystallization temperature and crystallization time significantly affected the melting behavior of PLLA/TBOD. The addition of TBOD could not change the thermal decomposition profile of the PLLA, but the thermal stability did not regularly decrease with increasing of TBOD concentration, indicating that there might exist intermolecular interaction between PLLA and TBOD.

Cell proliferation on PVA/sodium alginate and PVA/poly(γ-glutamic acid) electrospun fiber.

PubMed

Yang, Jen Ming; Yang, Jhe Hao; Tsou, Shu Chun; Ding, Chian Hua; Hsu, Chih Chin; Yang, Kai Chiang; Yang, Chun Chen; Chen, Ko Shao; Chen, Szi Wen; Wang, Jong Shyan

2016-09-01

To overcome the obstacles of easy dissolution of PVA nanofibers without crosslinking treatment and the poor electrospinnability of the PVA cross-linked nanofibers via electrospinning process, the PVA based electrospun hydrogel nanofibers are prepared with post-crosslinking method. To expect the electrospun hydrogel fibers might be a promising scaffold for cell culture and tissue engineering applications, the evaluation of cell proliferation on the post-crosslinking electrospun fibers is conducted in this study. At beginning, poly(vinyl alcohol) (PVA), PVA/sodium alginate (PVASA) and PVA/poly(γ-glutamic acid) (PVAPGA) electrospun fibers were prepared by electrospinning method. The electrospun PVA, PVASA and PVAPGA nanofibers were treated with post-cross-linking method with glutaraldehyde (Glu) as crosslinking agent. These electrospun fibers were characterized with thermogravimetry analysis (TGA) and their morphologies were observed with a scanning electron microscope (SEM). To support the evaluation and explanation of cell growth on the fiber, the study of 3T3 mouse fibroblast cell growth on the surface of pure PVA, SA, and PGA thin films is conducted. The proliferation of 3T3 on the electrospun fiber surface of PVA, PVASA, and PVAPGA was evaluated by seeding 3T3 fibroblast cells on these crosslinked electrospun fibers. The cell viability on electrospun fibers was conducted with water-soluble tetrazolium salt-1 assay (Cell Proliferation Reagent WST-1). The morphology of the cells on the fibers was also observed with SEM. The results of WST-1 assay revealed that 3T3 cells cultured on different electrospun fibers had similar viability, and the cell viability increased with time for all electrospun fibers. From the morphology of the cells on electrospun fibers, it is found that 3T3 cells attached on all electrospun fiber after 1day seeded. Cell-cell communication was noticed on day 3 for all electrospun fibers. Extracellular matrix (ECM) productions were found and cell-ECM adhesion was shown on day 7. The cell number was also increased on all of the crosslinked electrospun fibers. It seems that the PVA based electrospun hydrogel nanofibers prepared with post-crosslinking method can be used as scaffold for tissue engineering. Copyright © 2016 Elsevier B.V. All rights reserved.

Uniaxial drawing and mechanical properties of poly[(R)-3-hydroxybutyrate]/poly(L-lactic acid) blends.

PubMed

Park, Jun Wuk; Doi, Yoshiharu; Iwata, Tadahisa

2004-01-01

Blends of poly(L-lactic acid) (PLLA) with two kinds of poly[(R)-3-hydroxybutyrate] (PHB) having different molecular weights, commercial-grade bacterial PHB (bacterial-PHB) and ultrahigh molecular weight PHB (UHMW-PHB), were prepared by the solvent-casting method and uniaxially drawn at two drawing temperatures, around PHB's T(g) (2 degrees C) for PHB-rich blends and around PLLA's T(g) (60 degrees C) for PLLA-rich blends. Differential scanning calorimetry analysis showed that this system was immiscible over the entire composition range. Mechanical properties of all of the samples were improved in proportion to the draw ratio. Although PLLA domains in bacterial-PHB-rich blends remained almost unstretched during cold drawing, a good interfacial adhesion between two polymers and the reinforcing role of PLLA components led to enhanced mechanical properties proportionally to the PLLA content at the same draw ratio. On the contrary, in the case of UHMW-PHB-rich blends, the minor component PLLA was found to be also oriented by cold drawing in ice water due to an increase in the interfacial entanglements caused by the very long chain length of the matrix polymer. As a result, their mechanical properties were considerably improved with increasing PLLA content compared with the bacterial-PHB system. Scanning electron microscopy observations on the surface and cross-section revealed that a layered structure with uniformly oriented microporous in the interior was obtained by selectively removal of PLLA component after simple alkaline treatment.

Polymorphic Crystallization and Crystalline Reorganization of Poly(l-lactic acid)/Poly(d-lactic acid) Racemic Mixture Influenced by Blending with Poly(vinylidene fluoride).

PubMed

Yu, Chengtao; Han, Lili; Bao, Jianna; Shan, Guorong; Bao, Yongzhong; Pan, Pengju

2016-08-18

The effects of poly(vinylidene fluoride) (PVDF) on the crystallization kinetics, competing formations of homocrystallites (HCs) and stereocomplexes (SCs), polymorphic crystalline structure, and HC-to-SC crystalline reorganization of the poly(l-lactic acid)/poly(d-lactic acid) (PLLA/PDLA) racemic mixture were investigated. Even though the PLLA/PDLA/PVDF blends are immiscible, blending with PVDF enhances the crystallization rate and SC formation of PLLA/PDLA components at different temperatures that are higher or lower than the melting temperature of the PVDF component; it also facilitates the HC-to-SC melt reorganization upon heating. The crystallization rate and degree of SC crystallinity (Xc,SC) of PLLA/PDLA components in nonisothermal crystallization increase after immiscible blending with PVDF. At different isothermal crystallization temperatures, the crystallization half-time of PLLA/PDLA components decreases; its spherulitic growth rate and Xc,SC increase as the mass fraction of PVDF increases from 0 to 0.5 in the presence of either a solidified or a molten PVDF phase. The HCs formed in primary crystallization of PLLA/PDLA components melt and recrystallize into SCs upon heating; the HC-to-SC melt reorganization is promoted after blending with PVDF. We proposed that the PVDF-promoted crystallization, SC formation, and HC-to-SC melt reorganization of PLLA/PDLA components in PLLA/PDLA/PVDF blends stem from the enhanced diffusion ability of PLLA and PDLA chains.

Cytocompatibility of Siloxane-Containing Vaterite/Poly(l-lactic acid) Composite Coatings on Metallic Magnesium

PubMed Central

Yamada, Shinya; Maeda, Hirotaka; Obata, Akiko; Lohbauer, Ulrich; Yamamoto, Akiko; Kasuga, Toshihiro

2013-01-01

Poly(l-lactic acid)-based films which include 60 wt % of vaterite (V) or siloxane-containing vaterite (SiV) were coated on a pure magnesium substrate, denoted by PLLA/V or PLLA/SiV, respectively, to suppress early corrosion and improve its cytocompatibility. Both coating films adhered to the Mg substrate with 2.3–2.8 MPa of tensile bonding strength. Soaking test for 7 days in α-modified minimum essential medium revealed that the morphological instability of the PLLA/V film caused a higher amount of Mg2+ ion to be released from the coating sample. On the other hand, in the case of the coating with the PLLA/SiV film, no morphological change even after the soaking test was observed, owing to the suppression of the degradation rate. In cell culture tests, the proliferation of mouse osteoblast-like cell (MC3T3-E1) was significantly enhanced by both coatings, in comparison with the uncoated magnesium substrate. The cell morphology revealed that a few less-spread cells were observed on the PLLA/V film, while more elongated cells were done on the PLLA/SiV film. The cells on the PLLA/SiV film exhibited an extremely higher alkaline phosphatase activity after 21 days of incubation than that on the PLLA/V one. The PLLA/SiV film suppressed the early corrosion and enhanced cytocompatibility on metallic magnesium. PMID:28788425

Applications of molecular self-assembly in tissue engineering

NASA Astrophysics Data System (ADS)

Harrington, Daniel Anton

This thesis studied the application of three self-assembling molecular systems, as potential biomaterials for tissue engineering applications. Cholesteryl-(L-lactic acid)n molecules form thermotropic liquid crystals, which could be coated onto the inner and outer pores of biodegradable PLLA scaffolds, while retaining the lamellar order of the neat material. Primary bovine chondrocytes were cultured on these structures, demonstrating improved attachment and extended retention of phenotype on the C-LA-coated scaffolds. No difference in fibronectin adsorption to C-LA and PLLA surfaces was observed, suggesting a strong role for cholesterol in influencing cell phenotype. A family of peptide-amphiphiles, bearing the "RGD" adhesion sequence from fibronectin, was also assessed in the contexts of cartilage and bladder repair. These molecules self-assemble into one-dimensional fibers, with diameters of 6--8 nm, and lengths of 500 nm or greater. Chondrocytes were seeded and cultured on covalently-crosslinked PA gels and embedded within calcium-triggered PA gels. Cells became dormant over time, but remained viable, suggesting an inappropriate display of the adhesion sequence to cells. A family of "branched" PA molecules with lysine dendron headgroups was designed, in an effort to increase the spatial separation between molecules in the assembled state, and to theoretically improve epitope accessibility. These molecules coated reliably onto PGA fiber scaffolds, and dramatically increased the attachment of human bladder smooth muscle cells, possibly through better epitope display or electrostatic attraction. They also formed strong gels with several negatively-charged biologically-relevant macromolecules. In a third system, amphiphilic segmented dendrimers based on phenylene vinylene and L-lysine entered cells through an endocytic pathway with no discernible toxic effect on cell proliferation or morphology. These amphiphiles formed complex aggregates in aqueous solution, likely an equilibrium state of micelles (5--10 nm) and vesicles (25--35 nm). A pyrene analogue was shown to lyse cells, which correlated with the molecule's reduced propensity to form strong aggregates in aqueous solution. Other amino acid segments were substituted for L-lysine, and only those amphiphiles with basic residues were efficiently taken in by cells. For all three self-assembling systems, their nanoscale organization and their interaction with biological systems were directly related to the chemical nature of their constituent building blocks.

Preparation and characterization of biohybrid poly (3-hydroxybutyrate-co-3-hydroxyvalerate) based nanofibrous scaffolds

NASA Astrophysics Data System (ADS)

Kouhi, Monireh; Fathi, Mohammadhossein; Venugopal, Jayarama Reddy; Shamanian, Morteza; Ramakrishna, Seeram

2018-01-01

Development of bioengineered scaffolds for bone tissue regeneration is a growing area of research, especially those involving biodegradable electrospun nanofibers incorporated with ceramic nanoparticles, since they can mimic the extracellular matrix (ECM) of the native bone. In the current study, a biocomposite nanofibrous scaffolds consisting of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), fibrinogen (FIB) and bredigite (BR) nanoparticles was fabricated through electrospinning. The morphological, chemical and mechanical characteristics of the resultant scaffolds were studied by using field emission-scanning electron microscopy (FESEM), Fourier transform infrared spectroscopy (FTIR) and tensile tester, respectively. It was found that PHBV-FIB-BR scaffolds exhibited enhanced tensile strength and young modulus compared to PHBV and PHBV-FIB scaffolds. In addition, the measurements of the water contact angle suggested that incorporation of bredigite and fibrinogen into PHBV could improve the hydrophilicity of the composites. The results of bioactivity assessment performed in the simulated body fluid (SBF) demonstrated that the presence of the bredigite nanoparticles induced the nucleation and growth of apatite layer on the surface of PHBV-FIB-BR scaffold in SBF. Furthermore, the ion concentration changes of SBF solutions with composite scaffolds showed that PHBV-FIB-BR scaffolds released Ca and Si ions, which can stimulate osteoblast proliferation. The results of cell culture studies revealed the higher osteoblast proliferation, mineralization and differentiation on PHBV-FIB-BR and PHBV-FIB scaffolds than on PHBV. Our results suggest that PHBV-FIB-BR nanofibrous scaffold would be a promising candidate as a biocomposite nanofibrous scaffold material for tissue engineering applications.

Bacterial response to different surface chemistries fabricated by plasma polymerization on electrospun nanofibers.

PubMed

Abrigo, Martina; Kingshott, Peter; McArthur, Sally L

2015-12-06

Control over bacterial attachment and proliferation onto nanofibrous materials constitutes a major challenge for a variety of applications, including filtration membranes, protective clothing, wound dressings, and tissue engineering scaffolds. To develop effective devices, the interactions that occur between bacteria and nanofibers with different morphological and physicochemical properties need to be investigated. This paper explores the influence of fiber surface chemistry on bacterial behavior. Different chemical functionalities were generated on the surface of electrospun polystyrene nanofibers through plasma polymerization of four monomers (acrylic acid, allylamine, 1,7-octadiene, and 1,8-cineole). The interactions of Escherichia coli with the surface modified fibers were investigated through a combination of scanning electron microscopy and confocal laser scanning microscopy. Fiber wettability, surface charge, and chemistry were found to affect the ability of bacterial cells to attach and proliferate throughout the nanofiber meshes. The highest proportion of viable cells attachment occurred on the hydrophilic amine rich coating, followed by the hydrophobic octadiene. The acrylic acid coating rich in carboxyl groups showed a significantly lower attraction of bacterial cells. The 1,8-cineole retained the antibacterial activity of the monomer, resulting with a high proportion of dead isolated cells attached onto the fibers. Results showed that the surface chemistry properties of nanofibrous membranes can be strategically tuned to control bacterial behavior.

A novel basalt fiber-reinforced polylactic acid composite for hard tissue repair.

PubMed

Chen, Xi; Li, Yan; Gu, Ning

2010-08-01

A basalt fiber (BF) was, for the first time, introduced into a poly(l-lactic acid) (PLLA) matrix as innovative reinforcement to fabricate composite materials for hard tissue repair. Firstly, BF/PLLA composites and pure PLLA were produced by the methods of solution blending and freeze drying. The results showed that basalt fibers can be uniformly dispersed in the PLLA matrix and significantly improve the mechanical properties and hydrophilicity of the PLLA matrix. The presence of basalt fibers may retard the polymer degradation rate and neutralize the acid degradation from PLLA. Osteoblasts were cultured in vitro to evaluate the cytocompatibility of the composite. An MTT assay revealed that osteoblasts proliferated well for 7 days and there was little difference found in their viability on both PLLA and BF/PLLA films, which was consistent with the alkaline phosphatase (ALP) activity results. A fluorescent staining observation showed that osteoblasts grew well on the composites. SEM images displayed that osteoblasts tended to grow along the fiber axis. The formation of mineralized nodules was observed on the films by Alizarin red S staining. These results suggest that the presence of basalt fibers does not noticeably affect osteoblastic behavior and the designed composites are osteoblast compatible. It is concluded that basalt fibers, as reinforcing fibers, may have promising applications in hard tissue repair.

Piezoelectric Motion of Multilayer Film with Alternate Rows of Optical Isomers of Chiral Polymer Film

NASA Astrophysics Data System (ADS)

Yoshida, Tetsuo; Imoto, Kenji; Nakai, Takaaki; Uwami, Ryouta; Kataoka, Takuya; Inoue, Masataka; Fukumoto, Takahiro; Kamimura, Yuuki; Kato, Atsuko; Tajitsu, Yoshiro

2011-09-01

We realized a multilayer film laminated alternately with poly(L-lactic acid) (PLLA) and poly(D-lactic acid) (PDLA) films in order to improve the piezoelectric performance of the PLLA film. In the fabrication processes, the thicknesses of PLLA and PDLA films were reduced to improve the effective electric field, and a multilayer composed of more than 100 layers (PDLA/PLLA multilayer film) was realized to improve the piezoelectric performance. In general, a single PLLA film has a piezoelectric constant of about 5 pC/N, and it is difficult to observe the piezoelectric resonance in this film of centimeter-order size using a commercial impedance analyzer because of its small Q-value. In contrast, the PDLA/PLLA multilayer film of centimeter-order size has a piezoelectric performance equivalent to that of the piezoelectric material with a piezoelectric constant of 100 pC/N, and also, the piezoelectric resonance can be observed in this film. On the basis of these results, we confirmed that even an object of 259 g mass is made to vibrate under the piezoelectric resonance vibration of this PDLA/PLLA multilayer film. In other words, necessary quantities for actual work as an actuator could be obtained in the PDLA/PLLA multilayer film.

Biodegradable cellulose acetate nanofiber fabrication via electrospinning.

PubMed

Christoforou, Theopisti; Doumanidis, Charalabos

2010-09-01

Nanofiber manufacturing is one of the key advancements in nanotechnology today. Over the past few years, there has been a tremendous growth of research activities to explore electrospinning for nanofiber formation from a rich variety of materials. This quite simple and cost effective process operates on the principle that the solution is extracted under the action of a high electric field. Once the voltage is sufficiently high, a charged jet is ejected following a complicated looping trajectory. During its travel, the solvent evaporates leaving behind randomly oriented nanofibers accumulated on the collector. The combination of their nanoscale dimensionality, high surface area, porosity, flexibility and superior strength makes the electrospun fibers suitable for several value-added applications, such as filters, protecting clothes, high performance structures and biomedical devices. In this study biodegradable cellulose acetate (CA) nanofibrous membranes were produced using electrospinning. The device utilized consisted of a syringe equipped with a metal needle, a microdialysis pump, a high voltage supply and a collector. The morphology of the yielded fibers was determined using SEM. The effect of various parameters, including electric field strength, tip-to-collector distance, solution feed rate and composition on the morphological features of the electrospun fibers was examined. The optimum operating conditions for the production of uniform, non-beaded fibers with submicron diameter were also explored. The biodegradable CA nanofiber membranes are suitable as tissue engineering scaffolds and as reinforcements of biopolymer matrix composites in foils by ultrasonic welding methods.

Design of Nanostructured Biological Materials Through Self-Assembly of Peptides and Proteins

DTIC Science & Technology

2002-01-01

of applications, including scaffolding for tissue repair in regenerative medicine, drug delivery and biological surface engineering. Tirrell and...colleagues [2] designed artificial proteins that undergo self-assembly to form hydrogels responsive to pH and other environmental changes. Ghadiri and...showed that other β-sheet peptide systems can also undergo self-assembly into regular nanofiber structures. Although they share no sequence

In vivo guided vascular regeneration with a non-porous elastin-like polypeptide hydrogel tubular scaffold.

PubMed

Mahara, Atsushi; Kiick, Kristi L; Yamaoka, Tetsuji

2017-06-01

Herein, we demonstrate a new approach for small-caliber vascular reconstruction using a non-porous elastin-like polypeptide hydrogel tubular scaffold, based on the concept of guided vascular regeneration (GVR). The scaffolds are composed of elastin-like polypeptide, (Val-Pro-Gly-Ile-Gly) n , for compliance matching and antithrombogenicity and an Arg-Gly-Asp (RGD) motif for connective tissue regeneration. When the polypeptide was mixed with an aqueous solution of β-[Tris(hydroxymethyl)phosphino]propionic acid at 37°C, the polypeptide hydrogel was rapidly formed. The elastic modulus of the hydrogel was 4.4 kPa. The hydrogel tubular scaffold was formed in a mold and reinforced with poly(lactic acid) nanofibers. When tubular scaffolds with an inner diameter of 1 mm and length of 5 mm were implanted into rat abdominal aortae, connective tissue grew along the scaffold luminal surface from the flanking native tissues, resulting in new blood vessel tissue with a thickness of 200 μm in 1 month. In contrast, rats implanted with control scaffolds without the RGD motif died. These results indicate that the non-porous hydrogel tubular scaffold containing the RGD motif effectively induced rapid tissue regeneration and that GVR is a promising strategy for the regeneration of small-diameter blood vessels. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1746-1755, 2017. © 2017 Wiley Periodicals, Inc.

Piezoelectric antibacterial fabric comprised of poly(l-lactic acid) yarn

NASA Astrophysics Data System (ADS)

Ando, Masamichi; Takeshima, Satoshi; Ishiura, Yutaka; Ando, Kanako; Onishi, Osamu

2017-10-01

A lactic acid monomer has an asymmetric carbon in the molecule, so there are optical isomer l- and d-type. The most widely used poly(lactic acid) (PLA) for commercial applications is poly(l-lactic acid) (PLLA). PLLA is the polymerization product of l-lactide. Certain treatments of PLLA can yield a film that exhibits shear piezoelectricity. Thus, piezoelectric PLLA fiber can be generated by micro slitting piezoelectric PLLA films or by a melt spinning method. We prepared left-handed helical multi fiber yarn (S-yarn) and right-handed helical yarn (Z-yarn) using piezoelectric PLLA fiber. PLLA exhibited shear mode piezoelectricity, causing the electric polarity of the yarn surface to be reversed on the S-yarn and Z-yarn when tension was applied. An SZ-yarn was produced by combining the S-yarn and Z-yarn, and fabric was prepared using the SZ-yarn. This study demonstrated that the fabric has a strong antibacterial effect, which is thought to be due to the strong electric field between the yarns. The field is generated by a piezoelectric effect when the fabric was extended and contracted.

Crystallization, structural relaxation and thermal degradation in Poly(L-lactide)/cellulose nanocrystal renewable nanocomposites.

