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Sample records for hydroxyapatite scaffolds fabricated

  1. Characterization of the flow behavior of alginate/hydroxyapatite mixtures for tissue scaffold fabrication.

    PubMed

    Tian, X Y; Li, M G; Cao, N; Li, J W; Chen, X B

    2009-12-01

    Mixtures of alginate and hydroxyapatite (HA) are promising materials for biomedical applications such as the fabrication of tissue scaffolds. In this paper, the flow behavior of alginate/HA mixtures was investigated and determined to be dependent on the concentration of both alginate and HA, and temperature. The relationships were mathematically established and verified with experimental results. As applied to the tissue scaffold fabrication, the flow rate of the biomaterial solution was predicted from the established flow behavior and verified by experiments. On this basis, the moving speed of the needle was determined and used in the tissue scaffold fabrication. The results obtained show that the knowledge of the flow behavior is essential to the fabrication of tissue scaffolds with an interconnected microstructure.

  2. Fracture modes under uniaxial compression in hydroxyapatite scaffolds fabricated by robocasting.

    PubMed

    Miranda, Pedro; Pajares, Antonia; Saiz, Eduardo; Tomsia, Antoni P; Guiberteau, Fernando

    2007-12-01

    The fracture modes of hydroxyapatite (HA) scaffolds fabricated by direct-write assembly (robocasting) are analyzed in this work. Concentrated HA inks with suitable viscoelastic properties were developed to enable the fabrication of prototype structures consisting of a 3-D square mesh of interpenetrating rods. The fracture behavior of these model scaffolds under compressive stresses is determined from in situ uniaxial tests performed in two different directions: perpendicular to the rods and along one of the rod directions. The results are analyzed in terms of the stress field calculated by finite element modeling (FEM). This analysis provides valuable insight into the mechanical behavior of scaffolds for bone tissue engineering applications fabricated by robocasting. Copyright 2007 Wiley Periodicals, Inc.

  3. [Preparation of elastic porous cell scaffold fabricated with combined polydimethylsiloxane (PDMS) and hydroxyapatite (HA)].

    PubMed

    Yang, Yang; Lan, Ding; Huang, Yan; Li, Yanming; Wang, Yuren; Sun, Lianwen; Fan, Yubo

    2014-06-01

    Polydimethylsiloxane (PDMS) and hydroxyapatite (HA) were combined in our laboratory to fabricate an elastic porous cell scaffold with pore-forming agent, and then the scaffold was used as culture media for rat bone marrow derived mesenchymal stem cells (rBMSCs). Different porous materials (square and circular in shape) were prepared by different pore-forming agents (NaCl or paraffin spheres) with adjustable porosity (62%-76%). The HA crystals grew on the wall of hole when the material was exposed to SBF solutions, showing its biocompatibility and ability to support the cells to attach on the materials.

  4. Silk as a biocohesive sacrificial binder in the fabrication of hydroxyapatite load bearing scaffolds

    PubMed Central

    McNamara, Stephanie L.; Rnjak-Kovacina, Jelena; Schmidt, Daniel; Lo, Tim J.; Kaplan, David L.

    2014-01-01

    Limitations of current clinical methods for bone repair continue to fuel the demand for a high strength, bioactive bone replacement material. Recent attempts to produce porous scaffolds for bone regeneration have been limited by the intrinsic weakness associated with high porosity materials. In this study, ceramic scaffold fabrication techniques for potential use in load-bearing bone repairs have been developed using naturally derived silk from Bombyx mori. Silk was first employed for ceramic grain consolidation during green body formation, and later as a sacrificial polymer to impart porosity during sintering. These techniques allowed preparation of hydroxyapatite (HA) scaffolds that exhibited a wide range of mechanical and porosity profiles, with some displaying unusually high compressive strength up to 152.4 ± 9.1 MPa. Results showed that the scaffolds exhibited a wide range of compressive strengths and moduli (8.7 ± 2.7 MPa to 152.4 ± 9.1 MPa and 0.3 ± 0.1 GPa to 8.6 ± 0.3 GPa) with total porosities of up to 62.9 ± 2.7% depending on the parameters used for fabrication. Moreover, HA-silk scaffolds could be molded into large, complex shapes, and further machined post-sinter to generate specific three-dimensional geometries. Scaffolds supported bone marrow-derived mesenchymal stem cell attachment and proliferation, with no signs of cytotoxicity. Therefore, silk-fabricated HA scaffolds show promise for load bearing bone repair and regeneration needs. PMID:24881027

  5. In-vivo behavior of Si-hydroxyapatite/polycaprolactone/DMB scaffolds fabricated by 3D printing.

    PubMed

    Meseguer-Olmo, Luis; Vicente-Ortega, Vicente; Alcaraz-Baños, Miguel; Calvo-Guirado, José Luis; Vallet-Regí, María; Arcos, Daniel; Baeza, Alejandro

    2013-07-01

    Scaffolds made of polycaprolactone and nanocrystalline silicon-substituted hydroxyapatite have been fabricated by 3D printing rapid prototyping technique. To asses that the scaffolds fulfill the requirements to be considered for bone grafting applications, they were implanted in New Zealand rabbits. Histological and radiological studies have demonstrated that the scaffolds implanted in bone exhibited an excellent osteointegration without the interposition of fibrous tissue between bone and implants and without immune response after 4 months of implantation. In addition, we have evaluated the possibility of improving the scaffolds efficiency by incorporating demineralized bone matrix during the preparation by 3D printing. When demineralized bone matrix (DBM) is incorporated, the efficacy of the scaffolds is enhanced, as new bone formation occurs not only in the peripheral portions of the scaffolds but also within its pores after 4 months of implantation. This enhanced performance can be explained in terms of the osteoinductive properties of the DBM in the scaffolds, which have been assessed through the new bone tissue formation when the scaffolds are ectopically implanted.

  6. Fabrication of Porous Hydroxyapatite Scaffolds as Artificial Bone Preform and its Biocompatibility Evaluation

    PubMed Central

    2014-01-01

    In this study, a novel porous hydroxyapatite scaffold was designed and fabricated to imitate natural bone through a multipass extrusion process. The conceptual design manifested unidirectional microchannels at the exterior part of the scaffold to facilitate rapid biomineralization and a central canal that houses the bone marrow. External and internal fissures were minimized during microwave sintering at 1,100°C. No deformation was noted, and a mechanically stable scaffold was fabricated. Detailed microstructure of the fabricated artificial bone was examined by scanning electron microscope and X-ray diffractometer, and material properties like compressive strength were evaluated. The initial biocompatibility was examined by the cell proliferation of MG-63 osteoblast-like cells using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Preliminary in vivo investigation in a rabbit model after 4 weeks and 8 weeks of implantation showed full osteointegration of the scaffold with the native tissue, and formation of bone tissue within the pore network, as examined by microcomputed tomography analyses and histological staining. Osteon-like bone microarchitecture was observed along the unidirectional channel with microblood vessels. These confirm a biomimetic regeneration model in the implanted bone scaffold, which can be used as an artificial alternative for damaged bone. PMID:24399056

  7. Fabrication and characterization of drug-loaded nano-hydroxyapatite/polyamide 66 scaffolds modified with carbon nanotubes and silk fibroin

    PubMed Central

    Yao, Meng-Zhu; Huang-Fu, Ming-Yi; Liu, Hui-Na; Wang, Xia-Rong; Sheng, Xiaoxia; Gao, Jian-Qing

    2016-01-01

    Nano-hydroxyapatite/polyamide 66 (nHA/PA66) porous scaffolds were fabricated by a phase inversion method. Carbon nanotubes (CNTs) and silk fibroin (SF) were used to modify the surface of the nHA/PA66 scaffolds by freeze-drying and cross-linking. Dexamethasone was absorbed to the CNTs to promote the osteogenic differentiation of bone mesenchymal stem cells (BMSCs). The cell viability of BMSCs was investigated by changing the concentration of the CNT dispersion, and the most biocompatible scaffold was selected. In addition, the morphology and mechanical property of the scaffolds were investigated. The results showed that the nHA/PA66 scaffolds modified with CNTs and SF met the requirements of bone tissue engineering scaffolds. The dexamethasone-loaded CNT/SF-nHA/PA66 composite scaffold promoted the osteogenic differentiation of BMSCs, and the drug-loaded scaffolds are expected to function as effective bone tissue engineering scaffolds. PMID:27920525

  8. Fabrication and characterization of drug-loaded nano-hydroxyapatite/polyamide 66 scaffolds modified with carbon nanotubes and silk fibroin.

    PubMed

    Yao, Meng-Zhu; Huang-Fu, Ming-Yi; Liu, Hui-Na; Wang, Xia-Rong; Sheng, Xiaoxia; Gao, Jian-Qing

    Nano-hydroxyapatite/polyamide 66 (nHA/PA66) porous scaffolds were fabricated by a phase inversion method. Carbon nanotubes (CNTs) and silk fibroin (SF) were used to modify the surface of the nHA/PA66 scaffolds by freeze-drying and cross-linking. Dexamethasone was absorbed to the CNTs to promote the osteogenic differentiation of bone mesenchymal stem cells (BMSCs). The cell viability of BMSCs was investigated by changing the concentration of the CNT dispersion, and the most biocompatible scaffold was selected. In addition, the morphology and mechanical property of the scaffolds were investigated. The results showed that the nHA/PA66 scaffolds modified with CNTs and SF met the requirements of bone tissue engineering scaffolds. The dexamethasone-loaded CNT/SF-nHA/PA66 composite scaffold promoted the osteogenic differentiation of BMSCs, and the drug-loaded scaffolds are expected to function as effective bone tissue engineering scaffolds.

  9. Fabrication of hybrid nanocomposite scaffolds by incorporating ligand-free hydroxyapatite nanoparticles into biodegradable polymer scaffolds and release studies

    PubMed Central

    Farkas, Balazs; Rodio, Marina; Romano, Ilaria; Diaspro, Alberto; Intartaglia, Romuald

    2015-01-01

    Summary We report on the optical fabrication approach of preparing free-standing composite thin films of hydroxyapatite (HA) and biodegradable polymers by combining pulsed laser ablation in liquid and mask-projection excimer laser stereolithography (MPExSL). Ligand-free HA nanoparticles were prepared by ultrafast laser ablation of a HA target in a solvent, and then the nanoparticles were dispersed into the liquid polymer resin prior to the photocuring process using MPExSL. The resin is poly(propylene fumarate) (PPF), a photo-polymerizable, biodegradable material. The polymer is blended with diethyl fumarate in 7:3 w/w to adjust the resin viscosity. The evaluation of the structural and mechanical properties of the fabricated hybrid thin film was performed by means of SEM and nanoindentation, respectively, while the chemical and degradation studies were conducted through thermogravimetric analysis, and FTIR. The photocuring efficiency was found to be dependent on the nanoparticle concentration. The MPExSL process yielded PPF thin films with a stable and homogenous dispersion of the embedded HA nanoparticles. Here, it was not possible to tune the stiffness and hardness of the scaffolds by varying the laser parameters, although this was observed for regular PPF scaffolds. Finally, the gradual release of the hydroxyapatite nanoparticles over thin film biodegradation is reported. PMID:26734513

  10. Fabrication of hybrid nanocomposite scaffolds by incorporating ligand-free hydroxyapatite nanoparticles into biodegradable polymer scaffolds and release studies.

    PubMed

    Farkas, Balazs; Rodio, Marina; Romano, Ilaria; Diaspro, Alberto; Intartaglia, Romuald; Beke, Szabolcs

    2015-01-01

    We report on the optical fabrication approach of preparing free-standing composite thin films of hydroxyapatite (HA) and biodegradable polymers by combining pulsed laser ablation in liquid and mask-projection excimer laser stereolithography (MPExSL). Ligand-free HA nanoparticles were prepared by ultrafast laser ablation of a HA target in a solvent, and then the nanoparticles were dispersed into the liquid polymer resin prior to the photocuring process using MPExSL. The resin is poly(propylene fumarate) (PPF), a photo-polymerizable, biodegradable material. The polymer is blended with diethyl fumarate in 7:3 w/w to adjust the resin viscosity. The evaluation of the structural and mechanical properties of the fabricated hybrid thin film was performed by means of SEM and nanoindentation, respectively, while the chemical and degradation studies were conducted through thermogravimetric analysis, and FTIR. The photocuring efficiency was found to be dependent on the nanoparticle concentration. The MPExSL process yielded PPF thin films with a stable and homogenous dispersion of the embedded HA nanoparticles. Here, it was not possible to tune the stiffness and hardness of the scaffolds by varying the laser parameters, although this was observed for regular PPF scaffolds. Finally, the gradual release of the hydroxyapatite nanoparticles over thin film biodegradation is reported.

  11. Fabrication and Dynamic Mechanical Analysis of Hydroxyapatite Nanoparticle/Gelatin Porous Scaffolds

    NASA Astrophysics Data System (ADS)

    Ghossein, Hicham

    The application of engineered biomaterial scaffolds for hard tissue repair critically depends on the scaffold's internal architecture at various length scales. The pore size, shape, surface morphology, and pore connectivity are among the most important factors that affect the scaffold's mechanical properties and biointegration. Reported in this thesis are the results of the investigation of porous constructs fabricated by a freeze-drying process from synthetic nanosized hydroxyapatite / gelatin (nanoHA/Gel) dispersions with different nanoHA/Gel ratios (nanoHA loading was varied from 0 to 50 % by weight). The fabricated scaffolds had porosity up to 90% with pore size in the range of 100 - 500 im, and good distribution of HA nanoparticles within the gelatin matrix. Such porosity is considered to be close to optimal to promote a good cell adhesion in the potential applications of prepared constructs. The fabricated scaffolds have been investigated using X-ray diffraction (XRD), Fourier-Transform Infrared Spectroscopy (FTIR), and Dynamic Mechanical Analysis (DMA). Dynamic mechanical analysis of as-fabricated scaffolds revealed that the scaffolds achieved maximum bending and tensile moduli up to 1.28 GPa and 1.5 GPa, respectively, when nanoHA loading was around 30 % by weight. The bending modulus increases by a factor of 1.6, while the Tension modulus increased by a factor of 0.8 after the cross-linking of polymer. Higher nanoHA loading above 50 % by weight results in bending modulus of about 700 MPa and Tension modulus of about 200 MPa only. However, the cross-linking still enhanced the bending up to 1 GPa while it did not affect much the Tension modulus in 50% nanoHA/gelatin constructs. It has been shown that the cross-linking with glutaraldehyde solution improves the morphological structure of the scaffolds, while there was no apparent effect of the cross-linking on the chemical changes in both organic and inorganic content during the processing. The results of this

  12. In vitro and animal study of novel nano-hydroxyapatite/poly(epsilon-caprolactone) composite scaffolds fabricated by layer manufacturing process.

    PubMed

    Heo, Su-Jin; Kim, Seung-Eon; Wei, Jie; Kim, Dong Hwa; Hyun, Young-Taek; Yun, Hui-Suk; Kim, Hyung Keun; Yoon, Taek Rim; Kim, Su-Hyang; Park, Su-A; Shin, Ji Won; Shin, Jung-Woog

    2009-05-01

    The purpose of this study was to propose a computer-controllable scaffold structure made by a layer manufacturing process (LMP) with addition of nano- or micro-sized particles and to investigate the effects of particle size in vitro. In addition, the superiority of this LMP method over the conventional scaffolds made by salt leaching and gas forming process was investigated through animal study. Using the LMP, we have created a new nano-sized hydroxyapatite/poly(epsilon-caprolactone) composite (n-HPC) scaffold and a micro-sized hydroxyapatite/poly(epsilon-caprolactone) composite (m-HPC) scaffold for bone tissue engineering applications. The scaffold macropores were well interconnected, with a porosity of 73% and a pore size of 500 microm. The compressive modulus of the n-HPC and m-HPC scaffolds was 6.76 and 3.18 MPa, respectively. We compared the cellular responses to the two kinds of scaffolds. Both n-HPC and m-HPC exhibited good in vitro biocompatibility. Attachment and proliferation of mesenchymal stem cells were better on the n-HPC than on the m-HPC scaffold. Moreover, significantly higher alkaline phosphatase activity and calcium content were observed on the n-HPC than on the m-HPC scaffold. In an animal study, the LMP scaffolds enhanced bone formation, owing to their well-interconnected pores. Radiological and histological examinations confirmed that the new bony tissue had grown easily into the entire n-HPC scaffold fabricated by LMP. We suggest that the well-interconnected pores in the LMP scaffolds might encourage cell attachment, proliferation, and migration to stimulate cell functions, thus enhancing bone formation in the LMP scaffolds. This study shows that bioactive and biocompatible n-HPC composite scaffolds prepared using an LMP have potential applications in bone tissue engineering.

  13. Oxygen and nitrogen plasma etching of three-dimensional hydroxyapatite/chitosan scaffolds fabricated by additive manufacturing

    NASA Astrophysics Data System (ADS)

    Myung, Sung-Woon; Kim, Byung-Hoon

    2016-01-01

    Three-dimensional (3D) chitosan and hydroxyapatite (HAp)/chitosan (CH) scaffolds were fabricated by additive manufacturing, then their surfaces were etched with oxygen (O2) and nitrogen (N2) plasma. O2 and N2 plasma etching was performed to increase surface properties such as hydrophilicity, roughness, and surface chemistry on the scaffolds. After etching, hydroxyapatite was exposed on the surface of 3D HAp/CH scaffolds. The surface morphology and chemical properties were characterized by contact angle measurement, scanning electron microscopy, X-ray diffraction, and attenuated total reflection Fourier infrared spectroscopy. The cell viability of 3D chitosan scaffolds was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The differentiation of preosteoblast cells was evaluated by alkaline phosphatase assay. The cell viability was improved by O2 and N2 plasma etching of 3D chitosan scaffolds. The present fabrication process for 3D scaffolds might be applied to a potential tool for preparing biocompatible scaffolds.

  14. Hydrothermal fabrication of hydroxyapatite/chitosan/carbon porous scaffolds for bone tissue engineering.

    PubMed

    Long, Teng; Liu, Yu-Tai; Tang, Sha; Sun, Jin-Liang; Guo, Ya-Ping; Zhu, Zhen-An

    2014-11-01

    Porous carbon fiber felts (PCFFs) have great applications in orthopedic surgery because of the strong mechanical strength, low density, high stability, and porous structure, but they are biologically inert. To improve their biological properties, we developed, for the first time, the hydroxyapatite (HA)/chitosan/carbon porous scaffolds (HCCPs). HA/chitosan nanohybrid coatings have been fabricated on PCFFs according to the following stages: (i) deposition of chitosan/calcium phosphate precursors on PCFFs; and (ii) hydrothermal transformation of the calcium phosphate precursors in chitosan matrix into HA nanocrystals. The scanning electron microscopy images indicate that PCFFs are uniformly covered with elongated HA nanoplates and chitosan, and the macropores in PCFFs still remain. Interestingly, the calcium-deficient HA crystals exist as plate-like shapes with thickness of 10-18 nm, width of 30-40 nm, and length of 80-120 nm, which are similar to the biological apatite. The HA in HCCPs is similar to the mineral of natural bone in chemical composition, crystallinity, and morphology. As compared with PCFFs, HCCPs exhibit higher in vitro bioactivity and biocompatibility because of the presence of the HA/chitosan nanohybrid coatings. HCCPs not only promote the formation of bone-like apatite in simulated body fluid, but also improve the adhesion, spreading, and proliferation of human bone marrow stromal cells. Hence, HCCPs have great potentials as scaffold materials for bone tissue engineering and implantation. © 2014 Wiley Periodicals, Inc.

  15. Microwave-assisted fabrication of chitosan-hydroxyapatite superporous hydrogel composites as bone scaffolds.

    PubMed

    Beşkardeş, Işıl Gerçek; Demirtaş, Tuğrul Tolga; Durukan, Müge Dağlı; Gümüşderelioğlu, Menemşe

    2015-11-01

    In this study, a novel scaffold fabrication method was developed by combining microwave irradiation and gas foaming. Chitosan superporous hydrogels (SPHs) and chitosan-hydroxyapatite (HA) superporous hydrogel composites (SPHCs) were prepared by using this method in the presence of crosslinking agent, glyoxal, and a gas-blowing agent, NaHCO3. In order to examine the effect of HA on composite structure and cellular behaviour, two types of HA particles, i.e. spherical beads in 45-80 µm diameter and powder form, were used. While rapid heating with microwave irradiation enhances gas blowing, pH increment, which is accelerated by NaHCO3 decomposition, provides better crosslinking. Thus, interconnected and well-established macroporous hydrogels/hydrogel composites were produced easily and rapidly (~1 min). Cell culture studies, which were carried out under static and dynamic conditions with MC3T3-E1 pre-osteoblastic cells, indicated that chitosan-HA bead SPHCs supported cellular proliferation and osteoblastic differentiation better than chitosan SPHs and chitosan-HA powder SPHCs. In conclusion, simultaneous gas foaming and microwave crosslinking can be evaluated for the preparation of composite scaffolds which have superior properties for bone tissue engineering.

  16. Fabrication and characterization of layered chitosan/silk fibroin/nano-hydroxyapatite scaffolds with designed composition and mechanical properties.

    PubMed

    Zhou, Ting; Wu, Jingjing; Liu, Jiaoyan; Luo, Ying; Wan, Ying

    2015-07-30

    Chitosan/nano-hydroxyapatite (HA) composites were first prepared and then used together with chitosan and silk fibroin (SF) to produce a type of four-layer porous scaffold that is potentially applicable for articular cartilage repair. The bottom layer of the scaffold was built with the chitosan/HA composite and the other three layers of the scaffold were fabricated using chitosan/SF composites in which the content of the chitosan and SF was altered in a mutually reversed trend. The so-produced chitosan/SF/HA scaffolds were further crosslinked using tripolyphosphate to achieve enhanced mechanical properties. Interconnected porous microstructures throughout the scaffolds were constructed using a temperature gradient processing technique, and the resultant scaffolds were endowed with graded pore-sizes and porosities as well as porous interface zones between contiguous layers without visual clefts. The compressive modulus and stress at 10% strain of the scaffolds in wet state showed a gradient-changed trend which partially mimics the compressive mechanical properties of an articular cartilage matrix. Cell culture on some chitosan/SF/HA scaffolds for a period of time of up to 14 d showed that the scaffolds were able to well support the growth and infiltration of cells, suggesting that the presently developed chitosan/SF/HA scaffolds have promising potential for articular cartilage repair.

  17. Fabrication and characterization of novel nano-biocomposite scaffold of chitosan-gelatin-alginate-hydroxyapatite for bone tissue engineering.

    PubMed

    Sharma, Chhavi; Dinda, Amit Kumar; Potdar, Pravin D; Chou, Chia-Fu; Mishra, Narayan Chandra

    2016-07-01

    A novel nano-biocomposite scaffold was fabricated in bead form by applying simple foaming method, using a combination of natural polymers-chitosan, gelatin, alginate and a bioceramic-nano-hydroxyapatite (nHAp). This approach of combining nHAp with natural polymers to fabricate the composite scaffold, can provide good mechanical strength and biological property mimicking natural bone. Environmental scanning electron microscopy (ESEM) images of the nano-biocomposite scaffold revealed the presence of interconnected pores, mostly spread over the whole surface of the scaffold. The nHAp particulates have covered the surface of the composite matrix and made the surface of the scaffold rougher. The scaffold has a porosity of 82% with a mean pore size of 112±19.0μm. Swelling and degradation studies of the scaffold showed that the scaffold possesses excellent properties of hydrophilicity and biodegradability. Short term mechanical testing of the scaffold does not reveal any rupturing after agitation under physiological conditions, which is an indicative of good mechanical stability of the scaffold. In vitro cell culture studies by seeding osteoblast cells over the composite scaffold showed good cell viability, proliferation rate, adhesion and maintenance of osteoblastic phenotype as indicated by MTT assay, ESEM of cell-scaffold construct, histological staining and gene expression studies, respectively. Thus, it could be stated that the nano-biocomposite scaffold of chitosan-gelatin-alginate-nHAp has the paramount importance for applications in bone tissue-engineering in future regenerative therapies. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Improved biocomposite development of poly(vinyl alcohol) and hydroxyapatite for tissue engineering scaffold fabrication using selective laser sintering.

    PubMed

    Wiria, Florencia Edith; Chua, Chee Kai; Leong, Kah Fai; Quah, Zai Yan; Chandrasekaran, Margam; Lee, Mun Wai

    2008-03-01

    In scaffold guided tissue engineering (TE), temporary three-dimensional scaffolds are essential to guide and support cell proliferation. Selective Laser Sintering (SLS) is studied for the development of such scaffolds by eliminating pore spatial control problems faced in conventional scaffolds fabrication methods. SLS offers good user control over the scaffold's microstructures by adjusting its main processing parameters, namely the laser power, scan speed and part bed temperature. This research focuses on the improvements in the fabrication of TE scaffolds using SLS with powder biomaterials, namely hydroxyapatite (HA) and poly(vinyl alcohol) (PVA). Grinding of as-received PVA powder to varying particle sizes and two methods of mixing are investigated as the preparation process to determine a better mixing method that would enhance the mixture homogeneity. Suitable sintering conditions for the improved biocomposite are then achieved by varying the important process parameters such as laser power, scan speed and part bed temperature.SLS fabricated samples are characterized using Fourier Transform Infrared Spectrometer (FTIR) and Scanning Electron Microscope (SEM). FTIR results show that the grinding and sintering processes neither compromise the chemical composition of the PVA nor cause undue degradation. Visual analysis of the grinding, powder mixing and sintering effect are carried out with SEM. The SEM observations show improvements in the sintering effects. The favorable outcome ascertains PVA/HA biocomposite as a suitable material to be processed by SLS for TE scaffolds.

  19. Parameters optimization for the fabrication of phosphate glass/hydroxyapatite nanocomposite scaffold

    NASA Astrophysics Data System (ADS)

    Govindan, R.; Girija, E. K.

    2015-06-01

    Three-dimensional, highly porous, bioactive and biodegradable phosphate glass and nanohydroxyapatite (n-HA) composite scaffolds was fabricated by the polymer foam replication technique. Polyurethane foam (PU) and polyvinyl alcohol (PVA) were used as template and binder, respectively. Optimization of composition and sintering temperature is carried out for tissue engineering scaffold fabrication.

  20. Poly(lactide-co-glycolide)/hydroxyapatite nanofibrous scaffolds fabricated by electrospinning for bone tissue engineering.

    PubMed

    Lao, Lihong; Wang, Yingjun; Zhu, Yang; Zhang, Yuying; Gao, Changyou

    2011-08-01

    Poly(lactide-co-glycolide) (PLGA) nanofibrous composite scaffolds having nano-hydroxyapatite particles (HAp) in the fibers were prepared by electrospinning of PLGA and HAp with an average diameter of 266.6 ± 7.3 nm. Microscopy and spectroscopy characterizations confirmed integration of the crystalline HAp in the scaffolds. Agglomerates gradually appeared and increased on the fiber surface along with increase of the HAp concentration. In vitro mineralization in a 5 × simulated body fluid (SBF) revealed that the PLGA/HAp nanofibrous scaffolds had a stronger biomineralization ability than the control PLGA scaffolds. Biological performance of the nanofibrous scaffolds of the control PLGA and PLGA with 5 wt% HAp (PLGA/5HAp) was assessed by in vitro culture of neonatal mouse calvaria-derived MC3T3-E1 osteoblasts. Both types of the scaffolds could support cell proliferation and showed sharp increase of viability until 7 days, but the cells cultured on the PLGA/5HAp nanofibers showed a more spreading morphology. Despite the similar level of the cell viability and cell number at each time interval, the alkaline phosphatase secretion was significantly enhanced on the PLGA/5HAp scaffolds, indicating the higher bioactivity of the as-prepared nano-HAp and the success of the present method for preparing biomimetic scaffold for bone regeneration.

  1. Fabrication of novel Si-doped hydroxyapatite/gelatine scaffolds by rapid prototyping for drug delivery and bone regeneration.

    PubMed

    Martínez-Vázquez, F J; Cabañas, M V; Paris, J L; Lozano, D; Vallet-Regí, M

    2015-03-01

    Porous 3-D scaffolds consisting of gelatine and Si-doped hydroxyapatite were fabricated at room temperature by rapid prototyping. Microscopic characterization revealed a highly homogeneous structure, showing the pre-designed porosity (macroporosity) and a lesser in-rod porosity (microporosity). The mechanical properties of such scaffolds are close to those of trabecular bone of the same density. The biological behavior of these hybrid scaffolds is greater than that of pure ceramic scaffolds without gelatine, increasing pre-osteoblastic MC3T3-E1 cell differentiation (matrix mineralization and gene expression). Since the fabrication process of these structures was carried out at mild conditions, an antibiotic (vancomycin) was incorporated in the slurry before the extrusion of the structures. The release profile of this antibiotic was measured in phosphate-buffered saline solution by high-performance liquid chromatography and was adjusted to a first-order release kinetics. Vancomycin released from the material was also shown to inhibit bacterial growth in vitro. The implications of these results for bone tissue engineering applications are discussed. Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  2. Fabrication of hydroxyapatite-poly(epsilon-caprolactone) scaffolds by a combination of the extrusion and bi-axial lamination processes.

    PubMed

    Sun, Jong-Jae; Bae, Chang-Jun; Koh, Young-Hag; Kim, Hyoun-Ee; Kim, Hae-Won

    2007-06-01

    Hydroxyapatite (HA)/poly(epsilon-caprolactone) (PCL) composite scaffolds were fabricated using a combination of the extrusion and bi-axial lamination processes. Firstly, HA/PCL composites with various HA contents (0, 50, 60, 70 wt%) were prepared by mixing the HA powders and the molten PCL at 100 degrees C and then extruded through an orifice with dimensions of 600 x 600 microm to produce HA/PCL composite fibers. Isobutyl methacrylate (IBMA) polymer fiber was also prepared in a similar manner for use as a fugitive material. The 3-D scaffold was then produced by the bi-axial lamination of the HA/PCL and IBMA fibers, followed by solvent leaching to remove the IBMA. It was observed that the HA/PCL composites had a superior elastic modulus and biological properties, as compared to the pure PCL. The fabricated HA/PCL scaffold showed a controlled pore structure (porosity of approximately 49% and pore size of approximately 512 microm) and excellent welding between the HA/PCL fibers, as well as a high compressive strength of approximately 7.8 MPa.

  3. Study the bonding mechanism of binders on hydroxyapatite surface and mechanical properties for 3DP fabrication bone scaffolds.

    PubMed

    Wei, Qinghua; Wang, Yanen; Li, Xinpei; Yang, Mingming; Chai, Weihong; Wang, Kai; zhang, Yingfeng

    2016-04-01

    In 3DP fabricating artificial bone scaffolds process, the interaction mechanism between binder and bioceramics power determines the microstructure and macro mechanical properties of Hydroxyapatite (HA) bone scaffold. In this study, we applied Molecular Dynamics (MD) methods to investigating the bonding mechanism and essence of binders on the HA crystallographic planes for 3DP fabrication bone scaffolds. The cohesive energy densities of binders and the binding energies, PCFs g(r), mechanical properties of binder/HA interaction models were analyzed through the MD simulation. Additionally, we prepared the HA bone scaffold specimens with different glues by 3DP additive manufacturing, and tested their mechanical properties by the electronic universal testing machine. The simulation results revealed that the relationship of the binding energies between binders and HA surface is consistent with the cohesive energy densities of binders, which is PAM/HA>PVA/HA>PVP/HA. The PCFs g(r) indicated that their interfacial interactions mainly attribute to the ionic bonds and hydrogen bonds which formed between the polar atoms, functional groups in binder polymer and the Ca, -OH in HA. The results of mechanical experiments verified the relationship of Young׳s modulus for three interaction models in simulation, which is PVA/HA>PAM/HA>PVP/HA. But the trend of compressive strength is PAM/HA>PVA/HA>PVP/HA, this is consistent with the binding energies of simulation. Therefore, the Young׳s modulus of bone scaffolds are limited by the Young׳s modulus of binders, and the compressive strength is mainly decided by the viscosity of binder. Finally, the major reasons for differences in mechanical properties between simulation and experiment were found, the space among HA pellets and the incomplete infiltration of glue were the main reasons influencing the mechanical properties of 3DP fabrication HA bone scaffolds. These results provide useful information in choosing binder for 3DP fabrication

  4. Mechanical study of polycaprolactone-hydroxyapatite porous scaffolds created by porogen-based solid freeform fabrication method.

    PubMed

    Lu, Lin; Zhang, Qingwei; Wootton, David M; Chiou, Richard; Li, Dichen; Lu, Bingheng; Lelkes, Peter I; Zhou, Jack

    2014-12-30

    Polycaprolactone (PCL) and polycaprolactone-hydroxyapatite (PCL-HA) scaffolds with 600-µm pore size were fabricated by drop-on-demand printing (DDP) structured porogen method followed with injection molding. Specimens with special dimensions of 4.2×4.2×5.4 mm3 and 6.6×6.6×13.8 mm3 were designed and fabricated for compression and tensile tests, respectively. The mechanical study was performed on both solid and porous PCL and PCL-HA samples. The effect on mechanical properties of the HA content ratio in PCL-HA composites was investigated. Porous scaffold made of 80/20 PCL-HA composite had an ultimate compressive strength of 3.7±0.2 MPa and compression modulus of 61.4±3.4 MPa, which is in the range of reported trabecular bone's compressive strength. Increasing the concentration of HA in the composites raised compressive properties and stiffness significantly (P<0.05), which demonstrates that PCL-HA composites have the potential for application in bone regeneration. Tensile test of solid PCL and PCL-HA composites showed that the ultimate tensile strength and tensile modulus increased with increases of the concentration of HA in the composites. The tensile test was also conducted on PCL porous scaffold; the result indicated that the scaffold was slightly softer and weaker in tension compared with compression. Combining compression and tensile test results, our study may guide the possible application of these biomaterials in bone tissue engineering and support further development of microstructure-based models of scaffold mechanical properties.

  5. Hydroxyapatite-reinforced collagen tissue engineering scaffolds

    NASA Astrophysics Data System (ADS)

    Kane, Robert J.

    Scaffolds have been fabricated from a wide variety of materials and most have showed some success, either as bone graft substitutes or as tissue engineering scaffolds. However, all current scaffold compositions and architectures suffer from one or more flaws including poor mechanical properties, lack of biological response, nondegradability, or a scaffold architecture not conducive to osteointegration. Biomimetic approaches to scaffold design using the two main components of bone tissue, collagen and hydroxyapatite, resulted in scaffolds with superior biological properties but relatively poor mechanical properties and scaffold architecture. It was hypothesized that by optimizing scaffold composition and architecture, HA-collagen bone tissue engineering scaffolds could provide both an excellent biological response along with improved structural properties. The mechanical properties of freeze-dried HA-collagen scaffolds, the most common type of porous HA-collagen material, were first shown to be increased by the addition of HA reinforcements, but scaffold stiffness still fell far short of the desired range. Based on limitations inherent in the freeze-dried process, a new type of leached-porogen scaffold fabrication process was developed. Proof-of-concept scaffolds demonstrated the feasibility of producing leached-porogen HA-collagen materials, and the scaffold architecture was optimized though careful selection of porogen particle size and shape along with an improved crosslinking technique. The final scaffolds exhibited substantially increased compressive modulus compared to previous types HA-collagen scaffolds, while the porosity, pore size, and scaffold permeability were tailored to be suitable for bone tissue ingrowth. An in vitro study demonstrated the capacity of the leached-porogen scaffolds to serve as a substrate for the differentiation of osteoblasts and subsequent production of new bone tissue. The new leached-porogen scaffold HA-collagen scaffolds were

  6. 3D-Cultivation of bone marrow stromal cells on hydroxyapatite scaffolds fabricated by dispense-plotting and negative mould technique.

    PubMed

    Detsch, R; Uhl, F; Deisinger, U; Ziegler, G

    2008-04-01

    The main principle of a bone tissue engineering (BTE) strategy is to cultivate osteogenic cells in an osteoconductive porous scaffold. Ceramic implants for osteogenesis are based mainly on hydroxyapatite (HA), since this is the inorganic component of bone. Rapid Prototyping (RP) is a new technology in research for producing ceramic scaffolds. This technology is particularly suitable for the fabrication of individually and specially tailored single implants. For tissue engineering these scaffolds are seeded with osteoblast or osteoblast precursor cells. To supply the cultured osteoblastic cells efficiently with nutrition in these 3D-geometries a bioreactor system can be used. The aim of this study was to analyse the influence of differently fabricated HA-scaffolds on bone marrow stromal cells. For this, two RP-techniques, dispense-plotting and a negative mould method, were used to produce porous ceramics. The manufactured HA-scaffolds were then cultivated in a dynamic system (bioreactor) with an osteoblastic precursor cell line. In our study, the applied RP-techniques give the opportunity to design and process HA-scaffolds with defined porosity, interconnectivity and 3D pore distribution. A higher differentiation of bone marrow stromal cells could be detected on the negative mould fabricated scaffolds, while cell proliferation was higher on the dispense-plotted scaffolds. Nevertheless, both scaffold types can be used in tissue engineering applications.

  7. The osteogenesis of bone marrow stem cells on mPEG-PCL-mPEG/hydroxyapatite composite scaffold via solid freeform fabrication.

    PubMed

    Liao, Han-Tsung; Chen, Yo-Yu; Lai, Yu-Ting; Hsieh, Ming-Fa; Jiang, Cho-Pei

    2014-01-01

    The study described a novel bone tissue scaffold fabricated by computer-aided, air pressure-aided deposition system to control the macro- and microstructure precisely. The porcine bone marrow stem cells (PBMSCs) seeded on either mPEG-PCL-mPEG (PCL) or mPEG-PCL-mPEG/hydroxyapatite (PCL/HA) composite scaffold were cultured under osteogenic medium to test the ability of osteogenesis in vitro. The experimental outcomes indicated that both scaffolds possessed adequate pore size, porosity, and hydrophilicity for the attachment and proliferation of PBMSCs and the PBMSCs expressed upregulated genes of osteogensis and angiogenesis in similar manner on both scaffolds. The major differences between these two types of the scaffolds were the addition of HA leading to higher hardness of PCL/HA scaffold, cell proliferation, and VEGF gene expression in PCL/HA scaffold. However, the in vivo bone forming efficacy between PBMSCs seeded PCL and PCL/HA scaffold was different from the in vitro results. The outcome indicated that the PCL/HA scaffold which had bone-mimetic environment due to the addition of HA resulted in better bone regeneration and mechanical strength than those of PCL scaffold. Therefore, providing a bone-mimetic scaffold is another crucial factor for bone tissue engineering in addition to the biocompatibility, 3D architecture with high porosity, and interpored connection.

  8. The Osteogenesis of Bone Marrow Stem Cells on mPEG-PCL-mPEG/Hydroxyapatite Composite Scaffold via Solid Freeform Fabrication

    PubMed Central

    Liao, Han-Tsung; Jiang, Cho-Pei

    2014-01-01

    The study described a novel bone tissue scaffold fabricated by computer-aided, air pressure-aided deposition system to control the macro- and microstructure precisely. The porcine bone marrow stem cells (PBMSCs) seeded on either mPEG-PCL-mPEG (PCL) or mPEG-PCL-mPEG/hydroxyapatite (PCL/HA) composite scaffold were cultured under osteogenic medium to test the ability of osteogenesis in vitro. The experimental outcomes indicated that both scaffolds possessed adequate pore size, porosity, and hydrophilicity for the attachment and proliferation of PBMSCs and the PBMSCs expressed upregulated genes of osteogensis and angiogenesis in similar manner on both scaffolds. The major differences between these two types of the scaffolds were the addition of HA leading to higher hardness of PCL/HA scaffold, cell proliferation, and VEGF gene expression in PCL/HA scaffold. However, the in vivo bone forming efficacy between PBMSCs seeded PCL and PCL/HA scaffold was different from the in vitro results. The outcome indicated that the PCL/HA scaffold which had bone-mimetic environment due to the addition of HA resulted in better bone regeneration and mechanical strength than those of PCL scaffold. Therefore, providing a bone-mimetic scaffold is another crucial factor for bone tissue engineering in addition to the biocompatibility, 3D architecture with high porosity, and interpored connection. PMID:24868523

  9. 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.

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

    PubMed Central

    Viana, Tânia; Franco, Margarida; Malça, Cândida; Mateus, Artur; Moura, Carla; 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. PMID:27872844

  11. Facile fabrication of poly(L-lactic acid) microsphere-incorporated calcium alginate/hydroxyapatite porous scaffolds based on Pickering emulsion templates.

    PubMed

    Hu, Yang; Ma, Shanshan; Yang, Zhuohong; Zhou, Wuyi; Du, Zhengshan; Huang, Jian; Yi, Huan; Wang, Chaoyang

    2016-04-01

    In this study, we develop a facile one-pot approach to the fabrication of poly(L-lactic acid) (PLLA) microsphere-incorporated calcium alginate (ALG-Ca)/hydroxyapatite (HAp) porous scaffolds based on HAp nanoparticle-stabilized oil-in-water Pickering emulsion templates, which contain alginate in the aqueous phase and PLLA in the oil phase. The emulsion aqueous phase is solidified by in situ gelation of alginate with Ca(2+) released from HAp by decreasing pH with slow hydrolysis of D-gluconic acid δ-lactone (GDL) to produce emulsion droplet-incorporated gels, followed by freeze-drying to form porous scaffolds containing microspheres. The pore structure of porous scaffolds can be adjusted by varying the HAp or GDL concentration. The compressive tests show that the increase of HAp or GDL concentration is beneficial to improve the compressive property of porous scaffolds, while the excessive HAp can lead to the decrease in compressive property. Moreover, the swelling behavior studies display that the swelling ratios of porous scaffolds reduce with increasing HAp or GDL concentration. Furthermore, hydrophobic drug ibuprofen (IBU) and hydrophilic drug bovine serum albumin (BSA) are loaded into the microspheres and scaffold matrix, respectively. In vitro drug release results indicate that BSA has a rapid release while IBU has a sustained release in the dual drug-loaded scaffolds. In vitro cell culture experiments verify that mouse bone mesenchymal stem cells can proliferate on the porous scaffolds well, indicating the good biocompatibility of porous scaffolds. All these results demonstrate that the PLLA microsphere-incorporated ALG-Ca/HAp porous scaffolds have a promising potential for tissue engineering and drug delivery applications. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. The structural and biological properties of hydroxyapatite-modified titanate nanowire scaffolds.

    PubMed

    Zhao, Haixin; Dong, Wenjun; Zheng, Yingying; Liu, Aiping; Yao, Juming; Li, Chaorong; Tang, Weihua; Chen, Benyong; Wang, Ge; Shi, Zhan

    2011-09-01

    Hydroxyapatite-modified titanate nanowire scaffolds as alternative materials for tissue engineering have been developed via a titanate nanowire matrix assisted electrochemical deposition method. The macroporous titanate nanowire matrix on Ti metal was fabricated by a hydrothermal method, and then followed by an electrochemical synthesis of hydroxyapatite nanoparticles on titanate nanowire. The incorporation of titanate nanowire matrix with high oriented hydroxyapatite nanoparticles generates hierarchical scaffolds with highly osteogenic, structural integrity and excellent mechanical performance. As-prepared porous three dimensional interconnected hydroxyapatite-modified titanate nanowire scaffolds, mimicking the nature's extracellular matrix, could provide a suitable microenvironment for tissue cell ingrowth and differentiation. The ceramic titanate nanowire core with HA nanoparticle sheath structure displays superhydrophilicity, which facilitates the cell attachment and proliferation, and induces the in vitro tissue-engineered bone. Human osteoblast-like MG63 cells were cultured on the hydroxyapatite-modified titanate nanowire scaffolds, and the results showed that the scaffolds highly promote the bioactivity, osteoconductivity and osteoblast differentiation.

  13. 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

  14. Biocompatibility of hydroxyapatite scaffolds processed by lithography-based additive manufacturing.

    PubMed

    Tesavibul, Passakorn; Chantaweroad, Surapol; Laohaprapanon, Apinya; Channasanon, Somruethai; Uppanan, Paweena; Tanodekaew, Siriporn; Chalermkarnnon, Prasert; Sitthiseripratip, Kriskrai

    2015-01-01

    The fabrication of hydroxyapatite scaffolds for bone tissue engineering applications by using lithography-based additive manufacturing techniques has been introduced due to the abilities to control porous structures with suitable resolutions. In this research, the use of hydroxyapatite cellular structures, which are processed by lithography-based additive manufacturing machine, as a bone tissue engineering scaffold was investigated. The utilization of digital light processing system for additive manufacturing machine in laboratory scale was performed in order to fabricate the hydroxyapatite scaffold, of which biocompatibilities were eventually evaluated by direct contact and cell-culturing tests. In addition, the density and compressive strength of the scaffolds were also characterized. The results show that the hydroxyapatite scaffold at 77% of porosity with 91% of theoretical density and 0.36 MPa of the compressive strength are able to be processed. In comparison with a conventionally sintered hydroxyapatite, the scaffold did not present any cytotoxic signs while the viability of cells at 95.1% was reported. After 14 days of cell-culturing tests, the scaffold was able to be attached by pre-osteoblasts (MC3T3-E1) leading to cell proliferation and differentiation. The hydroxyapatite scaffold for bone tissue engineering was able to be processed by the lithography-based additive manufacturing machine while the biocompatibilities were also confirmed.

  15. Hydroxyapatite (HA) bone scaffolds with controlled macrochannel pores.

    PubMed

    Bae, Chang-Jun; Kim, Hae-Won; Koh, Young-Hag; Kim, Hyoun-Ee

    2006-06-01

    Hydroxyapatite (HA) macrochanneled porous scaffolds, with a controlled pore structure, were fabricated via a combination of the extrusion and lamination processes. The scaffold was architectured by aligning and laminating the extruded HA and carbon filaments. The macrochannel pores were formed by removing the carbon filaments after thermal treatments (binder removal and sintering). The porosity of the scaffolds was varied between 48 and 73% with a controlled pore size of approximately 450 microm, by adjusting the fractions of HA and carbon filaments. As the porosity was increased from 48 to 73%, the compressive strength decreased from 11.5 to 3.2 MPa. However, the osteoblast-like cell responses on the scaffold, such as the proliferation rate and alkaline phosphatase (ALP) activity, were significantly enhanced as the porosity was increased.

  16. Hydroxyapatite reinforced collagen scaffolds with improved architecture and mechanical properties.

    PubMed

    Kane, Robert J; Weiss-Bilka, Holly E; Meagher, Matthew J; Liu, Yongxing; Gargac, Joshua A; Niebur, Glen L; Wagner, Diane R; Roeder, Ryan K

    2015-04-01

    Hydroxyapatite (HA) reinforced collagen scaffolds have shown promise for synthetic bone graft substitutes and tissue engineering scaffolds. Freeze-dried HA-collagen scaffolds are readily fabricated and have exhibited osteogenicity in vivo, but are limited by an inherent scaffold architecture that results in a relatively small pore size and weak mechanical properties. In order to overcome these limitations, HA-collagen scaffolds were prepared by compression molding HA reinforcements and paraffin microspheres within a suspension of concentrated collagen fibrils (∼ 180 mg/mL), cross-linking the collagen matrix, and leaching the paraffin porogen. HA-collagen scaffolds exhibited an architecture with high porosity (85-90%), interconnected pores ∼ 300-400 μm in size, and struts ∼ 3-100 μm in thickness containing 0-80 vol% HA whisker or powder reinforcements. HA reinforcement enabled a compressive modulus of up to ∼ 1 MPa, which was an order of magnitude greater than unreinforced collagen scaffolds. The compressive modulus was also at least one order of magnitude greater than comparable freeze-dried HA-collagen scaffolds and two orders of magnitude greater than absorbable collagen sponges used clinically. Moreover, scaffolds reinforced with up to 60 vol% HA exhibited fully recoverable elastic deformation upon loading to 50% compressive strain for at least 100,000 cycles. Thus, the scaffold mechanical properties were well-suited for surgical handling, fixation, and bearing osteogenic loads during bone regeneration. The scaffold architecture, permeability, and composition were shown to be conducive to the infiltration and differentiation of adipose-derive stromal cells in vitro. Acellular scaffolds were demonstrated to induce angiogenesis and osteogenesis after subcutaneous ectopic implantation by recruiting endogenous cell populations, suggesting that the scaffolds were osteoinductive.

  17. Bilayer hydroxyapatite scaffolds for maxillofacial bone tissue engineering.

    PubMed

    Guda, Teja; Oh, Sunho; Appleford, Mark R; Ong, Joo L

    2012-01-01

    The frequency of alveolar ridge resorption and crestal bone loss emphasizes the clinical need for bone graft substitutes to improve local bone quality prior to dental implant placement. Microcomputed tomography has been extensively employed to estimate bone quality more objectively (ie, quantitatively) by relating it to architectural parameters. In the present study, the mechanical properties of open cellular fully interconnected bilayer hydroxyapatite scaffolds, which mimicked the cortical shell/trabecular core architecture of human bone, were investigated for suitability as bone graft substitutes for maxillofacial reconstruction. Hydroxyapatite scaffolds with different architectures were fabricated using polymeric template pore sizes of 450 or 340 μm for the inner trabecular cores and 200 or 250 μm for the outer cortical shells in three different core-to-shell volume ratios. The architectural and mechanical properties and fluid permeability of the scaffolds were compared to reported values for maxillofacial bone. Whereas the elastic moduli of the scaffolds were comparable, their compressive strength was observed to be in the lower range of human mandibular trabecular bone. The microcomputed tomography architectural indices for the scaffolds were comparable to those of human trabecular bone at different locations in the human body, including the maxilla and mandible. Scaffold compressive strength, elastic modulus, and fluid conductance were 0.3 to 2.3 MPa, 40.9 to 668.1 MPa, and 8.8 to 49.9 x 10-10 m3s-1Pa-1, respectively. Open-pore bilayer scaffolds can be fabricated to exhibit sufficient mechanical integrity for maxillofacial bone graft applications to match specific bone site architecture while providing sufficient permeability to sustain bone regeneration.

  18. Open-Porous Hydroxyapatite Scaffolds for Three-Dimensional Culture of Human Adult Liver Cells

    PubMed Central

    Schmelzer, Eva; Over, Patrick; Nettleship, Ian; Gerlach, Joerg C.

    2016-01-01

    Liver cell culture within three-dimensional structures provides an improved culture system for various applications in basic research, pharmacological screening, and implantable or extracorporeal liver support. Biodegradable calcium-based scaffolds in such systems could enhance liver cell functionality by providing endothelial and hepatic cell support through locally elevated calcium levels, increased surface area for cell attachment, and allowing three-dimensional tissue restructuring. Open-porous hydroxyapatite scaffolds were fabricated and seeded with primary adult human liver cells, which were embedded within or without gels of extracellular matrix protein collagen-1 or hyaluronan. Metabolic functions were assessed after 5, 15, and 28 days. Longer-term cultures exhibited highest cell numbers and liver specific gene expression when cultured on hydroxyapatite scaffolds in collagen-1. Endothelial gene expression was induced in cells cultured on scaffolds without extracellular matrix proteins. Hydroxyapatite induced gene expression for cytokeratin-19 when cells were cultured in collagen-1 gel while culture in hyaluronan increased cytokeratin-19 gene expression independent of the use of scaffold in long-term culture. The implementation of hydroxyapatite composites with extracellular matrices affected liver cell cultures and cell differentiation depending on the type of matrix protein and the presence of a scaffold. The hydroxyapatite scaffolds enable scale-up of hepatic three-dimensional culture models for regenerative medicine applications. PMID:27403430

  19. Open-Porous Hydroxyapatite Scaffolds for Three-Dimensional Culture of Human Adult Liver Cells.

    PubMed

    Finoli, Anthony; Schmelzer, Eva; Over, Patrick; Nettleship, Ian; Gerlach, Joerg C

    2016-01-01

    Liver cell culture within three-dimensional structures provides an improved culture system for various applications in basic research, pharmacological screening, and implantable or extracorporeal liver support. Biodegradable calcium-based scaffolds in such systems could enhance liver cell functionality by providing endothelial and hepatic cell support through locally elevated calcium levels, increased surface area for cell attachment, and allowing three-dimensional tissue restructuring. Open-porous hydroxyapatite scaffolds were fabricated and seeded with primary adult human liver cells, which were embedded within or without gels of extracellular matrix protein collagen-1 or hyaluronan. Metabolic functions were assessed after 5, 15, and 28 days. Longer-term cultures exhibited highest cell numbers and liver specific gene expression when cultured on hydroxyapatite scaffolds in collagen-1. Endothelial gene expression was induced in cells cultured on scaffolds without extracellular matrix proteins. Hydroxyapatite induced gene expression for cytokeratin-19 when cells were cultured in collagen-1 gel while culture in hyaluronan increased cytokeratin-19 gene expression independent of the use of scaffold in long-term culture. The implementation of hydroxyapatite composites with extracellular matrices affected liver cell cultures and cell differentiation depending on the type of matrix protein and the presence of a scaffold. The hydroxyapatite scaffolds enable scale-up of hepatic three-dimensional culture models for regenerative medicine applications.

  20. Hydroxyapatite reinforced inherent RGD containing silk fibroin composite scaffolds: Promising platform for bone tissue engineering.

    PubMed

    Behera, Sibaram; Naskar, Deboki; Sapru, Sunaina; Bhattacharjee, Promita; Dey, Tuli; Ghosh, Ananta K; Mandal, Mahitosh; Kundu, Subhas C

    2017-03-08

    Replacement and repair of ectopic bone defects and traumatized bone tissues are done using porous scaffolds and composites. The prerequisites for such scaffolds include high mechanical strength, osseoconductivity and cytocompatibility. The present work is designed to address such requirements by fabricating a reinforced cytocompatible scaffold. Biocompatible silk protein fibroin collected from tropical non-mulberry tasar silkworm (Antheraea mylitta) is used to fabricate fibroin-hydroxyapatite (HAp) nanocomposite particles using chemical precipitation method. In situ reinforcement of fibroin-HAp nanocomposite and external deposition of HAp particles on fibroin scaffold is carried out for comparative evaluations of bio-physical and biochemical characteristics. HAp deposited fibroin scaffolds provide greater mechanical strength and cytocompatibility, when compared with fibroin-HAp nanoparticles reinforced fibroin scaffolds. Minimal immune responses of both types of composite scaffolds are observed using osteoblast-macrophage co-culture model. Nanocomposite reinforced fibroin scaffold can be tailored further to accommodate different requirements depending on bone type or bone regeneration period.

  1. Scaffold development using selective laser sintering of polyetheretherketone-hydroxyapatite biocomposite blends.

    PubMed

    Tan, K H; Chua, C K; Leong, K F; Cheah, C M; Cheang, P; Abu Bakar, M S; Cha, S W

    2003-08-01

    In tissue engineering (TE), temporary three-dimensional scaffolds are essential to guide cell proliferation and to maintain native phenotypes in regenerating biologic tissues or organs. To create the scaffolds, rapid prototyping (RP) techniques are emerging as fabrication techniques of choice as they are capable of overcoming many of the limitations encountered with conventional manual-based fabrication processes. In this research, RP fabrication of solvent free porous polymeric and composite scaffolds was investigated. Biomaterials such as polyetheretherketone (PEEK) and hydroxyapatite (HA) were experimentally processed on a commercial selective laser sintering (SLS) RP system. The SLS technique is highly advantageous as it provides good user control over the microstructures of created scaffolds by adjusting the SLS process parameters. Different weight percentage (wt%) compositions of physically mixed PEEK/HA powder blends were sintered to assess their suitability for SLS processing. Microstructural assessments of the scaffolds were conducted using electron microscopy. The results ascertained the potential of SLS-fabricated TE scaffolds.

  2. Calcium silicate ceramic scaffolds toughened with hydroxyapatite whiskers for bone tissue engineering

    SciTech Connect

    Feng, Pei; Wei, Pingpin; Li, Pengjian; Gao, Chengde; Shuai, Cijun; Peng, Shuping

    2014-11-15

    Calcium silicate possessed excellent biocompatibility, bioactivity and degradability, while the high brittleness limited its application in load-bearing sites. Hydroxyapatite whiskers ranging from 0 to 30 wt.% were incorporated into the calcium silicate matrix to improve the strength and fracture resistance. Porous scaffolds were fabricated by selective laser sintering. The effects of hydroxyapatite whiskers on the mechanical properties and toughening mechanisms were investigated. The results showed that the scaffolds had a uniform and continuous inner network with the pore size ranging between 0.5 mm and 0.8 mm. The mechanical properties were enhanced with increasing hydroxyapatite whiskers, reached a maximum at 20 wt.% (compressive strength: 27.28 MPa, compressive Young's modulus: 156.2 MPa, flexural strength: 15.64 MPa and fracture toughness: 1.43 MPa·m{sup 1/2}) and then decreased by addition of more hydroxyapatite whiskers. The improvement of mechanical properties was due to whisker pull-out, crack deflection and crack bridging. Moreover, the degradation rate decreased with the increase of hydroxyapatite whisker content. A layer of bone-like apatite was formed on the scaffold surfaces after being soaked in simulated body fluid. Human osteoblast-like MG-63 cells spread well on the scaffolds and proliferated with increasing culture time. These findings suggested that the calcium silicate scaffolds reinforced with hydroxyapatite whiskers showed great potential for bone regeneration and tissue engineering applications. - Highlights: • HA whiskers were incorporated into CS to improve the properties. • The scaffolds were successfully fabricated by SLS. • Toughening mechanisms was whisker pull-out, crack deflection and bridging. • The scaffolds showed excellent apatite forming ability.

  3. [IN VIVO EVALUATION OF POLYCAPROLACTONE-HYDROXYAPATITE SCAFFOLD BIOCOMPATIBILITY].

    PubMed

    Ivanov, A N; Kozadaev, M N; Bogomolova, N V; Matveeva, O V; Puchinyan, D M; Norkin, I A; Sal'kovskii, Yu E; Lyubun, G P

    2015-01-01

    Biocompatibility is one of the main and very important properties for scaffolds. The aim of the present study was to investigate cells population dynamics in vivo in the process of original polycaprolactone-hydroxyapatite scaffold colonization, as well as tissue reactions to the implantation to assess the biocompatibility of the matrix. It has been found that tissue reactive changes in white rats subside completely up to the 21st day after subcutaneous polycaprolactone-hydroxyapatite scaffold implantation. Matrix was actively colonized by connective tissue cells in the period from the 7th to the 21st day of the experiment. However, intensive scaffold vascularization started from the 14th day after implantation. These findings suggest a high degree of the polycaprolactone-hydroxyapatite scaffold biocompatiblilitye.

  4. Recent Advances in Hydroxyapatite Scaffolds Containing Mesenchymal Stem Cells

    PubMed Central

    Michel, John; Penna, Matthew; Kochen, Juan; Cheung, Herman

    2015-01-01

    Modern day tissue engineering and cellular therapies have gravitated toward using stem cells with scaffolds as a dynamic modality to aid in differentiation and tissue regeneration. Mesenchymal stem cells (MSCs) are one of the most studied stem cells used in combination with scaffolds. These cells differentiate along the osteogenic lineage when seeded on hydroxyapatite containing scaffolds and can be used as a therapeutic option to regenerate various tissues. In recent years, the combination of hydroxyapatite and natural or synthetic polymers has been studied extensively. Due to the interest in these scaffolds, this review will cover the wide range of hydroxyapatite containing scaffolds used with MSCs for in vitro and in vivo experiments. Further, in order to maintain a progressive scope of the field this review article will only focus on literature utilizing adult human derived MSCs (hMSCs) published in the last three years. PMID:26106425

  5. Fabrication of in situ polymerized poly(butylene succinate-co-ethylene terephthalate)/hydroxyapatite nanocomposite to fibrous scaffolds for enhancement of osteogenesis.

    PubMed

    Shirali, Hadi; Rafizadeh, Mehdi; Afshar Taromi, Faramarz; Jabbari, Esmaiel

    2017-09-01

    A combination of elastic poly(butylene succinate-co-ethylene terephthalate) and rigid nano-hydroxyapatite were used to prepare an in-situ synthesized nanocomposite mimicing bone structure. The microstructure, morphology, and dispersion of nanoparticles in the nanocomposites were studied using proton nuclear magnetic resonance ((1) HNMR), scanning electron microscope (SEM), and transmission electron microscopy (TEM), respectively. Then, electrospinning method was used to produce nanofiber matrix with lowest fiber diameter. Presence of chemical bonds among nanoparticles and polymer leads to the excellent particle dispersion and solution phase stability. SEM results show that continuous and bead-free nanofibers were produced and incorporating nanoparticle slightly increased fibers diameter. Elastic modulus, tensile strength, crystallinity, hydrophilicity and hydrolytic degradability of resulted nanofiber increased with nanoparticle but elongation at break slightly decreased. Proliferation and osteogenic differentiation of human mesenchymal stem cell significantly improved by introducing nanoparticle which indicate that electrospun nanofibers could be used as scaffolds for bone tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2622-2631, 2017. © 2017 Wiley Periodicals, Inc.

  6. Mechanical properties of calcium phosphate scaffolds fabricated by robocasting.

    PubMed

    Miranda, Pedro; Pajares, Antonia; Saiz, Eduardo; Tomsia, Antoni P; Guiberteau, Fernando

    2008-04-01

    The mechanical behavior under compressive stresses of beta-tricalcium phosphate (beta-TCP) and hydroxyapatite (HA) scaffolds fabricated by direct-write assembly (robocasting) technique is analyzed. Concentrated colloidal inks prepared from beta-TCP and HA commercial powders were used to fabricate porous structures consisting of a 3-D tetragonal mesh of interpenetrating ceramic rods. The compressive strength and elastic modulus of these model scaffolds were determined by uniaxial testing to compare the relative performance of the selected materials. The effect of a 3-week immersion in simulated body fluid (SBF) on the strength of the scaffolds was also analyzed. The results are compared with those reported in the literature for calcium phosphate scaffolds and human bone. The robocast calcium phosphate scaffolds were found to exhibit excellent mechanical performances in terms of strength, especially the HA structures after SBF immersion, indicating a great potential of this type of scaffolds for use in load-bearing bone tissue engineering applications.

  7. Tissue response and biodegradation of composite scaffolds prepared from Thai silk fibroin, gelatin and hydroxyapatite.

    PubMed

    Tungtasana, Hathairat; Shuangshoti, Somruetai; Shuangshoti, Shanop; Kanokpanont, Sorada; Kaplan, David L; Bunaprasert, Tanom; Damrongsakkul, Siriporn

    2010-12-01

    This work aimed to investigate tissue responses and biodegradation, both in vitro and in vivo, of four types of Bombyx mori Thai silk fibroin based-scaffolds. Thai silk fibroin (SF), conjugated gelatin/Thai silk fibroin (CGSF), hydroxyapatite/Thai silk fibroin (SF4), and hydroxyapatite/conjugated gelatin/Thai silk fibroin (CGSF4) scaffolds were fabricated using salt-porogen leaching, dehydrothermal/chemical crosslinking and an alternate soaking technique for mineralization. In vitro biodegradation in collagenase showed that CGSF scaffolds had the slowest biodegradability, due to the double crosslinking by dehydrothermal and chemical treatments. The hydroxyapatite deposited from alternate soaking separated from the surface of the protein scaffolds when immersed in collagenase. From in vivo biodegradation studies, all scaffolds could still be observed after 12 weeks of implantation in subcutaneous tissue of Wistar rats and also following ISO10993-6: Biological evaluation of medical devices. At 2 and 4 weeks of implantation the four types of Thai silk fibroin based-scaffolds were classified as "non-irritant" to "slight-irritant", compared to Gelfoam(®) (control samples). These natural Thai silk fibroin-based scaffolds may provide suitable biomaterials for clinical applications.

  8. Robotic deposition of model hydroxyapatite scaffolds with multiple architectures and multiscale porosity for bone tissue engineering.

    PubMed

    Dellinger, Jennifer G; Cesarano, Joseph; Jamison, Russell D

    2007-08-01

    Model hydroxyapatite (HA) scaffolds with porosities spanning multiple length scales were fabricated by robocasting, a solid freeform fabrication technique based on the robotic deposition of colloidal pastes. Scaffolds of various architectures including periodic, radial, and superlattice structures were constructed. Macropores (100-600 microm) were designed by controlling the arrangement and spacing between rods of HA. Micropores (1-30 microm) and submicron pores (less than 1 microm) were produced within the rods by including polymer microsphere porogens in the HA pastes and by controlling the sintering of the scaffolds. These model scaffolds may be used to systematically study the effects of scaffold porosity on bone ingrowth processes both in vitro and in vivo.

  9. Three-dimensional plotted hydroxyapatite scaffolds with predefined architecture: comparison of stabilization by alginate cross-linking versus sintering.

    PubMed

    Kumar, Alok; Akkineni, Ashwini R; Basu, Bikramjit; Gelinsky, Michael

    2016-03-01

    Scaffolds for bone tissue engineering are essentially characterized by porous three-dimensional structures with interconnected pores to facilitate the exchange of nutrients and removal of waste products from cells, thereby promoting cell proliferation in such engineered scaffolds. Although hydroxyapatite is widely being considered for bone tissue engineering applications due to its occurrence in the natural extracellular matrix of this tissue, limited reports are available on additive manufacturing of hydroxyapatite-based materials. In this perspective, hydroxyapatite-based three-dimensional porous scaffolds with two different binders (maltodextrin and sodium alginate) were fabricated using the extrusion method of three-dimensional plotting and the results were compared in reference to the structural properties of scaffolds processed via chemical stabilization and sintering routes, respectively. With the optimal processing conditions regarding to pH and viscosity of binder-loaded hydroxyapatite pastes, scaffolds with parallelepiped porous architecture having up to 74% porosity were fabricated. Interestingly, sintering of the as-plotted hydroxyapatite-sodium alginate (cross-linked with CaCl2 solution) scaffolds led to the formation of chlorapatite (Ca9.54P5.98O23.8Cl1.60(OH)2.74). Both the sintered scaffolds displayed progressive deformation and delayed fracture under compressive loading, with hydroxyapatite-alginate scaffolds exhibiting a higher compressive strength (9.5 ± 0.5 MPa) than hydroxyapatite-maltodextrin scaffolds (7.0 ± 0.6 MPa). The difference in properties is explained in terms of the phase assemblage and microstructure.

  10. The mechanical properties and osteoconductivity of hydroxyapatite bone scaffolds with multi-scale porosity.

    PubMed

    Woodard, Joseph R; Hilldore, Amanda J; Lan, Sheeny K; Park, C J; Morgan, Abby W; Eurell, Jo Ann C; Clark, Sherrie G; Wheeler, Matthew B; Jamison, Russell D; Wagoner Johnson, Amy J

    2007-01-01

    The relative osteoconductivity and the change in the mechanical properties of hydroxyapatite (HA) scaffolds with multi-scale porosity were compared to scaffolds with a single pore size. Non-microporous (NMP) scaffolds contained only macroporosity (250-350 microm) and microporous (MP) scaffolds contained both macroporosity and microporosity (2-8 microm). Recombinant human bone morphogenetic protein-2 (rhBMP-2) was incorporated into all scaffolds via gelatin microspheres prior to implantation into the latissimus dorsi muscle of Yorkshire pigs. After 8 weeks, only the MP scaffolds contained bone. The result demonstrates the efficacy of the MP scaffolds as drug carriers. Implanted and as-fabricated scaffolds were compared using histology, microcomputed tomography, scanning electron microscopy, and compression testing. Implanted scaffolds exhibited a stress-strain response similar to that of cancellous bone with strengths between those of cancellous and cortical bone. The strength and stiffness of implanted NMP scaffolds decreased by 15% and 46%, respectively. Implanted MP scaffolds lost 30% of their strength and 31% of their stiffness. Bone arrested crack propagation effectively in MP scaffolds. The change in mechanical behavior is discussed and the study demonstrates the importance of scaffold microporosity on bone ingrowth and on the mechanical behavior of HA implant materials.

  11. Hydroxyapatite-silver nanoparticles coatings on porous polyurethane scaffold.

    PubMed

    Ciobanu, Gabriela; Ilisei, Simona; Luca, Constantin

    2014-02-01

    The present paper is focused on a study regarding the possibility of obtaining hydroxyapatite-silver nanoparticle coatings on porous polyurethane scaffold. The method applied is based on a combined strategy involving hydroxyapatite biomimetic deposition on polyurethane surface using a Supersaturated Calcification Solution (SCS), combined with silver ions reduction and in-situ crystallization processes on hydroxyapatite-polyurethane surface by sample immersing in AgNO3 solution. The morphology, composition and phase structure of the prepared samples were characterized by scanning electron microscopy coupled with energy dispersive X-ray spectroscopy (SEM-EDX), X-ray diffraction (XRD), UV-Vis spectroscopy and X-ray photoelectron spectroscopy (XPS) measurements. The data obtained show that a layer of hydroxyapatite was deposited on porous polyurethane support and the silver nanoparticles (average size 34.71 nm) were dispersed among and even on the hydroxyapatite crystals. Hydroxyapatite/polyurethane surface acts as a reducer and a stabilizing agent for silver ions. The surface plasmon resonance peak in UV-Vis absorption spectra showed an absorption maximum at 415 nm, indicating formation of silver nanoparticles. The hydroxyapatite-silver polyurethane scaffolds were tested against Staphylococcus aureus and Escherichia coli and the obtained data were indicative of good antibacterial properties of the materials.

  12. Hydroxyapatite scaffolds for bone tissue engineering made by 3D printing.

    PubMed

    Leukers, Barbara; Gülkan, Hülya; Irsen, Stephan H; Milz, Stefan; Tille, Carsten; Schieker, Matthias; Seitz, Hermann

    2005-12-01

    Nowadays, there is a significant need for synthetic bone replacement materials used in bone tissue engineering (BTE). Rapid prototyping and especially 3D printing is a suitable technique to create custom implants based on medical data sets. 3D printing allows to fabricate scaffolds based on Hydroxyapatite with complex internal structures and high resolution. To determine the in vitro behaviour of cells cultivated on the scaffolds, we designed a special test-part. MC3T3-E1 cells were seeded on the scaffolds and cultivated under static and dynamic setups. Histological evaluation was carried out to characterise the cell ingrowth. In summary, the dynamic cultivation method lead to a stronger population compared to the static cultivation method. The cells proliferated deep into the structure forming close contact to Hydroxyapatite granules.

  13. Chitosan-Based Bilayer Hydroxyapatite Nanorod Composite Scaffolds for Osteochondral Regeneration

    NASA Astrophysics Data System (ADS)

    Swanson, Shawn

    Osteochondral defects involve injury to bone and cartilage. As articular cartilage is worn down, bone in the joint begins to rub together, causing bone spurs. This is known as osteoarthritis, and is a common issue among the aging population. This problem presents an interesting opportunity for tissue engineering. Tissue engineering is an approach to treatment of tissue defects where synthetic, three dimensional (3-D) scaffolds are implanted in a defect to facilitate healing. The osteochondral scaffold consists of two regions in the form of a bilayer scaffold- one to mimic bone with osteoconductive properties, and one to mimic cartilage with biomimetic properties. One approach to improving the osteoconductivity of tissue engineering scaffolds is the addition of hydroxyapatite (HAp), the main mineral phase in bone. HAp with nanorod morphology is desirable because it is biomimetic for the calcium phosphate found in bone. Incorporating HAp nanorods in bone tissue engineering scaffolds to form a composite material may increase scaffold osteoconductivity. The cartilage scaffold is fabricated from chitosan and hyaluronic acid (HA). HA is a known component of cartilage and thus is biomimetic. The bilayer scaffolds were seeded with osteoblast-like MG-63 cells to investigate cell migration and were evaluated with Alamar Blue proliferation assay. The cells successfully migrated to the bone region of the scaffold, indicating that the bilayer scaffold provides a promising osteochondral scaffold.

  14. Microwave-induced biomimetic approach for hydroxyapatite coatings of chitosan scaffolds.

    PubMed

    Kaynak Bayrak, Gökçe; Demirtaş, T Tolga; Gümüşderelioğlu, Menemşe

    2017-02-10

    Simulated body fluid (SBF) can form calcium phosphates on osteoinductive materials, so it is widely used for coating of bone scaffolds to mimic natural extracellular matrix (ECM). However, difficulties of bulk coating in 3D scaffolds and the necessity of long process times are the common problems for coating with SBF. In the present study, a microwave-assisted process was developed for rapid and internal coating of chitosan scaffolds. The scaffolds were fabricated as superporous hydrogel (SPH) by combining microwave irradiation and gas foaming methods. Then, they were immersed into 10x  SBF-like solution and homogenous bone-like hydroxyapatite (HA) coating was achieved by microwave treatment at 600W without the need of any nucleating agent. Cell culture studies with MC3T3-E1 preosteoblasts showed that microwave-assisted biomimetic HA coating process could be evaluated as an efficient and rapid method to obtain composite scaffolds for bone tissue engineering.

  15. Precipitation of hydroxyapatite on electrospun polycaprolactone/aloe vera/silk fibroin nanofibrous scaffolds for bone tissue engineering.

    PubMed

    Shanmugavel, Suganya; Reddy, Venugopal Jayarama; Ramakrishna, Seeram; Lakshmi, B S; Dev, Vr Giri

    2014-07-01

    Advances in electrospun nanofibres with bioactive materials have enhanced the scope of fabricating biomimetic scaffolds for tissue engineering. The present research focuses on fabrication of polycaprolactone/aloe vera/silk fibroin nanofibrous scaffolds by electrospinning followed by hydroxyapatite deposition by calcium-phosphate dipping method for bone tissue engineering. Morphology, composition, hydrophilicity and mechanical properties of polycaprolactone/aloe vera/silk fibroin-hydroxyapatite nanofibrous scaffolds along with controls polycaprolactone and polycaprolactone/aloe vera/silk fibroin nanofibrous scaffolds were examined by field emission scanning electron microscopy, Fourier transform infrared spectroscopy, contact angle and tensile tests, respectively. Adipose-derived stem cells cultured on polycaprolactone/aloe vera/silk fibroin-hydroxyapatite nanofibrous scaffolds displayed highest cell proliferation, increased osteogenic markers expression (alkaline phosphatase and osteocalcin), osteogenic differentiation and increased mineralization in comparison with polycaprolactone control. The obtained results indicate that polycaprolactone/aloe vera/silk fibroin-hydroxyapatite nanofibrous scaffolds have appropriate physico-chemical and biological properties to be used as biomimetic scaffolds for bone tissue regeneration. © The Author(s) 2013 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  16. Fabrication of oriented hydroxyapatite film by RF magnetron sputtering

    NASA Astrophysics Data System (ADS)

    Hirata, Keishiro; Kubota, Takafumi; Koyama, Daisuke; Takayanagi, Shinji; Matsukawa, Mami

    2017-08-01

    Hydroxyapatite (HAp) is compatible with bone tissue and is used mainly as a bone prosthetic material, especially as the coating of implants. Oriented HAp film is expected to be a high-quality epitaxial scaffold of the neonatal bone. To fabricate highly oriented HAp thin films via the conventional plasma process, we deposited the HAp film on a Ti coated silica glass substrate using RF magnetron sputtering in low substrate temperature conditions. The X-ray diffraction pattern of the film sample consisted of an intense (002) peak, corresponding to the highly oriented HAp. The (002) peak in XRD diagrams can be attributed either to the monoclinic phase or the hexagonal phase. Pole figure analysis showed that the (002) plane grew parallel to the surface of the substrate, without inclination. Transmission Electron Microscope analysis also showed the fabrication of aligned HAp crystallites. The selected area diffraction patterns indicated the existence of monoclinic phase. The existence of hexagonal phase could not be judged. These results indicate the uniaxial films fabricated by this technique enable to be the epitaxial scaffold of the neonatal bone. This scaffold can be expected to promote connection with the surrounding bone tissue and recovery of the dynamic characteristics of the bone.

  17. Concentrated hydroxyapatite inks for direct-write assembly of 3-D periodic scaffolds.

    PubMed

    Michna, Sarah; Wu, Willie; Lewis, Jennifer A

    2005-10-01

    Hydroxyapatite (HA) scaffolds with a 3-D periodic architecture and multiscale porosity have been fabricated by direct-write assembly. Concentrated HA inks with tailored viscoelastic properties were developed to enable the construction of complex 3-D architectures comprised of self-supporting cylindrical rods in a layer-by-layer patterning sequence. By controlling their lattice constant and sintering conditions, 3-D periodic HA scaffolds were produced with a bimodal pore size distribution. Mercury intrusion porosimetry (MIP) was used to determine the characteristic pore size and volume associated with the interconnected pore channels between HA rods and the finer pores within the partially sintered HA rods.

  18. Freeze casting of hydroxyapatite scaffolds for bone tissue engineering.

    PubMed

    Deville, Sylvain; Saiz, Eduardo; Tomsia, Antoni P

    2006-11-01

    Although extensive efforts have been put into the development of porous scaffolds for bone regeneration, with encouraging results, all porous materials have a common limitation: the inherent lack of strength associated with porosity. Hence, the development of porous hydroxyapatite scaffolds has been hindered to non-load bearing applications. We report here how freeze casting can be applied to synthesize porous scaffolds exhibiting unusually high compressive strength, e.g. up to 145 MPa for 47% porosity and 65 MPa for 56% porosity. The materials are characterized by well-defined pore connectivity along with directional and completely open porosity. Various parameters affecting the porosity and compressive strength have been investigated, including initial slurry concentration, freezing rate, and sintering conditions. The implications and potential application as bone substitute are discussed. These results might open the way for hydroxyapatite-based materials designed for load-bearing applications. The biological response of these materials is yet to be tested.

  19. BMP-2 immobilized PLGA/hydroxyapatite fibrous scaffold via polydopamine stimulates osteoblast growth.

    PubMed

    Zhao, Xingyu; Han, Yu; Li, Jiawei; Cai, Bo; Gao, Hang; Feng, Wei; Li, Shuqiang; Liu, Jianguo; Li, Dongsong

    2017-09-01

    Combining biomaterials scaffolds with bone morphogenetic protein-2 (BMP-2) is currently used to promote the regeneration of bone tissue. However, the traditional strategies used to add BMP-2 into the polymer scaffolds directly suffer from limitations that can result in lower growth factor loading and damage the bioactivity of growth factors. In this study, we report the fabrication of poly(lactide-co-glycolide)/hydroxyapatite (PLGA/HA) composite fibrous scaffolds via melt-spinning method to mimic native extracellular matrix (ECM). In order to effectively immobilize BMP-2 on PLGA/HA composite fibrous scaffolds, the surface of the scaffold was modified with polydopamine (PDA) (PDA-PLGA/HA). PDA was chosen as an adhesive polymeric bridge-layer between PLGA/HA fibrous scaffolds and BMP-2. Analysis of the scaffold using scanning electron microscopy, Fourier transform infrared spectroscopy and X-ray photoelectron spectroscope revealed that the PDA coating was attached to the scaffold surface. Moreover, analysis of the scaffold using water contact angle demonstrated an increased hydrophilicity via PDA modification. Furthermore, the PDA coating effectively immobilized BMP-2 on the PDA-PLGA/HA fibrous scaffold and a sustained release profile of BMP-2 was achieved in the BMP-2-immobilized PLGA/HA fibrous scaffold. In vitro experiments showed that BMP-2-immobilized PLGA/HA fibrous scaffold significantly promoted the attachment and proliferation of MC3T3-E1 cells. More importantly, the ALP activity, mRNA expression of osteosis-related genes and calcium deposition in MC3T3-E1 cells cultured on BMP-2-immobilized PLGA/HA fibrous scaffold were significantly increased. These results collectively demonstrate that the BMP-2-immobilized PLGA/HA fibrous scaffold is a promising candidate for bone regeneration. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Robocasting nanocomposite scaffolds of poly(caprolactone)/hydroxyapatite incorporating modified carbon nanotubes for hard tissue reconstruction.

    PubMed

    Dorj, Biligzaya; Won, Jong-Eun; Kim, Joong-Hyun; Choi, Seong-Jun; Shin, Ueon Sang; Kim, Hae-Won

    2013-06-01

    Nanocomposite scaffolds with tailored 3D pore configuration are promising candidates for the reconstruction of bone. Here we fabricated novel nanocomposite bone scaffolds through robocasting. Poly(caprolactone) (PCL)-hydroxyapatite (HA) slurry containing ionically modified carbon nanotubes (imCNTs) was robotic-dispensed and structured layer-by-layer into macrochanneled 3D scaffolds under adjusted processing conditions. Homogeneous dispersion of imCNTs (0.2 wt % relative to PCL-HA) was achieved in acetone, aiding in the preparation of PCL-HA-imCNTs slurry with good mixing property. Incorporation of imCNTs into PCL-HA composition significantly improved the compressive strength and elastic modulus of the robotic-dispensed scaffolds (~1.5-fold in strength and ~2.5-fold in elastic modulus). When incubated in simulated body fluid (SBF), PCL-HA-imCNT nanocomposite scaffold induced substantial mineralization of apatite in a similar manner to the PCL-HA scaffold, which was contrasted in pure PCL scaffold. MC3T3-E1 cell culture on the scaffolds demonstrated that cell proliferation levels were significantly higher in both PCL-HA-imCNT and PCL-HA than in pure PCL, and no significant difference was found between the nanocomposite scaffolds. When the PCL-HA-imCNT scaffold was implanted into a rat subcutaneous tissue for 4 weeks, soft fibrous tissues with neo-blood vessels formed well in the pore channels of the scaffolds without any significant inflammatory signs. Tissue reactions in PCL-HA-imCNT scaffold were similar to those in PCL-HA scaffold, suggesting incorporated imCNT did not negate the beneficial biological roles of HA. While more long-term in vivo research in bone defect models is needed to confirm clinical availability, our results suggest robotic-dispensed PCL-HA-imCNT nanocomposite scaffolds can be considered promising new candidate matrices for bone regeneration.

  1. Hydroxyapatite whisker reinforced 63s glass scaffolds for bone tissue engineering.

    PubMed

    Shuai, Cijun; Cao, Yiyuan; Gao, Chengde; Feng, Pei; Xiao, Tao; Peng, Shuping

    2015-01-01

    Bioactive glass (BG) is widely used for bone tissue engineering. However, poor mechanical properties are the major shortcomings. In the study, hydroxyapatite nanowhisker (HANw) was used as a reinforcement to improve the mechanical properties. 63s glass/HANw scaffolds were successfully fabricated by selective laser sintering (SLS). It was found that the optimal compressive strength and fracture toughness were achieved when 10 wt.% HANw was added. This led to 36% increase in compressive strength and 83% increase in fracture toughness, respectively, compared with pure 63s glass scaffolds. Different reinforcement mechanisms were analyzed based on the microstructure investigation. Whisker bridging and whisker pulling-out were efficient in absorbing crack propagating energy, resulting in the improvement of the mechanical properties. Moreover, bioactivity and biocompatibility of the scaffolds were evaluated in vitro. The results showed that composite scaffolds with 10 wt.% HANw exhibited good apatite-forming ability and cellular affinity.

  2. Hydroxyapatite Whisker Reinforced 63s Glass Scaffolds for Bone Tissue Engineering

    PubMed Central

    Shuai, Cijun; Cao, Yiyuan; Gao, Chengde; Feng, Pei; Xiao, Tao; Peng, Shuping

    2015-01-01

    Bioactive glass (BG) is widely used for bone tissue engineering. However, poor mechanical properties are the major shortcomings. In the study, hydroxyapatite nanowhisker (HANw) was used as a reinforcement to improve the mechanical properties. 63s glass/HANw scaffolds were successfully fabricated by selective laser sintering (SLS). It was found that the optimal compressive strength and fracture toughness were achieved when 10 wt.% HANw was added. This led to 36% increase in compressive strength and 83% increase in fracture toughness, respectively, compared with pure 63s glass scaffolds. Different reinforcement mechanisms were analyzed based on the microstructure investigation. Whisker bridging and whisker pulling-out were efficient in absorbing crack propagating energy, resulting in the improvement of the mechanical properties. Moreover, bioactivity and biocompatibility of the scaffolds were evaluated in vitro. The results showed that composite scaffolds with 10 wt.% HANw exhibited good apatite-forming ability and cellular affinity. PMID:25821798

  3. Osteoinductive silk fibroin/titanium dioxide/hydroxyapatite hybrid scaffold for bone tissue engineering.

    PubMed

    Kim, Jung-Ho; Kim, Dong-Kyu; Lee, Ok Joo; Ju, Hyung Woo; Lee, Jung Min; Moon, Bo Mi; Park, Hyun Jung; Kim, Dong Wook; Lee, Jun Ho; Park, Chan Hum

    2016-01-01

    The present study demonstrated the fabrication that incorporation of titanium isopropoxide (TiO2) and hydroxyapatite (HA) nanoparticles into the silk fibroin (SF) scaffolds. In this process, we prepared TiO2 nanoparticles using sol-gel synthesis and the porous structure was developed by salt-leaching process. Homogeneous distribution of TiO2 and HA nanoparticles were confirmed by images of VP-FE-SEM and those equipped with energy dispersive X-ray spectrometer. Structural characteristics of the porous SF/TiO2/HA hybrid scaffold were also determined using FTIR analysis and X-ray diffractometer. In this study, the porous SF/TiO2/HA hybrid scaffold showed similar porosity, enhanced mechanical property, but decreased water binding abilities, compared with the porous SF scaffold. For evaluation of the osteogenic differentiation of rat bone marrow mesenchymal stem cells, alkaline phosphatase activity and osteogenic gene expression were employed. Our results revealed that the porous SF/TiO2/HA hybrid scaffold had improved osteoinductivity compared with the porous SF scaffold. These results suggest that the osteogenic property as well as mechanical property of the porous SF/TiO2/HA hybrid scaffold could be better than the porous SF scaffold. Therefore, the porous SF/TiO2/HA hybrid scaffold may be a good promising biomaterial for bone tissue engineering application.