PubMed

Lizundia, E; Vilas, J L; León, L M

2015-06-05

In this work, crystallization, structural relaxation and thermal degradation kinetics of neat Poly(L-lactide) (PLLA) and its nanocomposites with cellulose nanocrystals (CNC) and CNC-grafted-PLLA (CNC-g-PLLA) have been studied. Although crystallinity degree of nanocomposites remains similar to that of neat homopolymer, results reveal an increase on the crystallization rate by 1.7-5 times boosted by CNC, which act as nucleating agents during the crystallization process. In addition, structural relaxation kinetics of PLLA chains has been drastically reduced by 53% and 27% with the addition of neat and grafted CNC, respectively. The thermal degradation activation energy (E) has been determined from thermogravimetric analysis in the light of Kissinger's and Ozawa-Flynn-Wall theoretical models. Results reveal a reduction on the thermal stability when in presence of CNC-g-PLLA, while raw CNC slightly increases the thermal stability of PLLA. Fourier transform infrared spectroscopy and energy dispersive X-ray spectroscopy results confirm that the presence of residual catalyst in CNC-g-PLLA plays a pivotal role in the thermal degradation behavior of nanocomposites. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enhancement of the interface in poly(L-lactide) and poly(propylidene carbonate) blends by introducing of poly(L-lactide)-grafted graphene oxide to improve mechanical properties

NASA Astrophysics Data System (ADS)

Li, Qi; Qin, Shengxue; Tian, Xiujuan; Chen, Xueyang; Chen, Yunlei; Niu, Yanhua; Zhao, Lifen

2018-03-01

Enhancement of the interfacial structure has great significances in achieving polymer blends with high mechanical performance. To improve the mechanical properties of poly(L-lactide) (PLLA)/poly(propylidene carbonate) (PPC) blends, the covalent functionalized graphene oxide by PLLA chains (PLLA-g-GO) was synthesized by a two-step strategy. It could migrate from the thermally preferred PPC phase to the interfaces of PLLA and PPC and promote the formation of a network-like structure. As a consequence, the tensile strength and elongation at break were both improved. Furthermore, the PLLA-g-GO located at the interface could induce the crystallization at the boundary, which brought the significant improvement of the tensile strength and elongation at break. This result may be beneficial for designing high-performance PLLA materials.

Gamma (γ) irradiated and non-irradiated poly (L-lactide) carboxymethyl starch composite film

NASA Astrophysics Data System (ADS)

Yusof, Mohd Reusmaazran; Shamsudin, Roslinda; Abdullah, Yusof; Yaacob, Norzita

2018-04-01

A film of poly (L-lactide)(PLLA) and carboxymethyl starch (CMS) is prepared by casting evaporation method. The use of CMS blended with PLLA induces the porous film that is potentially used in tissue engineering applications. PLLA is blended with CMS in solution form and rolled on glass to produce a film. The film is then irradiated with gamma-ray (γ) at 10 and 80 kGy. FTIR analysis indicates weak interaction between PLLA and CMS at 10 kGy. Degradation and crosslinking are predicted to have occurred simultaneously at 10 kGy and massive degradation at 80 kGy as indicated in differential scanning calorimetry (DSC) curves. Mechanical analysis shows a higher strength at 10 kGy indicating that crosslinking has occured whereas degradation takes place at higher doses as shown in the reduction of mechanical strength for both PLLA and PLLA/CMS.

Advanced Scaffolds for Dental Pulp and Periodontal Regeneration.

PubMed

Bottino, Marco C; Pankajakshan, Divya; Nör, Jacques E

2017-10-01

No current therapy promotes root canal disinfection and regeneration of the pulp-dentin complex in cases of pulp necrosis. Antibiotic pastes used to eradicate canal infection negatively affect stem cell survival. Three-dimensional easy-to-fit antibiotic-eluting nanofibers, combined with injectable scaffolds, enriched or not with stem cells and/or growth factors, may increase the likelihood of achieving predictable dental pulp regeneration. Periodontitis is an aggressive disease that impairs the integrity of tooth-supporting structures and may lead to tooth loss. The latest advances in membrane biomodification to endow needed functionalities and technologies to engineer patient-specific membranes/constructs to amplify periodontal regeneration are presented. Copyright © 2017 Elsevier Inc. All rights reserved.

The Role of Electrospinning in the Emerging Field of Nanomedicine

PubMed Central

Chew, SY; Wen, Y; Dzenis, Y; Leong, KW

2008-01-01

The fact that in vivo the extracellular matrix or substratum with which cells interact often includes topography at the nanoscale underscores the importance of investigating cell-substrate interactions and performing cell culture at the submicron scale. An important and exciting direction of research in nanomedicine would be to gain an understanding and exploit the cellular response to nanostructures. Electrospinning is a simple and versatile technique that can produce a macroporous scaffold comprising randomly oriented or aligned nanofibers. It can also accommodate the incorporation of drug delivery function into the fibrous scaffold. Endowed with both topographical and biochemical signals such electrospun nanofibrous scaffolds may provide an optimal microenvironment for the seeded cells. This review covers the analysis and control of the electrospinning process, and describes the types of electrospun fibers fabricated for biomedical applications such as drug delivery and tissue engineering. PMID:17168776

Optimization of Polymer-ECM Composite Scaffolds for Tissue Engineering: Effect of Cells and Culture Conditions on Polymeric Nanofiber Mats

PubMed Central

Goyal, Ritu; Guvendiren, Murat; Freeman, Onyi; Mao, Yong; Kohn, Joachim

2017-01-01

The design of composite tissue scaffolds containing an extracellular matrix (ECM) and synthetic polymer fibers is a new approach to create bioactive scaffolds that can enhance cell function. Currently, studies investigating the effects of ECM-deposition and decellularization on polymer degradation are still lacking, as are data on optimizing the stability of the ECM-containing composite scaffolds during prolonged cell culture. In this study, we develop fibrous scaffolds using three polymer compositions, representing slow (E0000), medium (E0500), and fast (E1000) degrading materials, to investigate the stability, degradation, and mechanics of the scaffolds during ECM deposition and decellularization, and during the complete cellularization-decell-recell cycle. We report data on percent molecular weight (% Mw) retention of polymeric fiber mats, changes in scaffold stiffness, ECM deposition, and the presence of fibronectin after decellularization. We concluded that the fast degrading E1000 (Mw retention ≤ 50% after 28 days) was not sufficiently stable to allow scaffold handling after 28 days in culture, while the slow degradation of E0000 (Mw retention ≥ 80% in 28 days) did not allow deposited ECM to replace the polymer support. The scaffolds made from medium degrading E0500 (Mw retention about 60% at 28 days) allowed the gradual replacement of the polymer network with cell-derived ECM while maintaining the polymer network support. Thus, polymers with an intermediate rate of degradation, maintaining good scaffold handling properties after 28 days in culture, seem best suited for creating ECM-polymer composite scaffolds. PMID:28085047

[Biocompatibility of poly-L-lactic acid/Bioglass-guided bone regeneration membranes processed with oxygen plasma].

PubMed

Fang, Wei; Zeng, Shu-Guang; Gao, Wen-Feng

2015-04-01

To prepare and characterize a nano-scale fibrous hydrophilic poly-L-lactic acid/ Bioglass (PLLA/BG) composite membrane and evaluate its biocompatibility as a composite membrane for guiding bone regeneration (GBR). PLLA/BG-guided bone regeneration membrane was treated by oxygen plasma to improved its hydrophilicity. The growth of MG-63 osteoblasts on the membrane was observed using Hoechst fluorescence staining, and the biocompatibility of the membrane was evaluated by calculating the cells adhesion rate and proliferation rate. Osteogenesis of MG-63 cells was assessed by detecting alkaline phosphatase (ALP), and the formation of calcified nodules and cell morphology changes were observed using scanning electron microscope (SEM). The cell adhesion rates of PLLA/BG-guided bone regeneration membrane treated with oxygen plasma were (30.570±0.96)%, (47.27±0.78)%, and (66.78±0.69)% at 1, 3, and 6 h, respectively, significantly higher than those on PLLA membrane and untreated PLLA/BG membrane (P<0.01). The cell proliferation rates on the 3 membranes increased with time, but highest on oxygen plasma-treated PLLA/BG membrane (P<0.01). Hoechst fluorescence staining revealed that oxygen plasma treatment of the PLLA/BG membrane promoted cell adhesion. The membranes with Bioglass promoted the matrix secretion of the osteoblasts. Under SEM, the formation of calcified nodules and spindle-shaped cell morphology were observed on oxygen plasma-treated PLLA/BG membrane. Oxygen plasma-treated PLLA/BG composite membrane has good biocompatibility and can promote adhesion, proliferation and osteogenesis of the osteoblasts.

Synergistic effect of topography, surface chemistry and conductivity of the electrospun nanofibrous scaffold on cellular response of PC12 cells.

PubMed

Tian, Lingling; Prabhakaran, Molamma P; Hu, Jue; Chen, Menglin; Besenbacher, Flemming; Ramakrishna, Seeram

2016-09-01

Electrospun nanofibrous nerve implants is a promising therapy for peripheral nerve injury, and its performance can be tailored by chemical cues, topographical features as well as electrical properties. In this paper, a surface modified, electrically conductive, aligned nanofibrous scaffold composed of poly (lactic acid) (PLA) and polypyrrole (Ppy), referred to as o-PLAPpy_A, was fabricated for nerve regeneration. The morphology, surface chemistry and hydrophilicity of nanofibers were characterized by Scanning Electron Microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS) and water contact angle, respectively. The effects of these nanofibers on neuronal differentiation using PC12 cells were evaluated. A hydrophilic surface was created by Poly-ornithine coating, which was able to provide a better environment for cell attachment, and furthermore aligned fibers were proved to be able to guide PC12 cells grow along the fiber direction and be beneficial for neurite outgrowth. The cellular response of PC12 cells to pulsed electrical stimulation was evaluated by NF 200 and alpha tubulin expression, indicating that electrical stimulation with a voltage of 40mV could enhance the neurite outgrowth. The PC12 cells stimulated with electrical shock showed greater level of neurite outgrowth and smaller cell body size. Moreover, the PC12 cells under electrical stimulation showed better viability. In summary, the o-PLAPpy_A nanofibrous scaffold supported the attachment, proliferation and differentiation of PC12 cells in the absence of electrical stimulation, which could be potential candidate for nerve regeneration applications. Copyright © 2016 Elsevier B.V. All rights reserved.

Modified Filamentous Bacteriophage as a Scaffold for Carbon Nanofiber.

PubMed

Szot-Karpińska, Katarzyna; Golec, Piotr; Leśniewski, Adam; Pałys, Barbara; Marken, Frank; Niedziółka-Jönsson, Joanna; Węgrzyn, Grzegorz; Łoś, Marcin

2016-12-21

With the advent of nanotechnology, carbon nanomaterials such as carbon nanofibers (CNF) have aroused substantial interest in various research fields, including energy storage and sensing. Further improvement of their properties might be achieved via the application of viral particles such as bacteriophages. In this report, we present a filamentous M13 bacteriophage with a point mutation in gene VII (pVII-mutant-M13) that selectively binds to the carbon nanofibers to form 3D structures. The phage-display technique was utilized for the selection of the pVII-mutant-M13 phage from the phage display peptide library. The properties of this phage make it a prospective candidate for a scaffold material for CNFs. The results for binding of CNF by mutant phage were compared with those for maternal bacteriophage (pVII-M13). The efficiency of binding between pVII-mutant-M13 and CNF is about 2 orders of magnitude higher compared to that of the pVII-M13. Binding affinity between pVII-mutant-M13 and CNF was also characterized using atomic force microscopy, scanning electron microscopy, and transmission electron microscopy, which confirmed the specificity of the interaction of the phage pVII-mutant-M13 and the CNF; the binding occurs via the phage's ending, where the mutated pVII protein is located. No similar behavior has been observed for other carbon nanomaterials such as graphite, reduced graphene oxide, single-walled carbon nanotubes, and multiwalled carbon nanotubes. Infrared spectra confirmed differences in the interaction with CNF between the pVII-mutant-M13 and the pVII-M13. Basing on conducted research, we hypothesize that the interactions are noncovalent in nature, with π-π interactions playing the dominant role. Herein, the new bioconjugate material is introduced.

Hydroxyapatite-doped polycaprolactone nanofiber membrane improves tendon-bone interface healing for anterior cruciate ligament reconstruction.

PubMed

Han, Fei; Zhang, Peng; Sun, Yaying; Lin, Chao; Zhao, Peng; Chen, Jiwu

2015-01-01

Hamstring tendon autograft is a routine graft for anterior cruciate ligament (ACL) reconstruction. However, ways of improving the healing between the tendon and bone is often overlooked in clinical practice. This issue can be addressed by using a biomimetic scaffold. Herein, a biomimetic nanofiber membrane of polycaprolactone/nanohydroxyapatite/collagen (PCL/nHAp/Col) is fabricated that mimics the composition of native bone tissue for promoting tendon-bone healing. This membrane has good cytocompatibility, allowing for osteoblast cell adhesion and growth and bone formation. As a result, MC3T3 cells reveal a higher mineralization level in PCL/nHAp/Col membrane compared with PCL membrane alone. Further in vivo studies in ACL reconstruction in a rabbit model shows that PCL/nHAp/Col-wrapped tendon may afford superior tissue integration to nonwrapped tendon in the interface between the tendon and host bone as well as improved mechanical strength. This study shows that PCL/nHAp/Col nanofiber membrane wrapping of autologous tendon is effective for improving tendon healing with host bone in ACL reconstruction.

Improvement of β-TCP/PLLA biodegradable material by surface modification with stearic acid.

PubMed

Ma, Fengcang; Chen, Sai; Liu, Ping; Geng, Fang; Li, Wei; Liu, Xinkuan; He, Daihua; Pan, Deng

2016-05-01

Poly-L-lactide (PLLA) is a biodegradable polymer and used widely. Incorporation of beta tricalcium phosphate (β-TCP) into PLLA can enhance its osteoinductive properties. But the interfacial layer between β-TCP particles with PLLA matrix is easy to be destroyed due to inferior interfacial compatibility of the organic/inorganic material. In this work, a method of β-TCP surface modification with stearic acid was investigated to improve the β-TCP/PLLA biomaterial. The effects of surface modification on the β-TCP were investigated by FTIR, XPS, TGA and CA. It was found that the stearic acid reacted with β-TCP and oxhydryl was formed during the surface modification. Hydrophilicity of untreated or modified β-TCP/PLLA composite was increased by the addition of 10 wt.% β-TCP, but it decreased as the addition amount increased from 10 wt.% to 20 wt.%. Two models were suggested to describe the effect of β-TCP concentration on CA of the composites. Mechanical properties of β-TCP/PLLA composites were tested by bending and tensile tests. Fractures of the composites after mechanical test were observed by SEM. It was found that surface modification with stearic acid improved bending and tensile strengths of the β-TCP/PLLA composites obviously. The SEM results indicated that surface modification decreased the probability of interface debonding between fillers and matrix under load. Copyright © 2016 Elsevier B.V. All rights reserved.

Three-layer microfibrous peripheral nerve guide conduit composed of elastin-laminin mimetic artificial protein and poly(L-lactic acid)

NASA Astrophysics Data System (ADS)

Kakinoki, Sachiro; Nakayama, Midori; Moritan, Toshiyuki; Yamaoka, Tetsuji

2014-07-01

We developed a microfibrous poly(L-lactic acid) (PLLA) nerve conduit with a three-layered structure to simultaneously enhance nerve regeneration and prevent adhesion of surrounding tissue. The inner layer was composed of PLLA microfiber containing 25% elastin-laminin mimetic protein (AG73-(VPGIG)30) that promotes neurite outgrowth. The thickest middle layer was constructed of pure PLLA microfibers that impart the large mechanical stremgth to the conduit. A 10% poly(ethylene glycol) was added to the outer layer to prevent the adhesion with the surrounding tissue. The AG73-(VPGIG)30 composisting of an elastin-like repetitive sequence (VPGIG)30 and a laminin-derived sequence (RKRLQVQLSIRT: AG73) was biosynthesized using Escherichia coli. The PLLA microfibrous conduits were fabricated using an electrospinning procedure. AG73-(VPGIG)30 was successfully mixed in the PLLA microfibers, and the PLLA/AG73-(VPGIG)30 microfibers were stable under physiological conditions. The PLLA/AG73-(VPGIG)30 microfibers enhanced adhesion and neurite outgrowth of PC12 cells. The electrospun microfibrous conduit with a three-layered structure was implanted for bridging a 2.0-cm gap in the tibial nerve of a rabbit. Two months after implantation, no adhesion of surrounding tissue was observed, and the action potential was slightly improved in the nerve conduit with the PLLA/AG73-(VPGIG)30 inner layer.

Effects of protein-coated nanofibers on conformation of gingival fibroblast spheroids: potential utility for connective tissue regeneration.

PubMed

Kaufman, Gili; Whitescarver, Ryan A; Nunes, Laiz; Palmer, Xavier-Lewis; Skrtic, Drago; Tutak, Wojtek

2018-01-24

Deep wounds in the gingiva caused by trauma or surgery require a rapid and robust healing of connective tissues. We propose utilizing gas-brushed nanofibers coated with collagen and fibrin for that purpose. Our hypotheses are that protein-coated nanofibers will: (i) attract and mobilize cells in various spatial orientations, and (ii) regulate the expression levels of specific extracellular matrix (ECM)-associated proteins, determining the initial conformational nature of dense and soft connective tissues. Gingival fibroblast monolayers and 3D spheroids were cultured on ECM substrate and covered with gas-blown poly-(DL-lactide-co-glycolide) (PLGA) nanofibers (uncoated/coated with collagen and fibrin). Cell attraction and rearrangement was followed by F-actin staining and confocal microscopy. Thicknesses of the cell layers, developed within the nanofibers, were quantified by ImageJ software. The expression of collagen1α1 chain (Col1α1), fibronectin, and metalloproteinase 2 (MMP2) encoding genes was determined by quantitative reverse transcription analysis. Collagen- and fibrin- coated nanofibers induced cell migration toward fibers and supported cellular growth within the scaffolds. Both proteins affected the spatial rearrangement of fibroblasts by favoring packed cell clusters or intermittent cell spreading. These cell arrangements resembled the structural characteristic of dense and soft connective tissues, respectively. Within three days of incubation, fibroblast spheroids interacted with the fibers, and grew robustly by increasing their thickness compared to monolayers. While the ECM key components, such as fibronectin and MMP2 encoding genes, were expressed in both protein groups, Col1α1 was predominantly expressed in bundled fibroblasts grown on collagen fibers. This enhanced expression of collagen1 is typical for dense connective tissue. Based on results of this study, our gas-blown, collagen- and fibrin-coated PLGA nanofibers are viable candidates for engineering soft and dense connective tissues with the required structural characteristics and functions needed for wound healing applications. Rapid regeneration of these layers should enhance healing of open wounds in a harsh oral environment.

Gas Permeability and Permselectivity of Poly(L-Lactic Acid)/SiOx Film and Its Application in Equilibrium-Modified Atmosphere Packaging for Chilled Meat.

PubMed

Dong, Tungalag; Song, Shuxin; Liang, Min; Wang, Yu; Qi, Xiaojing; Zhang, Yuqin; Yun, Xueyan; Jin, Ye

2017-01-01

A layer of SiO x was deposited on the surface of poly(L-lactic acid) (PLLA) film to fabricate a PLLA/SiO x layered film, by plasma-enhanced chemical vapor deposition (PECVD) process. PLLA/SiO x film showed Young's modulus and tensile strength increased by 119.2% and 91.6%, respectively, over those of neat PLLA film. At 5 °C, the oxygen (O 2 ) and carbon dioxide (CO 2 ) permeability of PLLA/SiO x film decreased by 78.7% and 71.7%, respectively, and the CO 2 /O 2 permselectivity increased by 32.5%, compared to that of the neat PLLA film. When the PLLA/SiO x film was applied to the equilibrium-modified atmosphere packaging of chilled meat, the gas composition in packaging reached a dynamic equilibrium with 6% to 11% CO 2 and 8% to 13% O 2 . Combined with tea polyphenol pads, which effectively inhibited the microbial growth, the desirable color of meat was maintained and an extended shelf life of 52 d was achieved for the chilled meat. © 2016 Institute of Food Technologists®.

The effect of nanofibrous galactosylated chitosan scaffolds on the formation of rat primary hepatocyte aggregates and the maintenance of liver function.

PubMed

Feng, Zhang-Qi; Chu, Xuehui; Huang, Ning-Ping; Wang, Tao; Wang, Yichun; Shi, Xiaolei; Ding, Yitao; Gu, Zhong-Ze

2009-05-01

Liver tissue engineering requires a perfect extracellular matrix (ECM) for primary hepatocytes culture to maintain high level of liver-specific functions and desirable mechanical stability. The aim of this study was to develop a novel natural nanofibrous scaffold with surface-galactose ligands to enhance the bioactivity and mechanical stability of primary hepatocytes in culture. The nanofibrous scaffold was fabricated by electrospinning a natural material, galactosylated chitosan (GC), into nanofibers with an average diameter of approximately 160 nm. The GC nanofibrous scaffolds displayed slow degradation and suitable mechanical properties as an ECM for hepatocytes according to the evaluation of disintegration and Young's modulus testing. The results of morphology characterization, double-staining fluorescence assay and function detection showed that hepatocytes cultured on GC nanofibrous scaffold formed stably immobilized 3D flat aggregates and exhibited superior cell bioactivity with higher levels of liver-specific function maintenance in terms of albumin secretion, urea synthesis and cytochrome P-450 enzyme than 3D spheroid aggregates formed on GC films. These spheroid aggregates could be detached easily during culture period from the flat GC films. We suggest such GC-based nanofibrous scaffolds could be useful for various applications such as bioartificial liver-assist devices and tissue engineering for liver regeneration as primary hepatocytes culture substrates.

Bioactive Polymeric Materials for Tissue Repair

PubMed Central

Bienek, Diane R.; Tutak, Wojtek; Skrtic, Drago

2017-01-01

Bioactive polymeric materials based on calcium phosphates have tremendous appeal for hard tissue repair because of their well-documented biocompatibility. Amorphous calcium phosphate (ACP)-based ones additionally protect against unwanted demineralization and actively support regeneration of hard tissue minerals. Our group has been investigating the structure/composition/property relationships of ACP polymeric composites for the last two decades. Here, we present ACP’s dispersion in a polymer matrix and the fine-tuning of the resin affects the physicochemical, mechanical, and biological properties of ACP polymeric composites. These studies illustrate how the filler/resin interface and monomer/polymer molecular structure affect the material’s critical properties, such as ion release and mechanical strength. We also present evidence of the remineralization efficacy of ACP composites when exposed to accelerated acidic challenges representative of oral environment conditions. The utility of ACP has recently been extended to include airbrushing as a platform technology for fabrication of nanofiber scaffolds. These studies, focused on assessing the feasibility of incorporating ACP into various polymer fibers, also included the release kinetics of bioactive calcium and phosphate ions from nanofibers and evaluate the biorelevance of the polymeric ACP fiber networks. We also discuss the potential for future integration of the existing ACP scaffolds into therapeutic delivery systems used in the precision medicine field. PMID:28134776

In vitro degradation, hemolysis, and cytocompatibility of PEO/PLLA composite coating on biodegradable AZ31 alloy.