  4. A polycaprolactone/cuttlefish bone-derived hydroxyapatite composite porous scaffold for bone tissue engineering.

    PubMed

    Kim, Beom-Su; Yang, Sun-Sik; Lee, Jun

    2014-07-01

    Cuttlefish bone (CB) is an attractive natural biomaterial source to obtain hydroxyapatite (HAp). In this study, a porous polycaprolactone (PCL) scaffold incorporating CB-derived HAp (CB-HAp) powder was fabricated using the solvent casting and particulate leaching method. The presence of CB-HAp in PCL/CB-HAp scaffold was confirmed by X-ray diffraction (XRD). Scanning electron microscopy (SEM) and porosity analysis showed that the average pore dimension of the fabricated scaffold was approximately 200-300 μm, with ∼85% porosity, and that the compressive modulus increased after addition of CB-HAp powders. In vitro tests such as cell proliferation assay, cytotoxicity analysis, cell attachment observations, and alkaline phosphatase activity assays showed that the PCL/CB-HAp scaffold could improve the proliferation, viability, adherence, and osteoblast differentiation rate of MG-63 cells. When surgically implanted into rabbit calvarial bone defects, consistent with the in vitro results, PCL/CB-HAp scaffold implantation resulted in significantly higher new bone formation than did implantation of PCL alone. These findings suggest that addition of CB-HAp powder to the PCL scaffold can improve cellular response and that the PCL/CB-HAp composite scaffold has great potential for use in bone tissue engineering. © 2013 Wiley Periodicals, Inc.

  5. Extrusion-based 3D printing of poly(propylene fumarate) scaffolds with hydroxyapatite gradients.

    PubMed

    Trachtenberg, Jordan E; Placone, Jesse K; Smith, Brandon T; Fisher, John P; Mikos, Antonios G

    2017-04-01

    The primary focus of this work is to present the current challenges of printing scaffolds with concentration gradients of nanoparticles with an aim to improve the processing of these scaffolds. Furthermore, we address how print fidelity is related to material composition and emphasize the importance of considering this relationship when developing complex scaffolds for bone implants. The ability to create complex tissues is becoming increasingly relevant in the tissue engineering community. For bone tissue engineering applications, this work demonstrates the ability to use extrusion-based printing techniques to control the spatial deposition of hydroxyapatite (HA) nanoparticles in a 3D composite scaffold. In doing so, we combined the benefits of synthetic, degradable polymers, such as poly(propylene fumarate) (PPF), with osteoconductive HA nanoparticles that provide robust compressive mechanical properties. Furthermore, the final 3D printed scaffolds consisted of well-defined layers with interconnected pores, two critical features for a successful bone implant. To demonstrate a controlled gradient of HA, thermogravimetric analysis was carried out to quantify HA on a per-layer basis. Moreover, we non-destructively evaluated the tendency of HA particles to aggregate within PPF using micro-computed tomography (μCT). This work provides insight for proper fabrication and characterization of composite scaffolds containing particle gradients and has broad applicability for future efforts in fabricating complex scaffolds for tissue engineering applications.

  6. In vivo study on hydroxyapatite scaffolds with trabecular architecture for bone repair.

    PubMed

    Appleford, Mark R; Oh, Sunho; Oh, Namsik; Ong, Joo L

    2009-06-15

    The objective of this research was to investigate the bone formation and angio-conductive potential of hydroxyapatite (HA) scaffolds closely matched to trabecular bone in a canine segmental defect after 3 and 12 weeks post implantation. Histomorphometric comparisons were made between naturally forming trabecular bone (control) and defects implanted with scaffolds fabricated with micro-size (M-HA) and nano-size HA (N-HA) ceramic surfaces. Scaffold architecture was similar to trabecular bone formed in control defects at 3 weeks. No significant differences were identified between the two HA scaffolds; however, significant bone in-growth was observed by 12 weeks with 43.9 +/- 4.1% and 50.4 +/- 8.8% of the cross-sectional area filled with mineralized bone in M-HA and N-HA scaffolds, respectively. Partially organized, lamellar collagen fibrils were identified by birefringence under cross-polarized light at both 3 and 12 weeks post implantation. Substantial blood vessel infiltration was identified in the scaffolds and compared with the distribution and diameter of vessels in the surrounding cortical bone. Vessels were less numerous but significantly larger than native cortical Haversian and Volkmann canals reflecting the scaffold architecture where open spaces allowed interconnected channels of bone to form. This study demonstrated the potential of trabecular bone modeled, highly porous and interconnected, HA scaffolds for regenerative orthopedics.

  7. Fabrication of HA/PHBV composite scaffolds through the emulsion freezing/freeze-drying process and characterisation of the scaffolds.

    PubMed

    Sultana, Naznin; Wang, Min

    2008-07-01

    Biodegradable polymer-based scaffolds containing osteoconductive hydroxyapatite (HA) particles can be very useful for bone tissue engineering. In this investigation, HA nanoparticles were incorporated in poly(hydroxybutyrate-co-valerate) (PHBV) polymer to fabricate osteoconductive composite scaffolds. PHBV and HA/PHBV scaffolds were made using an emulsion freezing/freeze-drying technique. The scaffolds produced were subsequently characterized using several techniques. It was found that the scaffolds were highly porous and had interconnected porous structures. The pore size ranged from several microns to around 300 microm. The spherical HA nanoparticles which were produced in-house through a nanoemulsion process could be incorporated into composite scaffolds although some of these nanoparticles existed on the surface of pore walls when a relatively large amount of HA was used for composite scaffolds. The incorporation of HA nanoparticles also enhanced compressive mechanical properties of the scaffolds.

  8. Development of model hydroxyapatite bone scaffolds with multiscale porosity for potential load bearing applications

    NASA Astrophysics Data System (ADS)

    Dellinger, Jennifer Gwynne

    2005-11-01

    Model hydroxyapatite (HA) bone scaffolds consisting of a latticed pattern of rods were fabricated by a solid freeform fabrication (SFF) technique based on the robotic deposition of colloidal pastes. An optimal HA paste formulation for this method was developed. Local porosity, i.e. microporosity (1--30 mum) and sintering porosity (less than 1 mum), were produced by including polymer microsphere porogens in the HA pastes and by controlling the sintering of the scaffolds. Scaffolds with and without local porosity were evaluated with and without in vitro accelerated degradation. Percent weight loss of the scaffolds and calcium and phosphorus concentrations in solution increased with degradation time. After degradation, compressive strength and modulus decreased significantly for scaffolds with local porosity, but did not change significantly for scaffolds without local porosity. The compressive strength and modulus of scaffolds without local porosity were comparable to human cortical bone and were significantly greater than the scaffolds with local porosity. Micropores in HA disks caused surface pits that increased the surface roughness as compared to non-microporous HA disks. Mouse mesenchymal stem cells extended their cell processes into these microporous pits on HA disks in vitro. ALP expression was prolonged, cell attachment strength increased, and ECM production appeared greater on microporous HA disks compared to non-microporous HA disks and tissue culture treated polystyrene controls. Scaffolds with and without microporosity were implanted in goats bones. Microporous scaffolds with rhBMP-2 increased the percent of the scaffold filled with bone tissue compared to microporous scaffolds without rhBMP-2. Lamellar bone inside scaffolds was aligned near the rods junctions whereas lamellar bone was aligned in a more random configuration away from the rod junctions. Microporous scaffolds stained darkly with toluidine blue beneath areas of contact with new bone. This

  9. Injectable porous nano-hydroxyapatite/chitosan/tripolyphosphate scaffolds with improved compressive strength for bone regeneration.

    PubMed

    Uswatta, Suren P; Okeke, Israel U; Jayasuriya, Ambalangodage C

    2016-12-01

    In this study we have fabricated porous injectable spherical scaffolds using chitosan biopolymer, sodium tripolyphosphate (TPP) and nano-hydroxyapatite (nHA). TPP was primarily used as an ionic crosslinker to crosslink nHA/chitosan droplets. We hypothesized that incorporating nHA into chitosan could support osteoconduction by emulating the mineralized cortical bone structure, and improve the Ultimate Compressive Strength (UCS) of the scaffolds. We prepared chitosan solutions with 0.5%, 1% and 2% (w/v) nHA concentration and used simple coacervation and lyophilization techniques to obtain spherical scaffolds. Lyophilized spherical scaffolds had a mean diameter of 1.33mm (n=25). Further, portion from each group lyophilized scaffolds were soaked and dried to obtain Lyophilized Soaked and Dried (LSD) scaffolds. LSD scaffolds had a mean diameter of 0.93mm (n=25) which is promising property for the injectability. Scanning Electron Microscopy images showed porous surface morphology and interconnected pore structures inside the scaffolds. Lyophilized and LSD scaffolds had surface pores <10 and 2μm, respectively. 2% nHA/chitosan LSD scaffolds exhibited UCS of 8.59MPa compared to UCS of 2% nHA/chitosan lyophilized scaffolds at 3.93MPa. Standardize UCS values were 79.98MPa and 357MPa for 2% nHA/chitosan lyophilized and LSD particles respectively. One-way ANOVA results showed a significant increase (p<0.001) in UCS of 1% and 2% nHA/chitosan lyophilized scaffolds compared to 0% and 0.5% nHA/chitosan lyophilized scaffolds. Moreover, 2% nHA LSD scaffolds had significantly increased (p<0.005) their mean UCS by 120% compared to 2% nHA lyophilized scaffolds. In a drawback, all scaffolds have lost their mechanical properties by 95% on the 2nd day when fully immersed in phosphate buffered saline. Additionally live and dead cell assay showed no cytotoxicity and excellent osteoblast attachment to both lyophilized and LSD scaffolds at the end of 14th day of in vitro studies. 2% n

  10. Electrospun polyurethane/hydroxyapatite bioactive scaffolds for bone tissue engineering: the role of solvent and hydroxyapatite particles.

    PubMed

    Tetteh, G; Khan, A S; Delaine-Smith, R M; Reilly, G C; Rehman, I U

    2014-11-01

    Polyurethane (PU) is a promising polymer to support bone-matrix producing cells due to its durability and mechanical resistance. In this study two types of medical grade poly-ether urethanes Z3A1 and Z9A1 and PU-Hydroxyapatite (PU-HA) composites were investigated for their ability to act as a scaffold for tissue engineered bone. PU dissolved in varying concentrations of dimethylformamide (DMF) and tetrahydrofuran (THF) solvents were electrospun to attain scaffolds with randomly orientated non-woven fibres. Bioactive polymeric composite scaffolds were created using 15 wt% Z3A1 in a 70/30 DMF/THF PU solution and incorporating micro- or nano-sized HA particles in a ratio of 3:1 respectively, whilst a 25 wt% Z9A1 PU solution was doped in ratio of 5:1. Chemical properties of the resulting composites were evaluated by FTIR and physical properties by SEM. Tensile mechanical testing was carried out on all electrospun scaffolds. MLO-A5 osteoblastic mouse cells and human embryonic mesenchymal progenitor cells, hES-MPs were seeded on the scaffolds to test their biocompatibility and ability to support mineralised matrix production over a 28 day culture period. Cell viability was assayed by MTT and calcium and collagen deposition by Sirius red and alizarin red respectively. SEM images of both electrospun PU scaffolds and PU-HA composite scaffolds showed differences in fibre morphology with changes in solvent combinations and size of HA particles. Inclusion of THF eliminated the presence of beads in fibres that were present in scaffolds fabricated with 100% DMF solvent, and resulted in fibres with a more uniform morphology and thicker diameters. Mechanical testing demonstrated that the Young׳s Modulus and yield strength was lower at higher THF concentrations. Inclusion of both sizes of HA particles in PU-HA solutions reinforced the scaffolds leading to higher mechanical properties, whilst FTIR characterisation confirmed the presence of HA in all composite scaffolds. Although

  11. Hydroxyapatite/polylactide biphasic combination scaffold loaded with dexamethasone for bone regeneration.

    PubMed

    Son, Jun-Sik; Kim, Su-Gwan; Oh, Ji-Su; Appleford, Mark; Oh, Sunho; Ong, Joo L; Lee, Kyu-Bok

    2011-12-15

    This study presents a novel design of a ceramic/polymer biphasic combination scaffold that mimics natural bone structures and is used as a bone graft substitute. To mimic the natural bone structures, the outside cortical-like shells were composed of porous hydroxyapatite (HA) with a hollow interior using a polymeric template-coating technique; the inner trabecular-like core consisted of porous poly(D,L-lactic acid) (PLA) that was loaded with dexamethasone (DEX) and was directly produced using a particle leaching/gas forming technique to create the inner diameter of the HA scaffold. It was observed that the HA and PLA parts of the fabricated HA/PLA biphasic scaffold contained open and interconnected pore structures, and the boundary between both parts was tightly connected without any gaps. It was found that the structure of the combination scaffold was analogous to that of natural bone based on micro-computed tomography analysis. Additionally, the dense, uniform apatite layer was formed on the surface of the HA/PLA biphasic scaffold through a biomimetic process, and DEX was successfully released from the PLA of the biphasic scaffold over a 1-month period. This release caused human embryonic palatal mesenchyme cells to proliferate, differentiate, produce ECM, and form tissue in vitro. Therefore, it was concluded that this functionally graded scaffold is similar to natural bone and represents a potential bone-substitute material.

  12. The fabrication and characterization of biodegradable HA/PHBV nanoparticle-polymer composite scaffolds.

    PubMed

    Jack, Kevin S; Velayudhan, Shiny; Luckman, Paul; Trau, Matt; Grøndahl, Lisbeth; Cooper-White, Justin

    2009-09-01

    This study reports the fabrication and characterization of nano-sized hydroxyapatite (HA)/poly(hydroxyabutyrate-co-hydroxyvalerate) (PHBV) polymer composite scaffolds with high porosity and controlled pore architectures. These scaffolds were prepared using a modified thermally induced phase-separation technique. This investigation focuses on the effect of fabrication conditions on the overall pore architecture of the scaffolds and the dispersion of HA nanocrystals within the composite scaffolds. The morphologies, mechanical properties and in vitro bioactivity of the composite scaffolds were investigated. It was noted that the pore architectures could be manipulated by varying phase-separation parameters. The HA particles were dispersed in the pore walls of the scaffolds and were well bonded to the polymer. The introduction of HA greatly increased the stiffness and strength, and improved the in vitro bioactivity of the scaffolds. The results suggest these newly developed nano-HA/PHBV composite scaffolds may serve as an effective three-dimensional substrate in bone tissue engineering.

  13. Citric Acid-based Hydroxyapatite Composite Scaffolds Enhance Calvarial Regeneration

    PubMed Central

    Sun, Dawei; Chen, Yuhui; Tran, Richard T.; Xu, Song; Xie, Denghui; Jia, Chunhong; Wang, Yuchen; Guo, Ying; Zhang, Zhongmin; Guo, Jinshan; Yang, Jian; Jin, Dadi; Bai, Xiaochun

    2014-01-01

    Citric acid-based polymer/hydroxyapatite composites (CABP-HAs) are a novel class of biomimetic composites that have recently attracted significant attention in tissue engineering. The objective of this study was to compare the efficacy of using two different CABP-HAs, poly (1,8-octanediol citrate)-click-HA (POC-Click-HA) and crosslinked urethane-doped polyester-HA (CUPE-HA) as an alternative to autologous tissue grafts in the repair of skeletal defects. CABP-HA disc-shaped scaffolds (65 wt.-% HA with 70% porosity) were used as bare implants without the addition of growth factors or cells to renovate 4 mm diameter rat calvarial defects (n = 72, n = 18 per group). Defects were either left empty (negative control group), or treated with CUPE-HA scaffolds, POC-Click-HA scaffolds, or autologous bone grafts (AB group). Radiological and histological data showed a significant enhancement of osteogenesis in defects treated with CUPE-HA scaffolds when compared to POC-Click-HA scaffolds. Both, POC-Click-HA and CUPE-HA scaffolds, resulted in enhanced bone mineral density, trabecular thickness, and angiogenesis when compared to the control groups at 1, 3, and 6 months post-trauma. These results show the potential of CABP-HA bare implants as biocompatible, osteogenic, and off-shelf-available options in the repair of orthopedic defects. PMID:25372769

  14. Osseointegrative Properties of Electrospun Hydroxyapatite-Containing Nanofibrous Chitosan Scaffolds

    PubMed Central

    Frohbergh, Michael E.; Katsman, Anya; Mondrinos, Mark J.; Stabler, Collin T.; Hankenson, Kurt D.; Oristaglio, Jeffrey T.

    2015-01-01

    Our long-term goal is to develop smart biomaterials that can facilitate regeneration of critical-size craniofacial lesions. In this study, we tested the hypothesis that biomimetic scaffolds electrospun from chitosan (CTS) will promote tissue repair and regeneration in a critical size calvarial defect. To test this hypothesis, we first compared in vitro ability of electrospun CTS scaffolds crosslinked with genipin (CTS-GP) to those of mineralized CTS-GP scaffolds containing hydroxyapatite (CTS-HA-GP), by assessing proliferation/metabolic activity and alkaline phosphatase (ALP) levels of murine mesenchymal stem cells (mMSCs). The cells' metabolic activity exhibited a biphasic behavior, indicative of initial proliferation followed by subsequent differentiation for all scaffolds. ALP activity of mMSCs, a surrogate measure of osteogenic differentiation, increased over time in culture. After 3 weeks in maintenance medium, ALP activity of mMSCs seeded onto CTS-HA-GP scaffolds was approximately two times higher than that of cells cultured on CTS-GP scaffolds. The mineralized CTS-HA-GP scaffolds were also osseointegrative in vivo, as inferred from the enhanced bone regeneration in a murine model of critical size calvarial defects. Tissue regeneration was evaluated over a 3 month period by microCT and histology (Hematoxylin and Eosin and Masson's Trichrome). Treatment of the lesions with CTS-HA-GP scaffolds induced a 38% increase in the area of de novo generated mineralized tissue area after 3 months, whereas CTS-GP scaffolds only led to a 10% increase. Preseeding with mMSCs significantly enhanced the regenerative capacity of CTS-GP scaffolds (by ∼3-fold), to 35% increase in mineralized tissue area after 3 months. CTS-HA-GP scaffolds preseeded with mMSCs yielded 45% new mineralized tissue formation in the defects. We conclude that the presence of HA in the CTS-GP scaffolds significantly enhances their osseointegrative capacity and that mineralized chitosan-based scaffolds

  15. Hydroxyapatite porous scaffold engineered with biological polymer hybrid coating for antibiotic Vancomycin release.

    PubMed

    Kim, Hae-Won; Knowles, Jonathan C; Kim, Hyoun-Ee

    2005-03-01

    The purpose of this study is to improve hydroxyapatite (HA) porous scaffolds via coating with biological polymer-HA hybrids for use as wound healing and tissue regeneration. Highly porous HA scaffolds, fabricated by a polyurethane foam reticulate method, were coated with hybrid coating solution, consisting of poly(epsilon-caprolactone) (PCL), HA powders, and the antibiotic Vancomycin. The PCL to HA ratio was fixed at 1.5 and the drug amounts were varied [drug/(PCL + HA) = 0.02 and 0.04]. For the purpose of comparison, bare HA scaffold without the hybrid coating layer was also loaded with Vancomycin via an immersion-adsorption method. The hybrid coating structure and morphology were observed with Fourier transformed infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM). The effects of the hybrid coating on the compressive mechanical properties and the in vitro drug release of the scaffolds were investigated in comparison with bare HA scaffold. The PCL-HA hybrid coating altered the scaffold pore structure slightly, resulting in thicker stems and reduced porosity. With the hybrid coating, the HA scaffold responded to an applied compressive stress more effectively without showing a brittle failure. This was attributed to the shielding and covering of the framework surface by the coating layer. The encapsulated drugs within the coated scaffold was released in a highly sustained manner as compared to the rapid release of drugs directly adsorbed on the pure HA scaffold. These findings suggest that the coated HA scaffolds expand their applicability in hard tissue regeneration and wound healing substitutes delivering bioactive molecules.

  16. A functional agarose-hydroxyapatite scaffold for osteochondral interface regeneration

    PubMed Central

    Khanarian, Nora T.; Haney, Nora M.; Burga, Rachel A.; Lu, Helen H.

    2013-01-01

    Regeneration of the osteochondral interface is critical for integrative and functional cartilage repair. This study focuses on the design and optimization of a hydrogel-ceramic composite scaffold of agarose and hydroxyapatite (HA) for calcified cartilage formation. The first study objective was to compare the effects of HA on non-hypertrophic and hypertrophic chondrocytes cultured in the composite scaffold. Specifically, cell growth, biosynthesis, hypertrophy, and scaffold mechanical properties were evaluated. Next, the ceramic phase of the scaffold was optimized in terms of particle size (200 nm vs. 25 µm) and dose (0–6 w/v%). It was observed that while deep zone chondrocyte (DZC) biosynthesis and hypertrophy remained unaffected, hypertrophic chondrocytes measured higher matrix deposition and mineralization potential with the addition of HA. Most importantly, higher matrix content translated into significant increases in both compressive and shear mechanical properties. While cell hypertrophy was independent of ceramic size, matrix deposition was higher only with the addition of micron-sized ceramic particles. In addition, the highest matrix content, mechanical properties and mineralization potential were found in scaffolds with 3% micro-HA, which approximates both the mineral aggregate size and content of the native interface. These results demonstrate that the biomimetic hydrogel-ceramic composite is optimal for calcified cartilage formation and is a promising design strategy for osteochondral interface regeneration. PMID:22531222

  17. In vitro bioactivity of bioresorbable porous polymeric scaffolds incorporating hydroxyapatite microspheres.

    PubMed

    Li, L H; Kommareddy, K P; Pilz, C; Zhou, C R; Fratzl, P; Manjubala, I

    2010-07-01

    Biomimetic composites consisting of polymer and mineral components, resembling bone in structure and composition, were produced using a rapid prototyping technique for bone tissue engineering applications. Solid freeform fabrication, known as rapid prototyping (RP) technology, allows scaffolds to be designed with pre-defined and controlled external and internal architecture. Using the indirect RP technique, a three-component scaffold with a woodpile structure, consisting of poly-L-lactic acid (PLLA), chitosan and hydroxyapatite (HA) microspheres, was produced that had a macroporosity of more than 50% together with micropores induced by lyophilization. X-ray diffraction analysis indicated that the preparation and construction of the composite scaffold did not affect the phase composition of the HA. The compressive strength and elastic modulus (E) for the PLLA composites are 0.42 and 1.46 MPa, respectively, which are much higher than those of chitosan/HA composites and resemble the properties of cellular structure. These scaffolds showed excellent biocompatibility and ability for three-dimensional tissue growth of MC3T3-E1 pre-osteoblastic cells. The pre-osteoblastic cells cultured on these scaffolds formed a network on the HA microspheres and proliferated not only in the macropore channels but also in the micropores, as seen from the histological analysis and electron microscopy. The proliferating cells formed an extracellular matrix network and also differentiated into mature osteoblasts, as indicated by alkaline phosphatase enzyme activity. The properties of these scaffolds indicate that they can be used for non-load-bearing applications.

  18. Antibacterial chitosan coating on nano-hydroxyapatite/polyamide66 porous bone scaffold for drug delivery.

    PubMed

    Huang, Di; Zuo, Yi; Zou, Qin; Zhang, Li; Li, Jidong; Cheng, Lin; Shen, Juan; Li, Yubao

    2011-01-01

    This study describes a new drug-loaded coating scaffold applied in infection therapy during bone regeneration. Chitosan (CS) containing antibacterial berberine was coated on a nano-hydroxyapatite/polyamide66 (n-HA/PA66) scaffold to realize bone regeneration together with antimicrobial properties. The porous scaffold was fabricated using the phase-inversion method with a porosity of about 84% and macropore size of 400-600 μm. The morphology, mechanical properties and drug-release behavior were investigated at different ratios of chitosan to berberine. The results show that the elastic modulus and compressive strength of the coated scaffolds were improved to 35.4 MPa and 1.7 MPa, respectively, about 7 times and 3 times higher than the uncoated scaffolds. After a burst release of berberine within the first 3 h in PBS solution, a continuous berberine release can last 150 h, which is highly dependent on the coating concentration and suitable for antibacterial requirement of orthopaedic surgery. The bactericidal test confirms a strong antibiotic effect of the delivery system and the minimum inhibitory concentration of the drug is 0.02 mg/ml. Moreover, in vitro biological evaluation demonstrates that the coating scaffolds act as a good matrix for MG63 adhesion, crawl, growth and proliferation, suggesting that the antibacterial delivery system has no cytotoxicity. We expect the drug-delivery system to have a potential application in bone regeneration or defect repair.

  19. Effect of different hydroxyapatite incorporation methods on the structural and biological properties of porous collagen scaffolds for bone repair.

    PubMed

    Ryan, Alan J; Gleeson, John P; Matsiko, Amos; Thompson, Emmet M; O'Brien, Fergal J

    2015-12-01

    Scaffolds which aim to provide an optimised environment to regenerate bone tissue require a balance between mechanical properties and architecture known to be conducive to enable tissue regeneration, such as a high porosity and a suitable pore size. Using freeze-dried collagen-based scaffolds as an analogue of native ECM, we sought to improve the mechanical properties by incorporating hydroxyapatite (HA) in different ways while maintaining a pore architecture sufficient to allow cell infiltration, vascularisation and effective bone regeneration. Specifically we sought to elucidate the effect of different hydroxyapatite incorporation methods on the mechanical, morphological, and cellular response of the resultant collagen-HA scaffolds. The results demonstrated that incorporating either micron-sized (CHA scaffolds) or nano-sized HA particles (CnHA scaffolds) prior to freeze-drying resulted in moderate increases in stiffness (2.2-fold and 6.2-fold, respectively, vs. collagen-glycosaminoglycan scaffolds, P < 0.05, a scaffold known to support osteogenesis), while enabling good cell attachment, and moderate mesenchymal stem cell (MSC)-mediated calcium production after 28 days' culture (2.1-fold, P < 0.05, and 1.3-fold, respectively, vs. CG scaffolds). However, coating of collagen scaffolds with a hydroxyapatite precipitate after freeze-drying (CpHA scaffolds) has been shown to be a highly effective method to increase the compressive modulus (26-fold vs. CG controls, P < 0.001) of scaffolds while maintaining a high porosity (~ 98%). The coating of the ligand-dense collagen structure results in a lower cell attachment level (P < 0.05), although it supported greater cell-mediated calcium production (P < 0.0001) compared with other scaffold variants after 28 days' culture. The comparatively good mechanical properties of these high porosity scaffolds is obtained partially through highly crosslinking the scaffolds with both a physical (DHT) and chemical (EDAC) crosslinking

  20. Alternate Spray-coating for the Direct Fabrication of Hydroxyapatite Films without Crystal Growth Step in Solution.

    PubMed

    Watanabe, Satoshi; Kashiwagi, Rei; Matsumoto, Mutsuyoshi

    2017-03-01

    We discuss an alternate spray-coating technique for the direct fabrication of hydroxyapatite films using metal masks, suction-type spray nozzles and two calcification solutions of calcium hydroxide and phosphoric acid aqueous solutions. Hydroxyapatite films were formed only on the hydrophobic surface of the substrates. Scanning electron microscopy and energy dispersive X-ray spectroscopy showed that the spray-coated films consisted of hydroxyapatite nanoparticles. The Ca/P ratio was estimated to be about 1.26. X-ray diffraction patterns of the spray-coated films almost coincided with those of the hydroxyapatite powders, showing that the spray-coated films consisted of hydroxyapatite nanoparticles. Dot arrays of hydroxyapatite films at a diameter of 100 μm were formed by tuning the concentrations of calcium hydroxide and phosphoric acid aqueous solutions. This technique allows for the direct fabrication of the hydroxyapatite films without crystal growth process in hydroxyapatite precursors, the scaffolds of crystal growth such as biocompatibility SiO2-CaO glasses, or electrophoresis processes. By using this technique, large-area ceramic films with biocompatibility will be micropatterned with minimized material consumption, short fabrication time, and reduced equipment investments.

  1. Microcracks induce osteoblast alignment and maturation on hydroxyapatite scaffolds

    NASA Astrophysics Data System (ADS)

    Shu, Yutian

    Physiological bone tissue is a mineral/collagen composite with a hierarchical structure. The features in bone, such as mineral crystals, fibers, and pores can range from the nanometer to the centimeter in size. Currently available bone tissue scaffolds primarily address the chemical composition, pore size, and pore size distribution. While these design parameters are extensively investigated for mimicking bone function and inducing bone regeneration, little is known about microcracks, which is a prevalent feature found in fractured bone in vivo and associated with fracture healing and repair. Since the purpose of bone tissue engineering scaffold is to enhance bone regeneration, the coincidence of microcracks and bone densification should not be neglected but rather be considered as a potential parameter in bone tissue engineering scaffold design. The purpose of this study is to test the hypothesis that microcracks enhance bone healing. In vitro studies were designed to investigate the osteoblast (bone forming cells) response to microcracks in dense (94%) hydroxyapatite substrates. Microcracks were introduced using a well-established Vickers indentation technique. The results of our study showed that microcracks induced osteoblast alignment, enhanced osteoblast attachment and more rapid maturation. These findings may provide insight into fracture healing mechanism(s) as well as improve the design of bone tissue engineering orthopedic scaffolds for more rapid bone regeneration.

  2. Nanocomposite bone scaffolds based on biodegradable polymers and hydroxyapatite.

    PubMed

    Becker, Johannes; Lu, Lichun; Runge, M Brett; Zeng, Heng; Yaszemski, Michael J; Dadsetan, Mahrokh

    2015-08-01

    In tissue engineering, development of an osteoconductive construct that integrates with host tissue remains a challenge. In this work, the effect of bone-like minerals on maturation of pre-osteoblast cells was investigated using polymer-mineral scaffolds composed of poly(propylene fumarate)-co-poly(caprolactone) (PPF-co-PCL) and nano-sized hydroxyapatite (HA). The HA of varying concentrations was added to an injectable formulation of PPF-co-PCL and the change in thermal and mechanical properties of the scaffolds was evaluated. No change in onset of degradation temperature was observed due to the addition of HA, however compressive and tensile moduli of copolymer changed significantly when HA amounts were increased in composite formulation. The change in mechanical properties of copolymer was found to correlate well to HA concentration in the constructs. Electron microscopy revealed mineral nucleation and a change in surface morphology and the presence of calcium and phosphate on surfaces was confirmed using energy dispersive X-ray analysis. To characterize the effect of mineral on attachment and maturation of pre-osteoblasts, W20-17 cells were seeded on HA/copolymer composites. We demonstrated that cells attached more to the surface of HA containing copolymers and their proliferation rate was significantly increased. Thus, these findings suggest that HA/PPF-co-PCL composite scaffolds are capable of inducing maturation of pre-osteoblasts and have the potential for use as scaffold in bone tissue engineering.

  3. The drug release study of ceftriaxone from porous hydroxyapatite scaffolds.

    PubMed

    Al-Sokanee, Zeki N; Toabi, Abedl Amer H; Al-Assadi, Mohammed J; Alassadi, Erfan A S

    2009-01-01

    Hydroxyapatite (HAP) is an important biomedical material that is used for grafting osseous defects. It has an excellent bioactivity and biocompatibility properties. To isolate hydroxyapatite, pieces of cleaned cattle's bone were heated at different temperature range from 400 degrees C up to 1,200 degrees C. A reasonable yield of 60.32% w/w HAP was obtained at temperature range from 1,000 degrees C to 1,200 degrees C. Fourier transform infrared spectra and the thermogravimetric measurement showed a clear removal of organic at 600 degrees C as well as an excellent isolation of HAP from the bones which was achieved at 1,000-1,200 degrees C. This was also confirmed from X-ray diffraction of bone sample heated at 1,200 degrees C. The concentration ions were found to be sodium, potassium, lithium, zinc, copper, iron, calcium, magnesium, and phosphate present in bones within the acceptable limits for its role in the bioactivity property of HAP. Glucose powder was used as a porosifier. Glucose was novel and excellent as porogen where it was completely removed by heating, giving an efficient porosity in the used scaffolds. The results exhibited that the ceftriaxone drug release was increased with increasing the porosity. It was found that a faster, higher, and more regular drug release was obtained from the scaffold with a porosity of 10%.

  4. Integrating novel technologies to fabricate smart scaffolds.

    PubMed

    Moroni, L; de Wijn, J R; van Blitterswijk, C A

    2008-01-01

    Tissue engineering aims at restoring or regenerating a damaged tissue by combining cells, derived from a patient biopsy, with a 3D porous matrix functioning as a scaffold. After isolation and eventual in vitro expansion, cells are seeded on the 3D scaffolds and implanted directly or at a later stage in the patient's body. 3D scaffolds need to satisfy a number of requirements: (i) biocompatibility, (ii) biodegradability and/or bioresorbability, (iii) suitable mechanical properties, (iv) adequate physicochemical properties to direct cell-material interactions matching the tissue to be replaced and (v) ease in regaining the original shape of the damaged tissue and the integration with the surrounding environment. Still, it appears to be a challenge to satisfy all the aforementioned requisites with the biomaterials and the scaffold fabrication technologies nowadays available. 3D scaffolds can be fabricated with various techniques, among which rapid prototyping and electrospinning seem to be the most promising. Rapid prototyping technologies allow manufacturing scaffolds with a controlled, completely accessible pore network--determinant for nutrient supply and diffusion--in a CAD/CAM fashion. Electrospinning (ESP) allows mimicking the extracellular matrix (ECM) environment of the cells and can provide fibrous scaffolds with instructive surface properties to direct cell faith into the proper lineage. Yet, these fabrication methods have some disadvantages if considered alone. This review aims at summarizing conventional and novel scaffold fabrication techniques and the biomaterials used for tissue engineering and drug-delivery applications. A new trend seems to emerge in the field of scaffold design where different scaffolds fabrication technologies and different biomaterials are combined to provide cells with mechanical, physicochemical and biological cues at the macro-, micro- and nano-scale. If merged together, these integrated technologies may lead to the generation

  5. Morphological effects of porous poly-d,l-lactic acid/hydroxyapatite scaffolds produced by supercritical CO2 foaming on their mechanical performance.

    PubMed

    Rouholamin, Davood; van Grunsven, William; Reilly, Gwendolen C; Smith, Patrick J

    2016-08-01

    A novel supercritical CO2 foaming technique was used to fabricate scaffolds of controllable morphology and mechanical properties, with the potential to tailor the scaffolds to specific tissue engineering applications. Biodegradable scaffolds are widely used as temporary supportive structures for bone regeneration. The scaffolds must provide a sufficient mechanical support while allowing cell attachment and growth as well as metabolic activities. In this study, supercritical CO2 foaming was used to prepare fully interconnected porous scaffolds of poly-d,l-lactic acid and poly-d,l-lactic acid/hydroxyapatite. The morphological, mechanical and cell behaviours of the scaffolds were measured to examine the effect of hydroxyapatite on these properties. These scaffolds showed an average porosity in the range of 86%-95%, an average pore diameter of 229-347 µm and an average pore interconnection of 103-207 µm. The measured porosity, pore diameter, and interconnection size are suitable for cancellous bone regeneration. Compressive strength and modulus of up to 36.03 ± 5.90 and 37.97 ± 6.84 MPa were measured for the produced porous scaffolds of various compositions. The mechanical properties presented an improvement with the addition of hydroxyapatite to the structure. The relationship between morphological and mechanical properties was investigated. The matrices with different compositions were seeded with bone cells, and all the matrices showed a high cell viability and biocompatibility. The number of cells attached on the matrices slightly increased with the addition of hydroxyapatite indicating that hydroxyapatite improves the biocompatibility and proliferation of the scaffolds. The produced poly-d,l-lactic acid/hydroxyapatite scaffolds in this study showed a potential to be used as bone graft substitutes. © IMechE 2016.

  6. Selenium-Substituted Hydroxyapatite/Biodegradable Polymer/Pamidronate Combined Scaffold for the Therapy of Bone Tumour.

    PubMed

    Oledzka, Ewa; Sobczak, Marcin; Kolmas, Joanna; Nalecz-Jawecki, Grzegorz

    2015-09-14

    The present study evaluated a new concept of combined scaffolds as a promising bone replacement material for patients with a bone tumour or bone metastasis. The scaffolds were composed of hydroxyapatite doped with selenium ions and a biodegradable polymer (linear or branched), and contained an active substance-bisphosphonate. For this purpose, a series of biodegradable polyesters were synthesized through a ring-opening polymerization of ε-caprolactone or d,l-lactide in the presence of 2-hydroxyethyl methacrylate (HEMA) or hyperbranched 2,2-bis(hydroxymethyl)propionic acid polyester-16-hydroxyl (bis-MPA) initiators, substances often used in the synthesis of medical materials. The polymers were obtained with a high yield and a number-average molecular weight up to 45,300 (g/mol). The combined scaffolds were then manufactured by a direct compression of pre-synthesized hydroxyapatite doped with selenite or selenate ions, obtained polymer and pamidronate as a model drug. It was found that the kinetic release of the drug from the scaffolds tested in vitro under physiological conditions is strongly dependent on the physicochemical properties and average molecular weight of the polymers. Furthermore, there was good correlation with the hydrolytic biodegradation results of the scaffolds fabricated without drug. The preliminary findings suggest that the fabricated combined scaffolds could be effectively used for the sustained delivery of bioactive molecules at bone defect sites.

  7. Selenium-Substituted Hydroxyapatite/Biodegradable Polymer/Pamidronate Combined Scaffold for the Therapy of Bone Tumour

    PubMed Central

    Oledzka, Ewa; Sobczak, Marcin; Kolmas, Joanna; Nalecz-Jawecki, Grzegorz

    2015-01-01

    The present study evaluated a new concept of combined scaffolds as a promising bone replacement material for patients with a bone tumour or bone metastasis. The scaffolds were composed of hydroxyapatite doped with selenium ions and a biodegradable polymer (linear or branched), and contained an active substance—bisphosphonate. For this purpose, a series of biodegradable polyesters were synthesized through a ring-opening polymerization of ε-caprolactone or d,l-lactide in the presence of 2-hydroxyethyl methacrylate (HEMA) or hyperbranched 2,2-bis(hydroxymethyl)propionic acid polyester-16-hydroxyl (bis-MPA) initiators, substances often used in the synthesis of medical materials. The polymers were obtained with a high yield and a number-average molecular weight up to 45,300 (g/mol). The combined scaffolds were then manufactured by a direct compression of pre-synthesized hydroxyapatite doped with selenite or selenate ions, obtained polymer and pamidronate as a model drug. It was found that the kinetic release of the drug from the scaffolds tested in vitro under physiological conditions is strongly dependent on the physicochemical properties and average molecular weight of the polymers. Furthermore, there was good correlation with the hydrolytic biodegradation results of the scaffolds fabricated without drug. The preliminary findings suggest that the fabricated combined scaffolds could be effectively used for the sustained delivery of bioactive molecules at bone defect sites. PMID:26389884

  8. Poly(ε-caprolactone)/nano fluoridated hydroxyapatite scaffolds for bone tissue engineering: in vitro degradation and biocompatibility study.

    PubMed

    Johari, N; Fathi, M H; Golozar, M A; Erfani, E; Samadikuchaksaraei, A

    2012-03-01

    In this study, biodegradation and biocompatibility of novel poly(ε-caparolactone)/nano fluoridated hydroxyapatite (PCL-FHA) scaffolds were investigated. The FHA nanopowders were prepared via mechanical alloying method and had a chemical composition of Ca(10)(PO(4))(6)OH(2-x )F(x) (where x values were selected equal to 0.5 and 2.0). In order to fabricate PCL-FHA scaffolds, 10, 20, 30 and 40 wt% of the FHA were added to the PCL. The PCL-FHA scaffolds were produced by the solvent casting/particulate leaching using sodium chloride particles (with diameters of 300-500 μm) as the porogen. The phase structure, microstructure and morphology of the scaffolds were evaluated using X-ray diffraction, Fourier transform infrared spectroscopy and scanning electron microscopy techniques. Porosity of the scaffolds was measured using the Archimedes' Principle. In vitro degradation of PCL-FHA scaffolds was studied by incubating the samples in phosphate buffered saline at 37°C and pH 7.4 for 30 days. Moreover, biocompatibility was evaluated by MTT assay after seeding and culture of osteoblast-like cells on the scaffolds. Results showed that the osteoblast-like cells attached to and proliferated on PCL-FHA and increasing the porosity of the scaffolds increased the cell viability. Also, degradation rate of scaffolds were increased with increasing the fluorine content in scaffolds composition.

  9. Fabrication and characterization of bioactive wollastonite/PHBV composite scaffolds.

    PubMed

    Li, Haiyan; Chang, Jiang

    2004-11-01

    Composite scaffolds of polyhydroxybutyrate-polyhydroxyvalerate (PHBV) with bioactive wollastonite were fabricated by a compression moulding, thermal processing, and salt particulate leaching method. Structure and mechanical properties of the scaffolds were determined. The bioactivity of the composites was evaluated by soaking in a simulated body fluid (SBF), and the formation of the hydroxyapatite (HAp) layer was determined by Scanning Electron Microscope (SEM) and Energy-Dispersive Spectrometer (EDS). The results showed that the wollastonite/PHBV composites were bioactive as it induced the formation of HAp on the surface of the composite scaffolds after soaking in SBF for 14 days. In addition, the measurements of the water contact angles suggested that incorporation of wollastonite into PHBV could improve the hydrophilicity of the composites and the enhancement was dependent on the wollastonite content. Furthermore, the pH and ion concentration changes of SBF solutions with composite scaffolds showed that the composites released Ca and Si ions, which could neutralize the acidic by-products of the PHBV and stabilize the pH of the SBF solutions between 7.2 and 7.8 within a 3-week soaking period. All of these results suggest that the incorporation of wollastonite was a useful approach to obtain composite scaffolds with improved properties.

  10. Biomimetic hydroxyapatite coating on pore walls improves osteointegration of poly(L-lactic acid) scaffolds.

    PubMed

    Deplaine, H; Lebourg, M; Ripalda, P; Vidaurre, A; Sanz-Ramos, P; Mora, G; Prósper, F; Ochoa, I; Doblaré, M; Gómez Ribelles, J L; Izal-Azcárate, I; Gallego Ferrer, G

    2013-01-01

    Polymer-ceramic composites obtained as the result of a mineralization process hold great promise for the future of tissue engineering. Simulated body fluids (SBFs) are widely used for the mineralization of polymer scaffolds. In this work an exhaustive study with the aim of optimizing the mineralization process on a poly(L-lactic acid) (PLLA) macroporous scaffold has been performed. We observed that when an air plasma treatment is applied to the PLLA scaffold its hydroxyapatite nucleation ability is considerably improved. However, plasma treatment only allows apatite deposition on the surface of the scaffold but not in its interior. When a 5 wt % of synthetic hydroxyapatite (HAp) nanoparticles is mixed with PLLA a more abundant biomimetic hydroxyapatite layer grows inside the scaffold in SBF. The morphology, amount, and composition of the generated biomimetic hydroxyapatite layer on the pores' surface have been analyzed. Large mineralization times are harmful to pure PLLA as it rapidly degrades and its elastic compression modulus significantly decreases. Degradation is retarded in the composite scaffolds because of the faster and extensive biomimetic apatite deposition and the role of HAp to control the pH. Mineralized scaffolds, covered by an apatite layer in SBF, were implanted in osteochondral lesions performed in the medial femoral condyle of healthy sheep. We observed that the presence of biomimetic hydroxyapatite on the pore's surface of the composite scaffold produces a better integration in the subchondral bone, in comparison to bare PLLA scaffolds. Copyright © 2012 Wiley Periodicals, Inc.

  11. In vivo bone response to 3D periodic hydroxyapatite scaffolds assembled by direct ink writing.

    PubMed

    Simon, Joshua L; Michna, Sarah; Lewis, Jennifer A; Rekow, E Dianne; Thompson, Van P; Smay, James E; Yampolsky, Andrew; Parsons, J Russell; Ricci, John L

    2007-12-01

    The in vivo bone response of 3D periodic hydroxyapatite (HA) scaffolds is investigated. Two groups of HA scaffolds (11 mm diameter x 3.5 mm thick) are fabricated by direct-write assembly of a concentrated HA ink. The scaffolds consist of cylindrical rods periodically arranged into four quadrants with varying separation distances between rods. In the first group, HA rods (250 microm in diameter) are patterned to create pore channels, whose areal dimensions are 250 x 250 microm(2) in quadrant 1, 250 x 500 microm(2) in quadrants 2 and 4, and 500 x 500 microm(2) in quadrant 3. In the second group, HA rods (400 microm in diameter) are patterned to create pore channels, whose areal dimensions of 500 x 500 microm(2) in quadrant 1, 500 x 750 microm(2) in quadrants 2 and 4, and 750 x 750 microm(2) in quadrant 3. Each group of scaffolds is partially densified by sintering at 1200 degrees C prior to being implanted bilaterally in trephine defects of skeletally mature New Zealand White rabbits. Their tissue response is evaluated at 8 and 16 weeks using micro-computed tomography, histology, and scanning electron microscopy. New trabecular bone is conducted rapidly and efficiently across substantial distances within these patterned 3D HA scaffolds. Our observations suggest that HA rods are first coated with a layer of new bone followed by subsequent scaffold infilling via outward and inward radial growth of the coated regions. Direct-write assembly of 3D periodic scaffolds composed of micro-porous HA rods arrayed to produce macro-pores that are size-matched to trabecular bone may represent an optimal strategy for bone repair and replacement structures.

  12. 3D printing of porous hydroxyapatite scaffolds intended for use in bone tissue engineering applications.

    PubMed

    Cox, Sophie C; Thornby, John A; Gibbons, Gregory J; Williams, Mark A; Mallick, Kajal K

    2015-02-01

    A systematic characterisation of bone tissue scaffolds fabricated via 3D printing from hydroxyapatite (HA) and poly(vinyl)alcohol (PVOH) composite powders is presented. Flowability of HA:PVOH precursor materials was observed to affect mechanical stability, microstructure and porosity of 3D printed scaffolds. Anisotropic behaviour of constructs and part failure at the boundaries of interlayer bonds was highlighted by compressive strength testing. A trade-off between the ability to facilitate removal of PVOH thermal degradation products during sintering and the compressive strength of green parts was revealed. The ultimate compressive strength of 55% porous green scaffolds printed along the Y-axis and dried in a vacuum oven for 6h was 0.88 ± 0.02 MPa. Critically, the pores of 3D printed constructs could be user designed, ensuring bulk interconnectivity, and the imperfect packing of powder particles created an inherent surface roughness and non-designed porosity within the scaffold. These features are considered promising since they are known to facilitate osteoconduction and osteointegration in-vivo. Characterisation techniques utilised in this study include two funnel flow tests, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), compressive strength testing and computed tomography (CT). Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Direct writing of porous tissue scaffolds based on Vaseline-doped hydroxyapatite inks

    NASA Astrophysics Data System (ADS)

    Li, Ya-Yun; Li, Long-Tu; Li, Bo

    2015-05-01

    A novel type of 40 vol.% hydroxyapatite (HAp), Ca10(PO4)6(OH)2, suspension doped with Vaseline was developed, and porous three-dimensional (3D) scaffolds were fabricated by using a direct ink writing (DIW) method. The preparation of the HAp inks and the principles of the DIW technique were investigated. The microporosity of the scaffold wall increased after introducing the Vaseline, whereas macroporosity can be produced by varying the DIW technique. The micromorphology test results show that the samples sintered at 1150°C for 2 h formed ceramics with a set amount of pores, which benefit cell growth by providing more locations for cells to attach and proliferate. Under a microscope, the proliferations of human liver carcinoma cell line (HepG2) cells can be observed on the 3D HAp scaffolds. The DIW method has the advantages of a rapid process, ease of design and high precision control, potentially inspiring the design and application of biomaterials and scaffolds.

  14. Biomineralized hydroxyapatite nanoclay composite scaffolds with polycaprolactone for stem cell-based bone tissue engineering.

    PubMed

    Ambre, Avinash H; Katti, Dinesh R; Katti, Kalpana S

    2015-06-01

    Nanoclay modified with unnatural amino acid was used to design a nanoclay-hydroxyapatite (HAP) hybrid by mineralizing HAP in the nanoclay galleries mimicking biomineralization. This hybrid (in situ HAPclay) was used to fabricate polycaprolactone (PCL)/in situ HAPclay films and scaffolds for bone regeneration. Cell culture assays and imaging were used to study interactions between human mesenchymal stem cells (hMSCs) and PCL/in situ HAPclay composites (films and scaffolds). SEM imaging indicated MSC attachment, formation of mineralized extracellular (ECM) on PCL/in situ HAPclay films, and infiltration of MSCs to the interior of PCL/in situ HAPclay scaffolds. Mineralized ECM was formed by MSCs without use of osteogenic supplements. AFM imaging performed on this in vitro generated mineralized ECM on PCL/in situ HAPclay films revealed presence of components (collagen and mineral) of hierarchical organization reminiscent of natural bone. Cellular events observed during two-stage seeding experiments on PCL/in situ HAPclay films indicated similarities with events occurring during in vivo bone formation. PCL/in situ HAPclay films showed significantly increased (100-595% increase in elastic moduli) nanomechanical properties and PCL/in situ HAPclay scaffolds showed increased degradation. This work puts forth PCL/in situ HAPclay composites as viable biomaterials for bone tissue engineering. © 2014 Wiley Periodicals, Inc.

  15. Low-Temperature Additive Manufacturing of Biomimic Three-Dimensional Hydroxyapatite/Collagen Scaffolds for Bone Regeneration.

    PubMed

    Lin, Kai-Feng; He, Shu; Song, Yue; Wang, Chun-Mei; Gao, Yi; Li, Jun-Qin; Tang, Peng; Wang, Zheng; Bi, Long; Pei, Guo-Xian

    2016-03-23

    Low-temperature additive manufacturing (AM) holds promise for fabrication of three-dimensional (3D) scaffolds containing bioactive molecules and/or drugs. Due to the strict technical limitations of current approaches, few materials are suitable for printing at low temperature. Here, a low-temperature robocasting method was employed to print biomimic 3D scaffolds for bone regeneration using a routine collagen-hydroxyapatite (CHA) composite material, which is too viscous to be printed via normal 3D printing methods at low temperature. The CHA scaffolds had excellent 3D structure and maintained most raw material properties after printing. Compared to nonprinted scaffolds, printed scaffolds promoted bone marrow stromal cell proliferation and improved osteogenic outcome in vitro. In a rabbit femoral condyle defect model, the interconnecting pores within the printed scaffolds facilitated cell penetration and mineralization before the scaffolds degraded and enhanced repair, compared to nonprinted CHA scaffolds. Additionally, the optimal printing parameters for 3D CHA scaffolds were investigated; 600-μm-diameter rods were optimal in terms of moderate mechanical strength and better repair outcome in vivo. This low-temperature robocasting method could enable a variety of bioactive molecules to be incorporated into printed CHA materials and provides a method of bioprinting biomaterials without compromising their natural properties.

  16. CAD-CAM-generated hydroxyapatite scaffold to replace the mandibular condyle in sheep: preliminary results.

    PubMed

    Ciocca, Leonardo; Donati, Davide; Fantini, Massimiliano; Landi, Elena; Piattelli, Adriano; Iezzi, Giovanna; Tampieri, Anna; Spadari, Alessandro; Romagnoli, Noemi; Scotti, Roberto

    2013-08-01

    In this study, rapid CAD-CAM prototyping of pure hydroxyapatite to replace temporomandibular joint condyles was tested in sheep. Three adult animals were implanted with CAD-CAM-designed porous hydroxyapatite scaffolds as condyle substitutes. The desired scaffold shape was achieved by subtractive automated milling machining (block reduction). Custom-made surgical guides were created by direct metal laser sintering and were used to export the virtual planning of the bone cut lines into the surgical environment. Using the same technique, fixation plates were created and applied to the scaffold pre-operatively to firmly secure the condyles to the bone and to assure primary stability of the hydroxyapatite scaffolds during masticatory function. Four months post-surgery, the sheep were sacrificed. The hydroxyapatite scaffolds were explanted, and histological specimens were prepared. Different histological tissues penetrating the scaffold macropores, the sequence of bone remodeling, new apposition of bone and/or cartilage as a consequence of the different functional anatomic role, and osseointegration at the interface between the scaffold and bone were documented. This animal model was found to be appropriate for testing CAD-CAM customization and the biomechanical properties of porous, pure hydroxyapatite scaffolds used as joint prostheses.

  17. Porous Hydroxyapatite Scaffold with Three-Dimensional Localized Drug Delivery System Using Biodegradable Microspheres

    DTIC Science & Technology

    2011-03-21

    March 2011 Keywords: Hydroxyapatite Poly(lactic-co-glycolic acid ) Drug delivery system Scaffold Microsphere Dexamethasone In this study, ionic...immobilization of dexamethasone (DEX)-loaded poly(lactic-co-glycolic acid ) (PLGA) microspheres was performed on the hydroxyapatite (HAp) scaffold surfaces. It...polyurethane, HAp/ poly(lactic acid ), and HAp/poly(lactic-co-glycolic acid ) (PLGA). The goal has been to increase mechanical stability and improve tissue

  18. Fabrication of combinatorial polymer scaffold libraries.

    PubMed

    Simon, Carl G; Stephens, Jean S; Dorsey, Shauna M; Becker, Matthew L

    2007-07-01

    We have designed a novel combinatorial research platform to help accelerate tissue engineering research. Combinatorial methods combine many samples into a single specimen to enable accelerated experimentation and discovery. The platform for fabricating combinatorial polymer scaffold libraries can be used to rapidly identify scaffold formulations that maximize tissue formation. Many approaches for screening cell-biomaterial interactions utilize a two-dimensional format such as a film or surface to present test substrates to cells. However, cells in vivo exist in a three-dimensional milieu of extracellular matrix and cells in vitro behave more naturally when cultured in a three-dimensional environment than when cultured on a two-dimensional surface. Thus, we have designed a method for fabricating combinatorial biomaterial libraries where the materials are presented to cells in the form of three-dimensional, porous, salt-leached, polymer scaffolds. Many scaffold variations and compositions can be screened in a single experiment so that optimal scaffold formulations for tissue formation can be rapidly identified. In summary, we have developed a platform technology for fabricating combinatorial polymer scaffold libraries that can be used to screen cell response to materials in a three-dimensional, scaffold format.

  19. Fabrication of combinatorial polymer scaffold libraries

    NASA Astrophysics Data System (ADS)

    Simon, Carl G.; Stephens, Jean S.; Dorsey, Shauna M.; Becker, Matthew L.

    2007-07-01

    We have designed a novel combinatorial research platform to help accelerate tissue engineering research. Combinatorial methods combine many samples into a single specimen to enable accelerated experimentation and discovery. The platform for fabricating combinatorial polymer scaffold libraries can be used to rapidly identify scaffold formulations that maximize tissue formation. Many approaches for screening cell-biomaterial interactions utilize a two-dimensional format such as a film or surface to present test substrates to cells. However, cells in vivo exist in a three-dimensional milieu of extracellular matrix and cells in vitro behave more naturally when cultured in a three-dimensional environment than when cultured on a two-dimensional surface. Thus, we have designed a method for fabricating combinatorial biomaterial libraries where the materials are presented to cells in the form of three-dimensional, porous, salt-leached, polymer scaffolds. Many scaffold variations and compositions can be screened in a single experiment so that optimal scaffold formulations for tissue formation can be rapidly identified. In summary, we have developed a platform technology for fabricating combinatorial polymer scaffold libraries that can be used to screen cell response to materials in a three-dimensional, scaffold format.

  20. A review: fabrication of porous polyurethane scaffolds.

    PubMed

    Janik, H; Marzec, M

    2015-03-01

    The aim of tissue engineering is the fabrication of three-dimensional scaffolds that can be used for the reconstruction and regeneration of damaged or deformed tissues and organs. A wide variety of techniques have been developed to create either fibrous or porous scaffolds from polymers, metals, composite materials and ceramics. However, the most promising materials are biodegradable polymers due to their comprehensive mechanical properties, ability to control the rate of degradation and similarities to natural tissue structures. Polyurethanes (PUs) are attractive candidates for scaffold fabrication, since they are biocompatible, and have excellent mechanical properties and mechanical flexibility. PU can be applied to various methods of porous scaffold fabrication, among which are solvent casting/particulate leaching, thermally induced phase separation, gas foaming, emulsion freeze-drying and melt moulding. Scaffold properties obtained by these techniques, including pore size, interconnectivity and total porosity, all depend on the thermal processing parameters, and the porogen agent and solvents used. In this review, various polyurethane systems for scaffolds are discussed, as well as methods of fabrication, including the latest developments, and their advantages and disadvantages.

  1. Microsphere-Based Scaffolds Encapsulating Tricalcium Phosphate And Hydroxyapatite For Bone Regeneration

    PubMed Central

    Gupta, Vineet; Lyne, Dina V.; Barragan, Marilyn; Berkland, Cory J.; Detamore, Michael S.

    2016-01-01

    Bioceramic mixtures of tricalcium phosphate (TCP) and hydroxyapatite (HAp) are widely used for bone regeneration because of their excellent cytocompatibility, osteoconduction, and osteoinduction. Therefore, we hypothesized that incorporation of a mixture of TCP and HAp in microsphere-based scaffolds would enhance osteogenesis of rat bone marrow stromal cells (rBMSCs) compared to a positive control of scaffolds with encapsulated bone-morphogenic protein-2 (BMP-2). Poly(D,L-lactic-co-glycolic acid) (PLGA) microsphere-based scaffolds encapsulating TCP and HAp mixtures in two different ratios (7:3 and 1:1) were fabricated with the same net ceramic content (30 wt%) to evaluate how incorporation of these ceramic mixtures would affect the osteogenesis in rBMSCs. Encapsulation of TCP/HAp mixtures impacted microsphere morphologies and the compressive moduli of the scaffolds. Additionally, TCP/HAp mixtures enhanced the end-point secretion of extracellular matrix (ECM) components relevant to bone tissue compared to the “blank” (PLGA-only) microsphere-based scaffolds as evidenced by the biochemical, gene expression, histology, and immunohistochemical characterization. Moreover, the TCP/HAp mixture groups even surpassed the BMP-2 positive control group in some instances in terms of matrix synthesis and gene expression. Lastly, gene expression data suggested that the rBMSCs responded differently to different TCP/HAp ratios presented to them. Altogether, it can be concluded that TCP/HAp mixtures stimulated the differentiation of rBMSCs toward an osteoblastic phenotype, and therefore may be beneficial in gradient microsphere-based scaffolds for osteochondral regeneration. PMID:27272903

  2. [Effects of bore diameter of porous hydroxyapatite scaffolds on three-dimensional dynamic cultivation of osteoblasts].

    PubMed

    Wang, Hai; Zhang, Linghan; Qiu, Guixing; Huang, Wei; Zhou, Kui; Zhang, Xianglin; Wu, Zhihong

    2014-10-28

    To explore the relationship between bore diameter of porous hydroxyapatite (HA) scaffolds and the adhesion, proliferation and metabolism of osteoblasts by 3D cultivation. MC3T3-E1 cell suspension was separately dropped axially through different bore diameter scaffolds (Group A:150 µm, Group B:300 µm) to confirm initial seeding. Then scaffolds were transferred into a perfusion bioreactor of 5% CO2 at 37°C for 5 days with an average flow of 3.4 ml/min. After perfusion cultivation, cell proliferation between different groups of scaffolds was determined by methyl thiazolyl tetrazolium (MTT) assay and cell metabolic activities were determined by glucose consumption. Lastly cell adhesion and proliferation were observed directly by scanning electronic microscope (SEM). The results of MTT assay showed that the optical density/mass ratios were 1.31 ± 0.26 in group A and 1.51 ± 0.43 in group B. There was no significant difference (t test, P = 0.36). Glucose consumption in group A was significantly lower than that in group B [(162.38 ± 33.09) vs (217.97 ± 27.91) µmol/L, P = 0.01]. The adhesion, proliferation, pseudopodia and extracellular matrix of osteoblasts in internal part of scaffolds after perfusion cultivation were observed by SEM. With excellent biocompatibility, porous HA scaffolds are available for fabricating tissue engineering bones. There is no effect of bore diameter on the proliferation of osteoblasts. But it affects the metabolic activity of osteoblasts. So bore diameter may be increased within a feasible range on the premise of mechanical properties.

  3. Porous scaffolds of gelatin-hydroxyapatite nanocomposites obtained by biomimetic approach: characterization and antibiotic drug release.

    PubMed

    Kim, Hae-Won; Knowles, Jonathan C; Kim, Hyoun-Ee

    2005-08-01

    Gelatin-hydroxyapatite (HA) nanocomposite porous scaffolds were fabricated biomimetically, and their feasibility as a drug-delivery carrier for tissue-regeneration and wound-healing treatments was addressed. The composite sols were prepared by the precipitation of HA up to 30 wt % within a gelatin solution with the use of calcium and phosphate precursors, and the porous scaffold was obtained by casting the sols and further freeze drying. The obtained bodies were crosslinked with carbodiimide derivatives to retain chemical and thermal integrity. The apatite precipitates were observed to be a poorly crystallized carbonate-substituted HA. The nanocomposite scaffolds had porosities of approximately 89-92% and exhibited a bimodal pore distribution, that is, the macropores (approximately 300-500 microm) of the framework structure, and micropores (approximately 0.5-1 microm) formed on the framework surface. Transmission electron microscopy (TEM) observation revealed the precipitation of highly elongated HA nanocrystals on the gelatin network. The well-developed porous structure and organized nanocomposite configurations were in marked contrast to the directly mixed gelatin-HA powder conventional composites. For drug-release tests, tetracycline, an antibiotic drug, was entrapped within the scaffold, and the drug-release profile was examined with processing parameters, such as HA amount in gelatin, crosslinking degree, and initial drug addition. The drug entrapment decreased with increasing HA amount, but increased with increasing crosslinking degree and initial drug addition. The crosslinking of the gelatin was the prerequisite to sustaining and controlling the drug releases. Compared to pure gelatin, the gelatin-HA nanocomposites had lower drug releases, because of their lower water uptake and degradation. All the nanocomposite scaffolds released drugs in proportion to the initial drug addition, suggesting their capacity to deliver drugs in a controlled manner. Based on

  4. Lamination of microfibrous PLGA fabric by electrospinning a layer of collagen-hydroxyapatite composite nanofibers for bone tissue engineering.

    PubMed

    Kwon, Gi-Wan; Gupta, Kailash Chandra; Jung, Kyung-Hye; Kang, Inn-Kyu

    2017-01-01

    To mimic the muscle inspired cells adhesion through proteins secretion, the lamination of collagen-hydroxyapatite nanorod (nHA) composite nanofibers has been carried out successfully on polydopamine (PDA)-coated microfibrous polylactide-co-glycolide (PLGA) fabrics. The lamination of collagen-hydroxyapatite composite nanofibers on polydopamine-coated microfibrous PLGA fabrics was carried through electrospinning the solution of collagen containing L-glutamic acid-grafted hydroxyapatite nanorods (nHA-GA) at a flow rate of 1.5 mL/h and an applied voltage of 15 kV. In comparison to pristine PLGA, dopamine-coated PLGA and collagen-hydroxyapatite composite nanofiber lamination has produced more wettable surfaces and surface wettability is found to higher with dopamine-coated PLGA fabrics then pristine PLGA. The SEM micrographs have clearly indicated that the lamination of polydopamine-coated PLGA fabric with collagen-hydroxyapatite composite nanofibers has shown increased adhesion of MC3T3E1 cells in comparison to pristine PLGA fabrics. The results of these studies have clearly demonstrated that collagen-nHA composites fibers may be used to create bioactive 3D scaffolds using PLGA as an architectural support agent.

  5. Rapid Prototyping Amphiphilic Polymer/Hydroxyapatite Composite Scaffolds with Hydration-Induced Self-Fixation Behavior

    PubMed Central

    Kutikov, Artem B.; Gurijala, Anvesh

    2015-01-01

    Two major factors hampering the broad use of rapid prototyped biomaterials for tissue engineering applications are the requirement for custom-designed or expensive research-grade three-dimensional (3D) printers and the limited selection of suitable thermoplastic biomaterials exhibiting physical characteristics desired for facile surgical handling and biological properties encouraging tissue integration. Properly designed thermoplastic biodegradable amphiphilic polymers can exhibit hydration-dependent hydrophilicity changes and stiffening behavior, which may be exploited to facilitate the surgical delivery/self-fixation of the scaffold within a physiological tissue environment. Compared to conventional hydrophobic polyesters, they also present significant advantages in blending with hydrophilic osteoconductive minerals with improved interfacial adhesion for bone tissue engineering applications. Here, we demonstrated the excellent blending of biodegradable, amphiphilic poly(D,L-lactic acid)-poly(ethylene glycol)-poly(D,L-lactic acid) (PLA-PEG-PLA) (PELA) triblock co-polymer with hydroxyapatite (HA) and the fabrication of high-quality rapid prototyped 3D macroporous composite scaffolds using an unmodified consumer-grade 3D printer. The rapid prototyped HA-PELA composite scaffolds and the PELA control (without HA) swelled (66% and 44% volume increases, respectively) and stiffened (1.38-fold and 4-fold increases in compressive modulus, respectively) in water. To test the hypothesis that the hydration-induced physical changes can translate into self-fixation properties of the scaffolds within a confined defect, a straightforward in vitro pull-out test was designed to quantify the peak force required to dislodge these scaffolds from a simulated cylindrical defect at dry versus wet states. Consistent with our hypothesis, the peak fixation force measured for the PELA and HA-PELA scaffolds increased 6-fold and 15-fold upon hydration, respectively. Furthermore, we showed that

  6. Rapid prototyping amphiphilic polymer/hydroxyapatite composite scaffolds with hydration-induced self-fixation behavior.

    PubMed

    Kutikov, Artem B; Gurijala, Anvesh; Song, Jie

    2015-03-01

    Two major factors hampering the broad use of rapid prototyped biomaterials for tissue engineering applications are the requirement for custom-designed or expensive research-grade three-dimensional (3D) printers and the limited selection of suitable thermoplastic biomaterials exhibiting physical characteristics desired for facile surgical handling and biological properties encouraging tissue integration. Properly designed thermoplastic biodegradable amphiphilic polymers can exhibit hydration-dependent hydrophilicity changes and stiffening behavior, which may be exploited to facilitate the surgical delivery/self-fixation of the scaffold within a physiological tissue environment. Compared to conventional hydrophobic polyesters, they also present significant advantages in blending with hydrophilic osteoconductive minerals with improved interfacial adhesion for bone tissue engineering applications. Here, we demonstrated the excellent blending of biodegradable, amphiphilic poly(D,L-lactic acid)-poly(ethylene glycol)-poly(D,L-lactic acid) (PLA-PEG-PLA) (PELA) triblock co-polymer with hydroxyapatite (HA) and the fabrication of high-quality rapid prototyped 3D macroporous composite scaffolds using an unmodified consumer-grade 3D printer. The rapid prototyped HA-PELA composite scaffolds and the PELA control (without HA) swelled (66% and 44% volume increases, respectively) and stiffened (1.38-fold and 4-fold increases in compressive modulus, respectively) in water. To test the hypothesis that the hydration-induced physical changes can translate into self-fixation properties of the scaffolds within a confined defect, a straightforward in vitro pull-out test was designed to quantify the peak force required to dislodge these scaffolds from a simulated cylindrical defect at dry versus wet states. Consistent with our hypothesis, the peak fixation force measured for the PELA and HA-PELA scaffolds increased 6-fold and 15-fold upon hydration, respectively. Furthermore, we showed that

  7. Polyphosphazene/Nano-Hydroxyapatite Composite Microsphere Scaffolds for Bone Tissue Engineering

    PubMed Central

    Nukavarapu, Syam P.; Kumbar, Sangamesh G.; Brown, Justin L.; Krogman, Nicholas R.; Weikel, Arlin L.; Hindenlang, Mark D.; Nair, Lakshmi S.; Allcock, Harry R; Laurencin, Cato T.

    2009-01-01

    The non-toxic, neutral degradation products of amino acid ester polyphosphazenes make them ideal candidates for in vivo orthopaedic applications. The quest for new osteocompatible materials for load bearing tissue engineering applications has led us to investigate mechanically competent amino acid ester substituted polyphosphazenes. In this study, we have synthesized three biodegradable polyphosphazenes substituted with side groups namely leucine, valine and phenylalanine ethyl esters. Of these polymers, the phenylalanine ethyl ester substituted polyphosphazene showed the highest glass transition temperature (41.6 °C) and hence was chosen as a candidate material for forming composite microspheres with 100 nm sized hydroxyapatite (nHAp). The fabricated composite microspheres were sintered into a three-dimensional (3-D) porous scaffold by adopting a dynamic solvent sintering approach. The composite microsphere scaffolds showed compressive moduli of 46–81 MPa with mean pore diameters in the range of 86–145 µm. The three-dimensional polyphosphazene-nHAp composite microsphere scaffolds showed good osteoblast cell adhesion, proliferation and alkaline phosphatase expression, and are potential suitors for bone tissue engineering applications. PMID:18517248

  8. Selective laser sintering of hydroxyapatite reinforced polyethylene composites for bioactive implants and tissue scaffold development.

    PubMed

    Hao, L; Savalani, M M; Zhang, Y; Tanner, K E; Harris, R A

    2006-05-01

    Selective laser sintering (SLS) has been investigated for the production of bioactive implants and tissue scaffolds using composites of high-density polyethylene (HDPE) reinforced with hydroxyapatite (HA) with the aim of achieving the rapid manufacturing of customized implants. Single-layer and multilayer block specimens made of HA-HDPE composites with 30 and 40 vol % HA were sintered successfully using a CO2 laser sintering system. Laser power and scanning speed had a significant effect on the sintering behaviour. The degree of particle fusion and porosity were influenced by the laser processing parameters, hence control can be attained by varying these parameters. Moreover, the SLS processing allowed exposure of HA particles on the surface of the composites and thereby should provide bioactive products. Pores existed in the SLS-fabricated composite parts and at certain processing parameters a significant fraction of the pores were within the optimal sizes for tissue regeneration. The results indicate that the SLS technique has the potential not only to fabricate HA-HDPE composite products but also to produce appropriate features for their application as bioactive implants and tissue scaffolds.

  9. Gradient porous hydroxyapatite ceramics fabricated by freeze casting method

    NASA Astrophysics Data System (ADS)

    Zuo, Kai-hui; zhang, Yuan; Jiang, Dongliang; Zeng, Yu-Ping

    2011-04-01

    By controlling the cooling rates and the composition of slurries, the gradient porous hydroxyapatite ceramics are fabricated by the freeze casting method. According to the different cooling rate, the pores of HAP ceramics fabricated by gradient freeze casting are divided into three parts: one is lamellar pores, another is column pore and the last one is fine round pores. The laminated freeze casting is in favour of obtaining the gradient porous ceramics composed of different materials and the ceramics have unclear interfaces.

  10. Effect of Cell-seeded Hydroxyapatite Scaffolds on Rabbit Radius Bone Regeneration

    DTIC Science & Technology

    2013-06-22

    Library (wileyonlinelibrary.com). DOI: 10.1002/jbm.a.34834 Abstract: Highly porous hydroxyapatite (HA) scaffolds were developed as bone graft substitutes ...search for substitutes for autolo- gous bone grafting applications has led to the development of scaffolds composed of a variety of materials, including...also observed. (b) Three-dimensional tissue volume measurement from mCT data indicated comparable bone volume within the scaffolds in all three treatment

  11. Effect of freezing temperature in thermally induced phase separation method in hydroxyapatite/chitosan-based bone scaffold biomaterial

    NASA Astrophysics Data System (ADS)

    Albab, Muh Fadhil; Yuwono, Akhmad Herman; Sofyan, Nofrijon; Ramahdita, Ghiska

    2017-02-01

    In the current study, hydroxyapatite (HA)/chitosan-based bone scaffold has been fabricated using Thermally Induced Phase Separation (TIPS) method under freezing temperature variation of -20, -30, -40 and -80 °C. The samples with weight percent ratio of 70% HA and 30% chitosan were homogeneously mixed and subsequently dissolved in 2% acetic acid. The synthesized samples were further characterized using Fourier transform infrared (FTIR), compressive test and scanning electron microscope (SEM). The investigation results showed that low freezing temperature reduced the pore size and increased the compressive strength of the scaffold. In the freezing temperature of -20 °C, the pore size was 133.93 µm with the compressive strength of 5.9 KPa, while for -80 °C, the pore size declined to 60.55 µm with the compressive strength 29.8 KPa. Considering the obtained characteristics, HA/chitosan obtained in this work has potential to be applied as a bone scaffold.

  12. Comparison on mechanical properties of single layered and bilayered chitosan-gelatin coated porous hydroxyapatite scaffold prepared through freeze drying method

    NASA Astrophysics Data System (ADS)

    Effendi, M. D.; Gustiono, D.; Lukmana; Ayu, D.; Kurniawati, F.

    2017-02-01

    Biopolymer coated porous hydroxyapatite (HA) scaffolds were prepared for tissue engineering trough freeze drying method and impregnation. in this study, to mimic the mineral and organic component of natural bone, synthetic hydroxapatite (HA) scaffolds coated by polymer were prepared. Highly porous Hap scaffolds, fabricated by synthetic HA impregnation method on polyurethane foam, were coated with polymer coating solution, consisting of chitosan, Gelatin, and bilayered chitosan-gelatin prepared by aging and impregnating technique. For the purpose of comparison, The bare scaffolds without polymer coating layer were investigated. The Bare scaffolds were highly porous and interconnected with a pore size of around 150 µm–714 µm, has porosity at around 67,7% -85,7%, and has mechanical strength at around 0.06 Mpa - 0.071 Mpa, which is suitable for osteoblast cell Proliferation. Chitosan coated porous HA scaffold and gelatin coated porous HA scaffold had mechanical strength at around 0.81-0.85 Mpa, and 1.32-1.34 Mpa, respectively, with weight ratio of biopolymer and Hap was around 18%-22%. To compare these results, the coating on the bare scaffold with gelatin and chitosan had been conducted. Based on the result of FTIR, it could be concluded that coating procedure applied on porous hydroxy apatite (HA) coated by gelatin, chitosan coated HA scaffold, and bilayered Gelatin-chitosan coated porous HA scaffold, confirming that for allsampleshad no significant chemical effect on the coating structure. The compressive strength of bilayered Gelatin-chitosan coated HA scaffold had middle values between the rest, at around 1,06-1.2 Mpa for the samples at the same weight ratio of biopolymer: HA (around 18% - 22%). These results also confirming that coating by gelatin on porous hydroxyapatite was highest compresive strength and can be applied to improve mechanical properties of porous hydroxyapatite bare scaffold

  13. Hydroxyapatite/regenerated silk fibroin scaffold-enhanced osteoinductivity and osteoconductivity of bone marrow-derived mesenchymal stromal cells.

    PubMed

    Jiang, Jia; Hao, Wei; Li, Yuzhuo; Yao, Jinrong; Shao, Zhengzhong; Li, Hong; Yang, Jianjun; Chen, Shiyi

    2013-04-01

    A novel hydroxyapatite/regenerated silk fibroin scaffold was prepared and investigated for its potential to enhance both osteoinductivity and osteoconductivity of bone marrow-derived mesenchymal stromal cells in vitro. Approx. 12.4 ± 0.06 % (w/w) hydroxyapatite was deposited onto the scaffold, and cell viability and DNA content were significantly increased (18.5 ± 0.6 and 33 ± 1.2 %, respectively) compared with the hydroxyapatite scaffold after 14 days. Furthermore, alkaline phosphatase activity in the novel scaffold increased 41 ± 2.5 % after 14 days compared with the hydroxyapatite scaffold. The data indicate that this novel hydroxyapatite/regenerated silk fibroin scaffold has a positive effect on osteoinductivity and osteoconductivity, and may be useful for bone tissue engineering.

  14. 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.

  15. The effect of hydroxyapatite nanoparticles on mechanical behavior and biological performance of porous shape memory polyurethane scaffolds.

    PubMed

    Yu, Juhong; Xia, Hong; Teramoto, Akira; Ni, Qing-Qing

    2017-09-07

    The scaffold which provides space for cell growth, proliferation and differentiation, is a key factor in bone tissue engineering. However, improvements in scaffold design are needed to precisely match the irregular boundaries of bone defects as well as facilitate clinical application. In this study, controllable three-dimensional (3D) porous shape memory polyurethane/nano-hydroxyapatite (SMPU/nHAP) composite scaffold was successfully fabricated for bone defect reparation. Detailed studies were performed to evaluate its structure, apparent density, porosity and mechanical properties, emphasizing the contribution of nHAP particles on shape recovery behaviors and biological performance in vitro. The effect of nHAP particles in porous SMPU/nHAP composite scaffold was found to enhance the compression resistance by 37%, shorten the compression recovery time by 41%, reduce the tensile resistance by 78%, reach the shape recovery ratio of 99% and promote the cell proliferation by 13% after 7 days of culture. These results revealed that the 3D structure and aperture of as-prepared scaffold were controllable. And in minimally invasive surgery and bone repair surgery, this porous composite scaffold could significantly reduce the operative time and promote the bone cell growth. Therefore, this porous SMPU/nHAP composite scaffold design has potential applications for the bone tissue engineering. This article is protected by copyright. All rights reserved. © 2017 Wiley Periodicals, Inc.

  16. Polyurethane/fluor-hydroxyapatite nanocomposite scaffolds for bone tissue engineering. Part I: morphological, physical, and mechanical characterization

    PubMed Central

    Asefnejad, Azadeh; Behnamghader, Aliasghar; Khorasani, Mohammad Taghi; Farsadzadeh, Babak

    2011-01-01

    In this study, new nano-fluor-hydroxyapatite (nFHA)/polyurethane composite scaffolds were fabricated for potential use in bone tissue engineering. Polyester urethane samples were synthesized from polycaprolactone, hexamethylene diisocyanate, and 1,4-butanediol as chain extender. Nano fluor-hydroxyapatite (nFHA) was successfully synthesized by sol-gel method. The solid–liquid phase separation and solvent sublimation methods were used for preparation of the porous composites. Mechanical properties, chemical structure, and morphological characteristics of the samples were investigated by compressive test, Fourier transform infrared, and scanning electron microscopy (SEM) techniques, respectively. The effect of nFHA powder content on porosity and pore morphology was investigated. SEM images demonstrated that the scaffolds were constituted of interconnected and homogeneously distributed pores. The pore size of the scaffolds was in the range 50–250 μm. The result obtained in this research revealed that the porosity and pore average size decreased and compressive modulus increased with nFHA percentage. Considering morphological, physical, and mechanical properties, the scaffold with a higher ratio of nFHA has suitable potential use in tissue regeneration. PMID:21289986

  17. Polyurethane/fluor-hydroxyapatite nanocomposite scaffolds for bone tissue engineering. Part I: morphological, physical, and mechanical characterization.

    PubMed

    Asefnejad, Azadeh; Behnamghader, Aliasghar; Khorasani, Mohammad Taghi; Farsadzadeh, Babak

    2011-01-06

    In this study, new nano-fluor-hydroxyapatite (nFHA)/polyurethane composite scaffolds were fabricated for potential use in bone tissue engineering. Polyester urethane samples were synthesized from polycaprolactone, hexamethylene diisocyanate, and 1,4-butanediol as chain extender. Nano fluor-hydroxyapatite (nFHA) was successfully synthesized by sol-gel method. The solid-liquid phase separation and solvent sublimation methods were used for preparation of the porous composites. Mechanical properties, chemical structure, and morphological characteristics of the samples were investigated by compressive test, Fourier transform infrared, and scanning electron microscopy (SEM) techniques, respectively. The effect of nFHA powder content on porosity and pore morphology was investigated. SEM images demonstrated that the scaffolds were constituted of interconnected and homogeneously distributed pores. The pore size of the scaffolds was in the range 50-250 μm. The result obtained in this research revealed that the porosity and pore average size decreased and compressive modulus increased with nFHA percentage. Considering morphological, physical, and mechanical properties, the scaffold with a higher ratio of nFHA has suitable potential use in tissue regeneration.

  18. Evaluation of the novel three-dimensional porous poly (L-lactic acid)/nano-hydroxyapatite composite scaffold.

    PubMed

    Huang, Jianghong; Xiong, Jianyi; Liu, Jianquan; Zhu, Weimin; Chen, Jielin; Duan, Li; Zhang, Jufeng; Wang, Daping

    2015-01-01

    To determine the optimal ratio of nano-hydroxyapatite (n-HA) to polylactic acid (PLLA) in the novel three-dimensional porous PLLA/n-HA composite scaffolds, low-temperature rapid prototyping technology was employed to fabricate the composite materials with different n-HA contents. Mechanical properties and degradation behaviors of the composites were examined, and the scaffold microstructure and n-HA dispersion were observed by scanning electron microscope (SEM). Mechanical tests demonstrated that the tensile strength of the composite material gradually decreased with an increase in n-HA content. When the n-HA content reached 20 wt%, the bending strength of the composite material peaked at 138.5 MPa. SEM images demonstrated that the optimal content of n-HA was 20 wt% as the largest interconnected pore size that can be seen, with a porosity as high as 80%. In vitro degradation experiments demonstrated that the pH value of the material containing solution gradually decreased in a time-dependent manner, with a simultaneous weakening of the mechanical properties. In vitro study using rat osteoblast cells showed that the composite scaffolds were biocompatible; the 20 wt% n-HA scaffold offered particular improvement to rat osteoblast cell adhesion and proliferation compared to other compositions. It was therefore concluded that 20 wt% n-HA is the optimal nano-hydroxyapatite (n-HA) to polylactic acid (PLLA) ratio, with promise for bone tissue engineering.

  19. Evaluation of zinc-doped mesoporous hydroxyapatite microspheres for the construction of a novel biomimetic scaffold optimized for bone augmentation.

    PubMed

    Yu, Weilin; Sun, Tuan-Wei; Qi, Chao; Ding, Zhenyu; Zhao, Huakun; Zhao, Shichang; Shi, Zhongmin; Zhu, Ying-Jie; Chen, Daoyun; He, Yaohua

    2017-01-01

    Biomaterials with high osteogenic activity are desirable for sufficient healing of bone defects resulting from trauma, tumor, infection, and congenital abnormalities. Synthetic materials mimicking the structure and composition of human trabecular bone are of considerable potential in bone augmentation. In the present study, a zinc (Zn)-doped mesoporous hydroxyapatite microspheres (Zn-MHMs)/collagen scaffold (Zn-MHMs/Coll) was developed through a lyophilization fabrication process and designed to mimic the trabecular bone. The Zn-MHMs were synthesized through a microwave-hydrothermal method by using creatine phosphate as an organic phosphorus source. Zn-MHMs that consist of hydroxyapatite nanosheets showed relatively uniform spherical morphology, mesoporous hollow structure, high specific surface area, and homogeneous Zn distribution. They were additionally investigated as a drug nanocarrier, which was efficient in drug delivery and presented a pH-responsive drug release behavior. Furthermore, they were incorporated into the collagen matrix to construct a biomimetic scaffold optimized for bone tissue regeneration. The Zn-MHMs/Coll scaffolds showed an interconnected pore structure in the range of 100-300 μm and a sustained release of Zn ions. More importantly, the Zn-MHMs/Coll scaffolds could enhance the osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells. Finally, the bone defect repair results of critical-sized femoral condyle defect rat model demonstrated that the Zn-MHMs/Coll scaffolds could enhance bone regeneration compared with the Coll or MHMs/Coll scaffolds. The results suggest that the biomimetic Zn-MHMs/Coll scaffolds may be of enormous potential in bone repair and regeneration.