PubMed

Wei, Zhongling; Tian, Peng; Liu, Xuanyong; Zhou, Bangxin

2015-02-01

Magnesium and its alloys have large potential as degradable and absorbable biomaterials because of their mechanical properties and biocompatibility. However, their corrosion resistance is usually inadequate especially in physiological environment, which limits their broad applications in biomedical areas. In this work, plasma electrolytic oxidized/poly(l-lactide) (PEO/PLLA) composite coating was successfully fabricated on biodegradable AZ31 alloy by combing PEO process and sealing with PLLA. The microstructure, elemental composition, and phase composition of the PEO/PLLA composite coating were investigated. The in vitro degradation of the PEO/PLLA composite coating in simulated body fluid (SBF) was also systematically evaluated. The results revealed that the PEO/PLLA composite coating improved the corrosion resistance of AZ31 alloy significantly. The corrosion potential shifted from -1.663V to more positive position -1.317 V and the corrosion current density was reduced with six-order of magnitude. The Mg(2+) ions, hydrogen release, and pH value change of solution caused by degradation were all decreased significantly. Moreover, the PEO process played a critical role in sustaining the integrity of the implant in long-term service. The result of hemolysis test showed that the PEO/PLLA composite coating vested AZ31 alloy a low hemolysis ratio (0.806 ± 0.771)%, which is much lower than the safe value of 5% according to ISO 10993-4. For the cytocompatibility test, compared with bare AZ31 alloy and PEO coating, MC3T3-E1 cells showed much better adhesion and proliferation on the PEO/PLLA composite coating with nearly 4-fold increase of cells after 7-day cultivation, indicating that the PEO/PLLA composite coating has good biocompatibility for biomedical applications. © 2014 Wiley Periodicals, Inc.

Influence of pretreatment on the surface characteristics of PLLA fibers and subsequent hydroxyapatite coating.

PubMed

Peng, F; Olson, J R; Shaw, M T; Wei, M

2009-01-01

A fibrous precursor for bone repair composites was made by coating poly(L-lactide) (PLLA) fibers with hydroxyapatite (HA) using a biomimetic method. To enhance the bonding between the HA coating and the PLLA fiber, PLLA fibers were etched with either sodium hydroxide or sodium hypochlorite to generate carboxyl groups on fiber surfaces. The experiments were designed to determine the influence of etching on the fiber surface morphology and chemistry as well as the subsequent HA coating on the etched fiber surfaces. It was found that the etching pretreatment increased the roughness as well as the hydrophilicity of fibers, indicating that hydrolysis of PLLA chains had taken place on fiber surfaces. The etching pretreatment also promoted HA coating formation by introducing thicker coating on the surface of fibers with a longer etching time, a higher etching concentration, or with NaOCl as the etching agent. A mechanism of surface hydrolysis and oxidation of PLLA was proposed. (c) 2008 Wiley Periodicals, Inc.

Graphene oxide-stimulated myogenic differentiation of C2C12 cells on PLGA/RGD peptide nanofiber matrices

NASA Astrophysics Data System (ADS)

Shin, Y. C.; Lee, J. H.; Kim, M. J.; Hong, S. W.; Oh, J.-W.; Kim, C.-S.; Kim, B.; Hyun, J. K.; Kim, Y.-J.; Han, D.-W.

2015-07-01

During the last decade, much attention has been paid to graphene-based nanomaterials because they are considered as potential candidates for biomedical applications such as scaffolds for tissue engineering and substrates for the differentiation of stem cells. Until now, electrospun matrices composed of various biodegradable copolymers have been extensively developed for tissue engineering and regeneration; however, their use in combination with graphene oxide (GO) is novel and challenging. In this study, nanofiber matrices composed of poly(lactic-co-glycolic acid, PLGA) and M13 phage with RGD peptide displayed on its surface (RGD peptide-M13 phage) were prepared as extracellular matrix (ECM)-mimicking substrates. RGD peptide is a tripeptide (Arg-Gly-Asp) found on ECM proteins that promotes various cellular behaviors. The physicochemical properties of PLGA and RGD peptide-M13 phage (PLGA/RGD peptide) nanofiber matrices were characterized by atomic force microscopy, Fourier-transform infrared spectroscopy and thermogravimetric analysis. In addition, the growth of C2C12 mouse myoblasts on the PLGA/RGD peptide matrices was examined by measuring the metabolic activity. Moreover, the differentiation of C2C12 mouse myoblasts on the matrices when treated with GO was evaluated. The cellular behaviors, including growth and differentiation of C2C12 mouse myoblasts, were substantially enhanced on the PLGA/RGD peptide nanofiber matrices when treated with GO. Overall, these findings suggest that the PLGA/RGD peptide nanofiber matrices can be used in combination with GO as a novel strategy for skeletal tissue regeneration.

Fabrication and characterization of polycaprolactone-graphene powder electrospun nanofibers

NASA Astrophysics Data System (ADS)

Ginestra, Paola; Ghazinejad, Maziar; Madou, Marc; Ceretti, Elisabetta

2016-09-01

Porous fibrous membranes having multiple scales geometries and tailored properties have become attractive microfabrication materials in recent years. Due to the feasibility of incorporating graphene in electrospun nanofibres and the growing interest on these nanomaterials, the present paper focuses on the electrospinning of Poly (ɛ-Caprolactone) (PCL) solutions in the presence of different amounts of Graphene platelets. Electrospinning is a process whereby ultrafine fibers are formed in a high-voltage electrostatic field. The morphological appearance, fiber diameter, and structure of PCL nanofibers produced by the electrospinning process were studied in the presence of different concentration of graphene. Moreover, the effect of a successful incorporation of graphene nanosheets into PCL polymer nanofibers was analyzed. Scanning electron microscope micrographs of the electrospun fibers showed that the average fiber diameter increases in the presence of graphene. Furthermore, the intrinsic properties developed due to the interactions of graphene and PCL improved the mechanical properties of the nanofibers. The results reveal the effect of various graphene concentrations on PCL and the strong interfacial interactions between the graphene platelets phase and the polymer matrix. The functional complexity of the electrospun fibers provides significant advantages over other techniques and shows the promise of these fibers for many applications including air/water filters, sensors, organic solar cells, smart textiles, biocompatible scaffolds for tissue engineering and load-bearing applications. Optimizing deposition efficiency, however, is a necessary milestone for the widespread use of this technique.

Photosensitizer-Embedded Polyacrylonitrile Nanofibers as Antimicrobial Non-Woven Textile

PubMed Central

Stanley, Sarah L.; Scholle, Frank; Zhu, Jiadeng; Lu, Yao; Zhang, Xiangwu; Situ, Xingci; Ghiladi, Reza A.

2016-01-01

Toward the objective of developing platform technologies for anti-infective materials based upon photodynamic inactivation, we employed electrospinning to prepare a non-woven textile comprised of polyacrylonitrile nanofibers embedded with a porphyrin-based cationic photosensitizer; termed PAN-Por(+). Photosensitizer loading was determined to be 34.8 nmol/mg material; with thermostability to 300 °C. Antibacterial efficacy was evaluated against four bacteria belonging to the ESKAPE family of pathogens (Staphylococcus aureus; vancomycin-resistant Enterococcus faecium; Acinetobacter baumannii; and Klebsiella pneumonia), as well as Escherichia coli. Our results demonstrated broad photodynamic inactivation of all bacterial strains studied upon illumination (30 min; 65 ± 5 mW/cm2; 400–700 nm) by a minimum of 99.9996+% (5.8 log units) regardless of taxonomic classification. PAN-Por(+) also inactivated human adenovirus-5 (~99.8% reduction in PFU/mL) and vesicular stomatitis virus (>7 log units reduction in PFU/mL). When compared to cellulose-based materials employing this same photosensitizer; the higher levels of photodynamic inactivation achieved here with PAN-Por(+) are likely due to the combined effects of higher photosensitizer loading and a greater surface area imparted by the use of nanofibers. These results demonstrate the potential of photosensitizer-embedded polyacrylonitrile nanofibers to serve as scalable scaffolds for anti-infective or self-sterilizing materials against both bacteria and viruses when employing a photodynamic inactivation mode of action. PMID:28335205

Characterization of poly(L-lactide/Propylene glycol) based polyurethane films using ATR-FTIR spectroscopy

NASA Astrophysics Data System (ADS)

Manap, Siti Munirah; Ahmad, Azizan; Anuar, Farah Hannan

2016-11-01

A polyurethane films consisting of PLLA, PPG and PLLA-PPG were prepared using solution casting method. Three types of polyurethane were prepared: PPLA:PMDI, PPG:PMDI and PLLA-PPG:PMDI in the presence of polymeric diphenylmethane diisocyanate (PMDI) as the coupling agent and catalyst, Sn(Oct)2. The aim of this research was to improve the physicals properties of PLLA and PPG homopolymers through copolymerization between the two polymers. The homopolymers and polyurethane films were characterized using ATR-FTIR spectroscopy. Chemical reaction between PLLA, PPG and PMDI before and after the reaction were confirmed by observing the shifting of wavenumber for the carbonyl and ether group. Other than that, the additional band for N-H after the reaction indicated that the reaction was successful.

Film Sensor Device Fabricated by a Piezoelectric Poly(L-lactic acid) Film

NASA Astrophysics Data System (ADS)

Ando, Masamichi; Kawamura, Hideki; Kageyama, Keisuke; Tajitsu, Yoshiro

2012-09-01

Synthetic piezoelectric polymer films produced from petroleum feedstock have long been used as thin-film sensors and actuators. However, the fossil fuel requirements for synthetic polymer production and carbon dioxide emission from its combustion have raised concern about the environmental impact of its continued use. Eco-friendly biomass polymers, such as poly(L-lactic acid) (PLLA), are made from plant-based (vegetable starch) plastics and, thus, have a much smaller carbon footprint. Additionally, PLLA does not exhibit pyroelectricity or unnecessary poling. This suggests the usefulness of PLLA films for the human-machine interface (HMI). As an example of a new HMI, we have produced a TV remote control using a PLLA film. The intuitive operation provided by this PLLA device suggests that it is useful for the elderly or handicapped.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yue, Mengyao; Zhou, Baoming; Jiao, Kunyan

A switchable surface that promotes either hydrophobic or hydrophilic wettability of poly (L-lactide) (PLLA) microfibrous membranes is obtained by CF₄ microwave plasma treatment in this paper. The results indicated that both etching and grafting process occurred during the CF₄ plasma treatment and these two factors synergistically affected the final surface wettability of PLLA membranes. When plasma treatment was taken under a relatively low power, the surface wettability of PLLA membranes turned from hydrophobic to hydrophilic. Especially when CF₄ plasma treatment was taken under 100 W for 10 min and 150 W for 5 min, the water contact angle sharply decreasedmore » from 116 ± 3.0° to ~0°. According to Field-emission scanning electron microscopy (FESEM) results, the PLLA fibers were notably etched by CF₄ plasma treatment. Combined with the X-ray photoelectron spectroscopy (XPS) measurements, only a few fluorine-containing groups were grafted onto the surface, so the etching effect directly affected the surface wettability of PLLA membranes in low plasma power condition. However, with the plasma power increasing to 200 W, the PLLA membrane surface turned to hydrophobic again. In contrast, the morphology changes of PLLA fiber surfaces were not obvious while a large number of fluorine-containing groups grafted onto the surface. So the grafting effect gradually became the major factor for the final surface wettability.« less

Biodegradable polymer for sealing porous PEO layer on pure magnesium: An in vitro degradation study

NASA Astrophysics Data System (ADS)

Alabbasi, Alyaa; Mehjabeen, Afrin; Kannan, M. Bobby; Ye, Qingsong; Blawert, Carsten

2014-05-01

An attempt was made to seal the porous silicate-based plasma electrolytic oxidation (PEO) layer on pure magnesium (Mg) with a biodegradable polymer, poly(L-lactide) (PLLA), to delay the localized degradation of magnesium-based implants in body fluid for better in-service mechanical integrity. Firstly, a silicate-based PEO coating on pure magnesium was performed using a pulsed constant current method. In order to seal the pores in the PEO layer, PLLA was coated using a two-step spin coating method. The performance of the PEO-PLLA Mg was evaluated using electrochemical impedance spectroscopy (EIS) and potentiodynamic polarization. The EIS results showed that the polarization resistance (Rp) of the PEO-PLLA Mg was close to two orders of magnitude higher than that of the PEO Mg. While the corrosion current density (icorr) of the pure Mg was reduced by 65% with the PEO coating, the PEO-PLLA coating reduced the icorr by almost 100%. As expected, the Rp of the PEO-PLLA Mg decreased with increase in exposure time. However, it was noted that the Rp of the PEO-PLLA Mg even after 100 h was six times higher than that of the PEO Mg after 48 h exposure, and did not show any visible localized attack.

Healing improvement after rotator cuff repair using gelatin-grafted poly(L-lactide) electrospun fibrous membranes.

PubMed

Zhao, Song; Xie, Xiaoxing; Pan, Guoqing; Shen, Peng; Zhao, Jinzhong; Cui, Wenguo

2015-01-01

Rotator cuff tears (RCTs) are a common cause of shoulder pain and disability in middle and older age. Despite improvements in the understanding of this disease process and advances in surgical treatment, rotator cuff (RC) repair failure rates remain high. Insufficient healing capacity is likely the main factor for failure of reconstruction. We fabricated implantable biodegradable gelatin-grafted poly(L-lactide) (PLLA) fibrous membranes using electrospinning technology and evaluated them using in vitro cell proliferation assays. Then, we established chronic rat RCT models and randomly assigned rats into one of three groups. In group 1 (n = 48), the detached supraspinatus tendon was repaired to its anatomic footprint (transosseous repair). In groups 2 and 3, the rats underwent transosseous repair and were implanted with either pure PLLA membranes (n = 48) or gelatin-PLLA membranes (n = 48) to augment the repairs. The animals were killed at 2, 4, and 8 wk postoperatively, which was followed by histomorphometric and biomechanical evaluation. Histologic observations revealed that gelatin-PLLA membranes have excellent biocompatibility and biodegradability. At 2, 4, and 8 wk postoperatively, the gelatin-PLLA membranes significantly increased the area of glycosaminoglycan staining at the tendon-bone interface compared with the control group (P < 0.05) and significantly improved collagen organization, as measured by birefringence under polarized light at the healing enthesis compared with the control and PLLA groups (P < 0.05). Biomechanical testing revealed that the gelatin-PLLA group had a greater ultimate load to failure and stiffness than the control group at 4 and 8 wk (P < 0.05). The gelatin-PLLA membranes had the highest stress of the healing enthesis. Local application of gelatin-PLLA fibrous membranes to the healing tendon-bone interface after RC repair in a rat chronic RCT model was found to strengthen the healing enthesis, increase the area of fibrocartilage, and improve collagen organization compared with repair alone. Augmentation with gelatin-grafted PLLA may enhance healing after RC repair and might eventually lead to improvement of clinical surgical outcomes. Copyright © 2015 Elsevier Inc. All rights reserved.

Novel lactoferrin-conjugated amphiphilic poly(aminoethyl ethylene phosphate)/poly(L-lactide) copolymer nanobubbles for tumor-targeting ultrasonic imaging

PubMed Central

Luo, Binhua; Liang, Huageng; Zhang, Shengwei; Qin, Xiaojuan; Liu, Xuhan; Liu, Wei; Zeng, Fuqing; Wu, Yun; Yang, Xiangliang

2015-01-01

In the study reported here, a novel amphiphilic poly(aminoethyl ethylene phosphate)/poly(L-lactide) (PAEEP-PLLA) copolymer was synthesized by ring-opening polymerization reaction. The perfluoropentane-filled PAEEP-PLLA nanobubbles (NBs) were prepared using the O1/O2/W double-emulsion and solvent-evaporation method, with the copolymer as the shell and liquid perfluoropentane as the core of NBs. The prepared NBs were further conjugated with lactoferrin (Lf) for tumor-cell targeting. The resulting Lf-conjugated amphiphilic poly(aminoethyl ethylene phosphate)/poly(L-lactide) nanobubbles (Lf-PAEEP-PLLA NBs) were characterized by photon correlation spectroscopy, polyacrylamide gel electrophoresis, Fourier transform infrared spectroscopy, and transmission electron microscopy. The average size of the Lf-PAEEP-PLLA NBs was 328.4±5.1 nm, with polydispersity index of 0.167±0.020, and zeta potential of −12.6±0.3 mV. Transmission electron microscopy imaging showed that the Lf-PAEEP-PLLA NBs had a near-spherical structure, were quite monodisperse, and there was a clear interface between the copolymer shell and the liquid core inside the NBs. The Lf-PAEEP-PLLA NBs also exhibited good biocompatibility in cytotoxicity and hemolysis studies and good stability during storage. The high cellular uptake of Lf-PAEEP-PLLA NBs in C6 cells (low-density lipoprotein receptor-related protein 1-positive cells) at concentrations of 0–20 µg/mL indicated that the Lf provided effective targeting for brain-tumor cells. The in vitro acoustic behavior of Lf-PAEEP-PLLA NBs was evaluated using a B-mode clinical ultrasound imaging system. In vivo ultrasound imaging was performed on tumor-bearing BALB/c nude mice, and compared with SonoVue® microbubbles, a commercial ultrasonic contrast agent. Both in vitro and in vivo ultrasound imaging indicated that the Lf-PAEEP-PLLA NBs possessed strong, long-lasting, and tumor-enhanced ultrasonic contrast ability. Taken together, these results indicate that Lf-PAEEP-PLLA NBs represent a promising nano-sized ultrasonic contrast agent for tumor-targeting ultrasonic imaging. PMID:26396514

Novel lactoferrin-conjugated amphiphilic poly(aminoethyl ethylene phosphate)/poly(L-lactide) copolymer nanobubbles for tumor-targeting ultrasonic imaging.

PubMed

Luo, Binhua; Liang, Huageng; Zhang, Shengwei; Qin, Xiaojuan; Liu, Xuhan; Liu, Wei; Zeng, Fuqing; Wu, Yun; Yang, Xiangliang

2015-01-01

In the study reported here, a novel amphiphilic poly(aminoethyl ethylene phosphate)/poly(L-lactide) (PAEEP-PLLA) copolymer was synthesized by ring-opening polymerization reaction. The perfluoropentane-filled PAEEP-PLLA nanobubbles (NBs) were prepared using the O1/O2/W double-emulsion and solvent-evaporation method, with the copolymer as the shell and liquid perfluoropentane as the core of NBs. The prepared NBs were further conjugated with lactoferrin (Lf) for tumor-cell targeting. The resulting Lf-conjugated amphiphilic poly(aminoethyl ethylene phosphate)/poly(L-lactide) nanobubbles (Lf-PAEEP-PLLA NBs) were characterized by photon correlation spectroscopy, polyacrylamide gel electrophoresis, Fourier transform infrared spectroscopy, and transmission electron microscopy. The average size of the Lf-PAEEP-PLLA NBs was 328.4±5.1 nm, with polydispersity index of 0.167±0.020, and zeta potential of -12.6±0.3 mV. Transmission electron microscopy imaging showed that the Lf-PAEEP-PLLA NBs had a near-spherical structure, were quite monodisperse, and there was a clear interface between the copolymer shell and the liquid core inside the NBs. The Lf-PAEEP-PLLA NBs also exhibited good biocompatibility in cytotoxicity and hemolysis studies and good stability during storage. The high cellular uptake of Lf-PAEEP-PLLA NBs in C6 cells (low-density lipoprotein receptor-related protein 1-positive cells) at concentrations of 0-20 µg/mL indicated that the Lf provided effective targeting for brain-tumor cells. The in vitro acoustic behavior of Lf-PAEEP-PLLA NBs was evaluated using a B-mode clinical ultrasound imaging system. In vivo ultrasound imaging was performed on tumor-bearing BALB/c nude mice, and compared with SonoVue(®) microbubbles, a commercial ultrasonic contrast agent. Both in vitro and in vivo ultrasound imaging indicated that the Lf-PAEEP-PLLA NBs possessed strong, long-lasting, and tumor-enhanced ultrasonic contrast ability. Taken together, these results indicate that Lf-PAEEP-PLLA NBs represent a promising nano-sized ultrasonic contrast agent for tumor-targeting ultrasonic imaging.

Reinforcement of poly-l-lactic acid electrospun membranes with strontium borosilicate bioactive glasses for bone tissue engineering.