  20. Evaluation of zinc-doped mesoporous hydroxyapatite microspheres for the construction of a novel biomimetic scaffold optimized for bone augmentation

    PubMed Central

    Yu, Weilin; Sun, Tuan-Wei; Qi, Chao; Ding, Zhenyu; Zhao, Huakun; Zhao, Shichang; Shi, Zhongmin; Zhu, Ying-Jie; Chen, Daoyun; He, Yaohua

    2017-01-01

    Biomaterials with high osteogenic activity are desirable for sufficient healing of bone defects resulting from trauma, tumor, infection, and congenital abnormalities. Synthetic materials mimicking the structure and composition of human trabecular bone are of considerable potential in bone augmentation. In the present study, a zinc (Zn)-doped mesoporous hydroxyapatite microspheres (Zn-MHMs)/collagen scaffold (Zn-MHMs/Coll) was developed through a lyophilization fabrication process and designed to mimic the trabecular bone. The Zn-MHMs were synthesized through a microwave-hydrothermal method by using creatine phosphate as an organic phosphorus source. Zn-MHMs that consist of hydroxyapatite nanosheets showed relatively uniform spherical morphology, mesoporous hollow structure, high specific surface area, and homogeneous Zn distribution. They were additionally investigated as a drug nanocarrier, which was efficient in drug delivery and presented a pH-responsive drug release behavior. Furthermore, they were incorporated into the collagen matrix to construct a biomimetic scaffold optimized for bone tissue regeneration. The Zn-MHMs/Coll scaffolds showed an interconnected pore structure in the range of 100–300 μm and a sustained release of Zn ions. More importantly, the Zn-MHMs/Coll scaffolds could enhance the osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells. Finally, the bone defect repair results of critical-sized femoral condyle defect rat model demonstrated that the Zn-MHMs/Coll scaffolds could enhance bone regeneration compared with the Coll or MHMs/Coll scaffolds. The results suggest that the biomimetic Zn-MHMs/Coll scaffolds may be of enormous potential in bone repair and regeneration. PMID:28392688

  1. Improvement of mechanical strength and osteogenic potential of calcium sulfate-based hydroxyapatite 3-dimensional printed scaffolds by ε-polycarbonate coating.

    PubMed

    Kim, Beom-Su; Yang, Sun-Sik; Park, Ho; Lee, Se-Hwan; Cho, Young-Sam; Lee, Jun

    2017-09-01

    Powder-based three-dimensional (3D) printing is an excellent method to fabricate complex-shaped scaffolds for tissue engineering. However, their lower mechanical strength restricts their application in bone tissue engineering. Here, we created a 3D-printed scaffold coated with a ε-polycaprolactone (PCL) polymer solution (5 and 10 w/v %) to improve the mechanical strength of the scaffold. The 3D scaffold was fabricated from calcium sulfate hemihydrate powder (CaSO4-1/2 H2O), transformed into hydroxyapatite (HAp) by treatment with a hydrothermal reaction in an NH4H2PO4 solution. The surface properties and composition of the scaffold were evaluated using scanning electron microscopy and X-ray diffraction analysis. We demonstrated that the 3D scaffold coated with PCL had an improved mechanical modulus. Coating with 5 and 10% PCL increased the compressive strength significantly, by about 2-fold and 4-fold, respectively, compared with that of uncoated scaffolds. However, the porosity was reduced significantly by coating with 10% PCL. In vitro biological evaluation demonstrated that MG-63 cells adhered well and proliferated on the 3D scaffold coated with PCL, and the scaffold was not cytotoxic. In addition, alkaline phosphatase activity and real time polymerase chain reaction demonstrated that osteoblast differentiation also improved in the PCL-coated 3D scaffolds. These results indicated that PCL polymer coating could improve the compressive strength and biocompatibility of 3D HAp scaffolds for bone tissue engineering applications.

  2. In vivo study of chitosan-natural nano hydroxyapatite scaffolds for bone tissue regeneration.

    PubMed

    Lee, Jong Seo; Baek, Sang Dae; Venkatesan, Jayachandran; Bhatnagar, Ira; Chang, Hee Kyung; Kim, Hui Taek; Kim, Se-Kwon

    2014-06-01

    Significant development has been achieved with bioceramics and biopolymer scaffolds in the construction of artificial bone. In the present study, we have developed and compared chitosan-micro hydroxyapatite (chitosan-mHA) and chitosan-nano hydroxyapatite (chitosan-nHA) scaffolds as bone graft substitutes. The biocompatibility and cell proliferation of the prepared scaffolds were checked with preosteoblast (MC3T3-E1) cells. Total Volume (TV), bone volume (BV), bone surface (BS), trabecular thickness (Tb.Th), trabecular number (Tb.N) and trabecular separation (Tb.Sp) were found to be higher in chitosan-nHA than chitosan-mHA scaffold. Hence, we suggest that chitosan-nHA scaffold could be a promising biomaterial for bone tissue engineering. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. From Waste to Health: Synthesis of Hydroxyapatite Scaffolds From Fish Scales for Lead Ion Removal

    NASA Astrophysics Data System (ADS)

    Liu, Wen-Kuang; Liaw, Bor-Shuang; Chang, Haw-Kai; Wang, Yi-Feng; Chen, Po-Yu

    2017-02-01

    Fish scales are natural sources of collagen and hydroxyapatite, yet are typically considered waste. In this study, fish scale powder obtained from Tilapia fish (Oreochromis mossambicus) was used to synthesize scaffolds by freeze casting for the removal of lead ions from wastewater. The mineral hydroxyapatite was confirmed by x-ray diffraction. Scaffolds with a well-aligned lamellar microstructure (10-80-μm channel widths) and proper mechanical properties under compression were synthesized by controlling the solid loading, cooling rate, and sintering temperature. The maximum adsorption capacities (q max) were 208.3 mg/g and 344.8 mg/g in pH = 5 and pH = 2.2 solutions, respectively. More than 99.9% of the lead ion was removed after 20 min. The results show that fish-scale-derived hydroxyapatite scaffolds are promising for heavy metal ion removal and wastewater treatments.

  4. From Waste to Health: Synthesis of Hydroxyapatite Scaffolds From Fish Scales for Lead Ion Removal

    NASA Astrophysics Data System (ADS)

    Liu, Wen-Kuang; Liaw, Bor-Shuang; Chang, Haw-Kai; Wang, Yi-Feng; Chen, Po-Yu

    2017-04-01

    Fish scales are natural sources of collagen and hydroxyapatite, yet are typically considered waste. In this study, fish scale powder obtained from Tilapia fish ( Oreochromis mossambicus) was used to synthesize scaffolds by freeze casting for the removal of lead ions from wastewater. The mineral hydroxyapatite was confirmed by x-ray diffraction. Scaffolds with a well-aligned lamellar microstructure (10-80- μm channel widths) and proper mechanical properties under compression were synthesized by controlling the solid loading, cooling rate, and sintering temperature. The maximum adsorption capacities ( q max) were 208.3 mg/g and 344.8 mg/g in pH = 5 and pH = 2.2 solutions, respectively. More than 99.9% of the lead ion was removed after 20 min. The results show that fish-scale-derived hydroxyapatite scaffolds are promising for heavy metal ion removal and wastewater treatments.

  5. Effect of doping in carbon nanotubes on the viability of biomimetic chitosan-carbon nanotubes-hydroxyapatite scaffolds.

    PubMed

    Fonseca-García, Abril; Mota-Morales, Josué D; Quintero-Ortega, Iraís A; García-Carvajal, Zaira Y; Martínez-López, V; Ruvalcaba, Erika; Landa-Solís, Carlos; Solis, Lilia; Ibarra, Clemente; Gutiérrez, María C; Terrones, Mauricio; Sanchez, Isaac C; del Monte, Francisco; Velasquillo, María C; Luna-Bárcenas, G

    2014-10-01

    This work describes the preparation and characterization of biomimetic chitosan/multiwall carbon nanotubes/nano-hydroxyapatite (CTS/MWCNT/nHAp) scaffolds and their viability for bone tissue engineering applications. The cryogenic process ice segregation-induced self-assembly (ISISA) was used to fabricate 3D biomimetic CTS scaffolds. Proper combination of cryogenics, freeze-drying, nature and molecular ratio of solutes give rise to 3D porous interconnected scaffolds with clusters of nHAp distributed along the scaffold surface. The effect of doping in CNT (e.g. with oxygen and nitrogen atoms) on cell viability was tested. Under the same processing conditions, pore size was in the range of 20-150 μm and irrespective on the type of CNT. Studies on cell viability with scaffolds were carried out using human cells from periosteum biopsy. Prior to cell seeding, the immunophenotype of mesenchymal periosteum or periosteum-derived stem cells (MSCs-PCs) was characterized by flow cytometric analysis using fluorescence-activated and characteristic cell surface markers for MSCs-PCs. The characterized MSCs-PCs maintained their periosteal potential in cell cultures until the 2nd passage from primary cell culture. Thus, the biomimetic CTS/MWCNT/nHAp scaffolds demonstrated good biocompatibility and cell viability in all cases such that it can be considered as promising biomaterials for bone tissue engineering.

  6. Preparation and characterization of bionic bone structure chitosan/hydroxyapatite scaffold for bone tissue engineering.

    PubMed

    Zhang, Jiazhen; Nie, Jingyi; Zhang, Qirong; Li, Youliang; Wang, Zhengke; Hu, Qiaoling

    2014-01-01

    Three-dimensional oriented chitosan (CS)/hydroxyapatite (HA) scaffolds were prepared via in situ precipitation method in this research. Scanning electron microscopy (SEM) images indicated that the scaffolds with acicular nano-HA had the spoke-like, multilayer and porous structure. The SEM of osteoblasts which were polygonal or spindle-shaped on the composite scaffolds after seven-day cell culture showed that the cells grew, adhered, and spread well. The results of X-ray powder diffractometer and Fourier transform infrared spectrometer showed that the mineral particles deposited in the scaffold had phase structure similar to natural bone and confirmed that particles were exactly HA. In vitro biocompatibility evaluation indicated the composite scaffolds showed a higher degree of proliferation of MC3T3-E1 cell compared with the pure CS scaffolds and the CS/HA10 scaffold was the highest one. The CS/HA scaffold also had a higher ratio of adhesion and alkaline phosphate activity value of osteoblasts compared with the pure CS scaffold, and the ratio increased with the increase of HA content. The ALP activity value of composite scaffolds was at least six times of the pure CS scaffolds. The results suggested that the composite scaffolds possessed good biocompatibility. The compressive strength of CS/HA15 increased by 33.07% compared with the pure CS scaffold. This novel porous scaffold with three-dimensional oriented structure might have a potential application in bone tissue engineering.

  7. Development of Microspheres Covered with Hydroxyapatite Nanocrystals as Cell Scaffold for Angiogenesis

    NASA Astrophysics Data System (ADS)

    Iwamoto, Takashi; Terada, Takahiro; Kogai, Yasumichi; Okada, Masahiro; Fujii, Syuji; Furuzono, Tsutomu

    2012-06-01

    We prepared poly(L-lactide-co-glycolide) cell scaffolds coated with hydroxyapatite (HAp) nanocrystals with 50-100 nm in diameter via the Pickering emulsion method. Our cell scaffolds were composed of biodegradable polymers and HAp nanocrystals as a core and shell, respectively. The scaffolds were spherical but displayed uneven shapes when altering a shear speed of homogenization during syntheses. The surface coverage of HAp nanocrystals was examined because the HAp-coating ratio for the scaffolds was an important factor as cell scaffolds in order to enhance cell adhesion. On the basis of scanning electron microscopy observations and thermogravimetric analyses, it was found that the cell scaffolds showed distorted morphologies, and the HAp-coating ratio decreased with increasing the shear speed in the synthesis because shear stress influenced shapes of the scaffolds.

  8. Scaffolds for bone regeneration made of hydroxyapatite microspheres in a collagen matrix.

    PubMed

    Cholas, Rahmatullah; Kunjalukkal Padmanabhan, Sanosh; Gervaso, Francesca; Udayan, Gayatri; Monaco, Graziana; Sannino, Alessandro; Licciulli, Antonio

    2016-06-01

    Biomimetic scaffolds with a structural and chemical composition similar to native bone tissue may be promising for bone tissue regeneration. In the present work hydroxyapatite mesoporous microspheres (mHA) were incorporated into collagen scaffolds containing an ordered interconnected macroporosity. The mHA were obtained by spray drying of a nano hydroxyapatite slurry prepared by the precipitation technique. X-ray diffraction (XRD) analysis revealed that the microspheres were composed only of hydroxyapatite (HA) phase, and energy-dispersive x-ray spectroscopy (EDS) analysis revealed the Ca/P ratio to be 1.69 which is near the value for pure HA. The obtained microspheres had an average diameter of 6 μm, a specific surface area of 40 m(2)/g as measured by Brunauer-Emmett-Teller (BET) analysis, and Barrett-Joyner-Halenda (BJH) analysis showed a mesoporous structure with an average pore diameter of 16 nm. Collagen/HA-microsphere (Col/mHA) composite scaffolds were prepared by freeze-drying followed by dehydrothermal crosslinking. SEM observations of Col/mHA scaffolds revealed HA microspheres embedded within a porous collagen matrix with a pore size ranging from a few microns up to 200 μm, which was also confirmed by histological staining of sections of paraffin embedded scaffolds. The compressive modulus of the composite scaffold at low and high strain values was 1.7 and 2.8 times, respectively, that of pure collagen scaffolds. Cell proliferation measured by the MTT assay showed more than a 3-fold increase in cell number within the scaffolds after 15 days of culture for both pure collagen scaffolds and Col/mHA composite scaffolds. Attractive properties of this composite scaffold include the potential to load the microspheres for drug delivery and the controllability of the pore structure at various length scales.

  9. Biocompatibility and osteogenesis of biomimetic nano-hydroxyapatite/polyamide composite scaffolds for bone tissue engineering.

    PubMed

    Wang, Huanan; Li, Yubao; Zuo, Yi; Li, Jihua; Ma, Sansi; Cheng, Lin

    2007-08-01

    In this study, we prepared nano-hydroxyapatite/polyamide (n-HA/PA) composite scaffolds utilizing thermally induced phase inversion processing technique. The macrostructure and morphology as well as mechanical strength of the scaffolds were characterized. Mesenchymal stem cells (MSCs) derived from bone marrow of neonatal rabbits were cultured, expanded and seeded on n-HA/PA scaffolds. The MSC/scaffold constructs were cultured for up to 7 days and the adhesion, proliferation and differentiation of MSCs into osteoblastic phenotype were determined using MTT assay, alkaline phosphatase (ALP) activity and collagen type I (COL I) immunohistochemical staining and scanning electronic microscopy (SEM). The results confirm that n-HA/PA scaffolds are biocompatible and have no negative effects on the MSCs in vitro. To investigate the in vivo biocompatibility and osteogenesis of the composite scaffolds, both pure n-HA/PA scaffolds and MSC/scaffold constructs were implanted in rabbit mandibles and studied histologically and microradiographically. The results show that n-HA/PA composite scaffolds exhibit good biocompatibility and extensive osteoconductivity with host bone. Moreover, the introduction of MSCs to the scaffolds dramatically enhanced the efficiency of new bone formation, especially at the initial stage after implantation. In long term (more than 12 weeks implantation), however, the pure scaffolds show as good biocompatibility and osteogenesis as the hybrid ones. All these results indicate that the scaffolds fulfill the basic requirements of bone tissue engineering scaffold, and have the potential to be applied in orthopedic, reconstructive and maxillofacial surgery.

  10. Novel hydroxyapatite/carboxymethylchitosan composite scaffolds prepared through an innovative "autocatalytic" electroless coprecipitation route.

    PubMed

    Oliveira, J M; Costa, S A; Leonor, I B; Malafaya, P B; Mano, J F; Reis, R L

    2009-02-01

    A developmental composite scaffold for bone tissue engineering applications composed of hydroxyapatite (HA) and carboxymethylchitosan (CMC) was obtained using a coprecipitation method, which is based on the "autocatalytic" electroless deposition route. The results revealed that the pores of the scaffold were regular, interconnected, and possess a size in the range of 20-500 microm. Furthermore, the Fourier transform infra-red spectrum of the composite scaffolds exhibited all the characteristic peaks of apatite, and the appearance of typical bands from CMC, thus showing that coprecipitation of both organic and inorganic phases was effective. The X-ray diffraction pattern of composite scaffolds demonstrated that calcium-phosphates consisted of crystalline HA. From microcomputed tomography analysis, it was possible to determine that composite scaffolds possess a 58.9% +/- 6% of porosity. The 2D morphometric analysis demonstrated that on average the scaffolds consisted of 24% HA and 76% CMC. The mechanical properties were assessed using compressive tests, both in dry and wet states. Additionally, in vitro tests were carried out to evaluate the water-uptake capability, weight loss, and bioactive behavior of the composite scaffolds. The novel hydroxyapatite/carboxymethylchitosan composite scaffolds showed promise whenever degradability and bioactivity are simultaneously desired, as in the case of bone tissue-engineering scaffolding applications.

  11. Direct scaffolding of biomimetic hydroxyapatite-gelatin nanocomposites using aminosilane cross-linker for bone regeneration.

    PubMed

    Chiu, Chi-Kai; Ferreira, Joao; Luo, Tzy-Jiun M; Geng, Haixia; Lin, Feng-Chang; Ko, Ching-Chang

    2012-09-01

    Hydroxyapatite-gelatin modified siloxane (GEMOSIL) nanocomposite was developed by coating, kneading and hardening processes to provide formable scaffolding for alloplastic graft applications. The present study aims to characterize scaffolding formability and mechanical properties of GEMOSIL, and to test the in vitro and in vivo biocompatibility of GEMOSIL. Buffer Solution initiated formable paste followed by the sol-gel reaction led to a final hardened composite. Results showed the adequate coating of aminosilane, 11-19 wt%, affected the cohesiveness of the powders and the final compressive strength (69 MPa) of the composite. TGA and TEM results showed the effective aminosilane coating that preserves hydroxyapatite-gelatin nanocrystals from damage. Both GEMOSIL with and without titania increased the mineralization of preosteoblasts in vitro. Only did titania additives revealed good in vivo bone formation in rat calvarium defects. The scaffolding formability, due to cohesive bonding among GEMOSIL particles, could be further refined to fulfill the complicated scaffold processes.

  12. Direct Scaffolding of Biomimetic Hydroxyapatite-gelatin Nanocomposites using Aminosilane Cross-linker for Bone Regeneration

    PubMed Central

    Chiu, Chi-Kai; Ferreira, Joao; Luo, Tzy-Jiun M.; Geng, Haixia; Lin, Feng-Chang; Ko, Ching-Chang

    2012-01-01

    Hydroxyapatite-gelatin modified siloxane (GEMOSIL) nanocomposite was developed by coating, kneading and hardening processes to provide formable scaffolding for alloplastic graft applications. The present study aims to characterize scaffolding formability and mechanical properties of GEMOSIL, and to test the in vitro and in vivo biocompatibility of GEMOSIL. Buffer Solution initiated formable paste followed by the sol-gel reaction led to a final hardened composite. Results showed the adequate coating of aminosilane, 11–19 wt%, affected the cohesiveness of the powders and the final compressive strength (69 MPa) of the composite. TGA and TEM results showed the effective aminosilane coating that preserves hydroxyapatite-gelatin nanocrystals from damage. Both GEMOSIL with and without titania increased the mineralization of preosteoblasts in vitro. Only did titania additives revealed good in vivo bone formation in rat calvarium defects. The scaffolding formability, due to cohesive bonding among GEMOSIL particles, could be further refined to fulfill the complicated scaffold processes. PMID:22669282

  13. Chitosan-amylopectin/hydroxyapatite and chitosan-chondroitin sulphate/hydroxyapatite composite scaffolds for bone tissue engineering.

    PubMed

    Venkatesan, Jayachandran; Pallela, Ramjee; Bhatnagar, Ira; Kim, Se-Kwon

    2012-12-01

    Over the past few decades, artificial graft materials for bone tissue engineering are gaining much importance. In this study, tri-component scaffolds of chitosan/natural hydroxyapatite with chondroitin sulfate (chitosan-CS/HAp) and amylopectin (chitosan-AP/HAp) have been developed for the first time via freeze-drying method and were characterized physicochemically for bone grafting substitutes. Chemical interactions and dispersion of HAp, CS and AP in the chitosan matrix have been evaluated by various analytical techniques. The porosity and water uptake/retention ability of these composite scaffolds decreased whereas thermal stability increased when compared to the chitosan scaffold. The pore size of the chitosan/HAp, chitosan-CS/HAp and chitosan-AP/HAp scaffolds varied from 60 to 180 μm, 60 to 400 μm and 80 to 500 μm, respectively. Cell proliferation, alkaline phosphatase activity and type-1 collagen production was evaluated in vitro using MG-63 cell line, which was observed to be higher in the composite scaffolds. Excellent interconnected porosity, controlled biodegradation and enhanced cell proliferation of the novel chitosan-CS/HAp and chitosan-AP/HAp scaffolds suggests that these scaffolds are promising biomaterials for bone tissue engineering. Copyright © 2012 Elsevier B.V. All rights reserved.

  14. Fabrication of hydroxyapatite from fish bones waste using reflux method

    NASA Astrophysics Data System (ADS)

    Cahyanto, A.; Kosasih, E.; Aripin, D.; Hasratiningsih, Z.

    2017-02-01

    The aim of this present study was to investigate the fabrication of hydroxyapatites, which were synthesized from fish bone wastes using reflux method. The fish bone wastes collected from the restaurant were brushed and boiled at 100°C for 10 minutes to remove debris and fat. After drying, the fish bones were crushed, and ball milled into a fine powder. The fish bone wastes were then processed by refluxing using KOH and H3PO4 solutions. The samples were calcined at 900°C and characterized by X-Ray Diffraction (XRD) and Fourier Transform Infrared Spectrometry (FT-IR). The XRD pattern of samples after treatment revealed that the peak of hydroxyapatite was observed and the bands of OH- and PO4 3- were observed by FT-IR. The scanning electron microscope evaluation of sample showed the entangled crystal and porous structure of hydroxyapatite. In conclusion, the hydroxyapatite was successfully synthesized from fish bone wastes using reflux method.

  15. Bone regeneration of hydroxyapatite/alumina bilayered scaffold with 3 mm passage-like medullary canal in canine tibia model.

    PubMed

    Kim, Jong Min; Son, Jun Sik; Kang, Seong Soo; Kim, Gonhyung; Choi, Seok Hwa

    2015-01-01

    The aim of this study was to evaluate the bone regeneration of hydroxyapatite (HA)/alumina bilayered scaffold with a 3 mm passage-like medullary canal in a beagle tibia model. A porous HA/alumina scaffold was fabricated using a polymeric template-coating technique. HA/alumina scaffold dimensions were 10 mm in outer diameter, 20 mm in length, and with either a 3 mm passage or no passage. A 20 mm segmental defect was induced using an oscillating saw through the diaphysis of the beagle tibia. The defects of six beagles were filled with HA/alumina bilayered scaffolds with a 3 mm passage or without. The segmental defect was fixated using one bone plate and six screws. Bone regeneration within the HA/alumina scaffolds was observed at eight weeks after implantation. The evaluation of bone regeneration within the scaffolds after implantation in a beagle tibia was performed using radiography, computerized tomography (CT), micro-CT, and fluorescence microscopy. New bone successfully formed in the tibia defects treated with 3 mm passage HA/alumina scaffolds compared to without-passage HA/alumina scaffolds. It was concluded that the HA/alumina bilayered scaffold with 3 mm passage-like medullary canal was instrumental in inducing host-scaffold engraftment of the defect as well as distributing the newly formed bone throughout the scaffold at 8 weeks after implantation.

  16. Sol-gel method to fabricate CaP scaffolds by robocasting for tissue engineering.

    PubMed

    Houmard, Manuel; Fu, Qiang; Saiz, Eduardo; Tomsia, Antoni P

    2012-04-01

    Highly porous calcium phosphate (CaP) scaffolds for bone-tissue engineering were fabricated by combining a robocasting process with a sol-gel synthesis that mixed Calcium Nitrate Tetrahydrate and Triethyl Phosphite precursors in an aqueous medium. The resulting gels were used to print scaffolds by robocasting without the use of binder to increase the viscosity of the paste. X-ray diffraction analysis confirmed that the process yielded hydroxyapatite and β-tricalcium phosphate biphasic composite powders. Thus, the scaffold composition after crystallization of the amorphous structure could be easily modified by varying the initial Ca/P ratio during synthesis. The compressive strengths of the scaffolds are ~6 MPa, which is in the range of human cancellous bone (2-12 MPa). These highly porous scaffolds (~73 vol% porosity) are composed of macro-pores of ~260 μm in size; such porosity is expected to enable bone ingrowth into the scaffold for bone repair applications. The chemistry, porosity, and surface topography of such scaffolds can also be modified by the process parameters to favor bone formation. The studied sol-gel process can be used to coat these scaffolds by dip-coating, which induces a significant enhancement of mechanical properties. This can adjust scaffold properties such as composition and surface morphology, which consequently may improve their performances.

  17. Biofabrication and in vitro study of hydroxyapatite/mPEG-PCL-mPEG scaffolds for bone tissue engineering using air pressure-aided deposition technology.

    PubMed

    Jiang, Cho-Pei; Chen, Yo-Yu; Hsieh, Min-Fa

    2013-03-01

    The aims of this study were to fabricate biopolymer and biocomposite scaffolds for bone tissue engineering by an air pressure-aided deposition system and to carry out osteoblast cell culture tests to validate the biocompatibility of fabricated scaffolds. A mPEG-PCL-mPEG triblock copolymer was synthesized as a biopolymer material. Biocomposite material was composed of synthesized biopolymer and hydroxyapatite (HA) with a mean diameter of 100 μm. The weight ratio of HA added to the synthesized biopolymer was 0.1, 0.25, 0.5 and 1. The experimental results show that the maximum average compressive strength of biocomposite scaffolds, made of weight ratio 0.5, with mean pore size of 410 μm (porosity 81%) is 18.38 MPa which is two times stronger than that of biopolymer scaffolds. Osteoblast cells, MC3T3-E1, were seeded on both types of fabricated scaffolds to validate the biocompatibility using methylthianzol tetrazolium (MTT) assay and cell morphology observation. After 28 days of in vitro culturing, the seeded osteoblasts were well distributed in the interior of both types of scaffolds. Furthermore, MTT experimental results show that the cell viability of the biocomposite scaffold is higher than that of the biopolymer scaffold. This indicates that adding HA into synthesized biopolymer can enhance compressive strength and the proliferation of the osteoblast cell. Copyright © 2012 Elsevier B.V. All rights reserved.

  18. Fabrication of titania/hydroxyapatite composite granules for photo-catalyst

    SciTech Connect

    Ji Shidong Murakami, Setsuaki; Kamitakahara, Masanobu; Ioku, Koji

    2009-04-02

    The titania/hydroxyapatite composite granular photo-catalyst with novel microstructure was fabricated by the process based on the liquid immiscibility effect and followed by precalcination and hydrothermal treatment from commercially available powders of {alpha}-Tri-calcium phosphate and TiO{sub 2}. XRD, SEM, BET, optical microscopy and UV-vis spectrophotometer were applied to characterize the prepared photo-catalyst. Microstructure analysis indicated that the granule was weaved by rod-shaped hydroxyapatite crystals whose surface was covered by nano-sized TiO{sub 2}. In the composite granules, the active surface of anatase was retained effectively. With the hybridization of TiO{sub 2} and HAp, a 16-nm blue-shift of absorption edge could be observed and the crystallinity of anatase could be enhanced by precalcination. The granules with the rod-shaped hydroxyapatite crystals performing as scaffold work as three-dimensional high porous, size-controllable small reactor. The phase and microstructure transformation of the granule before and after hydrothermal treatment was investigated and its decomposition ability was evaluated by using Methylene blue as a target pollutant compound.

  19. Preparation and characterization of nano-sized hydroxyapatite/alginate/chitosan composite scaffolds for bone tissue engineering.

    PubMed

    Kim, Hye-Lee; Jung, Gil-Yong; Yoon, Jun-Ho; Han, Jung-Suk; Park, Yoon-Jeong; Kim, Do-Gyoon; Zhang, Miqin; Kim, Dae-Joon

    2015-09-01

    The aim of this study was to develop chitosan composite scaffolds with high strength and controlled pore structures by homogenously dispersed nano-sized hydroxyapatite (nano-HAp) powders. In the fabrication of composite scaffolds, nano-HAp powders distributed in an alginate (AG) solution with a pH higher than 10 were mixed with a chitosan (CS) solution and then freeze dried. While the HAp content increased up to 70 wt.%, the compressive strength and the elastic modulus of the composite scaffolds significantly increased from 0.27 MPa and 4.42 MPa to 0.68 MPa and 13.35 MPa, respectively. Higher content of the HAp also helped develop more differentiation and mineralization of the MC3T3-E1 cells on the composite scaffolds. The uniform pore structure and the excellent mechanical properties of the HAp/CS composite scaffolds likely resulted from the use of the AG solution at pH 10 as a dispersant for the nano-HAp powders.

  20. Fabrication of 13-93 bioactive glass scaffolds for bone tissue engineering using indirect selective laser sintering.

    PubMed

    Kolan, Krishna C R; Leu, Ming C; Hilmas, Gregory E; Brown, Roger F; Velez, Mariano

    2011-06-01

    Bioactive glasses are promising materials for bone scaffolds due to their ability to assist in tissue regeneration. When implanted in vivo, bioactive glasses can convert into hydroxyapatite, the main mineral constituent of human bone, and form a strong bond with the surrounding tissues, thus providing an advantage over polymer scaffold materials. Bone scaffold fabrication using additive manufacturing techniques can provide control over pore interconnectivity during fabrication of the scaffold, which helps in mimicking human trabecular bone. 13-93 glass, a third-generation bioactive material designed to accelerate the body's natural ability to heal itself, was used in the research described herein to fabricate bone scaffolds using the selective laser sintering (SLS) process. 13-93 glass mixed with stearic acid (as the polymer binder) by ball milling was used as the powder feedstock for the SLS machine. The fabricated green scaffolds underwent binder burnout to remove the stearic acid binder and were then sintered at temperatures between 675 °C and 695 °C. The sintered scaffolds had pore sizes ranging from 300 to 800 µm with 50% apparent porosity and an average compressive strength of 20.4 MPa, which is excellent for non-load bearing applications and among the highest reported for an interconnected porous scaffold fabricated with bioactive glasses using the SLS process. The MTT labeling experiment and measurements of MTT formazan formation are evidence that the rough surface of SLS scaffolds provides a cell-friendly surface capable of supporting robust cell growth.

  1. Biomimetic Collagen–Hydroxyapatite Composite Fabricated via a Novel Perfusion-Flow Mineralization Technique

    PubMed Central

    Antebi, Ben; Cheng, Xingguo; Harris, Jeffrey N.; Gower, Laurie B.; Chen, Xiao-Dong

    2013-01-01

    Prevalent three-dimensional scaffolds for bone tissue engineering are mineralized collagen–hydroxyapatite (Col/HA) composites. Conventional mineralization techniques are either to coat collagen scaffold surfaces with minerals or to simply mix collagen and mineral nanoparticles together. These conventional in vitro collagen mineralization methods are different from the in vivo bone formation process and often result in scaffolds that are not suitable for bone tissue engineering. In this study, a unique perfusion-flow (i.e., dynamic) in conjunction with a previously described polymer-induced liquid-precursor (PILP) method was used to fabricate a porous Col/HA composite. The dynamic flow emulated the physiological extracellular fluid flow containing the mineralization ions, while the PILP method facilitated the deposition of the HA crystals within the collagen fibrils (i.e., intrafibrillar mineralization). By utilizing a dynamic PILP technique to mimic the in vivo bone formation process, the resultant Col/HA composite has a similar structure and compositions like human trabecular bone. A comparison of the dynamic and static mineralization methods revealed that the novel dynamic technique facilitates more efficient and homogenous mineral deposition throughout the Col/HA composite. The dynamic intrafibrillar mineralization method generated stiff Col/HA composites with excellent surface property for cell attachment and growth. The human mesenchymal stem cells cultured on the Col/HA composites quickly remodeled the scaffolds and resulted in constructs with an extensive cell-derived extracellular matrix network. PMID:23157544

  2. A novel nano-hydroxyapatite - PMMA hybrid scaffolds adopted by conjugated thermal induced phase separation (TIPS) and wet-chemical approach: Analysis of its mechanical and biological properties.

    PubMed

    G, Radha; S, Balakumar; Venkatesan, Balaji; Vellaichamy, Elangovan

    2017-04-01

    In this study, we report the preparation of nano-hydroxyapatite (nHAp) incorporated poly(methyl methacrylate) (PMMA) scaffolds by conjugated thermal induced phase separation (TIPS) and wet-chemical approach, which essentially facilitates the enhancement of both mechanical as well as biological properties of the scaffolds. The dissolution of PMMA was accomplished by acetone (Ace scaffold), ethanol-water (E-W scaffold) and isopropanol-water (I-W scaffold) mixtures as solvents. The existence of nHAp in PMMA matrix was investigated systematically. The porosity of ~57.89% was achieved from Ace scaffold that was higher degree compared to both I-W and E-W scaffolds. On the other hand, the dense porous architecture of I-W scaffold exhibited superior hardness of ~65.6 HR 'D' than that of the Ace and E-W scaffolds. All the fabricated samples demonstrated enhanced in vitro bioactivity with respect to increasing immersion period as a result of flower-like in vitro apatite layer formation. The MTT assay was carried out for 1day and 3day culture using Saos-2 osteoblast-like cells, which showed better cell proliferation with increasing culture period owing to the interconnected pore architecture of scaffolds and the rational hemocompatibility as per the ASTM standard F756-00.

  3. PCL-coated hydroxyapatite scaffold derived from cuttlefish bone: morphology, mechanical properties and bioactivity.

    PubMed

    Milovac, Dajana; Gallego Ferrer, Gloria; Ivankovic, Marica; Ivankovic, Hrvoje

    2014-01-01

    In the present study, poly(ε-caprolactone)-coated hydroxyapatite scaffold derived from cuttlefish bone was prepared. Hydrothermal transformation of aragonitic cuttlefish bone into hydroxyapatite (HAp) was performed at 200°C retaining the cuttlebone architecture. The HAp scaffold was coated with a poly(ε-caprolactone) (PCL) using vacuum impregnation technique. The compositional and morphological properties of HAp and PCL-coated HAp scaffolds were studied by means of X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA) and scanning electron microscopy (SEM) with energy dispersive X-ray (EDX) analysis. Bioactivity was tested by immersion in Hank's balanced salt solution (HBSS) and mechanical tests were performed at compression. The results showed that PCL-coated HAp (HAp/PCL) scaffold resulted in a material with improved mechanical properties that keep the original interconnected porous structure indispensable for tissue growth and vascularization. The compressive strength (0.88MPa) and the elastic modulus (15.5MPa) are within the lower range of properties reported for human trabecular bones. The in vitro mineralization of calcium phosphate (CP) that produces the bone-like apatite was observed on both the pure HAp scaffold and the HAp/PCL composite scaffold. The prepared bioactive scaffold with enhanced mechanical properties is a good candidate for bone tissue engineering applications. © 2013.

  4. Controlled nucleation of hydroxyapatite on alginate scaffolds for stem cell-based bone tissue engineering.

    PubMed

    Suárez-González, Darilis; Barnhart, Kara; Saito, Eiji; Vanderby, Ray; Hollister, Scott J; Murphy, William L

    2010-10-01

    Current bone tissue engineering strategies aim to grow a tissue similar to native bone by combining cells and biologically active molecules with a scaffold material. In this study, a macroporous scaffold made from the seaweed-derived polymer alginate was synthesized and mineralized for cell-based bone tissue engineering applications. Nucleation of a bone-like hydroxyapatite mineral was achieved by incubating the scaffold in modified simulated body fluids (mSBF) for 4 weeks. Analysis using scanning electron microscopy and energy dispersive x-ray analysis indicated growth of a continuous layer of mineral primarily composed of calcium and phosphorous. X-ray diffraction analysis showed peaks associated with hydroxyapatite, the major inorganic constituent of human bone tissue. In addition to the mineral characterization, the ability to control nucleation on the surface, into the bulk of the material, or on the inner pore surfaces of scaffolds was demonstrated. Finally, human MSCs attached and proliferated on the mineralized scaffolds and cell attachment improved when seeding cells on mineral coated alginate scaffolds. This novel alginate- HAP composite material could be used in bone tissue engineering as a scaffold material to deliver cells, and perhaps also biologically active molecules.

  5. Osteogenic differentiation of equine cord blood multipotent mesenchymal stromal cells within coralline hydroxyapatite scaffolds in vitro.

    PubMed

    Figueroa, R J; Koch, T G; Betts, D H

    2011-01-01

    To investigate the osteogenic differentiation potential of equine umbilical cord blood-derived multipotent mesenchymal stromal cells (CB-MSC) within coralline hydroxyapatite scaffolds cultured in osteogenic induction culture medium. Scaffolds seeded with equine CB-MSC were cultured in cell expansion culture medium (control) or osteogenic induction medium (treatment). Cell viability and distribution were confirmed by the MTT cell viability assay and DAPI nuclear fluorescence staining, respectively. Osteogenic differentiation was evaluated after 10 days using reverse transcription polymerase chain reaction, alkaline phosphatase activity, and secreted osteocalcin concentration. Cell morphology and matrix deposition were assessed by scanning electron microscopy (SEM) after 14 days in culture. Cells showed viability and adequate distribution within the scaffold. Successful osteogenic differentiation within the scaffolds was demonstrated by the increased expression of osteogenic markers such as Runx2, osteopontin, osteonectin, collagen IA; increased levels of alkaline phosphatase activity; increased osteocalcin protein secretion and bone-like matrix presence in the scaffold pores upon SEM evaluation. These results demonstrate that equine CB-MSC maintain viability and exhibit osteogenic potential in coralline hydroxyapatite scaffolds when induced in vitro . Equine CB-MSC scaffold constructs deserve further investigation for their potential role as biologically active fillers to enhance bone-gap repair in the horse.

  6. Vaporizable Scaffolds for Fabricating Thermoelectric Modules

    NASA Technical Reports Server (NTRS)

    Sakamoto, Jeffrey; Yen, Shiao-pin; Fleurial, Jean-Pierre; Paik, Jong-Ah

    2006-01-01

    A process for fabricating thermoelectric modules with vacuum gaps separating the thermoelectric legs has been conceived, and the feasibility of some essential parts of the process has been demonstrated. The vacuum gaps are needed to electrically insulate the legs from each other. The process involves the use of scaffolding in the form of sheets of a polymer to temporarily separate the legs by the desired distance, which is typically about 0.5 mm. During a bonding subprocess that would take place in a partial vacuum at an elevated temperature, the polymer would be vaporized, thereby creating the vacuum gaps.

  7. Bone response to 3-D periodic hydroxyapatite scaffolds with and without tailored microporosity to deliver bone morphogenetic protein 2.

    SciTech Connect

    Eurell, Jo Ann; Dellinger, Jennifer Gwynne; Cesarano, Joseph, III; Jamison, Russell D.

    2005-06-01

    The in vivo bone response of 3D periodic hydroxyapatite (HA) scaffolds is investigated. Two groups of HA scaffolds (11 mm diameter x 3.5 mm thick) are fabricated by direct-write assembly of a concentrated HA ink. The scaffolds consist of cylindrical rods periodically arranged into four quadrants with varying separation distances between rods. In the first group, HA rods (250 {micro}m in diameter) are patterned to create pore channels, whose areal dimensions are 250 x 250 {micro}m{sup 2} in quadrant 1, 250 x 500 {micro}m{sup 2} in quadrants 2 and 4, and 500 x 500 {micro}m{sup 2} in quadrant 3. In the second group, HA rods (400 {micro}m in diameter) are patterned to create pore channels, whose areal dimensions of 500 x 500 {micro}m{sup 2} in quadrant 1, 500 x 750 {micro}m{sup 2} in quadrants 2 and 4, and 750 x 750 {micro}m{sup 2} in quadrant 3. Each group of scaffolds is partially densified by sintering at 1200 C prior to being implanted bilaterally in trephine defects of skeletally mature New Zealand White rabbits. Their tissue response is evaluated at 8 and 16 weeks using micro-computed tomography, histology, and scanning electron microscopy. New trabecular bone is conducted rapidly and efficiently across substantial distances within these patterned 3D HA scaffolds. Our observations suggest that HA rods are first coated with a layer of new bone followed by subsequent scaffold infilling via outward and inward radial growth of the coated regions. Direct-write assembly of 3D periodic scaffolds composed of micro-porous HA rods arrayed to produce macro-pores that are size-matched to trabecular bone may represent an optimal strategy for bone repair and replacement structures.

  8. Precision Extruding Deposition for Freeform Fabrication of PCL and PCL-HA Tissue Scaffolds

    NASA Astrophysics Data System (ADS)

    Shor, L.; Yildirim, E. D.; Güçeri, S.; Sun, W.

    Computer-aided tissue engineering approach was used to develop a novel Precision Extrusion Deposition (PED) process to directly fabricate Polycaprolactone (PCL) and composite PCL/Hydroxyapatite (PCL-HA) tissue scaffolds. The process optimization was carried out to fabricate both PCL and PCL-HA (25% concentration by weight of HA) with a controlled pore size and internal pore structure of the 0°/90° pattern. Two groups of scaffolds having 60 and 70% porosity and with pore sizes of 450 and 750 microns, respectively, were evaluated for their morphology and compressive properties using Scanning Electron Microscopy (SEM) and mechanical testing. The surface modification with plasma was conducted on PCL scaffold to increase the cellular attachment and proliferation. Our results suggested that inclusion of HA significantly increased the compressive modulus from 59 to 84 MPa for 60% porous scaffolds and from 30 to 76 MPa for 70% porous scaffolds. In vitro cell-scaffolds interaction study was carried out using primary fetal bovine osteoblasts to assess the feasibility of scaffolds for bone tissue engineering application. In addition, the results in surface hydrophilicity and roughness show that plasma surface modification can increase the hydrophilicity while introducing the nano-scale surface roughness on PCL surface. The cell proliferation and differentiation were calculated by Alamar Blue assay and by determining alkaline phosphatase activity. The osteoblasts were able to migrate and proliferate over the cultured time for both PCL as well as PCL-HA scaffolds. Our study demonstrated the viability of the PED process to the fabricate PCL and PCL-HA composite scaffolds having necessary mechanical property, structural integrity, controlled pore size and pore interconnectivity desired for bone tissue engineering.