PubMed

Fernandes, João S; Gentile, Piergiorgio; Martins, Margarida; Neves, Nuno M; Miller, Cheryl; Crawford, Aileen; Pires, Ricardo A; Hatton, Paul; Reis, Rui L

2016-10-15

Herein, for the first time, we combined poly-l-lactic acid (PLLA) with a strontium borosilicate bioactive glass (BBG-Sr) using electrospinning to fabricate a composite bioactive PLLA membrane loaded with 10% (w/w) of BBG-Sr glass particles (PLLA-BBG-Sr). The composites were characterised by scanning electron microscopy (SEM) and microcomputer tomography (μ-CT), and the results showed that we successfully fabricated smooth and uniform fibres (1-3μm in width) with a homogeneous distribution of BBG-Sr microparticles (<45μm). Degradation studies (in phosphate buffered saline) demonstrated that the incorporation of BBG-Sr glass particles into the PLLA membranes increased their degradability and water uptake with a continuous release of cations. The addition of BBG-Sr glass particles enhanced the membrane's mechanical properties (69% higher Young modulus and 36% higher tensile strength). Furthermore, cellular in vitro evaluation using bone marrow-derived mesenchymal stem cells (BM-MSCs) demonstrated that PLLA-BBG-Sr membranes promoted the osteogenic differentiation of the cells as demonstrated by increased alkaline phosphatase activity and up-regulated osteogenic gene expression (Alpl, Sp7 and Bglap) in relation to PLLA alone. These results strongly suggest that the composite PLLA membranes reinforced with the BBG-Sr glass particles have potential as an effective biomaterial capable of promoting bone regeneration. PLLA membranes were reinforced with 10% (w/w) of strontium-bioactive borosilicate glass microparticles, and their capacity to induce the osteogenic differentiation of bone marrow mesenchymal stem cells (BM-MSCs) was evaluated. These membranes presented an increased: degradability, water uptake, Young modulus and tensile strength. We also demonstrated that these membranes are non-cytotoxic and promote the attachment of BM-MSCs. The addition of the glass microparticles into the PLLA membranes promoted the increase of ALP activity (under osteogenic conditions), as well as the BM-MSCs osteogenic differentiation as shown by the upregulation of Alpl, Sp7 and Bglap gene expression. Overall, we demonstrated that the reinforcement of PLLA with glass microparticles results in a biomaterial with the appropriate properties for the regeneration of bone tissue. Copyright © 2016 Acta Materialia Inc. All rights reserved.

Spontaneous Differentiation of Human Mesenchymal Stem Cells on Poly-Lactic-Co-Glycolic Acid Nano-Fiber Scaffold.

PubMed

Sonomoto, Koshiro; Yamaoka, Kunihiro; Kaneko, Hiroaki; Yamagata, Kaoru; Sakata, Kei; Zhang, Xiangmei; Kondo, Masahiro; Zenke, Yukichi; Sabanai, Ken; Nakayamada, Shingo; Sakai, Akinori; Tanaka, Yoshiya

2016-01-01

Mesenchymal stem cells (MSCs) have immunosuppressive activity and can differentiate into bone and cartilage; and thus seem ideal for treatment of rheumatoid arthritis (RA). Here, we investigated the osteogenesis and chondrogenesis potentials of MSCs seeded onto nano-fiber scaffolds (NFs) in vitro and possible use for the repair of RA-affected joints. MSCs derived from healthy donors and patients with RA or osteoarthritis (OA) were seeded on poly-lactic-glycolic acid (PLGA) electrospun NFs and cultured in vitro. Healthy donor-derived MSCs seeded onto NFs stained positive with von Kossa at Day 14 post-stimulation for osteoblast differentiation. Similarly, MSCs stained positive with Safranin O at Day 14 post-stimulation for chondrocyte differentiation. Surprisingly, even cultured without any stimulation, MSCs expressed RUNX2 and SOX9 (master regulators of bone and cartilage differentiation) at Day 7. Moreover, MSCs stained positive for osteocalcin, a bone marker, and simultaneously also with Safranin O at Day 14. On Day 28, the cell morphology changed from a spindle-like to an osteocyte-like appearance with processes, along with the expression of dentin matrix protein-1 (DMP-1) and matrix extracellular phosphoglycoprotein (MEPE), suggesting possible differentiation of MSCs into osteocytes. Calcification was observed on Day 56. Expression of osteoblast and chondrocyte differentiation markers was also noted in MSCs derived from RA or OA patients seeded on NFs. Lactic acid present in NFs potentially induced MSC differentiation into osteoblasts. Our PLGA scaffold NFs induced MSC differentiation into bone and cartilage. NFs induction process resembled the procedure of endochondral ossification. This finding indicates that the combination of MSCs and NFs is a promising therapeutic technique for the repair of RA or OA joints affected by bone and cartilage destruction.

Fabrication of high-performance poly(l-lactic acid)/lignin-graft-poly(d-lactic acid) stereocomplex films.

PubMed

Liu, Rui; Dai, Lin; Hu, Li-Qiu; Zhou, Wen-Qin; Si, Chuan-Ling

2017-11-01

The need for green renewable alternatives such as lignin to traditional fillers has driven recent interest in polylactic acid blend materials. Herein, lignin-graft-polylactic acid copolymers (LG-g-PDLA, LG-g-PDLLA, and LG-g-PLLA) have been synthesized via ring-opening polymerization of d-, dl-, and l-lactic acid. Then poly(l-lactic acid)/lignin-graft-polylactic acid (PLLA/LG-g-PDLA, /LG-g-PDLLA, and /LG-g-PLLA) complex films have been prepared. The results showed that, compared with LG-g-PDLA and LG-g-PLLA, a small amount of LG-g-PDLA addition could improve the crystallization rate, reduce the glass transition temperature and cold crystallization temperature of PLLA due to the stereocomplex crystallites. The thermal stability, tensile strength and strain of the stereocomplex films were also enhanced. Moreover, the PLLA/LG-g-PDLA films have good ultraviolet resistance and excellent biocompatibility. This study provides a green approach to design advanced polylactic acid-based blends with renewable natural resources. Copyright © 2017 Elsevier B.V. All rights reserved.

Low temperature dielectric relaxation of poly (L-lactic acid) (PLLA) by Thermally Stimulated Depolarization Current

NASA Astrophysics Data System (ADS)

Mishra Patidar, Manju; Jain, Deepti; Nath, R.; Ganesan, V.

2016-10-01

Poly (L-lactic acid) (PLLA) is a biodegradable and biocompatible polyester that can be produced by renewable resources, like corn. Being non-toxic to human body, PLLA is used in biomedical applications, like surgical sutures, bone fixation devices, or controlled drug delivery. Besides its application studies, very few experiments have been done to study its dielectric relaxation in the low temperature region. Keeping this in mind we have performed a low temperature thermally stimulated depolarization current (TSDC) studies over the temperature range of 80K-400K to understand the relaxation phenomena of PLLA. We could observe a multi modal broad relaxation of small but significant intensity at low temperatures while a sharp and high intense peak around glass transition temperature, Tg∼ 333K, of PLLA has appeared. The fine structure of the low temperature TSDC peak may be attributed to the spherulites formation of crystallite regions inter twinned with the polymer as seen in AFM and appear to be produced due to an isothermal crystallization process. XRD analysis also confirms the semicrystalline nature of the PLLA film.

Synergistic effect of scaffold composition and dynamic culturing environment in multilayered systems for bone tissue engineering.

PubMed

Rodrigues, Márcia T; Martins, Albino; Dias, Isabel R; Viegas, Carlos A; Neves, Nuno M; Gomes, Manuela E; Reis, Rui L

2012-11-01

Bone extracellular matrix (ECM) is composed of mineralized collagen fibrils which support biological apatite nucleation that participates in bone outstanding properties. Understanding and mimicking bone morphological and physiological parameters at a biological scale is a major challenge in tissue engineering scaffolding. Using emergent (nano)technologies scaffold designing may be critically improved, enabling highly functional tissue substitutes for bone applications. This study aims to develop novel biodegradable composite scaffolds of tricalcium phosphate (TCPs) and electrospun nanofibers of poly(ϵ-caprolactone) (PCL), combining TCPs osteoconductivity with PCL biocompatibility and elasticity, mimicking bone structure and composition. We hypothesized that scaffolds with such structure/composition would stimulate the proliferation and differentiation of bone marrow stromal cells (BMSCs) towards the osteogenic phenotype. Composite scaffolds, developed by electrospining using consecutive stacked layers of PCL and TCPs, were characterized by FTIR spectroscopy, X-Ray diffraction and scanning electronic microscopy. Cellular behavior was assessed in goat BMSCs seeded onto composite scaffolds and cultured in static or dynamic conditions, using basal or osteogenic media during 7, 14 or 21 days. Cellular proliferation was quantified and osteogenic differentiation confirmed by alkaline phosphatase activity, alizarin red staining and immunocytochemistry for osteocalcin and collagen I. Results suggest that PCL-TCP scaffolds provide a 3D support for gBMSCs proliferation and osteogenic differentiation with production of ECM. TCPs positively stimulate the osteogenic process, especially under dynamic conditions, where PCL-TCP scaffolds are sufficient to promote osteogenic differentiation even in basal medium conditions. The enhancement of the osteogenic potential in dynamic conditions evidences the synergistic effect of scaffold composition and dynamic stimulation in gBMSCs osteogenic differentiation. Copyright © 2012 John Wiley & Sons, Ltd.

Switchable hydrophobic/hydrophilic surface of electrospun poly (l-lactide) membranes obtained by CF₄microwave plasma treatment

DOE PAGES

Yue, Mengyao; Zhou, Baoming; Jiao, Kunyan; ...

2014-11-29

A switchable surface that promotes either hydrophobic or hydrophilic wettability of poly (L-lactide) (PLLA) microfibrous membranes is obtained by CF₄ microwave plasma treatment in this paper. The results indicated that both etching and grafting process occurred during the CF₄ plasma treatment and these two factors synergistically affected the final surface wettability of PLLA membranes. When plasma treatment was taken under a relatively low power, the surface wettability of PLLA membranes turned from hydrophobic to hydrophilic. Especially when CF₄ plasma treatment was taken under 100 W for 10 min and 150 W for 5 min, the water contact angle sharply decreasedmore » from 116 ± 3.0° to ~0°. According to Field-emission scanning electron microscopy (FESEM) results, the PLLA fibers were notably etched by CF₄ plasma treatment. Combined with the X-ray photoelectron spectroscopy (XPS) measurements, only a few fluorine-containing groups were grafted onto the surface, so the etching effect directly affected the surface wettability of PLLA membranes in low plasma power condition. However, with the plasma power increasing to 200 W, the PLLA membrane surface turned to hydrophobic again. In contrast, the morphology changes of PLLA fiber surfaces were not obvious while a large number of fluorine-containing groups grafted onto the surface. So the grafting effect gradually became the major factor for the final surface wettability.« less

Enhancing the Biomechanical Performance of Anisotropic Nanofibrous Scaffolds in Tendon Tissue Engineering: Reinforcement with Cellulose Nanocrystals.

PubMed

Domingues, Rui M A; Chiera, Silvia; Gershovich, Pavel; Motta, Antonella; Reis, Rui L; Gomes, Manuela E

2016-06-01

Anisotropically aligned electrospun nanofibrous scaffolds based on natural/synthetic polymer blends have been established as a reasonable compromise between biological and biomechanical performance for tendon tissue engineering (TE) strategies. However, the limited tensile properties of these biomaterials restrict their application in this field due to the load-bearing nature of tendon/ligament tissues. Herein, the use of cellulose nanocrystals (CNCs) as reinforcing nanofillers in aligned electrospun scaffolds based on a natural/synthetic polymer blend matrix, poly-ε-caprolactone/chitosan (PCL/CHT) is reported. The incorporation of small amounts of CNCs (up to 3 wt%) into tendon mimetic nanofiber bundles has a remarkable biomaterial-toughing effect (85% ± 5%, p < 0.0002) and raises the scaffolds mechanical properties to tendon/ligament relevant range (σ = 39.3 ± 1.9 MPa and E = 540.5 ± 83.7 MPa, p < 0.0001). Aligned PCL/CHT/CNC nanocomposite fibrous scaffolds meet not only the mechanical requirements for tendon TE applications but also provide tendon mimetic extracellular matrix (ECM) topographic cues, a key feature for maintaining tendon cell's morphology and behavior. The strategy proposed here may be extended to other anisotropic aligned nanofibrous scaffolds based on natural/synthetic polymer blends and enable the full exploitation of the advantages provided by their tendon mimetic fibrous structures in tendon TE. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Fabrication and durable antibacterial properties of 3D porous wet electrospun RCSC/PCL nanofibrous scaffold with silver nanoparticles

NASA Astrophysics Data System (ADS)

Zhang, Mei; Lin, Han; Wang, Yilong; Yang, Guang; Zhao, He; Sun, Dahui

2017-08-01

Electrospunnanofibers are used as three-dimensional (3D) scaffold materials that can alter cell attachment and cell proliferation, change the antibacterial properties of materials, and can be used as wound dressings. But the fabrication of porous 3D scaffold structure and the antibacterial properties enhancing are challenges remained to improve. With the states here, a Ranachensinensis skin collagen (RCSC)/poly(ɛ-caprolactone) (PCL)AgNP-loaded3D nanofiber scaffold is fabricated as a wound dressing material by using an improved wet electrospinning method (blending). The nanoscale of the AgNPs is proved. The 3D porous morphologies of the materials with different AgNP loadings, are determined with field emission scanning electron microscopy (FESEM) and the presence and uniformity distribution of AgNPs is confirmed by Energy dispersive X-ray (EDX) spectroscopy. The silver-ion release rates, antibacterial properties, and cytotoxicities of dressing materials with different AgNP contents are evaluated using ICP-AES, the zone inhibition method, and MTT testing. These results showed that the improved wet electrospun is an effective way to fabricate AgNP loaded 3D scaffold materials with porous structure and nearly 90% porosity and the presence of AgNPs in dressing materials strengthen the antibacterial properties. The RCSC/PCL 3D scaffold materials containing 2.0%AgNP would be promising for dressing materials application nearly without cytotoxicities.

PEGylated graphene oxide-mediated quercetin-modified collagen hybrid scaffold for enhancement of MSCs differentiation potential and diabetic wound healing.

PubMed

Chu, Jing; Shi, Panpan; Yan, Wenxia; Fu, Jinping; Yang, Zhi; He, Chengmin; Deng, Xiaoyuan; Liu, Hanping

2018-05-24

Nanoscale delivery based on polyethylene glycol (PEG)ylated graphene oxide (GO-PEG) merits attention for biomedical applications owing to its functional surface modification, superior solubility/biocompatibility and controllable drug release capability. However, impaired skin regeneration in applications of these fascinating nanomaterials in diabetes is still limited, and critical issues need to be addressed regarding insufficient collagen hyperplasia and inadequate blood supply. Therefore, a high-performance tissue engineering scaffold with biocompatible and biodegradable properties is essential for diabetic wound healing. Natural and artificial acellular dermal matrix (ADM) scaffolds with spatially organized collagen fibers can provide a suitable architecture and environment for cell attachment and proliferation. Here, a novel collagen-nanomaterial-drug hybrid scaffold was constructed from GO-PEG-mediated quercetin (GO-PEG/Que)-modified ADM (ADM-GO-PEG/Que). The resulting unique and versatile hybrid scaffold exhibited multiple advantages, including the following: a biocompatible, cell-adhesive surface for accelerating mesenchymal stem cell (MSC) attachment and proliferation; superior stability and adjustability of the conduction potential of quercetin for inducing the differentiation of MSCs into adipocytes and osteoblasts; and a biodegradable nanofiber interface for promoting collagen deposition and angiogenesis in diabetic wound repair. This study provides new prospects for the design of innovative GO-PEG-based collagen hybrid scaffolds for application in efficient therapeutic drug delivery, stem cell-based therapies, tissue engineering and regenerative medicine.

Functionalization of PCL-3D Electrospun Nanofibrous Scaffolds for Improved BMP2-Induced Bone Formation.

PubMed

Miszuk, Jacob M; Xu, Tao; Yao, Qingqing; Fang, Fang; Childs, Josh D; Hong, Zhongkui; Tao, Jianning; Fong, Hao; Sun, Hongli

2018-03-01

Bone morphogenic protein 2 (BMP2) is a key growth factor for bone regeneration, possessing FDA approval for orthopedic applications. BMP2 is often required in supratherapeutic doses clinically, yielding adverse side effects and substantial treatment costs. Considering the crucial role of materials for BMPs delivery and cell osteogenic differentiation, we devote to engineering an innovative bone-matrix mimicking niche to improve low dose of BMP2-induced bone formation. Our previous work describes a novel technique, named thermally induced nanofiber self-agglomeration (TISA), for generating 3D electrospun nanofibrous (NF) polycaprolactone (PCL) scaffolds. TISA process could readily blend PCL with PLA, leading to increased osteogenic capabilities in vitro , however, these bio-inert synthetic polymers produced limited BMP2-induced bone formation in vivo. We therefore hypothesize that functionalization of NF 3D PCL scaffolds with bone-like hydroxyapatite (HA) and BMP2 signaling activator phenamil will provide a favorable osteogenic niche for bone formation at low doses of BMP2. Compared to PCL-3D scaffolds, PCL/HA-3D scaffolds demonstrated synergistically enhanced osteogenic differentiation capabilities of C2C12 cells with phenamil. Importantly, in vivo studies showed this synergism was able to generate significantly increased new bone in an ectopic mouse model, suggesting PCL/HA-3D scaffolds act as a favorable synthetic extracellular matrix for bone regeneration.

Polymeric biomaterials for nerve regeneration applications: From promoting cellular organization to the delivery of bioactive molecules

NASA Astrophysics Data System (ADS)

Delgado-Rivera, Roberto L.

Thousands of new cases of injury to the central nervous system (CNS) occur each year in the USA and all over the world. However, despite recent advances, at present there is no cure for the resulting paraplegia or quadriplegia. This research is directed towards engineering biomaterial platforms to promote cellular organization at the surface of polymer scaffolds that will be conducive to proper regeneration of injured CNS. In addition, the formulation of a delivery system for neuroactive molecules using polymer-based materials will be evaluated to establish its potential to treat CNS disorders. Initial studies involved the chemical modification of an electrospun nonwoven matrix of nanofibers with fibroblast growth factor 2 (FGF-2). Nanofibers alone up-regulated FGF-2, albeit to a lesser extent than nanofibers covalently modified with FGF-2. These results underscore the importance of both surface topography and growth factor presentation on cellular function. Moreover, that FGF-2 modified nanofibrillar scaffolds may demonstrate utility in tissue engineering applications for replacement and regeneration of damaged tissue following CNS injury or disease. Subsequent research efforts focused on a novel micropatterning technique called microscale plasma-initiated patterning (microPIP). This patterning method uses a polydimethylsiloxane (PDMS) stamp to selectively protect regions of an underlying substrate from oxygen plasma treatment resulting in hydrophobic and hydrophilic regions. FGF-2 and laminin-1 were applied to an electrospun polyamide nanofibrillar matrix following plasma treatment. In this work it, was possible to demonstrate that textured surfaces, such as nanofibrillar scaffolds, can be micropatterned to provide external chemical cues for cellular organization. Finally, a microsphere system capable of encapsulating proteins while minimizing the mechanisms of protein degradation and providing a controlled release was investigated. Microspheres were comprised of a salicylic-acid based poly(anhydride-ester) (PAE), a biodegradable polymer that incorporates salicylic acid into the polymer backbone (PolyAspirin). The use of microspheres formulated from PolyAspirin as a delivery vehicle can be advantageous due its ability of performing a dual delivery; biomolecule (protein) and drug. By combining these two properties, it will be possible to release neurotrophic factors to induce a biological response while mitigating inflammatory pathways due to the localized delivery of salicylic acid.

Engineering of near infrared fluorescent proteinoid-poly(L-lactic acid) particles for in vivo colon cancer detection.

PubMed

Kolitz-Domb, Michal; Grinberg, Igor; Corem-Salkmon, Enav; Margel, Shlomo

2014-08-12

The use of near-infrared (NIR) fluorescence imaging techniques has gained great interest for early detection of cancer owing to the negligible absorption and autofluorescence of water and other intrinsic biomolecules in this region. The main aim of the present study is to synthesize and characterize novel NIR fluorescent nanoparticles based on proteinoid and PLLA for early detection of colon tumors. The present study describes the synthesis of new proteinoid-PLLA copolymer and the preparation of NIR fluorescent nanoparticles for use in diagnostic detection of colon cancer. These fluorescent nanoparticles were prepared by a self-assembly process in the presence of the NIR dye indocyanine green (ICG), a FDA-approved NIR fluorescent dye. Anti-carcinoembryonic antigen antibody (anti-CEA), a specific tumor targeting ligand, was covalently conjugated to the P(EF-PLLA) nanoparticles through the surface carboxylate groups using the carbodiimide activation method. The P(EF-PLLA) nanoparticles are stable in different conditions, no leakage of the encapsulated dye into PBS containing 4% HSA was detected. The encapsulation of the NIR fluorescent dye within the P(EF-PLLA) nanoparticles improves significantly the photostability of the dye. The fluorescent nanoparticles are non-toxic, and the biodistribution study in a mouse model showed they evacuate from the body over 24 h. Specific colon tumor detection in a chicken embryo model and a mouse model was demonstrated for anti-CEA-conjugated NIR fluorescent P(EF-PLLA) nanoparticles. The results of this study suggest a significant advantage of NIR fluorescence imaging using NIR fluorescent P(EF-PLLA) nanoparticles over colonoscopy. In future work we plan to broaden this study by encapsulating cancer drugs such as paclitaxel and/or doxorubicin, within these biodegradable NIR fluorescent P(EF-PLLA) nanoparticles, for both detection and therapy of colon cancer.

Tissue-Engineered Nanofibrous Nerve Grafts for Enhancing the Rate of Nerve Regeneration

DTIC Science & Technology

2014-10-01

Acid/BSA, water, chitosan , and water. We used 30 µg of BSA in 50 µL of PBS for loading into each scaffold. 12 As seen from Fig. 10 the BSA had...publication we have shown a novel methodology and feasibility of electrospinning chitosan alone or in combination with synthetic polymers through...Awad, H. M., Nagarale, R. K., Kumbar, S.G., Smart Methodology to Fabricate Electrospun Chitosan Nanofiber Matrices for Regenerative Engineering

Efficient and reusable polyamide-56 nanofiber/nets membrane with bimodal structures for air filtration.

PubMed

Liu, Bowen; Zhang, Shichao; Wang, Xueli; Yu, Jianyong; Ding, Bin

2015-11-01

Nanofibrous media that both possess high airborne particle interception efficiency and robust air permeability would have broad technological implications for areas ranging from individual protection and industrial security to environmental governance; however, creating such filtration media has proved extremely challenging. Here we report a strategy to construct the bio-based polyamide-56 nanofiber/nets (PA-56 NFN) membranes with bimodal structures for effective air filtration via one-step electrospinning/netting. The PA-56 membranes are composed of completely covered two-dimensional (2D) ultrathin (∼20 nm) nanonets which are optimized by facilely regulating the solution concentration, and the bonded scaffold fibers constructed cavity structures which are synchronously created by using the CH3COOH inspiration. With integrated properties of small aperture, high porosity, and bonded scaffold, the resulting PA-56 NFN membranes exhibit high filtration efficiency of 99.995%, low pressure drop of 111 Pa, combined with large dust holding capacity of 49 g/m(2) and dust-cleaning regeneration ability, for filtrating ultrafine airborne particles in the most safe manner involving sieving principle and surface filtration. The successful synthesis of PA-56 NFN medium would not only make it a promising candidate for air filtration, but also provide new insights into the design and development of nanonet-based bimodal structures for various applications. Copyright © 2015 Elsevier Inc. All rights reserved.

Cell studies of hybridized carbon nanofibers containing bioactive glass nanoparticles using bone mesenchymal stromal cells

NASA Astrophysics Data System (ADS)

Zhang, Xiu-Rui; Hu, Xiao-Qing; Jia, Xiao-Long; Yang, Li-Ka; Meng, Qing-Yang; Shi, Yuan-Yuan; Zhang, Zheng-Zheng; Cai, Qing; Ao, Yin-Fang; Yang, Xiao-Ping

2016-12-01

Bone regeneration required suitable scaffolding materials to support the proliferation and osteogenic differentiation of bone-related cells. In this study, a kind of hybridized nanofibrous scaffold material (CNF/BG) was prepared by incorporating bioactive glass (BG) nanoparticles into carbon nanofibers (CNF) via the combination of BG sol-gel and polyacrylonitrile (PAN) electrospinning, followed by carbonization. Three types (49 s, 68 s and 86 s) of BG nanoparticles were incorporated. To understand the mechanism of CNF/BG hybrids exerting osteogenic effects, bone marrow mesenchymal stromal cells (BMSCs) were cultured directly on these hybrids (contact culture) or cultured in transwell chambers in the presence of these materials (non-contact culture). The contributions of ion release and contact effect on cell proliferation and osteogenic differentiation were able to be correlated. It was found that the ionic dissolution products had limited effect on cell proliferation, while they were able to enhance osteogenic differentiation of BMSCs in comparison with pure CNF. Differently, the proliferation and osteogenic differentiation were both significantly promoted in the contact culture. In both cases, CNF/BG(68 s) showed the strongest ability in influencing cell behaviors due to its fastest release rate of soluble silicium-relating ions. The synergistic effect of CNF and BG would make CNF/BG hybrids promising substrates for bone repairing.

Synthesis of cellulose diacetate based copolymer electrospun nanofibers for tissues scaffold

NASA Astrophysics Data System (ADS)

Liang, Wencheng; Hou, Jia; Fang, Xiangchen; Bai, Fudong; Zhu, Tonghe; Gao, Feifei; Wei, Chao; Mo, Xiumei; Lang, Meidong

2018-06-01

In this study, a novel cellulose diacetate based copolymer used as tissues scaffold, cellulose diacetate-graft-poly(ethylene terephthalate) (CDA-g-PET) was developed by "graft onto" strategy using 3-Isocyanatomethyl-3,5,5-trimethylcyc-lohexyl isocyanate (IPDI) as a coupling reagent of cellulose diacetate and poly(ethylene terephthalate), and using dibutyltin dilaurate (DBTDL) and 1-butyl-3-methylimidazolium chloride salt ([Bmim]Cl) as catalysts. CDA-g-PET copolymers with five different grafting ratios were obtained by the regulation of the reaction time. It was proved by the FT-IR spectra of the purified copolymers that PET had been successfully grafted onto CDA backbone. Afterwards, CDA-g-PET nanofibers were fabricated via electrospinning and further were cross-linked by means of treating in glutaraldehyde (25%wt) aqueous solution for 48 h. The uniform and smooth fiber morphology was proved by SEM and the diameter decreased with the increase of grafting ratio. Moreover, the value of TGA revealed that the grafting PET onto CDA backbone would improve heat-resistant quality of CDA and help to improve the ability of thermo processing. The graft of PET onto CDA significantly enhanced mechanical property of copolymer compared with CDA. The results of hemolysis ratio indicated that hemolysis ratio has decreased compared with CDA, highlighting the potential application in the field of contacting with blood. In vitro cell viability indicated that CDA-g-PET would enhance biocompatibility compared with CDA.

Virgin olive oil blended polyurethane micro/nanofibers ornamented with copper oxide nanocrystals for biomedical applications.

PubMed

Amna, Touseef; Hassan, M Shamshi; Yang, Jieun; Khil, Myung-Seob; Song, Ki-Duk; Oh, Jae-Don; Hwang, Inho

2014-01-01

Recently, substantial interest has been generated in using electrospun biomimetic nanofibers of hybrids, particularly organic/inorganic, to engineer different tissues. The present work, for the first time, introduced a unique natural and synthetic hybrid micronanofiber wound dressing, composed of virgin olive oil/copper oxide nanocrystals and polyurethane (PU), developed via facile electrospinning. The as-spun organic/inorganic hybrid micronanofibers were characterized by scanning electron microscopy (SEM), energy dispersive X-ray analysis, X-ray diffraction, electron probe microanalysis, and transmission electron microscopy. The interaction of cells with scaffold was studied by culturing NIH 3T3 fibroblasts on an as-spun hybrid micronanofibrous mat, and viability, proliferation, and growth were assessed. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay results and SEM observation showed that the hybrid micronanofibrous scaffold was noncytotoxic to fibroblast cell culture and was found to benefit cell attachment and proliferation. Hence our results suggest the potential utilization of as-spun micronanoscaffolds for tissue engineering. Copper oxide-olive oil/PU wound dressing may exert its positive beneficial effects at every stage during wound-healing progression, and these micronanofibers may serve diverse biomedical applications, such as tissue regeneration, damaged skin treatment, wound healing applications, etc. Conclusively, the fabricated olive oil-copper oxide/PU micronanofibers combine the benefits of virgin olive oil and copper oxide, and therefore hold great promise for biomedical applications in the near future.

Electrospinning as a powerful technique for biomedical applications: a critically selected survey.

PubMed

Villarreal-Gómez, Luis Jesús; Cornejo-Bravo, José Manuel; Vera-Graziano, Ricardo; Grande, Daniel

2016-01-01

Nowadays, electrospinning has become one of the most versatile, easy, and cost-effective techniques to engineer advanced materials used for many applications, especially in the biomedical and environmental areas. Like the numerous patents around the world, the increasing number of papers witnesses the huge potential of this simple process, and many companies have been emerged during the last years to exploit its innumerable applications. This article presents a critically selected overview of polymers that can be used to produce nanofibers, along with the biomedical applications of the resulting electrospun scaffolds. We have focused on about seven natural and synthetic polymers, but many more can be found in the literature, either as their pristine state or as composites with ceramics, metals, and other polymers. The description of some strategies for nanofiber production, and the characterization used to evaluate their optimization, has been discussed. Finally, several polymers have been recognized as highlights for future work.

Evaluation of Changes in Morphology and Function of Human Induced Pluripotent Stem Cell Derived Cardiomyocytes (HiPSC-CMs) Cultured on an Aligned-Nanofiber Cardiac Patch

PubMed Central

Khan, Mahmood; Xu, Yanyi; Hua, Serena; Johnson, Jed; Belevych, Andriy; Janssen, Paul M. L.; Gyorke, Sandor; Guan, Jianjun; Angelos, Mark G.

2015-01-01

Introduction Dilated cardiomyopathy is a major cause of progressive heart failure. Utilization of stem cell therapy offers a potential means of regenerating viable cardiac tissue. However, a major obstacle to stem cell therapy is the delivery and survival of implanted stem cells in the ischemic heart. To address this issue, we have developed a biomimetic aligned nanofibrous cardiac patch and characterized the alignment and function of human inducible pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) cultured on this cardiac patch. This hiPSC-CMs seeded patch was compared with hiPSC-CMs cultured on standard flat cell culture plates. Methods hiPSC-CMs were cultured on; 1) a highly aligned polylactide-co-glycolide (PLGA) nanofiber scaffold (~50 microns thick) and 2) on a standard flat culture plate. Scanning electron microscopy (SEM) was used to determine alignment of PLGA nanofibers and orientation of the cells on the respective surfaces. Analysis of gap junctions (Connexin-43) was performed by confocal imaging in both the groups. Calcium cycling and patch-clamp technique were performed to measure calcium transients and electrical coupling properties of cardiomyocytes. Results SEM demonstrated >90% alignment of the nanofibers in the patch which is similar to the extracellular matrix of decellularized rat myocardium. Confocal imaging of the cardiomyocytes demonstrated symmetrical alignment in the same direction on the aligned nanofiber patch in sharp contrast to the random appearance of cardiomyocytes cultured on a tissue culture plate. The hiPSC-CMs cultured on aligned nanofiber cardiac patches showed more efficient calcium cycling compared with cells cultured on standard flat surface culture plates. Quantification of mRNA with qRT-PCR confirmed that these cardiomyocytes expressed α-actinin, troponin-T and connexin-43 in-vitro. Conclusions Overall, our results demonstrated changes in morphology and function of human induced pluripotent derived cardiomyocytes cultured in an anisotropic environment created by an aligned nanofiber patch. In this environment, these cells better approximate normal cardiac tissue compared with cells cultured on flat surface and can serve as the basis for bioengineering of an implantable cardiac patch. PMID:25993466

Evaluation of Changes in Morphology and Function of Human Induced Pluripotent Stem Cell Derived Cardiomyocytes (HiPSC-CMs) Cultured on an Aligned-Nanofiber Cardiac Patch.

PubMed

Khan, Mahmood; Xu, Yanyi; Hua, Serena; Johnson, Jed; Belevych, Andriy; Janssen, Paul M L; Gyorke, Sandor; Guan, Jianjun; Angelos, Mark G

2015-01-01

Dilated cardiomyopathy is a major cause of progressive heart failure. Utilization of stem cell therapy offers a potential means of regenerating viable cardiac tissue. However, a major obstacle to stem cell therapy is the delivery and survival of implanted stem cells in the ischemic heart. To address this issue, we have developed a biomimetic aligned nanofibrous cardiac patch and characterized the alignment and function of human inducible pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) cultured on this cardiac patch. This hiPSC-CMs seeded patch was compared with hiPSC-CMs cultured on standard flat cell culture plates. hiPSC-CMs were cultured on; 1) a highly aligned polylactide-co-glycolide (PLGA) nanofiber scaffold (~50 microns thick) and 2) on a standard flat culture plate. Scanning electron microscopy (SEM) was used to determine alignment of PLGA nanofibers and orientation of the cells on the respective surfaces. Analysis of gap junctions (Connexin-43) was performed by confocal imaging in both the groups. Calcium cycling and patch-clamp technique were performed to measure calcium transients and electrical coupling properties of cardiomyocytes. SEM demonstrated >90% alignment of the nanofibers in the patch which is similar to the extracellular matrix of decellularized rat myocardium. Confocal imaging of the cardiomyocytes demonstrated symmetrical alignment in the same direction on the aligned nanofiber patch in sharp contrast to the random appearance of cardiomyocytes cultured on a tissue culture plate. The hiPSC-CMs cultured on aligned nanofiber cardiac patches showed more efficient calcium cycling compared with cells cultured on standard flat surface culture plates. Quantification of mRNA with qRT-PCR confirmed that these cardiomyocytes expressed α-actinin, troponin-T and connexin-43 in-vitro. Overall, our results demonstrated changes in morphology and function of human induced pluripotent derived cardiomyocytes cultured in an anisotropic environment created by an aligned nanofiber patch. In this environment, these cells better approximate normal cardiac tissue compared with cells cultured on flat surface and can serve as the basis for bioengineering of an implantable cardiac patch.

Effects of protein-coated nanofibers on conformation of gingival fibroblast spheroids: potential utility for connective tissues regeneration.

PubMed

Kaufman, Gili; Whitescarver, Ryan; Nunes, Laiz; Palmer, Xavier-Lewis; Skrtic, Drago; Tutak, Wojtek

2017-10-09

Deep wounds in the gingiva caused by trauma or surgery require a rapid and robust healing of connective tissues. We propose utilizing gas-brushed nanofibers coated with collagen and fibrin for that purpose. Our hypotheses are that protein-coated nanofibers will: (i) attract and mobilize cells in various spatial orientations, and (ii) regulate the expression levels of specific extracellular matrix (ECM)-associated proteins, determining the initial conformational nature of dense and soft connective tissues. Gingival fibroblast monolayers and 3D spheroids were cultured on ECM substrate and covered with gas-blown poly-(DL-lactide-co-glycolide) (PLGA) nanofibers (uncoated/coated with collagen and fibrin). Cell attraction and rearrangement was followed by F-actin staining and confocal microscopy. Thicknesses of the cell layers, developed within the nanofibers, were quantified by imageJ software. The expression of collagen1α1 chain (Col1α1), fibronectin, and metalloproteinase 2 (MMP2) encoding genes was determined by quantitative reverse transcription analysis. Collagen- and fibrin- coated nanofibers induced cell migration toward fibers and supported cellular growth within the scaffolds. Both proteins affected the spatial rearrangement of fibroblasts by favoring packed cell clusters or intermittent cell spreading. These cell arrangements resembled the structural characteristic of dense and soft connective tissues, respectively. Within 3 days of incubation, fibroblast spheroids interacted with the fibers and grew robustly by increasing their thickness compared to monolayers. While the ECM key components, such as fibronectin and MMP2 encoding genes, were expressed in both protein groups, Col1α1 was predominantly expressed in bundled fibroblasts grown on collagen fibers. This enhanced expression of collagen1 is typical for dense connective tissue. Based on results of this study, our gas-blown, collagen- and fibrin-coated PLGA nanofibers are viable candidates for engineering soft and dense connective tissues with the required structural characteristics and functions needed for wound healing applications. Rapid regeneration of these layers should enhance healing of open wounds in a harsh oral environment. © 2017 IOP Publishing Ltd.

Pickering-type water-in-oil-in-water multiple emulsions toward multihollow nanocomposite microspheres.

PubMed

Maeda, Hayata; Okada, Masahiro; Fujii, Syuji; Nakamura, Yoshinobu; Furuzono, Tsutomu

2010-09-07

Multihollow hydroxyapatite (HAp)/poly(L-lactic acid) (PLLA) nanocomposite microspheres were readily fabricated by solvent evaporation from a "Pickering-type" water-in-(dichloromethane solution of PLLA)-in-water multiple emulsion stabilized with HAp nanoparticles. The multiple emulsion was stabilized with the aid of PLLA molecules used as a wettability modifier for HAp nanoparticles, although HAp nanoparticles did not work solely as particulate emulsifiers for Pickering-type emulsions consisting of pure dichloromethane and water. The interaction between PLLA and HAp nanoparticles at the oil-water interfaces plays a crucial role toward the preparation of stable multiple emulsion and multihollow microspheres.

Application of nanosheets as an anti-adhesion barrier in partial hepatectomy.

PubMed

Niwa, Daisuke; Koide, Masatsugu; Fujie, Toshinori; Goda, Nobuhito; Takeoka, Shinji

2013-10-01

Postoperative adhesion often causes serious adverse effects such as bowl obstruction, chronic abdominal pain, pelvic pain, and infertility. We previously reported that a poly-L-lactic acid (PLLA) nanosheet can efficiently seal a surgical incision without scarring. In this report, we examined whether the PLLA nanosheet can form an effective anti-adhesion barrier in partial hepatectomy accompanied by severe hemorrhaging in rats. To evaluate the anti-adhesive property of the nanosheet, the liver wound surface was covered with TachoComb(®) , a well-known hemostat material used in clinical procedures, and then with the PLLA nanosheet. Dressing the wound surface with TachoComb(®) alone caused severe adhesion with omentum and/or residual parts of the liver. By contrast, combinational usage of TachoComb(®) and the PLLA nanosheet significantly reduced such adhesion, presumably by inhibiting the permeation of oozing blood cells and the infiltration of fibroblastic cells. Moreover, the nanosheet displayed low permeability against serum proteins as well as cells in vitro, supporting the notion that the PLLA nanosheet has anti-adhesive properties in vivo. These results strongly suggested that the PLLA nanosheet is a promising material for reducing unwanted postoperative adhesion. Copyright © 2013 Wiley Periodicals, Inc.

Advantages and challenges offered by biofunctional core-shell fiber systems for tissue engineering and drug delivery.

PubMed

Sperling, Laura E; Reis, Karina P; Pranke, Patricia; Wendorff, Joachim H

2016-08-01

Whereas highly porous scaffolds composed of electrospun nanofibers can mimick major features of the extracellular matrix in tissue engineering, they lack the ability to incorporate and release biocompounds (drugs, growth factors) safely in a controlled way. Here, electrospun core-shell fibers (core made from water and aqueous solutions of hydrophilic polymers and the shell from materials with well-defined release mechanisms) offer unique advantages in comparison with those that have helped make porous nanofibrillar scaffolds highly successful in tissue engineering. This review considers the preparation and biofunctionalization of such core-shell fibers as well as applications in various areas, including neural, vascular, cardiac, cartilage and bone tissue engineering, and touches on the topic of clinical trials. Copyright © 2016 Elsevier Ltd. All rights reserved.

Optimizing injectable poly-L-lactic acid administration for soft tissue augmentation: The rationale for three treatment sessions

PubMed Central

Bauer, Ute; Graivier, Miles H

2011-01-01

BACKGROUND: The availability and variety of different injectable modalities has led to a dramatic increase in soft tissue augmentation procedures in recent years. Injectable poly-L-lactic acid (PLLA) is a synthetic, biodegradable polymer device approved in the United States for use in immunocompetent patients as a single regimen of up to four treatment sessions for correction of shallow to deep nasolabial fold contour deficiencies and other facial wrinkles. Injectable PLLA is also approved for restoration and/or correction of signs of facial fat loss (lipoatrophy) in individuals with HIV. METHODS: The present article provides an overview of previous studies with injectable PLLA, and specifically focuses on the number of recommended treatment sessions and intervals between treatment sessions. The authors also provide two case studies to support their recommendations for an average of three treatment sessions. RESULTS: Although the specific mechanisms remain hypothetical, injections of PLLA are believed to cause a cascade of cellular events that lead to collagen repair and subsequent restoration of facial volume. Because the development of a response to injectable PLLA is gradual and its duration of effect is long lasting, sufficient time between treatment sessions should be allocated to avoid overcorrection. CONCLUSION: Studies of injectable PLLA support the hypothesized mode of operation, and the experience and clinical recommendations of the authors that suggest that three treatment sessions are an optimal regimen for use of injectable PLLA in the majority of patients. PMID:22942665

Effect of Mg content on the thermal stability and mechanical behaviour of PLLA/Mg composites processed by hot extrusion.

PubMed

Cifuentes, S C; Lieblich, M; López, F A; Benavente, R; González-Carrasco, J L

2017-03-01

In the field of bioabsorbable composites for biomedical applications, extrusion has been employed as a method to prepare homogeneous blends of polymeric matrices with bioactive ceramic fillers. In this work, the suitability of processing poly-l-lactic acid/Magnesium (PLLA/Mg) composites by hot extrusion has been assessed by a systematic characterization of PLLA/Mg composites containing different amounts of Mg particles up to 7wt%. The results show that extrusion causes a reduction of almost 20% in the viscosity average molecular weight of PLLA, which further decreases with increasing Mg content. Extrusion gave always rise to a homogeneous distribution of Mg particles within the PLLA matrix. This composite processing was not compromised by the degradation of the polymeric matrix because the processing temperature was always below the onset degradation temperature. In the processing conditions employed in the present work, degradation of the composite slightly increases as more Mg is added up to 5wt%, but is very high at 7wt%. This was also evident from the mechanical behaviour, so that Mg particles improved the stiffness and compression strength of neat PLLA until 5wt% of Mg content, which dropped drastically when the material had 7wt% of Mg. The filler strengthening factor decreases with the increment in Mg content. In order to obtain an optimised contribution of Mg particles, a balance between thermal degradation and mechanical resistance of PLLA must be achieved. Copyright © 2016 Elsevier B.V. All rights reserved.

Characterization of Silk/Poly 3-Hydroxybutyrate-chitosan-multi-walled Carbon Nanotube Micro-nano Scaffold: A New Hybrid Scaffold for Tissue Engineering Applications.

PubMed

Mirmusavi, Mohammad Hossein; Karbasi, Saeed; Semnani, Dariush; Kharazi, Anousheh Zargar

2018-01-01

Long-term healing tissue engineering scaffolds must hold its full mechanical strength at least for 12 weeks. Nano-micro scaffolds consist of electrospinning nanofibers and textile microfibers to support cell behavior and mechanical strength, respectively. The new nano-micro hybrid scaffold was fabricated by electrospinning poly 3-hydroxybutyrate-chitosan-multi-walled carbon nanotube (MWNT functionalized by COOH) solution on knitted silk in a random manner with different amounts of MWNT. The physical, mechanical, and biodegradation properties were assessed through scanning electron microscopy, Fourier-transform infrared (FTIR) spectroscopy, water contact angle test, tensile strength test, and weight loss test. The scaffold without MWNT was chosen as control sample. An increase in the amount of MWNT up to 1 wt% leads to better fiber diameter distribution, more hydrophilicity, biodegradation rate, and higher tensile strength in comparison with other samples. The porosity percentage of all scaffolds is more than 80%. According to FTIR spectra, the nanofibrous coat on knitted silk did not have any effect on silk fibroin crystallinity structures, and according to tensile strength test, the coat had a significant effect on tensile strength in comparison with pure knitted silk ( P ≤ 0.05). The average fiber diameter decreased due to an increase in electrical conductivity of the solution and fiber stretch in electrical field due to MWNTs. The scaffold containing 1 wt% MWNT was more hydrophilic due to the presence of many COOH groups of functionalized MWNT, thus an increase in the hydrolysis and degradation rate of this sample. High intrinsic tensile strength of MWNTs and improvement of nano-micro interface connection lead to an increase in tensile strength in scaffolds containing MWNT.