  9. Biologically inspired growth of hydroxyapatite crystals on bio-organics-defined scaffolds

    SciTech Connect

    Yang, Chunrong; Li, Yuli; Nan, Kaihui

    2013-03-15

    Graphical abstract: Petal-like crystals were observed to form on the surface of the BG/COL/ChS scaffolds. Highlights: ► Porous scaffolds were prepared using bioglass, collagen and chondroitin sulfate. ► Highly oriented HA crystals were grown on scaffolds using simulated body fluids ► The microstructure and orientation of HA were explained by molecular configuration. - Abstract: Several bio-organics-defined composite scaffolds were prepared using 58s-bioglass (BG), collagen (Col) and chondroitin sulfate (ChS). These scaffolds possess highly porous structure. X-ray diffraction of these scaffolds strongly indicated that hydroxyapatite (HA) crystals formed on their surfaces in simulated body fluids within 3 d, and similar formation process of crystals could be obtained on BG/Col and BG/Col/ChS scaffolds. The morphology and structure of the crystals were further examined by scanning electron microscopy. The results obtained indicate that an apatite with petal-like structure similar to that found on BG/Col scaffolds can be produced on BG/Col/ChS scaffolds through biomimetic synthesis, while that on BG/ChS scaffolds took place differently. The differences could be explained by self-assembly processes and the different macromolecular configurations of the Col and ChS fibrils which self-assemble spontaneously into their fibers. On the other hand, the bio-organics-defined composites have good cell biocompability. The results may be applicable to develop tailored biomaterials for peculiar bone substitute.

  10. Mechanical and cytotoxicity evaluation of nanostructured hydroxyapatite-bredigite scaffolds for bone regeneration.

    PubMed

    Eilbagi, Marjan; Emadi, Rahmatollah; Raeissi, Keyvan; Kharaziha, Mahshid; Valiani, Ali

    2016-11-01

    Despite the attractive characteristics of three-dimensional pure hydroxyapatite (HA) scaffolds, due to their weak mechanical properties, researches have focused on the development of composite scaffolds via introducing suitable secondary components. The aim of this study was to develop, for the first time, three-dimensional HA-bredigite (Ca7MgSi4O16) scaffolds containing various amounts of bredigite nanopowder (0, 5, 10 and 15wt.%) using space holder technique. Transmission electron microscopy, scanning electron microscopy, energy-dispersive X-ray spectroscopy and X-ray diffraction spectroscopy were applied in order to study the morphology, fracture surface and phase compositions of nanopowders and scaffolds. Furthermore, the effects of scaffold composition on the mechanical properties, bioactivity, biodegradability, and cytotoxicity were also evaluated. Results showed that the composite scaffolds with average pore size in the range of 220-310μm, appearance porosity of 63.1-75.9% and appearance density of 1.1±0.04g/cm(3) were successfully developed, depending on bredigite content. Indeed, the micropore size of the scaffolds reduced with increasing bredigite content confirming that the sinterability of the scaffolds was improved. Furthermore, the compression strength and modulus of the scaffolds significantly enhanced via incorporation of bredigite content from 0 to 15wt.%. The composite scaffolds revealed superior bioactivity and biodegradability with increasing bredigite content. Moreover, MTT assay confirmed that HA-15wt.% bredigite scaffold significantly promoted cell proliferation compared to tissue culture plate (control) and HA scaffold. Based on these results, three-dimensional HA-bredigite scaffolds could be promising replacements for HA scaffolds in bone regeneration.

  11. The efficacy of polycaprolactone/hydroxyapatite scaffold in combination with mesenchymal stem cells for bone tissue engineering.

    PubMed

    Chuenjitkuntaworn, Boontharika; Osathanon, Thanaphum; Nowwarote, Nunthawan; Supaphol, Pitt; Pavasant, Prasit

    2016-01-01

    Major drawbacks of using an autograft are the possibilities of insufficient bony source and patient's morbidity after operation. Bone tissue engineering technology, therefore, has been applied for repairing bony defects. Previous study showed that a novel fabricated 3D-Polycaprolactone/Hydroxyapatite (PCL/HAp) scaffold possessed a good biocompatibility for bone cells. This study aimed to determine the ability of PCL/HAp for supporting cell growth, gene expression, and osteogenic differentiation in three types of mesenchymal stem cells, including bone marrow-derived mesenchymal stem cells (BMSCs), dental pulp stem cells (DPSCs), and adiposed-derived mesenchymal stem cells (ADSCs). These were assessed by cell viability assay (MTT), reverse-transcription polymerase chain reaction (RT-PCR) analysis, alkaline phosphatase activity, and osteogenic differentiation by alizarin red-S staining. The results showed that PCL/HAp scaffold could support growth of all three types of mesenchymal stem cells. In addition, DPSCs with PCL/HAp showed the highest level of calcium deposition compared to other groups. In conclusion, DPSCs exhibited a better compatibility with these scaffolds compared to BMSCs and ADSCs. However, the PCL/HAp could be a good candidate scaffold for all tested mesenchymal stem cells in bone tissue engineering. © 2015 Wiley Periodicals, Inc.

  12. Engineered osteochondral grafts using biphasic composite solid free-form fabricated scaffolds.

    PubMed

    Schek, Rachel M; Taboas, Juan M; Segvich, Sharon J; Hollister, Scott J; Krebsbach, Paul H

    2004-01-01

    Tissue engineering has provided an alternative to traditional strategies to repair cartilage damaged by injury or degenerative disease. A successful strategy to engineer osteochondral tissue will mimic the natural contour of the articulating surface, achieve native mechanical properties and functional load-bearing ability, and lead to integration with host cartilage and underlying subchondral bone. Image-based design (IBD) and solid free-form (SFF) fabrication can be used to generate scaffolds that are load bearing and match articular geometry. The objective of this study was to utilize materials and biological factors in an integrated approach to regenerate a multitissue interface. Biphasic composite scaffolds manufactured by IBD and SFF fabrication were used to simultaneously generate bone and cartilage in discrete regions and provide for the development of a stable interface between cartilage and subchondral bone. Poly-L-lactic acid/hydroxyapatite composite scaffolds were differentially seeded with fibroblasts transduced with an adenovirus expressing bone morphogenetic protein 7 (BMP-7) in the ceramic phase and fully differentiated chondrocytes in the polymeric phase. After subcutaneous implantation into mice, the biphasic scaffolds promoted the simultaneous growth of bone, cartilage, and a mineralized interface tissue. Within the ceramic phase, the pockets of tissue generated included blood vessels, marrow stroma, and adipose tissue. This combination of IBD and SFF-fabricated biphasic scaffolds with gene and cell therapy is a promising approach to regenerate osteochondral defects.

  13. Conversion of borate-based glass scaffold to hydroxyapatite in a dilute phosphate solution.

    PubMed

    Liu, Xin; Pan, Haobo; Fu, Hailuo; Fu, Qiang; Rahaman, Mohamed N; Huang, Wenhai

    2010-02-01

    Porous scaffolds of a borate-based glass (composition in mol%: 6Na2O, 8K2O, 8MgO, 22CaO, 36B2O3, 18SiO2, 2P2O5), with interconnected porosity of approximately 70% and pores of size 200-500 microm, were prepared by a polymer foam replication technique. The degradation of the scaffolds and conversion to a hydroxyapatite-type material in a 0.02 M K2HPO4 solution (starting pH = 7.0) at 37 degrees C were studied by measuring the weight loss of the scaffolds, as well as the pH and the boron concentration of the solution. X-ray diffraction, scanning electronic microscopy and energy dispersive x-ray analysis showed that a hydroxyapatite-type material was formed on the glass surface within 7 days of immersion in the phosphate solution. Cellular response to the scaffolds was assessed using murine MLO-A5 cells, an osteogenic cell line. Scanning electron microscopy showed that the scaffolds supported cell attachment and proliferation during the 6 day incubation. The results indicate that this borate-based glass could provide a promising degradable scaffold material for bone tissue engineering applications.

  14. Three-dimensional printed PCL-hydroxyapatite scaffolds filled with CNTs for bone cell growth stimulation.

    PubMed

    Gonçalves, Elsa M; Oliveira, Filipe J; Silva, Rui F; Neto, Miguel A; Fernandes, M Helena; Amaral, Margarida; Vallet-Regí, María; Vila, Mercedes

    2016-08-01

    A three-phase [nanocrystalline hydroxyapatite (HA), carbon nanotubes (CNT), mixed in a polymeric matrix of polycaprolactone (PCL)] composite scaffold produced by 3D printing is presented. The CNT content varied between 0 and 10 wt % in a 50 wt % PCL matrix, with HA being the balance. With the combination of three well-known materials, these scaffolds aimed at bringing together the properties of all into a unique material to be used in tissue engineering as support for cell growth. The 3D printing technique allows producing composite scaffolds having an interconnected network of square pores in the range of 450-700 μm. The 2 wt % CNT scaffold offers the best combination of mechanical behaviour and electrical conductivity. Its compressive strength of ∼4 MPa is compatible with the trabecular bone. The composites show typical hydroxyapatite bioactivity, good cell adhesion and spreading at the scaffolds surface, this combination of properties indicating that the produced 3D, three-phase, scaffolds are promising materials in the field of bone regenerative medicine. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1210-1219, 2016. © 2015 Wiley Periodicals, Inc.

  15. In-situ hardening hydroxyapatite-based scaffold for bone repair.

    PubMed

    Zhang, Yu; Xu, Hockin H K; Takagi, Shozo; Chow, Laurence C

    2006-05-01

    Musculoskeletal conditions are becoming a major health concern because of an aging population and sports- and traffic-related injuries. While sintered hydroxyapatite implants require machining, calcium phosphate cement (CPC) bone repair material is moldable, self-hardens in situ, and has excellent osteoconductivity. In the present work, new approaches for developing strong and macroporous scaffolds of CPC were tested. Relationships were determined between scaffold porosity and strength, elastic modulus and fracture toughness. A biocompatible and biodegradable polymer (chitosan) and a water-soluble porogen (mannitol) were incorporated into CPC: Chitosan to make the material stronger, fast-setting and anti-washout; and mannitol to create macropores. Flexural strength, elastic modulus, and fracture toughness were measured as functions of mannitol mass fraction in CPC from 0% to 75%. After mannitol dissolution in a physiological solution, macropores were formed in CPC in the shapes of the original entrapped mannitol crystals, with diameters of 50 microm to 200 microm for cell infiltration and bone ingrowth. The resulting porosity in CPC ranged from 34.4% to 83.3% volume fraction. At 70.2% porosity, the hydroxyapatite scaffold possessed flexural strength (mean +/- sd; n = 6) of (2.5 +/- 0.2) MPa and elastic modulus of (0.71 +/- 0.10) GPa. These values were within the range for sintered porous hydroxyapatite and cancellous bone. Predictive equations were established by regression power-law fitting to the measured data (R(2) > 0.98) that described the relationships between scaffold porosity and strength, elastic modulus and fracture toughness. In conclusion, a new graft composition was developed that could be delivered during surgery in the form of a paste to harden in situ in the bone site to form macroporous hydroxyapatite. Compared to conventional CPC without macropores, the increased macroporosity of the new apatite scaffold may help facilitate implant fixation and

  16. PCL-coated hydroxyapatite scaffold derived from cuttlefish bone: in vitro cell culture studies.

    PubMed

    Milovac, Dajana; Gamboa-Martínez, Tatiana C; Ivankovic, Marica; Gallego Ferrer, Gloria; Ivankovic, Hrvoje

    2014-09-01

    In the present study, we examined the potential of using highly porous poly(ε-caprolactone) (PCL)-coated hydroxyapatite (HAp) scaffold derived from cuttlefish bone for bone tissue engineering applications. The cell culture studies were performed in vitro with preosteoblastic MC3T3-E1 cells in static culture conditions. Comparisons were made with uncoated HAp scaffold. The attachment and spreading of preosteoblasts on scaffolds were observed by Live/Dead staining Kit. The cells grown on the HAp/PCL composite scaffold exhibited greater spreading than cells grown on the HAp scaffold. DNA quantification and scanning electron microscopy (SEM) confirmed a good proliferation of cells on the scaffolds. DNA content on the HAp/PCL scaffold was significantly higher compared to porous HAp scaffolds. The amount of collagen synthesis was determined using a hydroxyproline assay. The osteoblastic differentiation of the cells was evaluated by determining alkaline phosphatase (ALP) activity and collagen type I secretion. Furthermore, cell spreading and cell proliferation within scaffolds were observed using a fluorescence microscope. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Fabrication of gelatin-strontium substituted calcium phosphate scaffolds with unidirectional pores for bone tissue engineering.

    PubMed

    Wu, Yu-Chun; Lin, Wei-Yu; Yang, Chyun-Yu; Lee, Tzer-Min

    2015-03-01

    This study fabricated homogeneous gelatin-strontium substituted calcium phosphate composites via coprecipitation in a gelatin solution. Unidirectional porous scaffolds with an oriented microtubular structure were then manufactured using freeze-drying technology. The resulting structure and pore alignment were determined using scanning electron microscopy. The pore size were in the range of 200-400 μm, which is considered ideal for the engineering of bone tissue. The scaffolds were further characterized using energy dispersive spectroscopy, Fourier transform infrared spectroscopy, and X-ray diffraction. Hydroxyapatite was the main calcium phosphate compound in the scaffolds, with strontium incorporated into the crystal structure. The porosity of the scaffolds decreased with increasing concentration of calcium-phosphate. The compressive strength in the longitudinal direction was two to threefold higher than that observed in the transverse direction. Our results demonstrate that the composite scaffolds degraded by approximately 20 % after 5 weeks. Additionally, in vitro results reveal that the addition of strontium significantly increased human osteoblastic cells proliferation. Scaffolds containing strontium with a Sr-CaP/(gelatin + Sr-CaP) ratio of 50 % provided the most suitable environment for cell proliferation, particularly under dynamic culture conditions. This study demonstrates the considerable potential of composite scaffolds composed of gelatin-strontium-substituted calcium phosphate for applications in bone tissue engineering.

  18. Improvement of dual-leached polycaprolactone porous scaffolds by incorporating with hydroxyapatite for bone tissue regeneration.

    PubMed

    Thadavirul, Napaphat; Pavasant, Prasit; Supaphol, Pitt

    2014-01-01

    Polycaprolactone (PCL)/hydroxyapatite (HA) composite scaffolds were prepared by combining solvent casting and salt particulate leaching with a polymer leaching technique. The hydrophilicity of the dual-leached scaffold was improved by alkaline (NaOH) treatment. Well-defined interconnected pores were detected by scanning electron microscopy. The water absorption capacity of the NaOH-treated PCL/HA dual-leached scaffold increased greatly, confirming that the hydrophilicity of the scaffold was improved by NaOH treatment. The compressive modulus of the PCL/HA dual-leached scaffold was greatly increased by the addition of HA particles. An indirect evaluation of the cytotoxicity of all PCL dual-leached scaffolds with mouse fibroblastic cells (L929) and mouse calvaria-derived pre-osteoblastic cells (MC3T3-E1) indicated that the PCL dual-leached scaffolds are non-toxic to cells. The ability of the scaffolds to support mouse calvaria-derived pre-osteoblastic cell (MC3T3-E1) attachment, proliferation, differentiation, and mineralization was also evaluated. Although the viability of cells was lower on the PCL/HA dual-leached scaffold than on the tissue-culture polystyrene plates (TCPS) and on the other substrates at early time points, both the PCL and NaOH-treated PCL/HA dual-leached scaffolds supported the attachment of MC3T3-E1 at significantly higher levels than TCPS. During the proliferation period (days 1-3), all of the PCL dual-leached scaffolds were able to support the proliferation of MC3T3-E1 at higher levels than the TCPS; in addition, the cells grown on NaOH-treated PCL/HA dual-leached scaffolds proliferated more rapidly. The cells cultured on the surfaces of NaOH-treated PCL/HA dual-leached scaffolds had the highest rate of mineral deposition.

  19. Osteogenic cell response to 3-D hydroxyapatite scaffolds developed via replication of natural marine sponges.

    PubMed

    Clarke, S A; Choi, S Y; McKechnie, Melanie; Burke, G; Dunne, N; Walker, G; Cunningham, E; Buchanan, F

    2016-02-01

    Bone tissue engineering may provide an alternative to autograft, however scaffold optimisation is required to maximize bone ingrowth. In designing scaffolds, pore architecture is important and there is evidence that cells prefer a degree of non-uniformity. The aim of this study was to compare scaffolds derived from a natural porous marine sponge (Spongia agaricina) with unique architecture to those derived from a synthetic polyurethane foam. Hydroxyapatite scaffolds of 1 cm(3) were prepared via ceramic infiltration of a marine sponge and a polyurethane (PU) foam. Human foetal osteoblasts (hFOB) were seeded at 1 × 10(5) cells/scaffold for up to 14 days. Cytotoxicity, cell number, morphology and differentiation were investigated. PU-derived scaffolds had 84-91% porosity and 99.99% pore interconnectivity. In comparison marine sponge-derived scaffolds had 56-61% porosity and 99.9% pore interconnectivity. hFOB studies showed that a greater number of cells were found on marine sponge-derived scaffolds at than on the PU scaffold but there was no significant difference in cell differentiation. X-ray diffraction and inductively coupled plasma mass spectrometry showed that Si ions were released from the marine-derived scaffold. In summary, three dimensional porous constructs have been manufactured that support cell attachment, proliferation and differentiation but significantly more cells were seen on marine-derived scaffolds. This could be due both to the chemistry and pore architecture of the scaffolds with an additional biological stimulus from presence of Si ions. Further in vivo tests in orthotopic models are required but this marine-derived scaffold shows promise for applications in bone tissue engineering.

  20. Improved mechanical properties of hydroxyapatite whisker-reinforced poly(L-lactic acid) scaffold by surface modification of hydroxyapatite.

    PubMed

    Fang, Zhou; Feng, Qingling

    2014-02-01

    To improve the mechanical properties of porous hydroxyapatite/poly(L-lactic acid) (HA/PLLA) composites, HA whiskers with high crystallinity and high aspect ratio were synthesized. HA whiskers were modified with γ-aminopropyltriethoxysilane (APTES) to improve the interface between HA whiskers and PLLA. The composite scaffold consists of a porous PLLA matrix with HA whiskers distributed homogeneously. The morphology and the distributions of pore sizes of PLLA scaffold was not influenced by introducing HA whiskers, while the mechanical properties were improved. Both the compressive strength and compressive modulus were increased with the weight ratio of APTES-modified HA whiskers up to 30 wt.%, but only up to 15 wt.% for non-modified HA whiskers. With more than 15 wt.% HA whiskers, the mechanical properties of HA/PLLA scaffold were better improved with APTES-modified HA whiskers than non-modified. The HA whisker/PLLA scaffold with high porosity and improved mechanical properties is attractive in the application of tissue engineering.

  1. 3D silicon doped hydroxyapatite scaffolds decorated with Elastin-like Recombinamers for bone regenerative medicine.

    PubMed

    Vila, Mercedes; García, Ana; Girotti, Alessandra; Alonso, Matilde; Rodríguez-Cabello, Jose Carlos; González-Vázquez, Arlyng; Planell, Josep A; Engel, Elisabeth; Buján, Julia; García-Honduvilla, Natalio; Vallet-Regí, María

    2016-11-01

    The current study reports on the manufacturing by rapid prototyping technique of three-dimensional (3D) scaffolds based on silicon substituted hydroxyapatite with Elastin-like Recombinamers (ELRs) functionalized surfaces. Silicon doped hydroxyapatite (Si-HA), with Ca10(PO4)5.7(SiO4)0.3(OH)1.7h0.3 nominal formula, was surface functionalized with two different types of polymers designed by genetic engineering: ELR-RGD that contain cell attachment specific sequences and ELR-SNA15/RGD with both hydroxyapatite and cells domains that interact with the inorganic phase and with the cells, respectively. These hybrid materials were subjected to in vitro assays in order to clarify if the ELRs coating improved the well-known biocompatible and bone regeneration properties of calcium phosphates materials. The in vitro tests showed that there was a total and homogeneous colonization of the 3D scaffolds by Bone marrow Mesenchymal Stromal Cells (BMSCs). In addition, the BMSCs were viable and able to proliferate and differentiate into osteoblasts. Bone tissue engineering is an area of increasing interest because its main applications are directly related to the rising life expectancy of the population, which promotes higher rates of several bone pathologies, so innovative strategies are needed for bone tissue regeneration therapies. Here we use the rapid prototyping technology to allow moulding ceramic 3D scaffolds and we use different bio-polymers for the functionalization of their surfaces in order to enhance the biological response. Combining the ceramic material (silicon doped hydroxyapatite, Si-HA) and the Elastin like Recombinamers (ELRs) polymers with the presence of the integrin-mediate adhesion domain alone or in combination with SNA15 peptide that possess high affinity for hydroxyapatite, provided an improved Bone marrow Mesenchymal Stromal Cells (BMSCs) differentiation into osteoblastic linkage. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights

  2. Assessment of protein entrapment in hydroxyapatite scaffolds by size exclusion chromatography.

    PubMed

    Espanol, Montserrat; Casals, Isidre; Lamtahri, Sarah; Valderas, Maria-Teresa; Ginebra, Maria-Pau

    2012-12-01

    Although it is well known that the textural properties of scaffolds play an important role in the process of tissue regeneration, the investigation of such effects remain difficult especially at the micro/nano level. Texture confers the material the additional ability to entrap/concentrate molecules circulating in the body fluid regardless of their binding affinity to the material. The goal of the present work is to isolate protein entrapment from protein adsorption phenomena in two macroporous hydroxyapatite scaffolds with identical chemical structure, similar macroporosity but different micro/nanoporosity using proteins of different sizes. This was achieved implementing size exclusion chromatography and using the scaffolds as chromatographic columns. The results showed that the larger the crystal size and the lower the packing density of the crystals composing the scaffold increased protein retention but decreased the protein dwelling time in the column. Differences in the amount of protein retained depended on the protein type.

  3. Biological evaluation of porous aliphatic polyurethane/hydroxyapatite composite scaffolds for bone tissue engineering.

    PubMed

    Yang, Wanxun; Both, Sanne K; Zuo, Yi; Birgani, Zeinab Tahmasebi; Habibovic, Pamela; Li, Yubao; Jansen, John A; Yang, Fang

    2015-07-01

    Biomaterial scaffolds meant to function as supporting structures to osteogenic cells play a pivotal role in bone tissue engineering. Recently, we synthesized an aliphatic polyurethane (PU) scaffold via a foaming method using non-toxic components. Through this procedure a uniform interconnected porous structure was created. Furthermore, hydroxyapatite (HA) particles were introduced into this process to increase the bioactivity of the PU matrix. To evaluate the biological performances of these PU-based scaffolds, their influence on in vitro cellular behavior and in vivo bone forming capacity of the engineered cell-scaffold constructs was investigated in this study. A simulated body fluid test demonstrated that the incorporation of 40 wt % HA particles significantly promoted the biomineralization ability of the PU scaffolds. Enhanced in vitro proliferation and osteogenic differentiation of the seeded mesenchymal stem cells were also observed on the PU/HA composite. Next, the cell-scaffold constructs were implanted subcutaneously in a nude mice model. After 8 weeks, a considerable amount of vascularized bone tissue with initial marrow stroma development was generated in both PU and PU/HA40 scaffold. In conclusion, the PU/HA composite is a potential scaffold for bone regeneration applications.

  4. The effects of hydroxyapatite/calcium phosphate glass scaffold and its surface modification with bovine serum albumin on 1-wall intrabony defects of beagle dogs: a preliminary study.

    PubMed

    Um, Yoo-Jung; Jung, Ui-Won; Chae, Gyung-Joon; Kim, Chang-Sung; Lee, Yong-Keun; Cho, Kyoo-Sung; Kim, Chong-Kwan; Choi, Seong-Ho

    2008-12-01

    The purpose of this study was to evaluate the effects of biphasic hydroxyapatite/calcium phosphate glass (HA/CPG) scaffold and its surface modification with bovine serum albumin (BSA) on periodontal regeneration. 1-wall intrabony defects were surgically created on five beagle dogs. HA/CPG scaffolds, with a hydroxyapatite (HA)/calcium phosphate glass (CPG) ratio of 95:5 by weight (%) and surface modification done by 2% bovine serum albumin, were used. The control group received surgical flap operation, and the experimental groups were filled with HA/CPG scaffolds and HA/CPG(BSA) scaffolds. The animals were sacrificed eight weeks after surgery. Histological findings revealed better space maintenance in the experimental groups than the control group, and showed new bone formation intermittently in between the residual material particles. The newly formed bone was mostly woven bone and the residual particles were undergoing resorption. Cementum regeneration was observed with limited root resorption in all the groups. Histometric analysis also revealed greater mean values in new bone formation, cementum regeneration and bone area than the control group in both experimental groups. However, similar findings were presented between HA/CPG and HA/CPG(BSA). The result of the present study revealed the newly fabricated HA/CPG scaffold to have a potential use as a bone substitute material.

  5. Coaxial electrospun aligned tussah silk fibroin nanostructured fiber scaffolds embedded with hydroxyapatite-tussah silk fibroin nanoparticles for bone tissue engineering.

    PubMed

    Shao, Weili; He, Jianxin; Sang, Feng; Ding, Bin; Chen, Li; Cui, Shizhong; Li, Kejing; Han, Qiming; Tan, Weilin

    2016-01-01

    The bone is a composite of inorganic and organic materials and possesses a complex hierarchical architecture consisting of mineralized fibrils formed by collagen molecules and coated with oriented hydroxyapatite. To regenerate bone tissue, it is necessary to provide a scaffold that mimics the architecture of the extracellular matrix in native bone. Here, we describe one such scaffold, a nanostructured composite with a core made of a composite of hydroxyapatite and tussah silk fibroin. The core is encased in a shell of tussah silk fibroin. The composite fibers were fabricated by coaxial electrospinning using green water solvent and were characterized using different techniques. In comparison to nanofibers of pure tussah silk, composite notably improved mechanical properties, with 90-fold and 2-fold higher initial modulus and breaking stress, respectively, obtained. Osteoblast-like MG-63 cells were cultivated on the composite to assess its suitability as a scaffold for bone tissue engineering. We found that the fiber scaffold supported cell adhesion and proliferation and functionally promoted alkaline phosphatase and mineral deposition relevant for biomineralization. In addition, the composite were more biocompatible than pure tussah silk fibroin or cover slip. Thus, the nanostructured composite has excellent biomimetic and mechanical properties and is a potential biocompatible scaffold for bone tissue engineering.

  6. Revitalization of open apex teeth with apical periodontitis using a collagen-hydroxyapatite scaffold.

    PubMed

    Nevins, Alan J; Cymerman, Jerome J

    2015-06-01

    An enhanced revision of the revitalization endodontic technique for immature teeth with apical periodontitis has been described. It includes the addition of collagen-hydroxyapatite scaffold to the currently practiced revascularization technique. Four cases treated in series are presented in this report, 1 case involving 2 teeth. Periapical diagnoses of immature teeth included "asymptomatic apical periodontitis," "symptomatic apical periodontitis," and "acute apical abscess." Additionally, 1 fully developed tooth that had undergone root canal treatment that failed had a periapical diagnosis of acute apical abscess. An established revascularization protocol was used for all teeth. In addition to stimulating blood clots, all teeth were filled with collagen-hydroxyapatite scaffolds. Periapical radiolucencies healed in all teeth, and diffuse radiopacity developed within the coronal portions of canal spaces. Root development with root lengthening occurred in the immature nonvital maxillary premolar that had not undergone prior treatment. The technique of adding a collagen-hydroxyapatite scaffold to the existing revitalization protocol has been described in which substantial hard tissue repair has occurred. This may leave teeth more fully developed and less likely to fracture.

  7. Indirect solid free form fabrication of local and global porous, biomimetic and composite 3D polymer-ceramic scaffolds.

    PubMed

    Taboas, J M; Maddox, R D; Krebsbach, P H; Hollister, S J

    2003-01-01

    Precise control over scaffold material, porosity, and internal pore architecture is essential for tissue engineering. By coupling solid free form (SFF) manufacturing with conventional sponge scaffold fabrication procedures, we have developed methods for casting scaffolds that contain designed and controlled locally porous and globally porous internal architectures. These methods are compatible with numerous bioresorbable and non-resorbable polymers, ceramics, and biologic materials. Phase separation, emulsion-solvent diffusion, and porogen leaching were used to create poly(L)lactide (PLA) scaffolds containing both computationally designed global pores (500, 600, or 800 microm wide channels) and solvent fashioned local pores (50-100 microm wide voids or 5-10 microm length plates). Globally porous PLA and polyglycolide/PLA discrete composites were made using melt processing. Biphasic scaffolds with mechanically interdigitated PLA and sintered hydroxyapatite regions were fabricated with 500 and 600 microm wide global pores. PLA scaffolds with complex internal architectures that mimicked human trabecular bone were produced. Our indirect fabrication using casting in SFF molds provided enhanced control over scaffold shape, material, porosity and pore architecture, including size, geometry, orientation, branching, and interconnectivity. These scaffolds that contain concurrent local and global pores, discrete material regions, and biomimetic internal architectures may prove valuable for multi-tissue and structural tissue interface engineering. Copyright 2002 Elsevier Science Ltd.

  8. Evaluation of chitosan-hydroxyapatite-collagen composite strength as scaffold material by immersion in simulated body fluid

    NASA Astrophysics Data System (ADS)

    Sari, N. K.; Indrani, D. J.; Johan, C.; Corputty, J. E. M.

    2017-08-01

    The reconstruction of bone tissue defects is a major challenge facing oral and maxillofacial surgeons. The essential elements needed for tissue engineering are cells, scaffolds (matrix), and stimulant molecules (growth factors). The mechanical properties of chitosan-hydroxyapatite-collagen scaffolds produced by BATAN, Jakarta, have not yet been studied. This study therefore analyzed the mechanical properties of chitosan-hydroxyapatite-collagen composite scaffolds prepared by BATAN, Jakarta, before and after immersion in simulated body fluid (SBF) for eight days. The compressive and tensile strengths of the chitosan-hydroxyapatite-collagen composite scaffolds were analyzed after immersion in SBF at 37°C for eight days. Each scaffold was removed and dried at room temperature on days 0, 2, 4, 6, and 8. The data obtained were processed and analyzed. Variations in the compressive strength and tensile strength were attributed to several aspects, such the specimen size, which was not uniform, the scaffold composition, scaffold pore size, which was also not uniform, and the degradation of the polymer. The chitosan-hydroxyapatite-collagen composite scaffold does not exhibit differences in the tensile strength and compressive strength before and after immersion in SBF.

  9. Influence of Fe3O4 Nanoparticles in Hydroxyapatite Scaffolds on Proliferation of Primary Human Fibroblast Cells

    NASA Astrophysics Data System (ADS)

    Maleki-Ghaleh, H.; Aghaie, E.; Nadernezhad, A.; Zargarzadeh, M.; Khakzad, A.; Shakeri, M. S.; Beygi Khosrowshahi, Y.; Siadati, M. H.

    2016-06-01

    Modern techniques for expanding stem cells play a substantial role in tissue engineering: the raw material that facilitates regeneration of damaged tissues and treats diseases. The environmental conditions and bioprocessing methods are the primary determinants of the rate of cultured stem cell proliferation. Bioceramic scaffolds made of calcium phosphate are effective substrates for optimal cell proliferation. The present study investigates the effects of two bioceramic scaffolds on proliferating cells in culture media. One scaffold was made of hydroxyapatite and the other was a mixture of hydroxyapatite and ferromagnetic material (Fe3O4 nanoparticles). Disk-shaped (10 mm × 2 mm) samples of the two scaffolds were prepared. Primary human fibroblast proliferation was 1.8- and 2.5-fold faster, respectively, when cultured in the presence of hydroxyapatite or ferrous nanoparticle/hydroxyapatite mixtures. Optical microscopy images revealed that the increased proliferation was due to enhanced cell-cell contact. The presence of magnetic Fe3O4 nanoparticles in the ceramic scaffolds significantly increased cell proliferation compared to hydroxyapatite scaffolds and tissue culture polystyrene.

  10. Biodegradable PCL/fibroin/hydroxyapatite porous scaffolds prepared by supercritical foaming for bone regeneration.

    PubMed

    Diaz-Gomez, Luis; García-González, Carlos A; Wang, Jiamian; Yang, Fang; Aznar-Cervantes, Salvador; Cenis, Jose Luis; Reyes, Ricardo; Delgado, Araceli; Évora, Carmen; Concheiro, Angel; Alvarez-Lorenzo, Carmen

    2017-07-15

    Regenerative medicine seeks advanced solutions for bone repair in the form of bioactive synthetic scaffolds by using simple and reproducible processing techniques. In this work, poly-ε-caprolactone (PCL)-based porous scaffolds with improved osteoconductive and osteoinductive properties were processed by supercritical foaming through a careful tuning of components and processing conditions. Composite scaffolds were prepared from various combinations of PCL, silk fibroin and nano-hydroxyapatite (nHA). The green and cost-effective supercritical CO2 foaming method applied rendered solid scaffolds with 67-70% porosity. The incorporation of fibroin and nHA in the scaffolds increased the compressive modulus, cellular adhesion and calcium deposition. The composite scaffolds were tested in vivo in a large-scale calvarial defect model, and bone regeneration was evaluated for up to 14 weeks after implantation. Histomorphometric results showed that all implanted constructs gave rise to the endochondral bone formation and unveiled the synergistic effect of silk fibroin and nHA on the bone repair extent. The information gathered may shed light on the design and processing criteria of bioactive bone scaffolds. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Graphene oxide nanoflakes incorporated gelatin-hydroxyapatite scaffolds enhance osteogenic differentiation of human mesenchymal stem cells

    NASA Astrophysics Data System (ADS)

    Nair, Manitha; Nancy, D.; Krishnan, Amit G.; Anjusree, G. S.; Vadukumpully, Sajini; Nair, Shantikumar V.

    2015-04-01

    In this study, graphene oxide (GO) nanoflakes (0.5 and 1 wt%) were incorporated into a gelatin-hydroxyapatite (GHA) matrix through a freeze drying technique and its effect to enhance mechanical strength and osteogenic differentiation was studied. The GHA matrix with GO demonstrated less brittleness in comparison to GHA scaffolds. There was no significant difference in mechanical strength between GOGHA0.5 and GOGHA1.0 scaffolds. When the scaffolds were immersed in phosphate buffered saline (to mimic physiologic condition) for 60 days, around 50-60% of GO was released in sustained and linear manner and the concentration was within the toxicity limit as reported earlier. Further, GOGHA0.5 scaffolds were continued for cell culture experiments, wherein the scaffold induced osteogenic differentiation of human adipose derived mesenchymal stem cells without providing supplements like dexamethasone, L-ascorbic acid and β glycerophosphate in the medium. The level of osteogenic differentiation of stem cells was comparable to those cultured on GHA scaffolds with osteogenic supplements. Thus biocompatible, biodegradable and porous GO reinforced gelatin-HA 3D scaffolds may serve as a suitable candidate in promoting bone regeneration in orthopaedics.

  12. Mechanical properties and in vitro evaluation of bioactivity and degradation of dexamethasone-releasing poly-D-L-lactide/nano-hydroxyapatite composite scaffolds.

    PubMed

    Chen, Ling; Tang, Chak Yin; Tsui, Chi Pong; Chen, Da Zhu

    2013-06-01

    The purpose of this study was to fabricate drug-release nano-composite scaffolds and perform in vitro evaluation of their mechanical properties, bioactivity, biodegradability and drug release behaviors. Porous drug-release poly-d-l-lactide (PDLLA) composite scaffolds filled with different amounts of nano-hydroxyapatite (nano-HAp) were prepared by a technique combining polymer coagulation, cold compression moulding, salt leaching and drug coating. Apatite detected on the scaffolds after exposure to a simulated body fluid showed improvement in bioactivity and the apatite formation ability through the addition of the nano-HAp content in the composites. Nano-HAp incorporation and apatite formation made a positive impact on the mechanical properties of the scaffolds; however, plasticization and degradation of PDLLA had a negative impact. The pH-compensation effect of the composite scaffolds can reduce the risk of chronic inflammation complications. The fabrication method in this study can produce scaffolds with controllable structure, appropriate mechanical properties and degradation rates for cancellous bone repair applications.

  13. Electrospun biomimetic scaffold of hydroxyapatite/chitosan supports enhanced osteogenic differentiation of mMSCs

    NASA Astrophysics Data System (ADS)

    Peng, Hongju; Yin, Zi; Liu, Huanhuan; Chen, Xiao; Feng, Bei; Yuan, Huihua; Su, Bo; Ouyang, Hongwei; Zhang, Yanzhong

    2012-12-01

    Engaging functional biomaterial scaffolds to regulate stem cell differentiation has drawn a great deal of attention in the tissue engineering and regenerative medicine community. In this study, biomimetic composite nanofibrous scaffolds of hydroxyapatite/chitosan (HAp/CTS) were prepared to investigate their capacity for inducing murine mesenchymal stem cells (mMSCs) to differentiate into the osteogenic lineage, in the absence and presence of an osteogenic supplementation (i.e., ascorbic acid, β-glycerol phosphate, and dexamethasone), respectively. Using electrospun chitosan (CTS) nanofibrous scaffolds as the control, cell morphology, growth, specific osteogenic genes expression, and quantified proteins secretion on the HAp/CTS scaffolds were sequentially examined and assessed. It appeared that the HAp/CTS scaffolds supported better attachment and proliferation of the mMSCs. Most noteworthy was that in the absence of the osteogenic supplementation, expression of osteogenic genes including collagen I (Col I), runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), and osteocalcin (OCN) were significantly upregulated in mMSCs cultured on the HAp/CTS nanofibrous scaffolds. Also increased secretion of the osteogenesis protein markers of alkaline phosphatase and collagen confirmed that the HAp/CTS nanofibrous scaffold markedly promoted the osteogenic commitment in the mMSCs. Moreover, the presence of osteogenic supplementation proved an enhanced efficacy of mMSC osteogenesis on the HAp/CTS nanofibrous scaffolds. Collectively, this study demonstrated that the biomimetic nanofibrous HAp/CTS scaffolds could support and enhance the adhesion, proliferation, and particularly osteogenic differentiation of the mMSCs. It also substantiated the potential of using biomimetic nanofibrous scaffolds of HAp/CTS for functional bone repair and regeneration applications.

  14. Biocompatibility of a porous alumina ceramic scaffold coated with hydroxyapatite and bioglass.

    PubMed

    Kido, Hueliton Wilian; Ribeiro, Daniel Araki; de Oliveira, Poliani; Parizotto, Nivaldo Antônio; Camilo, Claudia Cristiane; Fortulan, Carlos Alberto; Marcantonio, Elcio; da Silva, Victor Hugo Pereira; Renno, Ana Claudia Muniz

    2014-07-01

    This study aimed to evaluate the osteointegration and genotoxic potential of a bioactive scaffold, composed of alumina and coated with hydroxyapatite and bioglass, after their implantation in tibias of rats. For this purpose, Wistar rats underwent surgery to induce a tibial bone defect, which was filled with the bioactive scaffolds. Histology analysis (descriptive and morphometry) of the bone tissue and the single-cell gel assay (comet) in multiple organs (blood, liver, and kidney) were used to reach this aim after a period of 30, 60, 90, and 180 days of material implantation. The main findings showed that the incorporation of hydroxyapatite and bioglass in the alumina scaffolds produced a suitable environment for bone ingrowth in the tibial defects and did not demonstrate any genotoxicity in the organs evaluated in all experimental periods. These results clearly indicate that the bioactive scaffolds used in this study present osteogenic potential and still exhibit local and systemic biocompatibility. These findings are promising once they convey important information about the behavior of this novel biomaterial in biological system and highlight its possible clinical application.

  15. Collagen-Hydroxyapatite Scaffolds Induce Human Adipose Derived Stem Cells Osteogenic Differentiation In Vitro.

    PubMed

    Calabrese, Giovanna; Giuffrida, Raffaella; Fabbi, Claudia; Figallo, Elisa; Lo Furno, Debora; Gulino, Rosario; Colarossi, Cristina; Fullone, Francesco; Giuffrida, Rosario; Parenti, Rosalba; Memeo, Lorenzo; Forte, Stefano

    2016-01-01

    Mesenchymal stem cells (MSCs) play a crucial role in regulating normal skeletal homeostasis and, in case of injury, in bone healing and reestablishment of skeletal integrity. Recent scientific literature is focused on the development of bone regeneration models where MSCs are combined with biomimetic three-dimensional scaffolds able to direct MSC osteogenesis. In this work the osteogenic potential of human MSCs isolated from adipose tissue (hADSCs) has been evaluated in vitro in combination with collagen/Mg doped hydroxyapatite scaffolds. Results demonstrate the high osteogenic potential of hADSCs when cultured in specific differentiation induction medium, as revealed by the Alizarin Red S staining and gene expression profile analysis. In combination with collagen/hydroxyapatite scaffold, hADSCs differentiate into mature osteoblasts even in the absence of specific inducing factors; nevertheless, the supplement of the factors markedly accelerates the osteogenic process, as confirmed by the expression of specific markers of pre-osteoblast and mature osteoblast stages, such as osterix, osteopontin (also known as bone sialoprotein I), osteocalcin and specific markers of extracellular matrix maturation and mineralization stages, such as ALPL and osteonectin. Hence, the present work demonstrates that the scaffold per se is able to induce hADSCs differentiation, while the addition of osteo-inductive factors produces a significant acceleration of the osteogenic process. This observation makes the use of our model potentially interesting in the field of regenerative medicine for the treatment of bone defects.