Characterization of Silk/Poly 3-Hydroxybutyrate-chitosan-multi-walled Carbon Nanotube Micro-nano Scaffold: A New Hybrid Scaffold for Tissue Engineering Applications

PubMed Central

Mirmusavi, Mohammad Hossein; Karbasi, Saeed; Semnani, Dariush; Kharazi, Anousheh Zargar

2018-01-01

Background: Long-term healing tissue engineering scaffolds must hold its full mechanical strength at least for 12 weeks. Nano-micro scaffolds consist of electrospinning nanofibers and textile microfibers to support cell behavior and mechanical strength, respectively. Methods: The new nano-micro hybrid scaffold was fabricated by electrospinning poly 3-hydroxybutyrate-chitosan-multi-walled carbon nanotube (MWNT functionalized by COOH) solution on knitted silk in a random manner with different amounts of MWNT. The physical, mechanical, and biodegradation properties were assessed through scanning electron microscopy, Fourier-transform infrared (FTIR) spectroscopy, water contact angle test, tensile strength test, and weight loss test. The scaffold without MWNT was chosen as control sample. Results: An increase in the amount of MWNT up to 1 wt% leads to better fiber diameter distribution, more hydrophilicity, biodegradation rate, and higher tensile strength in comparison with other samples. The porosity percentage of all scaffolds is more than 80%. According to FTIR spectra, the nanofibrous coat on knitted silk did not have any effect on silk fibroin crystallinity structures, and according to tensile strength test, the coat had a significant effect on tensile strength in comparison with pure knitted silk (P ≤ 0.05). The average fiber diameter decreased due to an increase in electrical conductivity of the solution and fiber stretch in electrical field due to MWNTs. The scaffold containing 1 wt% MWNT was more hydrophilic due to the presence of many COOH groups of functionalized MWNT, thus an increase in the hydrolysis and degradation rate of this sample. Conclusions: High intrinsic tensile strength of MWNTs and improvement of nano-micro interface connection lead to an increase in tensile strength in scaffolds containing MWNT. PMID:29535924

In vitro fibroblast migration by sustained release of PDGF-BB loaded in chitosan nanoparticles incorporated in electrospun nanofibers for wound dressing applications.

PubMed

Piran, Mehrdad; Vakilian, Saeid; Piran, Mehran; Mohammadi-Sangcheshmeh, Abdollah; Hosseinzadeh, Simzar; Ardeshirylajimi, Abdolreza

2018-01-23

Migration of fibroblasts into wound area is a critical phenomenon in wound healing process. We used an appropriate system to fabricate an electrospun bioactive scaffold with controlled release of PDGF-BB in order to induce migration of primary skin fibroblast cells. First of all, protein-loaded chitosan nanoparticles based on ionic gelation interaction between chitosan and sodium tripolyphosphate were prepared. Then polycaprolactone electrospun fibers containing chitosan nanoparticles or PDGF-BB-loaded chitosan nanoparticles were prepared. Cellular attachment and morphology of cells seeded on scaffolds with or without PDGF-BB were evaluated by using a fluorescence microscope and scanning electron microscopy. Cells were well-oriented 72 h after seeding on the scaffolds containing PDGF-BB. The mean aspect ratio of populations on scaffold containing PDGF-BB-loaded chitosan nanoparticles was significantly greater than those on the scaffold containing chitosan nanoparticles but no PDGF-BB. Furthermore, the Arp2 gene, which is involved in cell protrusion formation, showed about three times more expression at mRNA level, in cells seeding on PDGF-BB-containing scaffold compared to cells seeding on scaffold containing only chitosan nanoparticles, using Real Time PCR test. Finally, under agarose migration assay results demonstrated that cells' chemotaxic behavior was more toward scaffold containing PDGF-BB compared to the PDGF-BB alone or FBS group. In addition, in terms of distance, the cell mass could grow faster, in response to scaffold containing PDGF-BB compared to FBS or PDGF-BB alone; however, the number of migrating cells might be the same or significantly higher in the latter groups.

The Pine-Needle-Inspired Structure of Zinc Oxide Nanorods Grown on Electrospun Nanofibers for High-Performance Flexible Supercapacitors.

PubMed

Sami, Syed Kamran; Siddiqui, Saqib; Shrivastava, Sajal; Lee, Nae-Eung; Chung, Chan-Hwa

2017-12-01

Flexible supercapacitors with high electrochemical performance and stability along with mechanical robustness have gained immense attraction due to the substantial advancements and rampant requirements of storage devices. To meet the exponentially growing demand of microsized energy storage device, a cost-effective and durable supercapacitor is mandatory to realize their practical applications. Here, in this work, the fabrication route of novel electrode materials with high flexibility and charge-storage capability is reported using the hybrid structure of 1D zinc oxide (ZnO) nanorods and conductive polyvinylidene fluoride-tetrafluoroethylene (P(VDF-TrFE)) electrospun nanofibers. The ZnO nanorods are conformably grown on conductive P(VDF-TrFE) nanofibers to fabricate the light-weighted porous electrodes for supercapacitors. The conductive nanofibers acts as a high surface area scaffold with significant electrochemical performance, while the addition of ZnO nanorods further enhances the specific capacitance by 59%. The symmetric cell with the fabricated electrodes presents high areal capacitance of 1.22 mF cm -2 at a current density of 0.1 mA cm -2 with a power density of more than 1600 W kg -1 . Furthermore, these electrodes show outstanding flexibility and high stability with 96% and 78% retention in specific capacitance after 1000 and 5000 cycles, respectively. The notable mechanical durability and robustness of the cell acquire both good flexibility and high performance. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Nanofibers of Human Tropoelastin-inspired peptides: Structural characterization and biological properties.

PubMed

Secchi, Valeria; Franchi, Stefano; Fioramonti, Marco; Polzonetti, Giovanni; Iucci, Giovanna; Bochicchio, Brigida; Battocchio, Chiara

2017-08-01

Regenerative medicine is taking great advantage from the use of biomaterials in the treatments of a wide range of diseases and injuries. Among other biomaterials, self-assembling peptides are appealing systems due to their ability to spontaneously form nanostructured hydrogels that can be directly injected into lesions. Indeed, self-assembling peptide scaffolds are expected to behave as biomimetic matrices able to surround cells, to promote specific interactions, and to control and modify cell behavior by mimicking the native environment as well. We selected three pentadecapeptides inspired by Human Tropoelastin, a natural protein of the extracellular matrix, expected to show high biocompatibility. Moreover, the here proposed self-assembling peptides (SAPs) are able to spontaneously aggregate in nanofibers in biological environment, as revealed by AFM (Atomic Force Microscopy). Peptides were characterized by XPS (X-ray Photoelectron Spectroscopy) and IRRAS (Infrared Reflection Absorption Spectroscopy) both as lyophilized (not aggregated) and as aggregated (nanofibers) samples in order to investigate some potential differences in their chemical composition and intermolecular interactions, and to analyze the surface and interface of nanofibers. Finally, an accurate investigation of the biological properties of the SAPs and of their interaction with cells was performed by culturing for the first time human Mesenchymal Stem Cells (hMSCs) in presence of SAPs. The final aim of this work was to assess if Human Tropoelastin-inspired nanostructured fibers could exert a cytotoxic effect and to evaluate their biocompatibility, cellular adhesion and proliferation. Copyright © 2017 Elsevier B.V. All rights reserved.

Optimization and characterization of bioactive glass nanofibers and nanocomposites

NASA Astrophysics Data System (ADS)

Scarber, Reginna E.

Disease affects different areas of the bone and can impact individuals of all pathologies and ethnicities. These bone diseases can result in weakening which leads to trauma during ordinary function, the need for reconstructive surgery, and eventual bone replacement. Tissue engineering can provide a less traumatic and more fundamental solution to the current therapies. Bioactive glasses are promising materials in tissue engineering applications because of their ability to form hydroxycarbonate apatite in the presence of simulated body fluid, support cell adhesion, growth, and differentiation, induce bone formation, and concentrate bone morphogenic proteins in vivo. The research in this dissertation will attempt to improve the quality, yield, and toughness of bioactive glass nanofibrous scaffolds. The three specific aims of this research include, (1) Optimization and Characterization of Surfactant Modified Bioactive Glass (2) Optimization of Direct Synthesis Bioactive glass Nanofibers from Sols (3) Mechanical Properties and In-vitro Biomineralization of Bioglass-loaded Polyglyconate Nanocomposites Created Using the Particulate Leaching Method. The purpose of the first specific aim was to optimize the processing of bioactive glass nanofibers, resulting in greater fiber uniformity with a reduction in beading. The increase in viscosity coupled with the ability of the surfactant to limit polymeric secondary bonding led to improved fiber quality. The focal point of the second specific aim is the production of sol-gel derived glass fibers with high bioactivity prepared by electrospinning without the use of any polymer carrier system. Advantages of this method include decreased processing time, increased production of fibers, and a decrease in the loss of material due to the calcining process. The solvent cast/ particulate leaching method was used to create a nanocomposite of bioglass and the co-polymer polyglyconate (MaxonRTM) for bone tissue scaffolds The biocompatibility of the composite foams was observed and calcium phosphate presence was quantified. The incorporation of bioglass into the polymer matrix improved the strength (modulus - 21.47 MPa) and biocompatibility of the polyglyconate foam. Keywords: Bioactive glass, Electrospinning, Solvent Casting/Particulate Leaching Method, Nanocomposites

Graphene oxide and hydroxyapatite as fillers of polylactic acid nanocomposites: preparation and characterization.

PubMed

Marques, Paula A A P; Gonçalves, Gil; Singh, Manoj K; Grácio, José

2012-08-01

Graphene and its derivatives have attracted great research interest for their potential applications in electronics, energy, materials and biomedical areas. When incorporated appropriately, these atomically thin carbon sheets are expected to improve physical properties of host polymers at extremely small loading. Herein, we report a novel two-step method for the preparation of PLLA/Hap/graphene oxide nanocomposites with augmented mechanical properties when compared to PLLA/Hap and neat PLLA. The presence of graphene oxide (GO) had a positive effect on the dispersion of hydroxyapatite particles on the polymeric matrix contributing for a good homogeneity of the final nanocomposite. PLLA nanocomposites prepared with 30% (w/w) of Hap and 1% (w/w) of GO showed the highest hardness and storage modulus values indicating an efficient load transfer between the fillers and the PLLA matrix. These materials may find interesting biomedical applications as for example bone screws. The following step on the study of these materials will be in vitro tests to access the biocompatibility of these new nanocomposites.

Transfer molding processes for nanoscale patterning of poly-L-lactic acid (PLLA) films

NASA Astrophysics Data System (ADS)

Dhakal, Rabin; Peer, Akshit; Biswas, Rana; Kim, Jaeyoun

2016-03-01

Nanoscale patterned structures composed of biomaterials exhibit great potential for the fabrication of functional biostructures. In this paper, we report cost-effective, rapid, and highly reproducible soft lithographic transfer-molding techniques for creating periodic micro- and nano-scale textures on poly (L-lactic acid) (PLLA) surface. These artificial textures can increase the overall surface area and change the release dynamics of the therapeutic agents coated on it. Specifically, we use the double replication technique in which the master pattern is first transferred to the PDMS mold and the pattern on PDMS is then transferred to the PLLA films through drop-casting as well as nano-imprinting. The ensuing comparison studies reveal that the drop-cast PLLA allows pattern transfer at higher levels of fidelity, enabling the realization of nano-hole and nano-cone arrays with pitch down to ~700 nm. The nano-patterned PLLA film was then coated with rapamycin to make it drug-eluting.

Mesenchymal cells condensation-inducible mesh scaffolds for cartilage tissue engineering.

PubMed

Kim, In Gul; Ko, Jaehoon; Lee, Hye Rim; Do, Sun Hee; Park, Kwideok

2016-04-01

Mesenchymal cells condensation is crucial in chondrogenic development. However current tissue-engineered scaffolds for chondrogenesis pay little attention to this phenomenon. In this study, we fabricate poly(l-lactide-co-glycolide) (PLGA)/poly(l-lactide) (PLLA) microfiber scaffolds and coat them with human fibroblast-derived matrix (hFDM) that is a decellularized extracellular matrix (ECM) obtained from in vitro cultured human lung fibroblasts (WI-38). Those scaffolds were then conjugated with heparin via EDC chemistry and subsequently immobilized with transforming growth factor (TGF)-β1. The amount of TGF-β1 was quantitatively measured and the release profile showed a continuous release of TGF-β1 for 4 weeks. Human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) were seeded in four different scaffolds; control, fibronectin (FN)-coated, hFDM-coated, hFDM/TGF-β1 and subjected to chondrogenic differentiation in vitro for up to 28 days. Both hFDM and hFDM/TGF-β1 groups exhibited significantly more synthesis of glycosaminoglycan (GAG) and much better upregulation of chondrogenic markers expression. Interestingly, MSCs condensation that led to cell aggregates was clearly observed with time in the two hFDM-coated groups and the quantitative difference was obvious compared to the control and FN group. A mechanistic study in gene and protein level indicated that focal adhesion kinase (FAK) was involved at the early stage of cell adhesion and cell-cell contact-related markers, N-cadherin and neural cell adhesion molecule (NCAM), were highly up-regulated at later time point. In addition histological analysis proved that hFDM/TGF-β1 group was the most effective in forming neocartilage tissue in a rabbit articular cartilage defect model. Taken together, this study demonstrates not only the positive effect of hFDM on chondrogenesis of MSCs and cartilage repair but also provides an important insight toward the significance of in vitro mesenchymal condensation on chondrogenic development of MSCs. Copyright © 2016 Elsevier Ltd. All rights reserved.

Improving Soldier Recovery from Catastrophic Bone Injuries: Developing an Animal Model for Standardizing the Bone Reparative Potential of Emerging Progenitor Cell Therapies

DTIC Science & Technology

2011-08-01

phase hydrogel or nanofiber mesh structures as a weight bearing scaffold for skeletal repair. This biocompatibility of this design will be first... delivery . If BMP is delivered into a site that is being filled with SMAA+ progenitors, it may hault the further inward migration by inducing...differentiation of the front wave of progenitor cells. This would predict that a shell of bone would be observed surrounding the delivery vehicle, but

Advanced polymeric matrix for valvular complications.

PubMed

Acharya, Gayathri; Hopkins, Richard A; Lee, Chi H

2012-05-01

Poly(L-lactic acid) (PLLA) matrix systems incorporated with poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) containing nitric oxide (NO) donors (DETA NONOate) were developed for prevention of heart valve complications through sustained and controlled release of NO. PLLA matrices were prepared using the salt leaching method and the properties and drug release profiles were characterized. For assessment of the effects of PLLA systems on the pharmacological responses and cytotoxicity, various factors, such as calcium content, alkaline phosphatase (ALP) activity, cyclic guanosine monophosphate (cGMP) expression, intercellular adhesion molecule (ICAM-1) expression and cell viability of porcine aortic valve interstitial cells (PAVICs), were evaluated. PLLA matrices embedded with PLGA- NPs demonstrated its usefulness in alleviating the calcification rate of the VICs. The cGMP levels under osteoblastic conditions significantly increased, supporting that anticalcification activity of NO is mediated through NO-cGMP signaling pathway. The level of ICAM-1 expression in cells exposed to NO was lowered, suggesting that NO has an inhibitory activity against tissue inflammation. NO releases from PLLA matrix embedded with PLGA NPs prevented valvular calcification and inflammation without causing any cytotoxic activities. PLLA matrix system loaded with NPs containing NO donors could provide a new platform for sustained and controlled delivery of NO, significantly reducing valvular complications. Copyright © 2012 Wiley Periodicals, Inc.

Innovative micro-textured hydroxyapatite and poly(l-lactic)-acid polymer composite film as a flexible, corrosion resistant, biocompatible, and bioactive coating for Mg implants.

PubMed

Kim, Sae-Mi; Kang, Min-Ho; Kim, Hyoun-Ee; Lim, Ho-Kyung; Byun, Soo-Hwan; Lee, Jong-Ho; Lee, Sung-Mi

2017-12-01

The utility of a novel ceramic/polymer-composite coating with a micro-textured microstructure that would significantly enhance the functions of biodegradable Mg implants is demonstrated here. To accomplish this, bioactive hydroxyapatite (HA) micro-dots can be created by immersing a Mg implant with a micro-patterned photoresist surface in an aqueous solution containing calcium and phosphate ions. The HA micro-dots can then be surrounded by a flexible poly(l-lactic)-acid (PLLA) polymer using spin coating to form a HA/PLLA micro-textured coating layer. The HA/PLLA micro-textured coating layer showed an excellent corrosion resistance when it was immersed in a simulated body fluid (SBF) solution and good biocompatibility, which was assessed by in vitro cell tests. In addition, the HA/PLLA micro-textured coating layer had high deformation ability, where no apparent changes in the coating layer were observed even after a 5% elongation, which would be unobtainable using HA and PLLA coating layers; furthermore, this allowed the mechanically-strained Mg implant with the HA/PLLA micro-textured coating layer to preserve its excellent corrosion resistance and biocompatibility in vitro. Copyright © 2017 Elsevier B.V. All rights reserved.

Production of GMP-grade radioactive holmium loaded poly(L-lactic acid) microspheres for clinical application.

PubMed

Zielhuis, S W; Nijsen, J F W; de Roos, R; Krijger, G C; van Rijk, P P; Hennink, W E; van het Schip, A D

2006-03-27

Radioactive holmium-166 loaded poly(L-lactic acid) microspheres are promising systems for the treatment of liver malignancies. The microspheres are loaded with holmium acetylacetonate (HoAcAc) and prepared by a solvent evaporation method. After preparation, the microspheres (Ho-PLLA-MS) are activated by neutron irradiation in a nuclear reactor. In this paper, the aspects of the production of a (relatively) large-scale GMP batch (4 g, suitable for treatment of 5-10 patients) of Ho-PLLA-MS are described. The critical steps of the Ho-PLLA-MS production process (sieving procedure, temperature control during evaporation and raw materials) were considered and the pharmaceutical quality of the microspheres was evaluated. The pharmaceutical characteristics (residual solvents, possible bacterial contaminations and endotoxins) of the produced Ho-PLLA-MS batches were in compliance with the requirements of the European Pharmacopoeia. Moreover, neutron irradiated Ho-PLLA-MS retained their morphological integrity and the holmium remained stably associated with the microspheres; it was observed that after 270h (10 times the half-life of Ho-166) only 0.3+/-0.1% of the loading was released from the microspheres in an aqueous solution. In conclusion, Ho-PLLA-MS which are produced as described in this paper, can be clinically applied, with respect to their pharmaceutical quality.

Increase the elongation at break of poly (lactic acid) composites for use in food packaging films

NASA Astrophysics Data System (ADS)

El-Hadi, Ahmed M.

2017-05-01

Poly (3-hydroxy butyrate) (PHB), cellulose nano crystal (CNC) and a plasticizer (TBC) are mixed together with PLLA with the aim to increase the elongation at break for use in the food packing sector. Spherical (CNC) and fibril nano crystal (CNF) were prepared by hydrolysis of microcrystalline cellulose (MCC) in distilled water, and then stirred using a magnetic stirrer for 15 days and ultrasonic treatment without using any acids as green method. The morphology, thermal, and mechanical properties were studied using POM, DSC, WAXD, SEM and tensile testing, respectively. DSC demonstrated that the addition of PHB, CNC and TBC to PLLA matrix lead to reduce Tg, TCC and Tm than pure PLLA. FT-IR verified that the carbonyl group C=O appeared broad and some peaks in the PLLA composites 5, 6 and 7 shifted from 3.98 × 108 to 4.07 × 108 Hz, at 3.54 × 108 to 3.44 × 108 Hz, at 3.19 × 108 to 3.13 × 108 Hz. Mechanical testing shows that pure PLLA is brittle, and the elongation at break of PLLA composites reaches up to 205%, making it suitable to use in food packaging.

Influence of purified multiwalled carbon nanotubes on the mechanical and morphological behavior in poly (L-lactic acid) matrix.

PubMed

Leal, C V; Martinez, D S T; Más, B A; Alves, O L; Duek, E A R

2016-06-01

Poly (L-latic acid) (PLLA) is a bioresorbable polymer widely used as a biomaterial, but its fragility can limit its use. An alternative is to produce polymer nanocomposites, which can enhance the mechanical properties of polymeric matrix, resulting in a material with differentiated properties. In this work, PLLA based nanocomposites containing 0.25, 0.5 and 1.0wt% of purified multiwalled carbon nanotubes (p-MWCNTs) were prepared by the solvent casting method. The morphology and mechanical properties results show an improvement in strain at break for 0.25 and 0.5wt% p-MWCNTs and an increase in stiffness and elastic modulus for all compositions. Nanocomposites presented a p-MWCNTs agglomeration; however, there was a good stress transfer between PLLA and p-MWCNTs, which was confirmed by the increase in the hardness and elastic modulus. Atomic force microscopy analysis indicated an increase in roughness after nanotube addition. The in vitro biological study showed that PLLA/p-MWCNTs nanocomposites are cytocompatible with osteoblasts cells. The capacity of PLLA nanocomposites to stimulate osteogenesis was investigated by alkaline phosphatase (ALP) activity assay. Higher ALP activity was found on osteoblasts cultured on nanocomposites with 0.25 and 0.5wt% p-MWCNT compared to neat PLLA, confirming that PLLA cytocompatibility was improved on these compositions. Finally, our results showed that by a simple and inexpensive solvent casting method, it is possible to manufacture biofunctional nanocomposites devices with potential for orthopedic applications. Copyright © 2016 Elsevier Ltd. All rights reserved.

β-Tricalcium phosphate/poly(glycerol sebacate) scaffolds with robust mechanical property for bone tissue engineering.