  16. Collagen-Hydroxyapatite Scaffolds Induce Human Adipose Derived Stem Cells Osteogenic Differentiation In Vitro

    PubMed Central

    Fabbi, Claudia; Figallo, Elisa; Lo Furno, Debora; Gulino, Rosario; Colarossi, Cristina; Fullone, Francesco; Giuffrida, Rosario; Parenti, Rosalba; Memeo, Lorenzo; Forte, Stefano

    2016-01-01

    Mesenchymal stem cells (MSCs) play a crucial role in regulating normal skeletal homeostasis and, in case of injury, in bone healing and reestablishment of skeletal integrity. Recent scientific literature is focused on the development of bone regeneration models where MSCs are combined with biomimetic three-dimensional scaffolds able to direct MSC osteogenesis. In this work the osteogenic potential of human MSCs isolated from adipose tissue (hADSCs) has been evaluated in vitro in combination with collagen/Mg doped hydroxyapatite scaffolds. Results demonstrate the high osteogenic potential of hADSCs when cultured in specific differentiation induction medium, as revealed by the Alizarin Red S staining and gene expression profile analysis. In combination with collagen/hydroxyapatite scaffold, hADSCs differentiate into mature osteoblasts even in the absence of specific inducing factors; nevertheless, the supplement of the factors markedly accelerates the osteogenic process, as confirmed by the expression of specific markers of pre-osteoblast and mature osteoblast stages, such as osterix, osteopontin (also known as bone sialoprotein I), osteocalcin and specific markers of extracellular matrix maturation and mineralization stages, such as ALPL and osteonectin. Hence, the present work demonstrates that the scaffold per se is able to induce hADSCs differentiation, while the addition of osteo-inductive factors produces a significant acceleration of the osteogenic process. This observation makes the use of our model potentially interesting in the field of regenerative medicine for the treatment of bone defects. PMID:26982592

  17. Fabrication, chemical composition change and phase evolution of biomorphic hydroxyapatite.

    PubMed

    Qian, Junmin; Kang, Yahong; Zhang, Wei; Li, Zhe

    2008-11-01

    Biomorphous, highly porous hydroxyapatite (HA) ceramics have been prepared by a combination of a novel biotemplating process and a sol-gel method, using natural plants like cane and pine as biotemplates. A HA sol was first synthesized from triethylphosphate and calcium nitrate used as the phosphorus and calcium precursors, respectively, and infiltrated into the biotemplates, and subsequently they were sintered at elevated temperatures to obtain porous HA ceramics. The microstructural changes, phase and chemical composition evolutions during the biotemplate-to-HA conversion were investigated by scanning electron microscopy (SEM), X-ray powder diffraction (XRD), and Fourier-transform infrared (FT-IR) spectroscopy. The XRD and FT-IR analysis revealed that the dominant phase of the product was HA, which contained a small amount of mixed A/B-type carbonated HA, closely resembling that of human bone apatite. Moreover, the appearance of a small amount of secondary phase CaCO(3) seemed unavoidable. The HA was not transformed to the other calcium phosphate phases up to 1400 degrees C, but it contained a trace amount of CaO when sintered at above 1100 degrees C. The possible transformation mechanism was proposed. The SEM observation and mechanical property test showed that as-produced HA ceramics retained the macro-/micro-porous structures of the biotemplates with high precision, and possessed acceptable mechanical strength, which is suggested to be potential scaffolds for bone tissue engineering.

  18. Development of a new carbon nanotube–alginate–hydroxyapatite tricomponent composite scaffold for application in bone tissue engineering

    PubMed Central

    Rajesh, Rajendiran; Dominic Ravichandran, Y

    2015-01-01

    In recent times, tricomponent scaffolds prepared from naturally occurring polysaccharides, hydroxyapatite, and reinforcing materials have been gaining increased attention in the field of bone tissue engineering. In the current work, a tricomponent scaffold with an oxidized multiwalled carbon nanotube (fMWCNT)–alginate–hydroxyapatite with the required porosity was prepared for the first time by a freeze-drying method and characterized using analytical techniques. The hydroxyapatite for the scaffold was isolated from chicken bones by thermal calcination at 800°C. The Fourier transform infrared spectra and X-ray diffraction data confirmed ionic interactions and formation of the fMWCNT–alginate–hydroxyapatite scaffold. Interconnected porosity with a pore size of 130–170 µm was evident from field emission scanning electron microscopy. The total porosity calculated using the liquid displacement method was found to be 93.85%. In vitro biocompatibility and cell proliferation on the scaffold was checked using an MG-63 cell line by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and cell attachment by Hoechst stain assay. In vitro studies showed better cell proliferation, cell differentiation, and cell attachment on the prepared scaffold. These results indicate that this scaffold could be a promising candidate for bone tissue engineering. PMID:26491303

  19. Development of a new carbon nanotube-alginate-hydroxyapatite tricomponent composite scaffold for application in bone tissue engineering.

    PubMed

    Rajesh, Rajendiran; Ravichandran, Y Dominic

    2015-01-01

    In recent times, tricomponent scaffolds prepared from naturally occurring polysaccharides, hydroxyapatite, and reinforcing materials have been gaining increased attention in the field of bone tissue engineering. In the current work, a tricomponent scaffold with an oxidized multiwalled carbon nanotube (fMWCNT)-alginate-hydroxyapatite with the required porosity was prepared for the first time by a freeze-drying method and characterized using analytical techniques. The hydroxyapatite for the scaffold was isolated from chicken bones by thermal calcination at 800°C. The Fourier transform infrared spectra and X-ray diffraction data confirmed ionic interactions and formation of the fMWCNT-alginate-hydroxyapatite scaffold. Interconnected porosity with a pore size of 130-170 µm was evident from field emission scanning electron microscopy. The total porosity calculated using the liquid displacement method was found to be 93.85%. In vitro biocompatibility and cell proliferation on the scaffold was checked using an MG-63 cell line by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and cell attachment by Hoechst stain assay. In vitro studies showed better cell proliferation, cell differentiation, and cell attachment on the prepared scaffold. These results indicate that this scaffold could be a promising candidate for bone tissue engineering.

  20. Porous Hydroxyapatite Bioceramic Scaffolds for Drug Delivery and Bone Regeneration

    NASA Astrophysics Data System (ADS)

    Loca, Dagnija; Locs, Janis; Salma, Kristine; Gulbis, Juris; Salma, Ilze; Berzina-Cimdina, Liga

    2011-10-01

    The conventional methods of supplying a patient with pharmacologic active substances suffer from being very poorly selective, so that damage can occurs to the healthy tissues and organs, different from the intended target. In addition, high drug doses can be required to achieve the desired effect. An alternative approach is based on the use of implantable delivery tools, able to release the active substance in a controlled way. In the current research local drug delivery devices containing 8mg of gentamicin sulphate were prepared using custom developed vacuum impregnation technique. In vitro dissolution tests showed that gentamicin release was sustained for 12h. In order to decrease gentamicin release rate, biopolymer coatings were applied and coating structure investigated. The results showed that gentamicin release can be sustained for more than 70h for poly(epsilon-caprolactone) coated calcium phosphate scaffolds. From poly lactic acid and polyvinyl alcohol coated scaffolds gentamicin was released within 20h and 50h, respectively.

  1. A novel one-pot process for near-net-shape fabrication of open-porous resorbable hydroxyapatite/protein composites and in vivo assessment.

    PubMed

    Mueller, Berit; Koch, Dietmar; Lutz, Rainer; Schlegel, Karl A; Treccani, Laura; Rezwan, Kurosch

    2014-09-01

    We present a mild one-pot freeze gelation process for fabricating near-net, complex-shaped hydroxyapatite scaffolds and to directly incorporate active proteins during scaffold processing. In particular, the direct protein incorporation enables a simultaneous adjustment and control of scaffold microstructure, porosity, resorbability and enhancement of initial mechanical and handling stability. Two proteins, serum albumin and lysozyme, are selected and their effect on scaffold stability and microstructure investigated by biaxial strength tests, electron microscopy, and mercury intrusion porosimetry. The resulting hydroxyapatite/protein composites feature adjustable porosities from 50% to 70% and a mechanical strength ranging from 2 to 6 MPa comparable to that of human spongiosa without any sintering step. Scaffold degradation behaviour and protein release are assessed by in vitro studies. A preliminary in vivo assessment of scaffold biocompatibility and resorption behaviour in adult domestic pigs is discussed. After implantation, composites were resorbed up to 50% after only 4 weeks and up to 65% after 8 weeks. In addition, 14% new bone formation after 4 weeks and 37% after 8 weeks were detected. All these investigations demonstrate the outstanding suitability of the one-pot-process to create, in a customisable and reliable way, biocompatible scaffolds with sufficient mechanical strength for handling and surgical insertion, and for potential use as biodegradable bone substitutes and versatile platform for local drug delivery.

  2. Carbon Nanofiber/Polycaprolactone/Mineralized Hydroxyapatite Nanofibrous Scaffolds for Potential Orthopedic Applications.

    PubMed

    Elangomannan, Shinyjoy; Louis, Kavitha; Dharmaraj, Bhagya Mathi; Kandasamy, Venkata Saravanan; Soundarapandian, Kannan; Gopi, Dhanaraj

    2017-02-22

    Hydroxyapatite (Ca10 (PO4)6(OH)2, HAP), a multimineral substituted calcium phosphate is one of the most substantial bone mineral component that has been widely used as bone replacement materials because of its bioactive and biocompatible properties. However, the use of HAP as bone implants is restricted due to its brittle nature and poor mechanical properties. To overcome this defect and to generate suitable bone implant material, HAP is combined with biodegradable polymer (polycaprolactone, PCL). To enhance the mechanical property of the composite, carbon nanofibers (CNF) is incorporated to the composite, which has long been considered for hard and soft tissue implant due to its exceptional mechanical and structural properties. It is well-known that nanofibrous scaffold are the most-prominent material for the bone reconstruction. We have developed a new remarkable CNF/PCL/mineralized hydroxyapatite (M-HAP) nanofibrous scaffolds on titanium (Ti). The as-developed coatings were characterized by various techniques. The results indicate the formation and homogeneous distribution of components in the nanofibrous scaffolds. Incorporation of CNF into the PCL/M-HAP composite significantly improves the adhesion strength and elastic modulus of the scaffolds. Furthermore, the responses of human osteosarcoma (HOS MG63) cells cultured onto the scaffolds demonstrate that the viability of cells were considerably high for CNF-incorporated PCL/M-HAP than for PCL/M-HAP. In vivo analysis show the presence of soft fibrous tissue growth without any significant inflammatory signs, which suggests that incorporated CNF did not counteract the favorable biological roles of HAP. For load-bearing applications, research in various bone models is needed to substantiate the clinical availability. Thus, from the obtained results, we suggest that CNF/PCL/M-HAP nanofibrous scaffolds can be considered as potential candidates for orthopedic applications.

  3. Degradation and biocompatibility of porous nano-hydroxyapatite/polyurethane composite scaffold for bone tissue engineering

    NASA Astrophysics Data System (ADS)

    Dong, Zhihong; Li, Yubao; Zou, Qin

    2009-04-01

    Porous scaffold containing 30 wt% nano-hydroxyapatite (n-HA) and 70 wt% polyurethane (PU) from castor oil was prepared by a foaming method and investigated by X-ray diffraction (XRD), Fourier transform infrared absorption (FTIR), scanning electron microscopy (SEM) techniques. The results show that n-HA particles disperse homogeneously in the PU matrix. The porous scaffold has not only macropores of 100-800 μm in size but also a lot of micropores on the walls of macropores. The porosity and compressive strength of scaffold are 80% and 271 kPa, respectively. After soaking in simulated body fluid (SBF), hydrolysis and deposition partly occur on the scaffold. The biological evaluation in vitro and in vivo shows that the n-HA/PU scaffold is non-cytotoxic and degradable. The porous structure provides a good microenvironment for cell adherence, growth and proliferation. The n-HA/PU composite scaffold can be satisfied with the basic requirement for tissue engineering, and has the potential to be applied in repair and substitute of human menisci of the knee-joint and articular cartilage.

  4. Ribose mediated crosslinking of collagen-hydroxyapatite hybrid scaffolds for bone tissue regeneration using biomimetic strategies.

    PubMed

    Krishnakumar, Gopal Shankar; Gostynska, Natalia; Campodoni, Elisabetta; Dapporto, Massimiliano; Montesi, Monica; Panseri, Silvia; Tampieri, Anna; Kon, Elizaveta; Marcacci, Maurilio; Sprio, Simone; Sandri, Monica

    2017-08-01

    This study explores for the first time the application of ribose as a highly biocompatible agent for the crosslinking of hybrid mineralized constructs, obtained by bio-inspired mineralization of self-assembling Type I collagen matrix with magnesium-doped-hydroxyapatite nanophase, towards a biomimetic mineralized 3D scaffolds (MgHA/Coll) with excellent compositional and structural mimicry of bone tissue. To this aim, two different crosslinking mechanisms in terms of pre-ribose glycation (before freeze drying) and post-ribose glycation (after freeze drying) were investigated. The obtained results explicate that with controlled freeze-drying, highly anisotropic porous structures with opportune macro-micro porosity are obtained. The physical-chemical features of the scaffolds characterized by XRD, FTIR, ICP and TGA demonstrated structural mimicry analogous to the native bone. The influence of ribose greatly assisted in decreasing solubility and increased enzymatic resistivity of the scaffolds. In addition, enhanced mechanical behaviour in response to compressive forces was achieved. Preliminary cell culture experiments reported good cytocompatibility with extensive cell adhesion, proliferation and colonization. Overall, scaffolds developed by pre-ribose glycation process are preferred, as the related crosslinking technique is more facile and robust to obtain functional scaffolds. As a proof of concept, we have demonstrated that ribose crosslinking is cost-effective, safe and functionally effective. This study also offers new insights and opportunities in developing promising scaffolds for bone tissue engineering. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Porous hydroxyapatite/gelatine scaffolds with ice-designed channel-like porosity for biomedical applications.

    PubMed

    Landi, Elena; Valentini, Federica; Tampieri, Anna

    2008-11-01

    A cryogenic process, including freeze-casting and drying has been performed to obtain hydroxyapatite (HA) scaffolds (approx. diameter 10 mm, height 20 mm) with completely lamellar morphology due to preferentially aligned channel-like pores. Changing the process parameters that influence the cold transmission efficiency from the bottom to the top of the poured HA slurry, lamellar ice crystals with different thickness grew throughout the samples. After sintering, scaffolds with porosity features nearly resembling the ice ones were obtained. The interconnection of pores and the ability of the scaffolds to be rapidly penetrated by synthetic body fluid has been proven. Biohybrid HA/gel composites were prepared, infiltrating HA lamellar scaffolds (45-55 vol.% of porosity) with a 10wt.% solution of gelatine. Colouring genipine was used to cross-link gelatine and clearly show the distribution of the protein in the composite. The compressive mechanical properties of lamellar scaffolds improved with the addition of gelatine: the strength increased up to 5-6 times, while the elastic modulus and strain approximately doubled. The effectiveness of the cross-linkage has been preliminarily verified following scaffold degradation in synthetic body fluid.

  6. Effect of cell-seeded hydroxyapatite scaffolds on rabbit radius bone regeneration.

    PubMed

    Rathbone, C R; Guda, T; Singleton, B M; Oh, D S; Appleford, M R; Ong, J L; Wenke, J C

    2014-05-01

    Highly porous hydroxyapatite (HA) scaffolds were developed as bone graft substitutes using a template coating process, characterized, and seeded with bone marrow-derived mesenchymal stem cells (BMSCs). To test the hypothesis that cell-seeded HA scaffolds improve bone regeneration, HA scaffolds without cell seeding (HA-empty), HA scaffolds with 1.5 × 10(4) BMSCs (HA-low), and HA scaffolds with 1.5 × 10(6) BMSCs (HA-high) were implanted in a 10-mm rabbit radius segmental defect model for 4 and 8 weeks. Three different fluorochromes were administered at 2, 4, and 6 weeks after implantation to identify differences in temporal bone growth patterns. It was observed from fluorescence histomorphometry analyses that an increased rate of bone infiltration occurred from 0 to 2 weeks (p < 0.05) of implantation for the HA-high group (2.9 ± 0.5 mm) as compared with HA-empty (1.8 ± 0.8 mm) and HA-low (1.3 ± 0.2 mm) groups. No significant differences in bone formation within the scaffold or callus formation was observed between all groups after 4 weeks, with a significant increase in bone regenerated for all groups from 4 to 8 weeks (28.4% across groups). Although there was no difference in bone formation within scaffolds, callus formation was significantly higher in HA-empty scaffolds (100.9 ± 14.1 mm(3) ) when compared with HA-low (57.8 ± 7.3 mm(3) ; p ≤ 0.003) and HA-high (69.2 ± 10.4 mm(3) ; p ≤ 0.02) after 8 weeks. These data highlight the need for a better understanding of the parameters critical to the success of cell-seeded HA scaffolds for bone regeneration.

  7. Improving osteointegration and osteogenesis of three-dimensional porous Ti6Al4V scaffolds by polydopamine-assisted biomimetic hydroxyapatite coating.

    PubMed

    Li, Yong; Yang, Wei; Li, Xiaokang; Zhang, Xing; Wang, Cairu; Meng, Xiangfei; Pei, Yifeng; Fan, Xiangli; Lan, Pingheng; Wang, Chunhui; Li, Xiaojie; Guo, Zheng

    2015-03-18

    Titanium alloys with various porous structures can be fabricated by advanced additive manufacturing techniques, which are attractive for use as scaffolds for bone defect repair. However, modification of the scaffold surfaces, particularly inner surfaces, is critical to improve the osteointegration of these scaffolds. In this study, a biomimetic approach was employed to construct polydopamine-assisted hydroxyapatite coating (HA/pDA) onto porous Ti6Al4V scaffolds fabricated by the electron beam melting method. The surface modification was characterized with the field emission scanning electron microscopy, energy dispersive spectroscopy, water contact angle measurement, and confocal laser scanning microscopy. Attachment and proliferation of MC3T3-E1 cells on the scaffold surface were significantly enhanced by the HA/pDA coating compared to the unmodified surfaces. Additionally, MC3T3-E1 cells grown on the HA/pDA-coated Ti6Al4V scaffolds displayed significantly higher expression of runt-related transcription factor-2, alkaline phosphatase, osteocalcin, osteopontin, and collagen type-1 compared with bare Ti6Al4V scaffolds after culture for 14 days. Moreover, microcomputed tomography analysis and Van-Gieson staining of histological sections showed that HA/pDA coating on surfaces of porous Ti6Al4V scaffolds enhanced osteointegration and significantly promoted bone regeneration after implantation in rabbit femoral condylar defects for 4 and 12 weeks. Therefore, this study provides an alternative to biofunctionalized porous Ti6Al4V scaffolds with improved osteointegration and osteogenesis functions for orthopedic applications.

  8. Fabrication of dopamine modified polylactide-poly(ethylene glycol) scaffolds with adjustable properties.

    PubMed

    Shen, Jiali; Shi, Dongjian; Shi, Chang; Li, Xiaojie; Chen, Mingqing

    2017-12-01

    Bio-based polymers have been widely used to be as scaffolds for repairing the bone defects. However, the polymer scaffolds are generally lack of bioactivity and cell recognition site. Seeking effective ways to improve the bioactivity and interaction between materials and tissue or cells is clinically important for long-term performance of bone repair materials. In this work, polylactide-b-poly(ethylene glycol)-b-polylactide (PLA-PEG-PLA, PLEL) tri-block copolymers were firstly synthesized by ring-opening polymerization of lactide using PEG with various molecular weights. Inspired by excellent adhesion of dopamine (DA), a facile and effective method was developed to fabricate polydopamine (PDA) and polydopamine/nano-hydroxyapatite (PDA/n-HA) modified PLEL scaffolds by deposition of PDA and PDA/n-HA coating. The surface structure, degradation rates and mineralization of the modified PLEL scaffolds were investigated, and obviously improved after immobilization of PDA and PDA/n-HA coatings. Moreover, the biocompatible results showed a significant increase in cells viability and adhesion. Therefore, the surface modification with PDA and PDA/n-HA could not only adjust the properties of scaffolds, but also reinforce the interfacial adhesion between the PLEL and cells.

  9. Composition of chitosan-hydroxyapatite-collagen composite scaffold evaluation after simulated body fluid immersion as reconstruction material

    NASA Astrophysics Data System (ADS)

    Verisqa, F.; Triaminingsih, S.; Corputty, J. E. M.

    2017-08-01

    Hydroxyapatite (HA) formation is one of the most important aspects of bone regeneration. Because domestically made chitosan-hydroxyapatite-collagen composite scaffolding from crab shell and bovine bone and tendon has potential as a maxillofacial reconstruction material, the material’s HA-forming ability requires evaluation. The aim of this research is to investigate chitosan-hydroxyapatite-collagen composite scaffold’s potential as a maxillofacial reconstruction material by observing the scaffold’s compositional changes. Scaffold specimens were immersed in 37°C simulated body fluid (SBF) for periods of 2, 4, 6, and 8 days. Scaffold composition was then evaluated by using energy dispersive spectroscopy (EDS). The calcium (Ca) and phosphorus (P) percentages of the scaffold were found to increase following SBF immersion. The high Ca/P ratio (3.82) on the scaffold indicated HA formation. Ion exchange played a significant role in the increased percentages of Ca and P, which led to new HA layer formation. The scaffold’s HA acted as a nucleation site of Ca and P from the SBF, with collagen and chitosan as the scaffold’s matrix. Chitosan-hydroxyapatite-collagen composite scaffold shows potential as a maxillofacial reconstruction material, since its composition favors HA formation.

  10. Biocomposite scaffolds for bone regeneration: Role of chitosan and hydroxyapatite within poly-3-hydroxybutyrate-co-3-hydroxyvalerate on mechanical properties and in vitro evaluation.

    PubMed

    Zhang, Sai; Prabhakaran, Molamma P; Qin, Xiaohong; Ramakrishna, Seeram

    2015-11-01

    Bio-engineered scaffolds for bone tissue regeneration is an exploding area of research mainly because they can satisfy the essential demands and current challenges in bone replacement therapies, by imitating the extracellular matrix (ECM) of the native bone. We fabricated bio-composite nanofibrous scaffolds with a blend of poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV), chitosan (CTS) and hydroxyapatite (HA) during this study. Morphological evaluation confirmed the fiber diameters of PHBV, PHBV/CTS (90:10), PHBV/CTS/HA4 (85.5:9.5:5) and PHBV/CTS/HA8 (81:9:10) as 405 ± 74 nm, 334 ± 82 nm, 316 ± 103 nm and 256 ± 110 nm, respectively. The PHBV/CTS/HA4 and PHBV/CTS/HA8 scaffolds were capable of enduring the long term culture of human fetal osteoblasts (hFOB) with ultimate tensile strength of 3.55 ± 0.22 MPa and 4.19 ± 0.19 MPa, respectively. The proliferation of osteoblasts on PHBV/CTS/HA8 scaffold was found 34.10% higher than that on PHBV scaffold on day 20. Cell maturation identified by alkaline phosphatase activity on day 20 was significantly higher on PHBV/CTS/HA8 scaffold than that on PHBV scaffold. The cells on PHBV/CTS/HA8 scaffold also acquired higher mineral deposition (25.79%) than the mineral deposition on PHBV scaffold by day 20, confirmed by EDX analysis. Based on the results, we concluded that the electrospun PHBV/CTS/HA8 scaffolds hold great potential to promote the regeneration of bone tissue due to the synergistic effect of chitosan and HA, whereby chitosan provided cell recognition sites while HA acted as a chelating agent for organizing the apatite-like mineralization.

  11. Fabrication of polymeric scaffolds with a controlled distribution of pores.

    PubMed

    Capes, J S; Ando, H Y; Cameron, R E

    2005-12-01

    The design of tissue engineering scaffolds must take into account many factors including successful vascularisation and the growth of cells. Research has looked at refining scaffold architecture to promote more directed growth of tissues through well-defined anisotropy in the pore structure. In many cases it is also desirable to incorporate therapeutic ingredients, such as growth factors, into the scaffold so that their release occurs as the scaffold degrades. Therefore, scaffold fabrication techniques must be found to precisely control, not only the overall porosity of scaffolds, but also the pore size, shape and spatial distribution. This work describes the use of a regularly shaped porogen, sugar spheres, to manufacture polymeric scaffolds. Results show that pre-assembling the spheres created scaffolds with a constant porosity of 60%, but with varying pores sizes from 200-800 microm, leading to a variation in the surface area and likely degradation rate of the scaffolds. Employing different polymer impregnation techniques tailored the number of pores present with a diameter of less than 100 microm to suit different functions, and altering the packing structure of the sugar spheres created scaffolds with novel layered porosity. Replacing sugar spheres with sugar strands formed scaffolds with pores aligned in one direction.

  12. Low-pressure foaming: a novel method for the fabrication of porous scaffolds for tissue engineering.

    PubMed

    Chung, Eun Ji; Sugimoto, Matthew; Koh, Jason L; Ameer, Guillermo A

    2012-02-01

    Scaffolds for tissue engineering applications must incorporate porosity for optimal cell seeding, tissue ingrowth, and vascularization, but common fabrication methods for achieving porosity are incompatible with a variety of polymers, limiting widespread use. In this study, porous scaffolds consisting of poly(1,8-octanediol-co-citrate) (POC) containing hydroxyapatite nanocrystals (HA) were fabricated using low-pressure foaming (LPF). LPF is a novel method of fabricating an interconnected, porous scaffold with relative ease. LPF takes advantage of air bubbles that act as pore nucleation sites during a polymer mixing step. Vacuum is applied to expand the nucleation sites into interconnected pores that are stabilized through cross-linking. POC was combined with 20%, 40%, and 60% by weight HA, and the effect of increasing HA particle content on porosity, mechanical properties, and alkaline phosphatase (ALP) activity of human mesenchymal stem cells (hMSC) was evaluated. The effect of the prepolymer viscosity on porosity and the mechanical properties of POC with 40% by weight HA (POC-40HA) were also assessed. POC-40HA scaffolds were also implanted in an osteochondral defect of a rabbit model, and the explants were assessed at 6 weeks using histology. With increasing HA content, the pore size of POC-HA scaffolds can be varied (85 to 1,003 μm) and controlled to mimic the pore size of native trabecular bone. The compression modulus increased with greater HA content under dry conditions and were retained to a greater extent than with porous scaffolds fabricated using salt-leaching under wet conditions. Furthermore, all POC-HA scaffolds prepared using LPF supported hMSC attachment, and an increase in ALP activity correlated with an increase in HA content. An increase in the prepolymer viscosity resulted in increased compression modulus, greater distance between pores, and less porosity. After 6 weeks in vivo, cell and tissue infiltration was present throughout the scaffold

  13. Three-dimensional polycaprolactone-hydroxyapatite scaffolds combined with bone marrow cells for cartilage tissue engineering.

    PubMed

    Wei, Bo; Yao, Qingqiang; Guo, Yang; Mao, Fengyong; Liu, Shuai; Xu, Yan; Wang, Liming

    2015-08-01

    The goal of this study was to investigate the chondrogenic potential of three-dimensional polycaprolactone-hydroxyapatite (PCL-HA) scaffolds loaded with bone marrow cells in vitro and the effect of PCL-HA scaffolds on osteochondral repair in vivo. Here, bone marrow was added to the prepared PCL-HA scaffolds and cultured in chondrogenic medium for 10 weeks. Osteochondral defects were created in the trochlear groove of 29 knees in 17 New Zealand white rabbits, which were then divided into four groups that underwent: implantation of PCL-HA scaffolds (left knee, n = 17; Group 1), microfracture (right knee, n = 6; Group 2), autologous osteochondral transplantation (right knee, n = 6; Group 3), and no treatment (right knee, n = 5; Control). Extracellular matrix produced by bone marrow cells covered the surface and filled the pores of PCL-HA scaffolds after 10 weeks in culture. Moreover, many cell-laden cartilage lacunae were observed, and cartilage matrix was concentrated in the PCL-HA scaffolds. After a 12-week repair period, Group 1 showed excellent vertical and lateral integration with host bone, but incomplete cartilage regeneration and matrix accumulation. An uneven surface of regenerated cartilage and reduced distribution of cartilage matrix were observed in Group 2. In addition, abnormal bone growth and unstable integration between repaired and host tissues were detected. For Group 3, the integration between transplanted and host cartilage was interrupted. Our findings indicate that the PCL-HA scaffolds loaded with bone marrow cells improved chondrogenesis in vitro and implantation of PCL-HA scaffolds for osteochondral repairenhanced integration with host bone. However, cartilage regeneration remained unsatisfactory. The addition of trophic factors or the use of precultured cell-PCL-HA constructs for accelerated osteochondral repair requires further investigation.

  14. Dissolution characteristics of extrusion freeformed hydroxyapatite-tricalcium phosphate scaffolds.

    PubMed

    Yang, H Y; Thompson, I; Yang, S F; Chi, X P; Evans, J R G; Cook, R J

    2008-11-01

    The dissolution behaviour of calcium phosphate filaments made by extrusion freeforming for hard tissue scaffolds was measured. The solubility of filaments with different HA/beta-TCP ratios sintered at temperatures from 1,100 to 1,300 degrees C was measured under simulated physiological conditions (tris buffer solution: tris(hydroxyl) methyl-aminomethane-HCl), pH 7.4, 37 degrees C). Calcium and phosphate concentrations were measured separately by inductively coupled plasma (ICP) atomic emission spectroscopy. Surface morphologies and composition before and after immersion were analyzed by SEM and EDS. The results clearly show that as the beta-TCP content increased, the dissolution increased. Higher sintering temperatures, with consequent closure of surface pores, resulted in lower dissolution. Examination of the surface suggested dissolution on preferred sites by pitting.

  15. Investigation of fabrication and environmental effects on bioceramic bone scaffolds

    NASA Astrophysics Data System (ADS)

    Vivanco Morales, Juan Francisco

    2011-12-01

    Bioactive ceramic materials like tricalcium phosphates (TCP) have been emerging as viable material alternatives to the current therapies of bone scaffolding to target fracture healing and osteoporosis. Once scaffolds are implanted at the defect site they should provide mechanical and biological functions, ultimately serving to facilitate with surrounding native tissue. Optimal osteogenic signal expression and subsequent differentiation of cells seeded on the scaffold in both in vivo and in vitro conditions is known to be influenced by scaffold properties and biomechanical environmental conditions. Thus, the objective of this research was to investigate the effect of fabrication and environmental variables on the properties of bioceramic scaffolds for bone tissue engineering applications. Specifically, the effect of sintering temperature in the range of 950°C -1150°C of a cost-effective on a large scale manufacturing process, on the physical and mechanical properties of bioceramic bone scaffolds, was investigated. In addition, the effect of a controlled environment was investigated by implementing a bioreactor and bone loading system to study the response of ex vivo trabecular bone to compressive load while perfused with culture medium. Collectively, this thesis demonstrates that: (1) the sintering temperature to fabricate bioceramic scaffolds can be tuned to structural properties, and (2) the use of a controlled mechanical and biochemical environment can enhance bone tissue development. These findings support the development of clinically successful bioceramic scaffolds that may stimulate bone regeneration and scaffold integration while providing structural integrity.

  16. Electrospun fibrous scaffolds combined with nanoscale hydroxyapatite induce osteogenic differentiation of human periodontal ligament cells.

    PubMed

    Wu, Xiaonan; Miao, Leiying; Yao, Yingfang; Wu, Wenlei; Liu, Yu; Chen, Xiaofeng; Sun, Weibin

    2014-01-01

    Periodontal repair is a complex process in which regeneration of alveolar bone is a vital component. The aim of this study was to develop a biodegradable scaffold with good biocompatibility and osteoinductive ability. Two types of composite fibrous scaffolds were produced by electrospinning, ie, type I collagen/poly(ε-caprolactone) (COL/PCL) and type I collagen/poly(ε-caprolactone)/nanoscale hydroxyapatite (COL/PCL/nHA) with an average fiber diameter of about 377 nm. After a simulated body fluid (SBF) immersion test, the COL/PCL/nHA-SBF scaffold developed a rough surface because of the calcium phosphate deposited on the fibers, suggesting that the presence of nHA promoted the mineralization potential of the scaffold. Energy dispersive X-ray spectroscopy clearly showed the calcium and phosphorus content in the COL/PCL/nHA and COL/PCL/nHA-SBF scaffolds, confirming the findings of nHA and calcium phosphate precipitation on scanning electron micrographs. Water contact analysis revealed that nHA could improve the hydrophilic nature of the COL/PCL/nHA-SBF scaffold. The morphology of periodontal ligament cells cultured on COL/PCL-SBF and COL/PCL/nHA-SBF was evaluated by scanning electron microscopy. The results showed that cells adhered to either type of scaffold and were slightly spindle-shaped in the beginning, then extended gradually with stretched filopodia, indicating an ability to fill the fiber pores. A Cell Counting Kit-8 assay showed that both scaffolds supported cell proliferation. However, real-time quantitative polymerase chain reaction analysis showed that expression of the bone-related markers, alkaline phosphatase and osteocalcin, was upregulated only on the COL/PCL/nHA-SBF scaffold, indicating that this scaffold had the ability to induce osteogenic differentiation of periodontal ligament cells. In this study, COL/PCL/nHA-SBF produced by electrospinning followed by biomimetic mineralization had combined electrospun fibers with nHA in it. This scaffold has

  17. Electrospun fibrous scaffolds combined with nanoscale hydroxyapatite induce osteogenic differentiation of human periodontal ligament cells

    PubMed Central

    Wu, Xiaonan; Miao, Leiying; Yao, Yingfang; Wu, Wenlei; Liu, Yu; Chen, Xiaofeng; Sun, Weibin

    2014-01-01

    Periodontal repair is a complex process in which regeneration of alveolar bone is a vital component. The aim of this study was to develop a biodegradable scaffold with good biocompatibility and osteoinductive ability. Two types of composite fibrous scaffolds were produced by electrospinning, ie, type I collagen/poly(ε-caprolactone) (COL/PCL) and type I collagen/poly(ε-caprolactone)/nanoscale hydroxyapatite (COL/PCL/nHA) with an average fiber diameter of about 377 nm. After a simulated body fluid (SBF) immersion test, the COL/PCL/nHA-SBF scaffold developed a rough surface because of the calcium phosphate deposited on the fibers, suggesting that the presence of nHA promoted the mineralization potential of the scaffold. Energy dispersive X-ray spectroscopy clearly showed the calcium and phosphorus content in the COL/PCL/nHA and COL/PCL/nHA-SBF scaffolds, confirming the findings of nHA and calcium phosphate precipitation on scanning electron micrographs. Water contact analysis revealed that nHA could improve the hydrophilic nature of the COL/PCL/nHA-SBF scaffold. The morphology of periodontal ligament cells cultured on COL/PCL-SBF and COL/PCL/nHA-SBF was evaluated by scanning electron microscopy. The results showed that cells adhered to either type of scaffold and were slightly spindle-shaped in the beginning, then extended gradually with stretched filopodia, indicating an ability to fill the fiber pores. A Cell Counting Kit-8 assay showed that both scaffolds supported cell proliferation. However, real-time quantitative polymerase chain reaction analysis showed that expression of the bone-related markers, alkaline phosphatase and osteocalcin, was upregulated only on the COL/PCL/nHA-SBF scaffold, indicating that this scaffold had the ability to induce osteogenic differentiation of periodontal ligament cells. In this study, COL/PCL/nHA-SBF produced by electrospinning followed by biomimetic mineralization had combined electrospun fibers with nHA in it. This scaffold has

  18. Comparison of the osteoblastic activity conferred on Si-doped hydroxyapatite scaffolds by different osteostatin coatings.

    PubMed

    Manzano, Miguel; Lozano, Daniel; Arcos, Daniel; Portal-Núñez, Sergio; Orden, Carlos Lopez la; Esbrit, Pedro; Vallet-Regí, María

    2011-10-01

    Parathyroid hormone-related protein (107-111) (osteostatin) induces osteogenic effects in osteoblasts in vitro and in regenerating bone in mice and rabbits. In this study we used osteoblastic MC3T3-E1 cell cultures to evaluate and compare the bioactivity of this peptide either adsorbed or covalently bound (by its C-terminus) to Si-doped hydroxyapatite (Si-HA) scaffolds after organic (-NH(2)) functionalization. By these means osteostatin can be locally released or kept anchored to the scaffold surface. This was confirmed by chemical analysis and by testing the efficiency of osteostatin-loaded Si-HA scaffolds (placed in Transwell chambers) in healing a scratch wound in mouse pluripotent mesenchymal C3H10T1/2 cells. Our results show that exposure of MC3T3-E1 cell monolayers to Si-HA scaffolds with both types of osteostatin coating (deliverable or immobilized), in contrast to those without peptide, similarly stimulated cell growth and matrix mineralization. These findings demonstrate that osteostatin release from Si-HA scaffolds is not essential to promote osteoblastic growth and function in vitro, and lend credence to considering osteostatin a bone regenerating factor. Copyright © 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  19. Bone regeneration in a rabbit critical femoral defect by means of magnetic hydroxyapatite macroporous scaffolds.

    PubMed

    Russo, A; Bianchi, M; Sartori, M; Boi, M; Giavaresi, G; Salter, D M; Jelic, M; Maltarello, M C; Ortolani, A; Sprio, S; Fini, M; Tampieri, A; Marcacci, M

    2017-02-15

    Magnetic scaffolds have recently attracted significant attention in tissue engineering due to the prospect of improving bone tissue formation by conveying soluble factors such as growth factors, hormones, and polypeptides directly to the site of implantation, as well as to the possibility of improving implant fixation and stability. The objective of this study was to compare bone tissue formation in a preclinical rabbit model of critical femoral defect treated either with a hydroxyapatite (HA)/magnetite (90/10 wt %) or pure HA porous scaffolds at 4 and 12 weeks after implantation. The biocompatibility and osteogenic activity of the novel magnetic constructs was assessed with analysis of the amount of newly formed bone tissue and its nanomechanical properties. The osteoconductive properties of the pure HA were confirmed. The HA/magnetite scaffold was able to induce and support bone tissue formation at both experimental time points without adverse tissue reactions. Biomechanically, similar properties were obtained from nanoindentation analysis of bone formed following implantation of magnetic and control scaffolds. The results indicate that the osteoconductive properties of an HA scaffold are maintained following inclusion of a magnetic component. These provide a basis for future studies investigating the potential benefit in tissue engineering of applying magnetic stimuli to enhance bone formation. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2017.

  20. Graphene oxide as an interface phase between polyetheretherketone and hydroxyapatite for tissue engineering scaffolds

    PubMed Central

    Peng, Shuping; Feng, Pei; Wu, Ping; Huang, Wei; Yang, Youwen; Guo, Wang; Gao, Chengde; Shuai, Cijun

    2017-01-01

    The poor bonding strength between biopolymer and bioceramic has remained an unsolved issue. In this study, graphene oxide (GO) was introduced as an interface phase to improve the interfacial bonding between polyetheretherketone (PEEK) and hydroxyapatite (HAP) for tissue engineering scaffolds. On the one hand, the conjugated structure of GO could form strong π-π stacking interaction with the benzene rings in PEEK. On the other hand, GO with a negatively charge resulting from oxygen functional groups could adsorb the positively charged calcium atoms (C sites) of HAP. Consequently, the dispersibility and compatibility of HAP in the PEEK matrix increased with increasing GO content up to 1 wt%. At this time, the compressive strength and modulus of scaffolds increased by 79.45% and 42.07%, respectively. Furthermore, the PEEK-HAP with GO (PEEK-HAP/GO) scaffolds possessed the ability to induce formation of bone-like apatite. And they could support cellular adhesion, proliferation as well as osteogenic differentiation. More importantly, in vivo bone defect repair experiments showed that new bone formed throughout the scaffolds at 60 days after implantation. All these results suggested that the PEEK-HAP/GO scaffolds have a promising potential for bone tissue engineering application. PMID:28425470

  1. Magnetic bioinspired hybrid nanostructured collagen-hydroxyapatite scaffolds supporting cell proliferation and tuning regenerative process.

    PubMed

    Tampieri, Anna; Iafisco, Michele; Sandri, Monica; Panseri, Silvia; Cunha, Carla; Sprio, Simone; Savini, Elisa; Uhlarz, Marc; Herrmannsdörfer, Thomas

    2014-09-24

    A bioinspired mineralization process was applied to develop biomimetic hybrid scaffolds made of (Fe(2+)/Fe(3+))-doped hydroxyapatite nanocrystals nucleated on self-assembling collagen fibers and endowed with super-paramagnetic properties, minimizing the formation of potentially cytotoxic magnetic phases such as magnetite or other iron oxide phases. Magnetic composites were prepared at different temperatures, and the effect of this parameter on the reaction yield in terms of mineralization degree, morphology, degradation, and magnetization was investigated. The influence of scaffold properties on cells was evaluated by seeding human osteoblast-like cells on magnetic and nonmagnetic materials, and differences in terms of viability, adhesion, and proliferation were studied. The synthesis temperature affects mainly the chemical-physical features of the mineral phase of the composites influencing the degradation, the microstructure, and the magnetization values of the entire scaffold and its biological performance. In vitro investigations indicated the biocompatibility of the materials and that the magnetization of the super-paramagnetic scaffolds, induced applying an external static magnetic field, improved cell proliferation in comparison to the nonmagnetic scaffold.

  2. Addition of MgO nanoparticles and plasma surface treatment of three-dimensional printed polycaprolactone/hydroxyapatite scaffolds for improving bone regeneration.

    PubMed

    Roh, Hee-Sang; Lee, Chang-Min; Hwang, Young-Hyoun; Kook, Min-Suk; Yang, Seong-Won; Lee, Donghun; Kim, Byung-Hoon

    2017-05-01

    Magnesium (Mg) plays an important role in the body in mediating cell-extracellular matrix interactions and controlling bone apatite structure and density. Hydroxyapatite (HAp) has been used for osteoconductive bone replacement because of its good compressive strength and biocompatibility. The object of this study is to investigate the effects of adding Magnesium oxide (MgO) nanoparticles to polycaprolactone (PCL)/HAp composites and treating PCL/HAp/MgO scaffolds with oxygen and nitrogen plasma. The 3D PCL/HAp/MgO scaffolds were fabricated using a 3D bioextruder. PCL was mixed with 1-15wt% of MgO and HAp. The scaffolds were treated with oxygen and nitrogen plasma under anisotropic etching conditions to improve the bioactivity. The plasma-treated surfaces were analyzed by X-ray photoelectron spectroscopy, scanning electron microscopy, and atomic force microscopy. In addition, the proliferation and differentiation of pre-osteoblast (MC3T3-E1) cells were examined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and alkaline phosphatase activity. Cell mineralization within the produced scaffolds was analyzed by the quantification of alizarin stainings. The addition of MgO/HAp nanoparticles and plasma treatment enhanced the adhesion, proliferation, and differentiation of MC3T3-E1 cells in the PCL scaffolds. Hence, changes in physical surface morphology and surface chemical properties of the 3D scaffold by plasma treatment can affect the behavior of MC3T3-E1 cells.