PubMed

Yang, Kai; Zhang, Jing; Ma, Xiaoyu; Ma, Yifan; Kan, Chao; Ma, Haiyan; Li, Yulin; Yuan, Yuan; Liu, Changsheng

2015-11-01

Despite good biocompatibility and osteoconductivity, porous β-TCP scaffolds still lack the structural stability and mechanical robustness, which greatly limit their application in the field of bone regeneration. The hybridization of β-TCP with conventional synthetic biodegradable PLA and PCL only produced a limited toughening effect due to the plasticity of the polymers in nature. In this study, a β-TCP/poly(glycerol sebacate) scaffold (β-TCP/PGS) with well interconnected porous structure and robust mechanical property was prepared. Porous β-TCP scaffold was first prepared with polyurethane sponge as template and then impregnated into PGS pre-polymer solution with moderate viscosity, followed by in situ heat crosslinking and freezing-drying process. The results indicated that the freezing-drying under vacuum process could further facilitate crosslinking of PGS and formation of Ca(2+)-COO(-) ionic complexing and thus synergistically improved the mechanical strength of the β-TCP/PGS with in situ heat crosslinking. Particularly, the β-TCP/PGS with 15% PGS content after heat crosslinking at 130°C and freezing-drying at -50°C under vacuum exhibited an elongation at break of 375±25% and a compressive strength of 1.73MPa, 3.7-fold and 200-fold enhancement compared to the β-TCP, respectively. After the abrupt drop of compressive load, the β-TCP/PGS scaffolds exhibited a full recovery of their original shape. More importantly, the PGS polymer in the β-TCP/PGS scaffolds could direct the biomineralization of Ca/P from particulate shape into a nanofiber-interweaved structure. Furthermore, the β-TCP/PGS scaffolds allowed for cell penetration and proliferation, indicating a good cytobiocompatibility. It is believed that β-TCP/PGS scaffolds have great potential application in rigid tissue regeneration. Copyright © 2015 Elsevier B.V. All rights reserved.

Piezoelectric Characteristics of Chiral Polymer Composite Films Obtained under Strong Magnetic Field

NASA Astrophysics Data System (ADS)

Nakiri, Takuo; Okuno, Masaki; Maki, Nobuyuki; Kanasaki, Masayoshi; Morimoto, Yu; Okamoto, Satoshi; Ishizuka, Masayuki; Fukuda, Kazuyuki; Takaki, Toshihiko; Tajitsu, Yoshiro

2005-09-01

It is difficult to obtain a drawn chiral polymer/inorganic material composite membrane with shear piezoelectricity by the conventional method because the chiral polymer/inorganic material composite membrane breaks during the drawing process by which shear piezoelectricity is realized. Using a strong magnetic field, we propose to manufacture a drawn composite membrane of poly-l-lactic acid (PLLA), a chiral polymer, and hydroxyapatite (Hap), an inoroganic material (PLLA/Hap composite membrane). The manufacturing method used here is effective for obtaining a drawn PLLA/Hap composite membrane with a large uniform area. Also, the shear piezoelectric constant of the drawn PLLA/Hap composite membrane is about 20 pC/N. This value is large for piezoelectric polymers.

An electrospun scaffold integrating nucleic acid delivery for treatment of full thickness wounds

PubMed Central

Kobsa, Serge; Kristofik, Nina J.; Sawyer, Andrew J.; Bothwell, Alfred L.M.; Kyriakides, Themis R.; Saltzman, W. Mark

2013-01-01

We developed a multi-functional construct capable of controlled delivery of bioactive substances that can improve wound repair by supporting the intrinsic ability of the skin to heal. We synthesized electrospun scaffolds—composed of a blend of the degradable polymers poly(L-lactide) (PLA) or polycaprolactone (PCL)—that produce highly efficient non-viral in vivo gene delivery to cells in the wound bed, provide a protective barrier during early wound healing, and support cell migration and growth. This multi-functional material was tested for its influence on wound healing: scaffolds were loaded with plasmids encoding keratinocyte growth factor (KGF) and applied to full thickness wounds in mice. Compared to scaffolds with control plasmids, animals receiving the KGF plasmid-loaded scaffold produced significant enhancements in wound healing, which was quantified by improvements in the rate of wound re-epithelialization, keratinocyte proliferation, and granulation response. Further, we quantified the expression level of endogenous and plasmid-derived KGF in wound samples: qRT-PCR on wound sections revealed a correlation between the levels of plasmid-derived protein expression and histological analysis of wound healing, revealing an inverse relationship between the expression level of exogenous KGF and the size of the unhealed epithelial layer in wounds. Our findings suggest that engineered nanofiber PLA/PCL scaffolds are capable of highly efficient controlled DNA delivery and are promising materials for treatment of cutaneous wounds. PMID:23453058

An acid-free water-born quaternized chitosan/montmorillonite loaded into an innovative ultra-fine bead-free water-born nanocomposite nanofibrous scaffold; in vitro and in vivo approaches.

PubMed

Dastjerdi, Roya; Sharafi, Mahsa; Kabiri, Kourosh; Mivehi, Leila; Samadikuchaksaraei, Ali

2017-07-26

An acid-free water-born chitosan derivative/montmorillonite has been successfully synthesized. A natural-based biopolymer, N-(2-hydroxy) propyl-3-trimethyl ammonium chitosan chloride, was synthesized, and its structure confirmed by Fourier transform infrared microscopy and conductometric titration. It was applied to the cationic ion-exchange reaction of montmorillonite. Then, the synthesized materials were used to produce water-born composite scaffolds for tissue engineering applications and formed an ultra-fine bead-free multicomponent nanofibrous scaffold. The scaffold was subjected to in vitro and in vivo investigations. The effects of both acidic and neutral reaction media on the efficiency of the cationic ion-exchange reaction of montmorillonite were investigated. A mechanism has been suggested for the more efficient cationic ion-exchange reaction achieved in the absence of the acid. In in vitro studies, the modified montmorillonite showed synergistic biocompatibility and cell growth with enhanced bioactivity compared to unmodified clay and even chitosan and the chitosan derivative. Scanning electron microscopy showed ultra-fine bead-free nanocomposite nanofibers. Improved biocompatibility, cell attachment, and cell growth were observed for the nanofibrous scaffolds compared to the individual components. In vivo experiments showed complete restoration of a critical-sized full-thickness wound without infection in 21 d. The technique provides a guideline to achieve chitosan nanofibrous morphology for multifunctional biomedical applications.

Electrospun Nanocomposite Materials, A Novel Synergy of Polyurethane and Bovine Derived Hydroxyapatite

NASA Astrophysics Data System (ADS)

Bozkurt, Y.; Sahin, A.; Sunulu, A.; Aydogdu, M. O.; Altun, E.; Oktar, F. N.; Ekren, N.; Gunduz, O.

2017-04-01

Polyurethane (PU) is a synthetic polymer that is used for construction of scaffold in tissue engineering applications in order to obtain desirable mechanical, physical and chemical properties like elasticity and durability. Bovine derived hydroxyapatite (BHAp) is a ceramic based natural polymer that is used as the most preferred implant material in orthopedics and dentistry due to their chemically and biologically similarity to the mineral phase found in the human bone structure. PU and bovine derived hydroxyapatite (BHAp) solutions with different concentrations were prepared with dissolving polyurethane and BHAp in Dimethylformamide (DMF) and Tetrahydrofuran (THF) solutions. Blended PU-BHAp solutions in different concentrations were used for electrospinning technique to create nanofiber scaffolds and new biocomposite material together. SEM, FTIR and physical analysis such as viscosity, electrical conductivity, density measurement and tensile strength measurement tests were carried out after production process.

Peripheral Nerve Repair in Rats Using Composite Hydrogel-Filled Aligned Nanofiber Conduits with Incorporated Nerve Growth Factor

PubMed Central

Jin, Jenny; Limburg, Sonja; Joshi, Sunil K.; Landman, Rebeccah; Park, Michelle; Zhang, Qia; Kim, Hubert T.

2013-01-01

Repair of peripheral nerve defects with current synthetic, tubular nerve conduits generally shows inferior recovery when compared with using nerve autografts, the current gold standard. We tested the ability of composite collagen and hyaluronan hydrogels, with and without the nerve growth factor (NGF), to stimulate neurite extension on a promising aligned, nanofiber poly-L-lactide-co-caprolactone (PLCL) scaffold. In vitro, the hydrogels significantly increased neurite extension from dorsal root ganglia explants. Consistent with these results, the addition of hydrogels as luminal fillers within aligned, nanofiber tubular PLCL conduits led to improved sensory function compared to autograft repair in a critical-size defect in the sciatic nerve in a rat model. Sensory recovery was assessed 3 and 12 weeks after repair using a withdrawal assay from thermal stimulation. The addition of hydrogel did not enhance recovery of motor function in the rat model. The NGF led to dose-dependent improvements in neurite out-growth in vitro, but did not have a significant effect in vivo. In summary, composite collagen/hyaluronan hydrogels enhanced sensory neurite outgrowth in vitro and sensory recovery in vivo. The use of such hydrogels as luminal fillers for tubular nerve conduits may therefore be useful in assisting restoration of protective sensation following peripheral nerve injury. PMID:23659607

Basal Lamina Mimetic Nanofibrous Peptide Networks for Skeletal Myogenesis

NASA Astrophysics Data System (ADS)

Yasa, I. Ceren; Gunduz, Nuray; Kilinc, Murat; Guler, Mustafa O.; Tekinay, Ayse B.

2015-11-01

Extracellular matrix (ECM) is crucial for the coordination and regulation of cell adhesion, recruitment, differentiation and death. Therefore, equilibrium between cell-cell and cell-matrix interactions and matrix-associated signals are important for the normal functioning of cells, as well as for regeneration. In this work, we describe importance of adhesive signals for myoblast cells’ growth and differentiation by generating a novel ECM mimetic peptide nanofiber scaffold system. We show that not only structure but also composition of bioactive signals are important for cell adhesion, growth and differentiation by mimicking the compositional and structural properties of native skeletal muscle basal lamina. We conjugated laminin-derived integrin binding peptide sequence, “IKVAV”, and fibronectin-derived well known adhesive sequence, “RGD”, into peptide nanostructures to provide adhesive and myogenic cues on a nanofibrous morphology. The myogenic and adhesive signals exhibited a synergistic effect on model myoblasts, C2C12 cells. Our results showed that self-assembled peptide nanofibers presenting laminin derived epitopes support adhesion, growth and proliferation of the cells and significantly promote the expression of skeletal muscle-specific marker genes. The functional peptide nanofibers used in this study present a biocompatible and biodegradable microenvironment, which is capable of supporting the growth and differentiation of C2C12 myoblasts into myotubes.

Basal Lamina Mimetic Nanofibrous Peptide Networks for Skeletal Myogenesis

PubMed Central

Yasa, I. Ceren; Gunduz, Nuray; Kilinc, Murat; Guler, Mustafa O.; Tekinay, Ayse B.

2015-01-01

Extracellular matrix (ECM) is crucial for the coordination and regulation of cell adhesion, recruitment, differentiation and death. Therefore, equilibrium between cell-cell and cell-matrix interactions and matrix-associated signals are important for the normal functioning of cells, as well as for regeneration. In this work, we describe importance of adhesive signals for myoblast cells’ growth and differentiation by generating a novel ECM mimetic peptide nanofiber scaffold system. We show that not only structure but also composition of bioactive signals are important for cell adhesion, growth and differentiation by mimicking the compositional and structural properties of native skeletal muscle basal lamina. We conjugated laminin-derived integrin binding peptide sequence, “IKVAV”, and fibronectin-derived well known adhesive sequence, “RGD”, into peptide nanostructures to provide adhesive and myogenic cues on a nanofibrous morphology. The myogenic and adhesive signals exhibited a synergistic effect on model myoblasts, C2C12 cells. Our results showed that self-assembled peptide nanofibers presenting laminin derived epitopes support adhesion, growth and proliferation of the cells and significantly promote the expression of skeletal muscle-specific marker genes. The functional peptide nanofibers used in this study present a biocompatible and biodegradable microenvironment, which is capable of supporting the growth and differentiation of C2C12 myoblasts into myotubes. PMID:26555958

Thermosensitive nanospheres with a gold layer revealed as low-cytotoxic drug vehicles.

PubMed

Qin, Jian; Jo, Yun Suk; Ihm, Jong Eun; Kim, Do Kyung; Muhammed, Mamoun

2005-09-27

In this paper, the positive effect of a gold layer on cell viability is demonstrated by examining the results given by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfop henyl)-2H-tetrazolium (MTS) assay and two-color cell fluorescence viability (TCCV) assay. These cytotoxicity tests were performed with human cervical adenocarcinoma cells (HeLa cell line) and transformed African green monkey kidney fibroblast cells (Cos-7 cell line). To fabricate the nanostructures as drug vehicles, first, poly(l,l-lactide-co-ethylene glycol) (PLLA-PEG) and poly(N-isopropylacrylamide-co-D,D-lactide) (PNIPAAm-PDLA) were synthesized, and then two kinds of thermosensitive nanospheres comprising "shell-in-shell" structures without a gold layer (PLLA-PEG@PNIPAAm-PDLA) and with a gold layer (Au@PLLA-PEG@PNIPAAm-PDLA) were constructed by a modified double-emulsion method (MDEM). Both of them displayed a unique thermosensitive character exhibiting the lower critical solubility temperature (LCST) at 36.7 degrees C which was confirmed by UV-vis spectroscopy and differential scanning calorimetry (DSC). The release profiles of entrapped bovine serum albumin (BSA) were monitored at 22 and 37 degrees C, respectively, to reveal the thermal dependence on the release rate. In cell viability tests, both PLLA-PEG@PNIPAAm-PDLA and Au@PLLA-PEG@PNIPAAm-PDLA showed excellent cell viability, and furthermore, Au@PLLA-PEG@PNIPAAm-PDLA, particularly at high doses, exhibited more enhanced cell viability than PLLA-PEG@PNIPAAm-PDLA. This effect is mainly attributed to the gold layer which binds the protein molecules first and consequently facilitates transmembrane uptake of essential nutrients in the cell media, resulting in favorable cell proliferation.

Development of Magnesium and Siloxane-Containing Vaterite and Its Composite Materials for Bone Regeneration

PubMed Central

Yamada, Shinya; Obata, Akiko; Maeda, Hirotaka; Ota, Yoshio; Kasuga, Toshihiro

2015-01-01

Development of novel biomaterials with Mg2+, Ca2+, and silicate ions releasability for bone regeneration is now in progress. Several inorganic ions have been reported to stimulate bone-forming cells. We featured Ca2+, silicate, and especially, Mg2+ ions as growth factors for osteoblasts. Various biomaterials, such as ceramic powders and organic–inorganic composites, that release the ions, have been developed and investigated for their cytocompatibilities in our previous work. Through the investigation, providing the three ions was found to be effective to activate osteogenic cells. Magnesium and siloxane-­containing vaterite was prepared by a carbonation process as an inorganic particle that can has the ability to simultaneously release Ca2+, silicate, and Mg2+ ions to biodegradable polymers. Poly (l-lactic acid) (PLLA)- and bioactive PLLA-based composites containing vaterite coatings were discussed regarding their degradability and cytocompatibility using a metallic Mg substrate as Mg2+ ion source. PLLA/SiV composite film, which has a releasability of silicate ions besides Ca2+ ion, was coated on a pure Mg substrate to be compared with the PLLA/V coating. The degradability and releasability of inorganic ions were morphologically and quantitatively monitored in a cell culture medium. The bonding strength between the coatings and Mg substrates was one of the key factors to control Mg2+ ion release from the substrates. The cell culture tests were conducted using mouse osteoblast-like cells (MC3T3-E1 cells); cellular morphology, proliferation, and differentiation on the materials were evaluated. The PLLA/V and PLLA/SiV coatings on Mg substrates were found to enhance the proliferation, especially the PLLA/SiV coating possessed a higher ability to induce the osteogenic differentiation of the cells. PMID:26697421

Coffee Grounds to Multifunctional Quantum Dots: Extreme Nanoenhancers of Polymer Biocomposites.

PubMed

Xu, Huan; Xie, Lan; Li, Jinlai; Hakkarainen, Minna

2017-08-23

Central to the design and execution of nanocomposite strategies is the invention of polymer-affinitive and multifunctional nanoreinforcements amenable to economically viable processing. Here, a microwave-assisted approach enabled gram-scale fabrication of polymer-affinitive luminescent quantum dots (QDs) from spent coffee grounds. The ultrasmall dimensions (approaching 20 nm), coupled with richness of diverse oxygen functional groups, conferred the zero-dimensional QDs with proper exfoliation and uniform dispersion in poly(l-lactic acid) (PLLA) matrix. The unique optical properties of QDs were inherited by PLLA nanocomposites, giving intensive luminescence and high visible transparency, as well as nearly 100% UV-blocking ratio in the full-UV region at only 0.5 wt % QDs. The strong anchoring of PLLA chains at the nanoscale surfaces of QDs facilitated PLLA crystallization, which was accompanied by substantial improvements in thermomechanical and tensile properties. With 1 wt % QDs, for example, the storage modulus at 100 °C and tensile strength increased over 2500 and 69% compared to those of pure PLLA (4 and 57.3 MPa), respectively. The QD-enabled energy-dissipating and flexibility-imparting mechanisms upon tensile deformation, including the generation of numerous shear bands, crazing, and nanofibrillation, gave an unusual combination of elasticity and extensibility for PLLA nanocomposites. This paves the way to biowaste-derived nanodots with high affinity to polymer for elegant implementation of distinct light management and extreme nanoreinforcements in an ecofriendly manner.

Dynamic Tensile Loading Improves the Functional Properties of Mesenchymal Stem Cell-Laden Nanofiber-Based Fibrocartilage

PubMed Central

Baker, Brendon M.; Shah, Roshan P.; Huang, Alice H.

2011-01-01

Fibrocartilaginous tissues such as the meniscus serve critical load-bearing roles, relying on arrays of collagen fibers to resist tensile loads experienced with normal activity. As these structures are frequently injured and possess limited healing capacity, there exists great demand for tissue-engineered replacements. Toward recreating the structural features of these anisotropic tissues in vitro, we employ scaffolds composed of co-aligned nanofibers that direct mesenchymal stem cell (MSC) orientation and the formation of organized extracellular matrix (ECM). Concomitant with ECM synthesis, the mechanical properties of constructs increase with free-swelling culture, but ultimately failed to achieve equivalence with meniscal fibrocartilage. As mechanical forces are essential to the development and maintenance of musculoskeletal tissues, this work examined the effect of cyclic tensile loading on MSC-laden nanofibrous constructs. We hypothesized that loading would modulate the transcriptional behavior of MSCs, spur the deposition of ECM, and lead to enhancements in construct mechanical properties compared to free-swelling controls. Fiber-aligned scaffolds were seeded with MSCs and dynamically loaded daily in tension or maintained as nonloaded controls for 4 weeks. With mechanical stimulation, fibrous gene expression increased, collagen deposition increased, and the tensile modulus increased by 16% relative to controls. These results show that dynamic tensile loading enhances the maturation of MSC-laden aligned nanofibrous constructs, suggesting that recapitulation of the structural and mechanical environment of load-bearing tissues results in increases in functional properties that can be exploited for tissue engineering applications. PMID:21247342

Dynamic tensile loading improves the functional properties of mesenchymal stem cell-laden nanofiber-based fibrocartilage.

PubMed

Baker, Brendon M; Shah, Roshan P; Huang, Alice H; Mauck, Robert L

2011-05-01

Fibrocartilaginous tissues such as the meniscus serve critical load-bearing roles, relying on arrays of collagen fibers to resist tensile loads experienced with normal activity. As these structures are frequently injured and possess limited healing capacity, there exists great demand for tissue-engineered replacements. Toward recreating the structural features of these anisotropic tissues in vitro, we employ scaffolds composed of co-aligned nanofibers that direct mesenchymal stem cell (MSC) orientation and the formation of organized extracellular matrix (ECM). Concomitant with ECM synthesis, the mechanical properties of constructs increase with free-swelling culture, but ultimately failed to achieve equivalence with meniscal fibrocartilage. As mechanical forces are essential to the development and maintenance of musculoskeletal tissues, this work examined the effect of cyclic tensile loading on MSC-laden nanofibrous constructs. We hypothesized that loading would modulate the transcriptional behavior of MSCs, spur the deposition of ECM, and lead to enhancements in construct mechanical properties compared to free-swelling controls. Fiber-aligned scaffolds were seeded with MSCs and dynamically loaded daily in tension or maintained as nonloaded controls for 4 weeks. With mechanical stimulation, fibrous gene expression increased, collagen deposition increased, and the tensile modulus increased by 16% relative to controls. These results show that dynamic tensile loading enhances the maturation of MSC-laden aligned nanofibrous constructs, suggesting that recapitulation of the structural and mechanical environment of load-bearing tissues results in increases in functional properties that can be exploited for tissue engineering applications.

Virgin olive oil blended polyurethane micro/nanofibers ornamented with copper oxide nanocrystals for biomedical applications

PubMed Central

Amna, Touseef; Hassan, M Shamshi; Yang, Jieun; Khil, Myung-Seob; Song, Ki-Duk; Oh, Jae-Don; Hwang, Inho

2014-01-01

Recently, substantial interest has been generated in using electrospun biomimetic nanofibers of hybrids, particularly organic/inorganic, to engineer different tissues. The present work, for the first time, introduced a unique natural and synthetic hybrid micronanofiber wound dressing, composed of virgin olive oil/copper oxide nanocrystals and polyurethane (PU), developed via facile electrospinning. The as-spun organic/inorganic hybrid micronanofibers were characterized by scanning electron microscopy (SEM), energy dispersive X-ray analysis, X-ray diffraction, electron probe microanalysis, and transmission electron microscopy. The interaction of cells with scaffold was studied by culturing NIH 3T3 fibroblasts on an as-spun hybrid micronanofibrous mat, and viability, proliferation, and growth were assessed. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay results and SEM observation showed that the hybrid micronanofibrous scaffold was noncytotoxic to fibroblast cell culture and was found to benefit cell attachment and proliferation. Hence our results suggest the potential utilization of as-spun micronanoscaffolds for tissue engineering. Copper oxide–olive oil/PU wound dressing may exert its positive beneficial effects at every stage during wound-healing progression, and these micronanofibers may serve diverse biomedical applications, such as tissue regeneration, damaged skin treatment, wound healing applications, etc. Conclusively, the fabricated olive oil–copper oxide/PU micronanofibers combine the benefits of virgin olive oil and copper oxide, and therefore hold great promise for biomedical applications in the near future. PMID:24611006

Electrospinning fundamentals: optimizing solution and apparatus parameters.

PubMed

Leach, Michelle K; Feng, Zhang-Qi; Tuck, Samuel J; Corey, Joseph M

2011-01-21

Electrospun nanofiber scaffolds have been shown to accelerate the maturation, improve the growth, and direct the migration of cells in vitro. Electrospinning is a process in which a charged polymer jet is collected on a grounded collector; a rapidly rotating collector results in aligned nanofibers while stationary collectors result in randomly oriented fiber mats. The polymer jet is formed when an applied electrostatic charge overcomes the surface tension of the solution. There is a minimum concentration for a given polymer, termed the critical entanglement concentration, below which a stable jet cannot be achieved and no nanofibers will form - although nanoparticles may be achieved (electrospray). A stable jet has two domains, a streaming segment and a whipping segment. While the whipping jet is usually invisible to the naked eye, the streaming segment is often visible under appropriate lighting conditions. Observing the length, thickness, consistency and movement of the stream is useful to predict the alignment and morphology of the nanofibers being formed. A short, non-uniform, inconsistent, and/or oscillating stream is indicative of a variety of problems, including poor fiber alignment, beading, splattering, and curlicue or wavy patterns. The stream can be optimized by adjusting the composition of the solution and the configuration of the electrospinning apparatus, thus optimizing the alignment and morphology of the fibers being produced. In this protocol, we present a procedure for setting up a basic electrospinning apparatus, empirically approximating the critical entanglement concentration of a polymer solution and optimizing the electrospinning process. In addition, we discuss some common problems and troubleshooting techniques.

Electrochemical biosensor based on functional composite nanofibers for detection of K-ras gene via multiple signal amplification strategy.

PubMed

Wang, Xiaoying; Shu, Guofang; Gao, Chanchan; Yang, Yu; Xu, Qian; Tang, Meng

2014-12-01

An electrochemical biosensor based on functional composite nanofibers for hybridization detection of specific K-ras gene that is highly associated with colorectal cancer via multiple signal amplification strategy has been developed. The carboxylated multiwalled carbon nanotubes (MWCNTs) doped nylon 6 (PA6) composite nanofibers (MWCNTs-PA6) was prepared using electrospinning, which served as the nanosized backbone for thionine (TH) electropolymerization. The functional composite nanofibers [MWCNTs-PA6-PTH, where PTH is poly(thionine)] used as supporting scaffolds for single-stranded DNA1 (ssDNA1) immobilization can dramatically increase the amount of DNA attachment and the hybridization sensitivity. Through the hybridization reaction, a sandwich format of ssDNA1/K-ras gene/gold nanoparticle-labeled ssDNA2 (AuNPs-ssDNA2) was fabricated, and the AuNPs offered excellent electrochemical signal transduction. The signal amplification was further implemented by forming network-like thiocyanuric acid/gold nanoparticles (TA/AuNPs). A significant sensitivity enhancement was obtained; the detection limit was down to 30fM, and the discriminations were up to 54.3 and 51.9% between the K-ras gene and the one-base mismatched sequences including G/C and A/T mismatched bases, respectively. The amenability of this method to the analyses of K-ras gene from the SW480 colorectal cancer cell lysates was demonstrated. The results are basically consistent with those of the K-ras Kit (HRM: high-resolution melt). The method holds promise for the diagnosis and management of cancer. Copyright © 2014 Elsevier Inc. All rights reserved.

Fabrication, characterization and biomedical application of two-nozzle electrospun polycaprolactone/zein-calcium lactate composite nonwoven mat.

PubMed

Liao, Nina; Joshi, Mahesh Kumar; Tiwari, Arjun Prasad; Park, Chan-Hee; Kim, Cheol Sang

2016-07-01

The objective of the current work is to incorporate calcium lactate (CL) into polycaprolactone (PCL)/zein composite micro/nanofibrous scaffolds via electrospinning to engineer bone tissue. In this study, a composite micro/nano fibrous scaffold was fabricated using a single two-nozzle electrospinning system to combine indicative nanofibers from a blended solution of zein-CL and micro-sized fibers from a PCL solution. Incorporation of the CL into the PCL/zein fibers were shown to improve the wettability, tensile strength and biological activity of the composite mats. Moreover, the composite mats have a high efficiency to nucleate calcium phosphate from simulated body fluid (SBF) solution. An in vitro cell culture with osteoblast cells demonstrated that the electrospun composite mats possessed improved biological properties, including a better cell adhesion, spread and proliferation. This study has demonstrated that the PCL/zein-CL composite provides a simple platform to fabricate a new biomimetic scaffold for bone tissue engineering, which can recapitulate both the morphology of extracellular matrix and composition of the bone. Copyright © 2016 Elsevier Ltd. All rights reserved.

Stereocomplexation of low molecular weight poly(L-lactic acid) and high molecular weight poly(D-lactic acid), radiation crosslinking PLLA/PDLA stereocomplexes and their characterization

NASA Astrophysics Data System (ADS)

Quynh, Tran Minh; Mai, Hoang Hoa; Lan, Pham Ngoc

2013-02-01

Poly(L-lactic acid)s (PLLAx) were synthesized from L-lactic acid by polycondensation. Different stereocomplexes were also obtained with equimolar mixtures of synthesized PLLAx and a commercial PDLA. The stereocomplexes were crosslinked with triallyl isocyanurate (TAIC) by gamma irradiation. Crosslinking density increased with radiation doses, the heavier the crosslinking network, the lower its swelling degree. The crosslinking structures were introduced in the stereocomplexes inhibiting the mobility for crystallization of PLLA molecules. Thermal and mechanical properties of PLA stereocomplexes were remarkably enhanced by radiation induced crosslinking. PLA stereocomplex does not seem to be degraded by PLLA degrading microorganisms existing in compost at room temperature, but the synthesized PLLA was significantly degraded.

Fabrication of core-shell nanofibers for controlled delivery of bromelain and salvianolic acid B for skin regeneration in wound therapeutics.

PubMed

Shoba, Ekambaram; Lakra, Rachita; Syamala Kiran, Manikantan; Korrapati, Purna Sai

2017-06-05

The physiological and pathological complexity of the wound healing process makes it more challenging to design an ideal tissue regeneration scaffold. Precise scaffolding with high drug loading efficiency, efficient intracellular efficacy for therapeutic delivery, minimal nonspecific cellular and blood protein binding, and maximum biocompatibility forms the basis for an ideal delivery system. This paper describes a combinational multiphasic delivery system, where biomolecules are delivered through the fabrication of coaxial electrospinning of different biocompatible polymers. The ratio and specificity of polymers for specific biofunction are optimized and the delivery system is completely characterized with reference to the mechanical property and structural integrity of bromelain (debridement enzyme) and salvianolic acid B (pro-angiogenesis and re-epithelialization). The in vitro release profile illustrated the sustained release of debriding protease and bioactive component in a timely fashion. The fabricated scaffold showed angiogenic potential through in vitro migration of endothelial cells and increased new capillaries from the existing blood vessel in response to an in ovo chicken chorioallantoic membrane assay. In addition, in vivo studies confirm the efficacy of the fabricated scaffold. Our results therefore open up a new avenue for designing a bioactive combinational multiphasic delivery system to enhance wound healing.

Fabrication of porous chitosan/poly(vinyl alcohol) reinforced single-walled carbon nanotube nanocomposites for neural tissue engineering.

PubMed

Shokrgozar, Mohammad Ali; Mottaghitalab, Fatemeh; Mottaghitalab, Vahid; Farokhi, Mehdi

2011-04-01

With the ability to form a nano-sized fibrous structure with large pore sizes mimicking the extracellular matrix (ECM), electrospinning was used to fabricate chitosan/poly(vinyl alcohol) nanofibers reinforced by single-walled carbon nanotube (SWNT-CS/PVA) for potential use in neural tissue engineering. Moreover, ultrasonication was performed to fabricate highly dispersed SWNT/CS solution with 7%, 12%, and 17% SWNT content prior to electrospinning process. In the present study, a number of properties of CS/PVA reinforced SWNTs nanocomposites were evaluated. The in vitro biocompatibility of the electrospun fiber mats was also assessed using human brain-derived cells and U373 cell lines. The results have shown that SWNTs as reinforcing phase can augment the morphology, porosity, and structural properties of CS/PVA nanofiber composites and thus benefit the proliferation rate of both cell types. In addition, the cells exhibit their normal morphology while integrating with surrounding fibers. The results confirmed the potential of SWNT-CS/PVA nanocomposites as scaffold for neural tissue engineering.

Recent progress concerning the production of controlled highly oriented electrospun nanofibrous arrays

NASA Astrophysics Data System (ADS)

Manea, L. R.; Hristian, L.; Leon, A. L.; Popa, A.

2016-08-01

Among the foreground domains of all the research-development programs at national and international level, a special place is occupied by that concerning the nanosciences, nanotechnologies, new materials and technologies. Electrospinning found a well-deserved place in this space, offering the preparation of nanomaterials with distinctive properties and applications in medicine, environment, photonic sensors, filters, etc. These multiple applications are generated by the fact that the electrospinning technology makes available the production of nanofibers with controllable characteristics (length, porosity, density, and mechanical characteristics), complexity and architecture. The apparition of 3D printing technology favors the production of complex nanofibrous structures, controlled assembly, self-assembly of electrospun nanofibers for the production of scaffolds used in various medical applications. The architecture of fibrous deposits has a special influence on the subsequent development of the cells of the reconstructed organism. The present work proposes to study of recent progress concerning the production of controlled highly oriented electrospun nanofibrous arrays and progress in research on the production of complex 2D and 3D structures.

RGD peptide-displaying M13 bacteriophage/PLGA nanofibers as cell-adhesive matrices for smooth muscle cells

NASA Astrophysics Data System (ADS)

Shin, Yong Cheol; Lee, Jong Ho; Jin, Oh Seong; Lee, Eun Ji; Jin, Lin Hua; Kim, Chang-Seok; Hong, Suck Won; Han, Dong-Wook; Kim, Chuntae; Oh, Jin-Woo

2015-01-01

Extracellular matrices (ECMs) are network structures that play an essential role in regulating cellular growth and differentiation. In this study, novel nanofibrous matrices were fabricated by electrospinning M13 bacteriophage and poly(lactic- co-glycolic acid) (PLGA) and were shown to be structurally and functionally similar to natural ECMs. A genetically-engineered M13 bacteriophage was constructed to display Arg-Gly-Asp (RGD) peptides on its surface. The physicochemical properties of RGD peptide-displaying M13 bacteriophage (RGD-M13 phage)/PLGA nanofibers were characterized by using scanning electron microscopy and Fourier-transform infrared spectroscopy. We used immunofluorescence staining to confirm that M13 bacteriophages were homogenously distributed in RGD-M13 phage/PLGA matrices. Furthermore, RGD-M13 phage/PLGA nanofibrous matrices, having excellent biocompatibility, can enhance the behaviors of vascular smooth muscle cells. This result suggests that RGD-M13 phage/PLGA nanofibrous matrices have potentials to serve as tissue engineering scaffolds.

Facile Fabrication of Nanofibrillated Chitin/Ag2O Heterostructured Aerogels with High Iodine Capture Efficiency.

PubMed

Gao, Runan; Lu, Yun; Xiao, Shaoliang; Li, Jian

2017-06-27

Nanofibrillated chitin/Ag 2 O aerogels were fabricated for radioiodine removal. Chitin was first fabricated into nanofibers with abundant acetyl amino groups (-NHCOCH 3 ) on the surface. Then, highly porous chitin nanofiber (ChNF) aerogels were obtained via freeze-drying. The ChNF aerogels exhibited a low bulk density of 2.19 mg/cm 3 and a high specific surface area of 179.71 m 2 /g. Ag 2 O nanoparticles were evenly anchored on the surfaces of ChNF scaffolds via strong interactions with -NHCOCH 3 groups, subsequently yielding Ag 2 O@ChNF heterostructured aerogels. The composites were used as efficient absorbents to remove radioiodine anions from water and capture a high amount of I 2 vapor in the forms of AgI and iodine molecules. The adsorption capacity of the composite monoliths can reach up to 2.81 mmol/g of I - anions. The high adsorbability of the composite monolithic aerogel signifies its potential applications in radioactive waste disposal.

Study of poly(L-lactide) microparticles based on supercritical CO2.

PubMed

Chen, Ai-Zheng; Pu, Xi-Ming; Kang, Yun-Qing; Liao, Li; Yao, Ya-Dong; Yin, Guang-Fu

2007-12-01

Poly(L-lactide) (PLLA) microparticles were prepared in supercritical anti-solvent process. The effects of several key factors on surface morphology, and particle size and particle size distribution were investigated. These factors included initial drops size, saturation ratio of PLLA solution, pressure, temperature, concentration of the organic solution, the flow rate of the solution and molecular weight of PLLA. The results indicated that the saturation ratio of PLLA solution, concentration of the organic solution and flow rate of the solution played important roles on the properties of products. Various microparticles with the mean particle size ranging from 0.64 to 6.64 microm, could be prepared by adjusting the operational parameters. Fine microparticles were obtained in a process namely solution-enhanced dispersion by supercritical fluids (SEDS) process with dichloromethane/acetone mixture as solution.

Effect of Adipose-Derived Stromal Cells and BMP12 on Intrasynovial Tendon Repair: A Biomechanical, Biochemical, and Proteomics Study

PubMed Central

Gelberman, Richard H.; Shen, Hua; Kormpakis, Ioannis; Rothrauff, Benjamin; Yang, Guang; Tuan, Rocky S.; Xia, Younan; Sakiyama-Elbert, Shelly; Silva, Matthew J.; Thomopoulos, Stavros

2016-01-01

The outcomes of flexor tendon repair are highly variable. As recent efforts to improve healing have demonstrated promise for growth factor- and cell-based therapies, the objective of the current study was to enhance repair via application of autologous adipose derived stromal cells (ASCs) and the tenogenic growth factor bone morphogenetic protein (BMP) 12. Controlled delivery of cells and growth factor was achieved in a clinically relevant canine model using a nanofiber/fibrin-based scaffold. Control groups consisted of repair-only (no scaffold) and acellular scaffold. Repairs were evaluated after 28 days of healing using biomechanical, biochemical, and proteomics analyses. Range of motion was reduced in the groups that received scaffolds compared to normal. There was no effect of ASC+BMP12 treatment for range of motion or tensile properties outcomes versus repair-only. Biochemical assays demonstrated increased DNA, glycosaminoglycans, and crosslink concentration in all repair groups compared to normal, but no effect of ASC+BMP12. Total collagen was significantly decreased in the acellular scaffold group compared to normal and significantly increased in the ASC+BMP12 group compared to the acellular scaffold group. Proteomics analysis comparing healing tendons to uninjured tendons revealed significant increases in proteins associated with inflammation, stress response, and matrix degradation. Treatment with ASC+BMP12 amplified these unfavorable changes. In summary, the treatment approach used in this study induced a negative inflammatory reaction at the repair site leading to poor healing. Future approaches should consider cell and growth factor delivery methods that do not incite negative local reactions. PMID:26445383

Nitric Oxide-Releasing Silica Nanoparticle-Doped Polyurethane Electrospun Fibers

PubMed Central

Koh, Ahyeon; Carpenter, Alexis W.; Slomberg, Danielle L.; Schoenfisch, Mark H.

2013-01-01

Electrospun polyurethane fibers doped with nitric oxide (NO)-releasing silica particles are presented as novel macromolecular scaffolds with prolonged NO-release and high porosity. Fiber diameter (119–614 nm) and mechanical strength (1.7–34.5 MPa of modulus) were varied by altering polyurethane type and concentration, as well as the NO-releasing particle composition, size, and concentration. The resulting NO-releasing electrospun nanofibers exhibited ~83% porosity with flexible plastic or elastomeric behavior. The use of N-diazeniumdiolate- or S-nitrosothiol-modified particles yielded scaffolds exhibiting a wide range of NO release totals and durations (7.5 nmol mg−1–0.12 μmol mg−1 and 7 h to 2 weeks, respectively). The application of NO-releasing porous materials as coating for subcutaneous implants may improve tissue biocompatibility by mitigating the foreign body response and promoting cell integration. PMID:23915047

The toxicity of rifampicin polylactic acid nanoparticles against Mycobacterium bovis BCG and human macrophage THP-1 cell line

NASA Astrophysics Data System (ADS)

Erokhina, M.; Rybalkina, E.; Barsegyan, G.; Onishchenko, G.; Lepekha, L.

2015-11-01

Tuberculosis is rapidly becoming a major health problem. The rise in tuberculosis incidence stimulates efforts to develop more effective delivery systems for the existing antituberculous drugs while decreasing the side effects. The nanotechnology may provide novel drug delivery tools allowing controlled drug release. Rifampicin is one of the main antituberculous drugs, characterized by high toxicity, and Poly (L-lactic acid) (PLLA) is a biodegradable polymer used for the preparation of encapsulated drugs. The aim of our work was to evaluate the toxicity of rifampicin-PLLA nanoparticles against Mycobacterium bovis BCG using human macrophage THP-1 cell line. Our data demonstrate that rifampicin-PLLA is effective against M. bovis BCG in the infected macrophages. The drug is inducing the dysfunction of mitochondria and apoptosis in the macrophages and is acting as a potential substrate of Pgp thereby modulating cell chemosensitivity. The severity of the toxic effects of the rifampicin-PLLA nanoparticles is increasing in a dose-dependent manner. We suggest that free rifampicin induces death of M. bovis BCG after PLLA degradation and diffusion from phago-lysosomes to cytoplasm causing mitochondria dysfunction and affecting the Pgp activity.

Morphology characterization and biocompatibility study of PLLA (Poly-L-Llactid-Acid) coating chitosan as stent for coronary heart disease

NASA Astrophysics Data System (ADS)

Widiyanti, Prihartini; Paramadini, Adanti W.; Jabbar, Hajria; Fatimah, Inas; Nisak, Fadila N. K.; Puspitasari, Rahma A.

2016-03-01

Cardiovascular disease is a global disease with high urgency. In the severe case of coronary heart disease while a blockage in the coronary arteries reach 75% or more, the patient required stent implantation. Stents are made of metal which has many limitations that can lead to blood clots and stent incompatibility toward the size of the blood vessels. There is a metal stent replacement solution that made from polymer material which is biocompatible. PLLA also has biocompatibility and good mechanical strength. PLLA stent will be coated with chitosan as a candidate for drug-coated stents which is able to work as a drug carrier. The aim of this study is to know the morphology information and biocompability status of PLLA coating chitosan as candidate of heart stent. Morphological results using SEM showed a smooth surface structure which reinforced clinical standard of stent material. Results of cytotoxicity test by MTT Assay method showed that the result of four samples in this experiment living cells is reached 90% which is non toxic and safe to use in the human body. %). The conclusion of this study is PLLA is polymer has potency to be used as stent material.

Tough and Sustainable Graft Block Copolymer Thermoplastics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Jiuyang; Li, Tuoqi; Mannion, Alexander M.

Fully sustainable poly[HPMC-g-(PMVL-b-PLLA)] graft block copolymer thermoplastics were prepared from hydroxypropyl methylcellulose (HPMC), β-methyl-δ-valerolactone (MVL), and l-lactide (LLA) using a facile two-step sequential addition approach. In these materials, rubbery PMVL functions as a bridge between the semirigid HPMC backbone and the hard PLLA end blocks. This specific arrangement facilitates PLLA crystallization, which induces microphase separation and physical cross-linking. By changing the backbone molar mass or side chain composition, these thermoplastic materials can be easily tailored to access either plastic or elastomeric behavior. Moreover, the graft block architecture can be utilized to overcome the processing limitations inherent to linear block polymers.more » Good control over molar mass and composition enables the deliberate design of HPMC-g-(PMVL-b-PLLA) samples that are incapable of microphase separation in the melt state. These materials are characterized by relatively low zero shear viscosities in the melt state, an indication of easy processability. The simple and scalable synthetic procedure, use of inexpensive and renewable precursors, and exceptional rheological and mechanical properties make HPMC-g-(PMVL-b-PLLA) polymers attractive for a broad range of applications.« less

One-Step Method to Prepare PLLA Porous Microspheres in a High-Voltage Electrostatic Anti-Solvent Process

PubMed Central

Wang, Ying; Zhu, Li-Hui; Chen, Ai-Zheng; Xu, Qiao; Hong, Yu-Juan; Wang, Shi-Bin

2016-01-01

A one-step method using a high-voltage electrostatic anti-solvent process was employed to fabricate poly-l-lactide (PLLA) porous microspheres (PMs). To address the simplification and control of the preparation process, a 24 full factorial experiment was performed to optimize the operating process and analyze the effect of the factors on the morphology and aerodynamic properties of the PLLA PMs, and various characterization tests were also performed. The resulting PLLA PMs exhibited an even and porous morphology with a density less than 0.4 g/cm3, a geometric mean diameter (Dg) of 10–30 μm, an aerodynamic diameter (Da) of 1–5 μm, a fine particle fraction (FPF) of 56.3%, and a porosity of 76.2%, meeting the requirements for pulmonary drug delivery. The physicochemical characterizations reveal that no significant chemical change occurred in the PLLA during the process. An investigation of its in vitro cytotoxicity and pulmonary toxicity shows no obvious toxic response, indicating good biosafety. This study indicates that the one-step method using a high-voltage electrostatic anti-solvent process has great potential in developing an inhalable drug carrier for pulmonary drug delivery. PMID:28773489