  3. Incorporation of polymeric microparticles into collagen-hydroxyapatite scaffolds for the delivery of a pro-osteogenic peptide for bone tissue engineering

    NASA Astrophysics Data System (ADS)

    López-Noriega, Adolfo; Quinlan, Elaine; Celikkin, Nehar; O'Brien, Fergal J.

    2015-01-01

    Collagen-hydroxyapatite scaffolds are outstanding materials for bone tissue engineering as they are biocompatible, bioresorbable, osteoconductive, and osteoinductive. The objective of the present work was to assess the potential of increasing their regenerative capacity by functionalising the scaffolds for therapeutic delivery. This was achieved by the utilization of polymeric drug carriers. With this purpose, alginate, chitosan, gelatine, and poly(lactic-co-glycolic acid) (PLGA) microparticles eluting PTHrP 107-111, an osteogenic pentapeptide, were fabricated and tested by incorporating them into the scaffolds. Among them, PLGA microparticles show the most promising characteristics for use as drug delivery devices. Following the incorporation of the microparticles, the scaffolds maintained their interconnected porous structure and the mechanical properties of the materials were not adversely affected. In addition, the microparticles released all their PTHrP 107-111 cargo. Most importantly, the delivered peptide proved to be bioactive and promoted enhanced osteogenesis as assessed by alkaline phosphatase production and osteocalcin and osteopontin gene expression when pre-osteoblastic cells were seeded on the scaffolds. While the focus was on bone repair, the release system described in this study can be used for the delivery of therapeutics for healing and regeneration of a variety of tissue types depending on the type of collagen scaffold chosen.

  4. Fabrication and characterization of multiscale electrospun scaffolds for cartilage regeneration.

    PubMed

    Levorson, Erica J; Raman Sreerekha, Perumcherry; Chennazhi, Krishna Prasad; Kasper, F Kurtis; Nair, Shantikumar V; Mikos, Antonios G

    2013-02-01

    Recently, scaffolds for tissue regeneration purposes have been observed to utilize nanoscale features in an effort to reap the cellular benefits of scaffold features resembling extracellular matrix (ECM) components. However, one complication surrounding electrospun nanofibers is limited cellular infiltration. One method to ameliorate this negative effect is by incorporating nanofibers into microfibrous scaffolds. This study shows that it is feasible to fabricate electrospun scaffolds containing two differently scaled fibers interspersed evenly throughout the entire construct as well as scaffolds containing fibers composed of two discrete materials, specifically fibrin and poly(ε-caprolactone). In order to accomplish this, multiscale fibrous scaffolds of different compositions were generated using a dual extrusion electrospinning setup with a rotating mandrel. These scaffolds were then characterized for fiber diameter, porosity and pore size and seeded with human mesenchymal stem cells to assess the influence of scaffold architecture and composition on cellular responses as determined by cellularity, histology and glycosaminoglycan (GAG) content. Analysis revealed that nanofibers within a microfiber mesh function to maintain scaffold cellularity under serum-free conditions as well as aid the deposition of GAGs. This supports the hypothesis that scaffolds with constituents more closely resembling native ECM components may be beneficial for cartilage regeneration.

  5. Immobilization of salvianolic acid B-loaded chitosan microspheres distributed three-dimensionally and homogeneously on the porous surface of hydroxyapatite scaffolds.

    PubMed

    Li, Jinyu; Wang, Qin; Zhi, Wei; Wang, Jianxin; Feng, Bo; Qu, Shuxin; Mu, Yandong; Weng, Jie

    2016-10-07

    Porous hydroxyapatite (HA) scaffolds combined with a drug delivery system have attracted much attention for bone tissue engineering. In this study, an easy and highly efficient method was developed to immobilize salvianolic acid B (Sal B)-loaded chitosan (CS) microspheres three dimensionally and homogeneously on the surface of HA scaffolds pre-coated with alginate. Porous HA scaffolds were prepared via a template-leaching process and CS microspheres (used as drug carriers) were fabricated by an emulsion method. To improve adhesion between the microspheres and HA scaffolds, alginate was used to pre-coat the porous surface of the HA scaffolds. Various concentrations of alginate were used to optimize the adhesion of Sal B-loaded CS microspheres to the scaffold surface. During the adherence process, coated HA scaffolds were immersed in an aqueous solution containing Sal B-loaded CS microspheres, followed by standing or shaking at 37 °C for a certain time. The results showed that the microspheres were solidly and homogeneously distributed on the porous surface of the alginate pre-coated HA scaffolds via electrostatic interactions. Few microspheres detached from the porous surface, even after the HA scaffolds with microspheres were treated by shaking in distilled water for as long as 7 d. Compared with the static condition, the distribution of Sal B-loaded CS microspheres on the porous surface of pre-coated HA scaffolds in the shaken condition was more homogeneous and almost unaggregated. Additionally, the compressive strength of the scaffolds coated with alginate was obviously improved. The optimal alginate coating concentration was 1% (i.e. the microstructure of the porous surfaces of the HA scaffolds was almost unchanged). The release profile of Sal B over a 30 d immersion found an initial burst release followed by a sustained release. The result of cell culture in vitro was that 1% alginate-coated scaffolds with Sal B-loaded CS microspheres obviously promoted cell

  6. Gold and Hydroxyapatite Nano-Composite Scaffolds for Anterior Cruciate Ligament Reconstruction: In Vitro Characterization.

    PubMed

    Smith, S E; White, R A; Grant, D A; Grant, S A

    2016-01-01

    Current anterior cruciate ligament (ACL) graft replacement materials often fail due to the lack of biological integration. While many newly developed extracellular matrix based scaffolds show good biocompatibility they often do not entice cellular remodeling and the rebuilding of a functional ligament. We have proposed the conjugation of gold nanoparticles (AuNP) and hydroxyapatite nanoparticles (nano-HAp) to acellular tissue to enhance cell attachment and proliferation while maintaining an improved degradation resistance and open microstructure. We are the first to investigate the double conjugation of AuNP and nano-HAp onto decellularized tissue to improve the tissue remodeling response. Decellularized porcine diaphragm was crosslinked with two types of nano-HAp and amine-functionalized AuNP with 1-ethyl-3-(3-dimethlaminopropyl) carbodiimide (EDC) crosslinker. Scaffolds were characterized using electron microscopy, differential scanning calorimetry, and fibroblast assays. Results demonstrated that scaffolds with nano-HAp have increased thermal stability at low levels of crosslinking. The open microstructure of the scaffold was not compromised allowing for cell migration while still providing increased degradation resistance. The addition of < 200 nm nano-HAp decreased cell viability compared to scaffolds without nanoparticles, but the addition of AuNP to scaffolds showed enhanced cell viability in the presence of < 200 nm nano-HAp. The addition of < 40 nm nano-HAp showed an increase in cell viability compared to scaffolds crosslinked without nanoparticles. It is concluded that attaching AuNP and < 40nm nano-HAp to extracellular matrices may improve overall properties.

  7. Hydroxyapatite/gelatin/gellan sponges as nanocomposite scaffolds for bone reconstruction.

    PubMed

    Barbani, Niccoletta; Guerra, Giulio D; Cristallini, Caterina; Urciuoli, Patrizia; Avvisati, Riccardo; Sala, Alessandro; Rosellini, Elisabetta

    2012-01-01

    The aim of this work was the morphological, physicochemical, mechanical and biological characterization of a new composite system, based on gelatin, gellan and hydroxyapatite, and mimicking the composition of natural bone. Porous scaffolds were prepared by freeze-drying technique, under three different conditions of freezing. The morphological analysis showed a homogeneous porosity, with well interconnected pores, for the sample which underwent a more rapid freezing. The elastic modulus of the same sample was close to that of the natural bone. The presence of interactions among the components was demonstrated through the physicochemical investigation. In addition, the infrared chemical imaging analysis pointed out the similarity among the composite scaffold and the natural bone, in terms of chemical composition, homogeneity, molecular interactions and structural conformation. Preliminary biological characterization showed a good adhesion and proliferation of human mesenchymal stem cells.

  8. Multiscale fabrication of biomimetic scaffolds for tympanic membrane tissue engineering.

    PubMed

    Mota, Carlos; Danti, Serena; D'Alessandro, Delfo; Trombi, Luisa; Ricci, Claudio; Puppi, Dario; Dinucci, Dinuccio; Milazzo, Mario; Stefanini, Cesare; Chiellini, Federica; Moroni, Lorenzo; Berrettini, Stefano

    2015-05-07

    The tympanic membrane (TM) is a thin tissue able to efficiently collect and transmit sound vibrations across the middle ear thanks to the particular orientation of its collagen fibers, radiate on one side and circular on the opposite side. Through the combination of advanced scaffolds and autologous cells, tissue engineering (TE) could offer valuable alternatives to autografting in major TM lesions. In this study, a multiscale approach based on electrospinning (ES) and additive manufacturing (AM) was investigated to fabricate scaffolds, based on FDA approved copolymers, resembling the anatomic features and collagen fiber arrangement of the human TM. A single scale TM scaffold was manufactured using a custom-made collector designed to confer a radial macro-arrangement to poly(lactic-co-glycolic acid) electrospun fibers during their deposition. Dual and triple scale scaffolds were fabricated combining conventional ES with AM to produce poly(ethylene oxide terephthalate)/poly(butylene terephthalate) block copolymer scaffolds with anatomic-like architecture. The processing parameters were optimized for each manufacturing method and copolymer. TM scaffolds were cultured in vitro with human mesenchymal stromal cells, which were viable, metabolically active and organized following the anisotropic character of the scaffolds. The highest viability, cell density and protein content were detected in dual and triple scale scaffolds. Our findings showed that these biomimetic micro-patterned substrates enabled cell disposal along architectural directions, thus appearing as promising substrates for developing functional TM replacements via TE.

  9. Influence of Starting Powders on Hydroxyapatite Coatings Fabricated by Room Temperature Spraying Method.

    PubMed

    Seo, Dong Seok; Lee, Jong Kook; Hwang, Kyu Hong; Hahn, Byung Dong; Yoon, Seog Young

    2015-08-01

    Three types of raw materials were used for the fabrication of hydroxyapatite coatings by using the room temperature spraying method and their influence on the microstructure and in vitro characteristics were investigated. Starting hydroxyapatite powders for coatings on titanium substrate were prepared by a heat treatment at 1100 °C for 2 h of bovine bone, bone ash, and commercial hydroxyapatite powders. The phase compositions and Ca/P ratios of the three hydroxyapatite coatings were similar to those of the raw materials without decomposition or formation of a new phase. All hydroxyapatite coatings showed a honeycomb structure, but their surface microstructures revealed different features in regards to surface morphology and roughness, based on the staring materials. All coatings consisted of nano-sized grains and had dense microstructure. Inferred from in vitro experiments in pure water, all coatings have a good dissolution-resistance and biostability in water.

  10. Scaffolds with a standardized macro-architecture fabricated from several calcium phosphate ceramics using an indirect rapid prototyping technique

    PubMed Central

    Wilson, C. E.; van Blitterswijk, C. A.; Verbout, A. J.; de Bruijn, J. D.

    2010-01-01

    Calcium phosphate ceramics, commonly applied as bone graft substitutes, are a natural choice of scaffolding material for bone tissue engineering. Evidence shows that the chemical composition, macroporosity and microporosity of these ceramics influences their behavior as bone graft substitutes and bone tissue engineering scaffolds but little has been done to optimize these parameters. One method of optimization is to place focus on a particular parameter by normalizing the influence, as much as possible, of confounding parameters. This is difficult to accomplish with traditional fabrication techniques. In this study we describe a design based rapid prototyping method of manufacturing scaffolds with virtually identical macroporous architectures from different calcium phosphate ceramic compositions. Beta-tricalcium phosphate, hydroxyapatite (at two sintering temperatures) and biphasic calcium phosphate scaffolds were manufactured. The macro- and micro-architectures of the scaffolds were characterized as well as the influence of the manufacturing method on the chemistries of the calcium phosphate compositions. The structural characteristics of the resulting scaffolds were remarkably similar. The manufacturing process had little influence on the composition of the materials except for the consistent but small addition of, or increase in, a beta-tricalcium phosphate phase. Among other applications, scaffolds produced by the method described provide a means of examining the influence of different calcium phosphate compositions while confidently excluding the influence of the macroporous structure of the scaffolds. PMID:21069558

  11. The effect of processing parameters and solid concentration on the mechanical and microstructural properties of freeze-casted macroporous hydroxyapatite scaffolds.

    PubMed

    Farhangdoust, S; Zamanian, A; Yasaei, M; Khorami, M

    2013-01-01

    The design and fabrication of macroporous hydroxyapatite scaffolds, which could overcome current bone tissue engineering limitations, have been considered in recent years. In the current study, controlled unidirectional freeze-casting at different cooling rates was investigated. In the first step, different slurries with initial hydroxyapatite concentrations of 7-37.5 vol.% were prepared. In the next step, different cooling rates from 2 to 14 °C/min were applied to synthesize the porous scaffold. Additionally, a sintering temperature of 1350 °C was chosen as an optimum temperature. Finally, the phase composition (by XRD), microstructure (by SEM), mechanical characteristics, and the porosity of sintered samples were assessed. The porosity of the sintered samples was in a range of 45-87% and the compressive strengths varied from 0.4 MPa to 60 MPa. The mechanical strength of the scaffolds increased as a function of initial concentration, cooling rate, and sintering temperature. With regards to mechanical strength and pore size, the samples with the initial concentration and the cooling rate of 15 vol.% and 5 °C/min, respectively, showed better results.

  12. Alginate/nanohydroxyapatite scaffolds with designed core/shell structures fabricated by 3D plotting and in situ mineralization for bone tissue engineering.

    PubMed

    Luo, Yongxiang; Lode, Anja; Wu, Chengtie; Chang, Jiang; Gelinsky, Michael

    2015-04-01

    Composite scaffolds, especially polymer/hydroxyapatite (HAP) composite scaffolds with predesigned structures, are promising materials for bone tissue engineering. Various methods including direct mixing of HAP powder with polymers or incubating polymer scaffolds in simulated body fluid for preparing polymer/HAP composite scaffolds are either uncontrolled or require long times of incubation. In this work, alginate/nano-HAP composite scaffolds with designed pore parameters and core/shell structures were fabricated using 3D plotting technique and in situ mineralization under mild conditions (at room temperature and without the use of any organic solvents). Light microscopy, scanning electron microscopy, microcomputer tomography, X-ray diffraction, and Fourier transform infrared spectroscopy were applied to characterize the fabricated scaffolds. Mechanical properties and protein delivery of the scaffolds were evaluated, as well as the cell response to the scaffolds by culturing human bone-marrow-derived mesenchymal stem cells (hBMSC). The obtained data indicate that this method is suitable to fabricate alginate/nano-HAP composite scaffolds with a layer of nano-HAP, coating the surface of the alginate strands homogeneously and completely. The surface mineralization enhanced the mechanical properties and improved the cell attachment and spreading, as well as supported sustaining protein release, compared to pure alginate scaffolds without nano-HAP shell layer. The results demonstrated that the method provides an interesting option for bone tissue engineering application.

  13. PHBV/PLLA-based composite scaffolds fabricated using an emulsion freezing/freeze-drying technique for bone tissue engineering: surface modification and in vitro biological evaluation.

    PubMed

    Sultana, Naznin; Wang, Min

    2012-03-01

    Tissue engineering combines living cells with biodegradable materials and/or bioactive components. Composite scaffolds containing biodegradable polymers and nanosized osteoconductive bioceramic with suitable properties are promising for bone tissue regeneration. In this paper, based on blending two biodegradable and biocompatible polymers, namely poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and poly(L-lactic acid) (PLLA) with incorporated nano hydroxyapatite (HA), three-dimensional composite scaffolds with controlled microstructures and an interconnected porous structure, together with high porosity, were fabricated using an emulsion freezing/freeze-drying technique. The influence of various parameters involved in the emulsion freezing/freeze-drying technique was studied for the fabrication of good-quality polymer scaffolds based on PHBV polymers. The morphology, mechanical properties and crystallinity of PHBV/PLLA and HA in PHBV/PLLA composite scaffolds and PHBV polymer scaffolds were studied. The scaffolds were coated with collagen in order to improve wettability. During in vitro biological evaluation study, it was observed that SaOS-2 cells had high attachment on collagen-coated scaffolds. Significant improvement in cell proliferation and alkaline phosphatase activity for HA-incorporated composite scaffolds was observed due to the incorporation of HA. After 3 and 7 days of culture on all scaffolds, SaOS-2 cells also had normal morphology and growth. These results indicated that PHBV/PLLA-based scaffolds fabricated via an emulsion freezing/freeze-drying technique were favorable sites for osteoblastic cells and are promising for the applications of bone tissue engineering.

  14. Towards the synthesis of hydroxyapatite/protein scaffolds with controlled porosities: bulk and interfacial shear rheology of a hydroxyapatite suspension with protein additives.

    PubMed

    Maas, Michael; Bodnar, Pedro Marcus; Hess, Ulrike; Treccani, Laura; Rezwan, Kurosch

    2013-10-01

    The synthesis of porous hydroxyapatite scaffolds is essential for biomedical applications such as bone tissue engineering and replacement. One way to induce macroporosity, which is needed to support bone in-growth, is to use protein additives as foaming agents. Another reason to use protein additives is the potential to introduce a specific biofunctionality to the synthesized scaffolds. In this work, we study the rheological properties of a hydroxyapatite suspension system with additions of the proteins bovine serum albumin (BSA), lysozyme (LSZ) and fibrinogen (FIB). Both the rheology of the bulk phase as well as the interfacial shear rheology are studied. The bulk rheological data provides important information on the setting behavior of the thixotropic suspension, which we find to be faster with the addition of FIB and LSZ and much slower with BSA. Foam bubble stabilization mechanisms can be rationalized via interfacial shear rheology and we show that it depends on the growth of interfacial films at the suspension/air interface. These interfacial films support the stabilization of bubbles within the ceramic matrix and thereby introduce macropores. Due to the weak interaction of the protein molecules with the hydroxyapatite particles of the suspension, we find that BSA forms the most stable interfacial films, followed by FIB. LSZ strongly interacts with the hydroxyapatite particles and thus only forms thin films with very low elastic moduli. In summary, our study provides fundamental rheological insights which are essential for tailoring hydroxyapatite/protein suspensions in order to synthesize scaffolds with controlled porosities. Copyright © 2013 Elsevier Inc. All rights reserved.

  15. Three dimensional printed macroporous polylactic acid/hydroxyapatite composite scaffolds for promoting bone formation in a critical-size rat calvarial defect model

    PubMed Central

    Zhang, Haifeng; Mao, Xiyuan; Du, Zijing; Jiang, Wenbo; Han, Xiuguo; Zhao, Danyang; Han, Dong; Li, Qingfeng

    2016-01-01

    Abstract We have explored the applicability of printed scaffold by comparing osteogenic ability and biodegradation property of three resorbable biomaterials. A polylactic acid/hydroxyapatite (PLA/HA) composite with a pore size of 500 μm and 60% porosity was fabricated by three-dimensional printing. Three-dimensional printed PLA/HA, β-tricalcium phosphate (β-TCP) and partially demineralized bone matrix (DBM) seeded with bone marrow stromal cells (BMSCs) were evaluated by cell adhesion, proliferation, alkaline phosphatase activity and osteogenic gene expression of osteopontin (OPN) and collagen type I (COL-1). Moreover, the biocompatibility, bone repairing capacity and degradation in three different bone substitute materials were estimated using a critical-size rat calvarial defect model in vivo. The defects were evaluated by micro-computed tomography and histological analysis at four and eight weeks after surgery, respectively. The results showed that each of the studied scaffolds had its own specific merits and drawbacks. Three-dimensional printed PLA/HA scaffolds possessed good biocompatibility and stimulated BMSC cell proliferation and differentiation to osteogenic cells. The outcomes in vivo revealed that 3D printed PLA/HA scaffolds had good osteogenic capability and biodegradation activity with no difference in inflammation reaction. Therefore, 3D printed PLA/HA scaffolds have potential applications in bone tissue engineering and may be used as graft substitutes in reconstructive surgery. PMID:27877865

  16. Three dimensional printed macroporous polylactic acid/hydroxyapatite composite scaffolds for promoting bone formation in a critical-size rat calvarial defect model.

    PubMed

    Zhang, Haifeng; Mao, Xiyuan; Du, Zijing; Jiang, Wenbo; Han, Xiuguo; Zhao, Danyang; Han, Dong; Li, Qingfeng

    2016-01-01

    We have explored the applicability of printed scaffold by comparing osteogenic ability and biodegradation property of three resorbable biomaterials. A polylactic acid/hydroxyapatite (PLA/HA) composite with a pore size of 500 μm and 60% porosity was fabricated by three-dimensional printing. Three-dimensional printed PLA/HA, β-tricalcium phosphate (β-TCP) and partially demineralized bone matrix (DBM) seeded with bone marrow stromal cells (BMSCs) were evaluated by cell adhesion, proliferation, alkaline phosphatase activity and osteogenic gene expression of osteopontin (OPN) and collagen type I (COL-1). Moreover, the biocompatibility, bone repairing capacity and degradation in three different bone substitute materials were estimated using a critical-size rat calvarial defect model in vivo. The defects were evaluated by micro-computed tomography and histological analysis at four and eight weeks after surgery, respectively. The results showed that each of the studied scaffolds had its own specific merits and drawbacks. Three-dimensional printed PLA/HA scaffolds possessed good biocompatibility and stimulated BMSC cell proliferation and differentiation to osteogenic cells. The outcomes in vivo revealed that 3D printed PLA/HA scaffolds had good osteogenic capability and biodegradation activity with no difference in inflammation reaction. Therefore, 3D printed PLA/HA scaffolds have potential applications in bone tissue engineering and may be used as graft substitutes in reconstructive surgery.

  17. Fabrication of porous hydroxyapatite bodies by a new direct consolidation method: starch consolidation.

    PubMed

    Rodríguez-Lorenzo, L M; Vallet-Regí, M; Ferreira, J M F

    2002-05-01

    A new direct consolidation method known as "starch consolidation" was developed especially for the fabrication of porous ceramics, as adopted to prepare hydroxyapatite (OHAp) bodies with different pore sizes and pore volume fractions. The method is based on the swelling ability of starch when it is heated to 80 degrees C in the presence of water. An OHAp powder prepared by a precipitation method and heat treated at 900 degrees C to obtain an appropriate specific surface area for colloidal processing was used in the present work. Wet ball milling was required to deagglomerate the powder and improve its processing ability. Different starch volume fractions and starch particle sizes were added to the OHAp slurries, which were then heat treated to promote consolidation by starch gelification. A linear relationship between the planned and measured porosity values was observed. SEM pictures reveal the presence of interconnected pores with shape and sizes corresponding to starch granules used. Flexural strength varied from about 2 to 15 MPa, for pore volume fractions of approximately 70 and 45%, respectively. A degradation study in an acellular synthetic body fluid (SBF) has shown that the prepared bodies keep their integrity under physiological conditions during the studied time. The method offers possibilities of manufacturing materials that can be used as scaffolds engineering and/or systems for controlled delivery of drugs. Copyright 2002 John Wiley & Sons, Inc. J Biomed Mater Res 60: 232--240, 2002; DOI 10.1002/jbm.10036

  18. 3D-Printed Atsttrin-Incorporated Alginate/Hydroxyapatite Scaffold Promotes Bone Defect Regeneration with TNF/TNFR Signaling Involvement.

    PubMed

    Wang, Quan; Xia, Qingqing; Wu, Yan; Zhang, Xiaolei; Wen, Feiqiu; Chen, Xiaowen; Zhang, Shufang; Heng, Boon Chin; He, Yong; Ouyang, Hong-Wei

    2015-08-05

    High expression levels of pro-inflammatory tumor necrosis factor (TNF)-α within bone defects can decelerate and impair bone regeneration. However, there are few available bone scaffolds with anti-inflammatory function. The progranulin (PGRN)-derived engineered protein, Atsttrin, is known to exert antagonistic effects on the TNF-α function. Hence, this study investigates whether 3D-printed Atsttrin-incorporated alginate(Alg)/hydroxyapatite(nHAp) scaffolds can facilitate bone healing through affecting the TNF/TNFR signaling. A 3D bioprinting system is used to fabricate Atsttrin-Alg/nHAp composite scaffolds, and the Atsttrin release from this scaffold is characterized, followed by evaluation of its efficacy on bone regeneration both in vitro and in vivo. The 3D-printed Atsttrin-Alg/nHAp scaffold exhibits a precisely defined structure, can sustain Atsttrin release for at least 5 days, has negligible cytotoxicity, and supports cell adhesion. Atsttrin can also attenuate the suppressive effects of TNF-α on BMP-2-induced osteoblastic differentiation in vitro. The 3D-printed Atsttrin-Alg/nHAp scaffold significantly reduces the number of TNF-α positive cells within wound sites, 7 days after post-calvarial defect surgery. Additionally, histological staining and X-ray scanning results also show that the 3D-printed Atsttrin-Alg/nHAp scaffold enhances the regeneration of mice calvarial bone defects. These findings thus demonstrate that the precise structure and anti-inflammatory properties of 3D-printed Atsttrin-Alg/nHAp scaffolds may promote bone defect repair.

  19. Preparation and Properties of Bamboo Fiber/Nano-hydroxyapatite/Poly(lactic-co-glycolic) Composite Scaffold for Bone Tissue Engineering.

    PubMed

    Jiang, Liuyun; Li, Ye; Xiong, Chengdong; Su, Shengpei; Ding, Haojie

    2017-02-08

    In this study, bamboo fiber was first designed to incorporate into nano-hydroxyapatite/poly(lactic-co-glycolic) to obtain a new composite scaffold of bamboo fiber/nano-hydroxyapatite/poly(lactic-co- glycolic) (BF/n-HA/PLGA) by freeze-drying method. The effect of their components and some factors consisting of different freeze temperatures, concentrations, and pore-forming agents on the porous morphology, porosity, and compressive properties of the scaffold were investigated by scanning electron microscope, modified liquid displacement method, and electromechanical universal testing machine. The results indicated that the 5% BF/30% n-HA/PLGA composite scaffold, prepared with 5% (w/v) high concentration and frozen at -20 °C without pore-forming agent, had the best ideal porous structure and porosity as well as compressive properties, which far exceed those of n-HA/PLGA composite scaffold. In addition, the in vitro simulated body fluids soaking and cell culture experiment showed the addition of BF into the scaffold accelerated the BF/n-HA/PLGA composite scaffolds degradation and exhibited good cytocompatibility, including attachment and proliferation. All the results of the study show that BF has improved the properties of n-HA/PLGA composite scaffolds and BF/n-HA/PLGA might have a great potential for bone tissue engineering scaffold.

  20. Vertical ridge augmentation of the atrophic posterior mandible with custom-made, computer-aided design/computer-aided manufacturing porous hydroxyapatite scaffolds.

    PubMed

    Figliuzzi, Michele; Mangano, Francesco Guido; Fortunato, Leonzio; De Fazio, Rossella; Macchi, Aldo; Iezzi, Giovanna; Piattelli, Adriano; Mangano, Carlo

    2013-05-01

    The present study describes a new protocol for the manufacturing of custom-made hydroxyapatite scaffolds using computer-aided design/computer-aided manufacturing (CAD/CAM), to augment posterior mandibular bone and minimize surgery when severe atrophy is present. Computed tomographic images of an atrophic posterior mandible were acquired and modified into a 3-dimensional (3D) reconstruction model. This model was transferred as a stereolithographic file to a CAD program, where virtual 3D reconstructions of the alveolar ridge were performed, drawing 2 anatomically shaped, custom-made scaffolds. Computer-aided-manufacturing software generated a set of tool-paths for manufacture on a computer-numerical-control milling machine into the exact shape of the 3D projects. Clinically sized, anatomically shaped scaffolds were generated from commercially available porous hydroxyapatite blocks. The custom-made scaffolds well matched the shape of the bone defects and could be easily implanted during surgery. This matching of the shape helped to reduce the time for the operation and contributed to the good healing of the defects. At the 6-month recall, a newly formed and well-integrated bone was observed, completely filling the mandibular posterior defects, and implants were placed, with good primary stability. At the 1-year follow-up examination, the implant-supported restorations showed a good functional and esthetic integration. Although this is an interim report, this study demonstrates that anatomically shaped custom-made scaffolds can be fabricated by combining computed tomographic scans and CAD/CAM techniques. Further studies are needed to confirm these results.

  1. Synthesis and characterization of nanocomposite scaffolds based on triblock copolymer of L-lactide, ε-caprolactone and nano-hydroxyapatite for bone tissue engineering.

    PubMed

    Torabinejad, Bahman; Mohammadi-Rovshandeh, Jamshid; Davachi, Seyed Mohammad; Zamanian, Ali

    2014-09-01

    The employment of biodegradable polymer scaffolds is one of the main approaches for achieving a tissue engineered construct to reproduce bone tissues, which provide a three dimensional template to regenerate desirable tissues for different applications. The main goal of this study is to design a novel triblock scaffold reinforced with nano-hydroxyapatite (nHA) for hard tissue engineering using gas foaming/salt leaching method with minimum solvent usage. With this end in view, the biodegradable triblock copolymers of l-lactide and ε-caprolactone with different mol% were synthesized by ring-opening polymerization method in the presence of Sn(Oct)2 catalyst as initiator and ethylene glycol as co-initiator. The chemical compositions of biodegradable copolymers were characterized by means of FTIR and NMR. The thermal and crystallization behaviors of copolymers were characterized using TGA and DSC thermograms. Moreover, nano-hydroxyapatite was synthesized by the chemical precipitation process and was thoroughly characterized by FTIR, XRD and TEM. Additionally, the nanocomposites with different contents of nHA were prepared by mixing triblock copolymer with nHA. Mechanical properties of the prepared nanocomposites were evaluated by stress-strain measurements. It was found that the nanocomposite with 30% of nHA showed the optimum result. Therefore, nanocomposite scaffolds with 30% nHA were fabricated by gas foaming/salt leaching method and SEM images were used to observe the microstructure and morphology of nanocomposites and nanocomposite scaffolds before and after cell culture. The in-vitro and cell culture tests were also carried out to further evaluate the biological properties. The results revealed that the porous scaffolds were biocompatible to the osteoblast cells because the cells spread and grew well. The resultant nanocomposites could be considered as good candidates for use in bone tissue engineering.

  2. Excavating the Role of Aloe Vera Wrapped Mesoporous Hydroxyapatite Frame Ornamentation in Newly Architectured Polyurethane Scaffolds for Osteogenesis and Guided Bone Regeneration with Microbial Protection.

    PubMed

    Selvakumar, M; Pawar, Harpreet Singh; Francis, Nimmy K; Das, Bodhisatwa; Dhara, Santanu; Chattopadhyay, Santanu

    2016-03-09

    Guided bone regeneration (GBR) scaffolds are unsuccessful in many clinical applications due to a high incidence of postoperative infection. The objective of this work is to fabricate GBR with an anti-infective electrospun scaffold by ornamenting segmented polyurethane (SPU) with two-dimensional Aloe vera wrapped mesoporous hydroxyapatite (Al-mHA) nanorods. The antimicrobial characteristic of the scaffold has been retrieved from the prepared Al-mHA frame with high aspect ratio (∼14.2) via biosynthesis route using Aloe vera (Aloe barbadensis miller) extract. The Al-mHA frame was introduced into an unprecedented SPU matrix (solution polymerized) based on combinatorial soft segments of poly(ε-caprolactone) (PCL), poly(ethylene carbonate) (PEC), and poly(dimethylsiloxane) (PDMS), by an in situ technique followed by electrospinning to fabricate scaffolds. For comparison, pristine mHA nanorods are also ornamented into it. An enzymatic ring-opening polymerization technique was adapted to synthesize soft segment of (PCL-PEC-b-PDMS). Structure elucidation of the synthesized polymers is established by nuclear magnetic resonance spectroscopy. Sparingly, Al-mHA ornamented scaffolds exhibit tremendous improvement (175%) in the mechanical properties with promising antimicrobial activity against various human pathogens. After confirmation of high osteoconductivity, improved biodegradation, and excellent biocompatibility against osteoblast-like MG63 cells (in vitro), the scaffolds were implanted in rabbits as an animal model by subcutaneous and intraosseous (tibial) sites. Improved in vivo biocompatibilities, biodegradation, osteoconductivity, and the ability to provide an adequate biomimetic environment for biomineralization for GBR of the scaffolds (SPU and ornamented SPUs) have been found from the various histological sections. Early cartilage formation, endochondral ossification, and rapid bone healing at 4 weeks were found in the defects filled with Al-mHA ornamented

  3. Fabrication of Bioceramic Bone Scaffolds for Tissue Engineering

    NASA Astrophysics Data System (ADS)

    Liu, Fwu-Hsing

    2014-10-01

    In this study, microhydroxyapatite and nanosilica sol were used as the raw materials for fabrication of bioceramic bone scaffold using selective laser sintering technology in a self-developed 3D Printing apparatus. When the fluidity of ceramic slurry is matched with suitable laser processing parameters, a controlled pore size of porous bone scaffold can be fabricated under a lower laser energy. Results shown that the fabricated scaffolds have a bending strength of 14.1 MPa, a compressive strength of 24 MPa, a surface roughness of 725 nm, a pore size of 750 μm, an apparent porosity of 32%, and a optical density of 1.8. Results indicate that the mechanical strength of the scaffold can be improved after heat treatment at 1200 °C for 2 h, while simultaneously increasing surface roughness conducive to osteoprogenitor cell adhesion. MTT method and SEM observations confirmed that bone scaffolds fabricated under the optimal manufacturing process possess suitable biocompatibility and mechanical properties, allowing smooth adhesion and proliferation of osteoblast-like cells. Therefore, they have great potential for development in the field of tissue engineering.

  4. Needle-like ion-doped hydroxyapatite crystals influence osteogenic properties of PCL composite scaffolds.

    PubMed

    Guarino, V; Veronesi, F; Marrese, M; Giavaresi, G; Ronca, A; Sandri, M; Tampieri, A; Fini, M; Ambrosio, Luigi

    2016-02-29

    Surface topography and chemistry both play a crucial role on influencing cell response in 3D porous scaffolds in terms of osteogenesis. Inorganic materials with peculiar morphology and chemical functionalities may be proficiently used to improve scaffold properties-in the bulk and along pore surface-promoting in vitro and in vivo osseous tissue in-growth. The present study is aimed at investigating how bone regenerative properties of composite scaffolds made of poly(Ɛ-caprolactone) (PCL) can be augmented by the peculiar properties of Mg(2+) ion doped hydroxyapatite (dHA) crystals, mainly emphasizing the role of crystal shape on cell activities mediated by microstructural properties. At the first stage, the study of mechanical response by crossing experimental compression tests and theoretical simulation via empirical models, allow recognizing a significant contribution of dHA shape factor on scaffold elastic moduli variation as a function of the relative volume fraction. Secondly, the peculiar needle-like shape of dHA crystals also influences microscopic (i.e. crystallinity, adhesion forces) and macroscopic (i.e. roughness) properties with relevant effects on biological response of the composite scaffold: differential scanning calorimetry (DSC) analyses clearly indicate a reduction of crystallization heat-from 66.75 to 43.05 J g(-1)-while atomic force microscopy (AFM) ones show a significant increase of roughness-from (78.15  ±  32.71) to (136.13  ±  63.21) nm-and of pull-off forces-from 33.7% to 48.7%. Accordingly, experimental studies with MG-63 osteoblast-like cells show a more efficient in vitro secretion of alkaline phosphatase (ALP) and collagen I and a more copious in vivo formation of new bone trabeculae, thus suggesting a relevant role of dHA to support the main mechanisms involved in bone regeneration.

  5. Control of the pore architecture in three-dimensional hydroxyapatite-reinforced hydrogel scaffolds.

    PubMed

    Román, Jesús; Cabañas, María Victoria; Peña, Juan; Vallet-Regí, María

    2011-08-01

    Hydrogels (gellan or agarose) reinforced with nanocrystalline carbonated hydroxyapatite (nCHA) were prepared by the GELPOR3D technique. This simple method is characterized by compositional flexibility; it does not require expensive equipment, thermal treatment, or aggressive or toxic solvents, and yields a three-dimensional (3D) network of interconnected pores 300-900 μm in size. In addition, an interconnected porosity is generated, yielding a hierarchical porous architecture from the macro to the molecular scale. This porosity depends on both the drying/preservation technology (freeze drying or oven drying at 37 (○)C) and on the content and microstructure of the reinforcing ceramic. For freeze-dried samples, the porosities were approximately 30, 66 and below 3% for pore sizes of 600-900 μm, 100-200 μm and 50-100 nm, respectively. The pore structure depends much on the ceramic content, so that higher contents lead to the disappearance of the characteristic honeycomb structure observed in low-ceramic scaffolds and to a lower fraction of the 100-200-μm-sized pores. The nature of the hydrogel did not affect the pore size distribution but was crucial for the behavior of the scaffolds in a hydrated medium: gellan-containing scaffolds showed a higher swelling degree owing to the presence of more hydrophilic groups.

  6. Assisted deposition of nano-hydroxyapatite onto exfoliated carbon nanotube oxide scaffolds

    NASA Astrophysics Data System (ADS)

    Zanin, H.; Rosa, C. M. R.; Eliaz, N.; May, P. W.; Marciano, F. R.; Lobo, A. O.

    2015-05-01

    Electrodeposited nano-hydroxyapatite (nHAp) is more similar to biological apatite in terms of microstructure and dimension than apatites prepared by other processes. Reinforcement with carbon nanotubes (CNTs) enhances its mechanical properties and increases adhesion of osteoblasts. Here, we carefully studied nHAp deposited onto vertically aligned multi-walled CNT (VAMWCNT) scaffolds by electrodeposition and soaking in a simulated body fluid (SBF). VAMWCNTs are porous biocompatible scaffolds with nanometric porosity and exceptional mechanical and chemical properties. The VAMWCNT films were prepared on a Ti substrate by a microwave plasma chemical vapour deposition method, and then oxidized and exfoliated by oxygen plasma etching (OPE) to produce graphene oxide (GO) at the VAMWCNT tips. The attachment of oxygen functional groups was found to be crucial for nHAp nucleation during electrodeposition. A thin layer of plate-like and needle-like nHAp with high crystallinity was formed without any need for thermal treatment. This composite (henceforth referred to as nHAp-VAMWCNT-GO) served as the scaffold for in vitro biomineralization when soaked in the SBF, resulting in the formation of both carbonate-rich and carbonate-poor globular-like nHAp. Different steps in the deposition of biological apatite onto VAMWCNT-GO and during the short-term biomineralization process were analysed. Due to their unique structure and properties, such nano-bio-composites may become useful in accelerating in vivo bone regeneration processes.

  7. Evaluation of nanoparticles of hydroxyapatite and MWCNT’s in scaffolds of poly lactic acid

    NASA Astrophysics Data System (ADS)

    Román-Doval, R.; Morales-Corona, J.; Olayo, R.; Escamilla-Rivera, V.; Uribe-Ramírez, M.; Ortega-López, M.

    2016-12-01

    In the tissue engineering, the cytotoxicity test is an important part of the biomaterials performance. This research reports the production and characterization of polylactic acid (PLA)-supported hydroxyapatite (HA) and multiwalled carbon nanotubes (MWCNT) scaffolds as a bone graft material. Samples containing different HA/MWCNT wt% ratios were prepared by electrospinning. The obtained samples displayed valuable characteristics for the cell adhesion because of their porous-spongy bone-like morphology. The Fourier transforms infrared and Raman analyses indicated no chemical interaction of HA and MWCNT with PLA molecules, but they appear to be only embedded into the PLA fibers. As indicated by x-ray diffraction, crystalline HA and MWCNT’s are supported in the amorphous PLA fibers. Under tensile stress, scaffolds display a Young’s Modulus about 86 MPa, whilst the scaffolds resistance increases with the HA-MWCNT’s ratio. However, the MTS in-vitro assays using the hFOB 1.19 (ATCC CRL-11372) cells, for cell exposure time of 24 and 48 h, revealed that viability reduces for HA-MWCNT’s ratio values over 25 wt%. Our results suggest that a maximum HA/MWCNT’s ratio of 19:1 could be acceptable for cell proliferation while maintaining HA at 200 mg.

  8. Control of the pore architecture in three-dimensional hydroxyapatite-reinforced hydrogel scaffolds

    PubMed Central

    Román, Jesús; Cabañas, María Victoria; Peña, Juan; Vallet-Regí, María

    2011-01-01

    Hydrogels (gellan or agarose) reinforced with nanocrystalline carbonated hydroxyapatite (nCHA) were prepared by the GELPOR3D technique. This simple method is characterized by compositional flexibility; it does not require expensive equipment, thermal treatment, or aggressive or toxic solvents, and yields a three-dimensional (3D) network of interconnected pores 300–900 μm in size. In addition, an interconnected porosity is generated, yielding a hierarchical porous architecture from the macro to the molecular scale. This porosity depends on both the drying/preservation technology (freeze drying or oven drying at 37 ○C) and on the content and microstructure of the reinforcing ceramic. For freeze-dried samples, the porosities were approximately 30, 66 and below 3% for pore sizes of 600–900 μm, 100–200 μm and 50–100 nm, respectively. The pore structure depends much on the ceramic content, so that higher contents lead to the disappearance of the characteristic honeycomb structure observed in low-ceramic scaffolds and to a lower fraction of the 100–200-μm-sized pores. The nature of the hydrogel did not affect the pore size distribution but was crucial for the behavior of the scaffolds in a hydrated medium: gellan-containing scaffolds showed a higher swelling degree owing to the presence of more hydrophilic groups. PMID:27877422

  9. Levofloxacin loaded mesoporous silica microspheres/nano-hydroxyapatite/polyurethane composite scaffold for the treatment of chronic osteomyelitis with bone defects.

    PubMed

    Wang, Qi; Chen, Cheng; Liu, Wen; He, Xiaoqiang; Zhou, Nian; Zhang, Dongli; Gu, Hongchen; Li, Jidong; Jiang, Jiaxing; Huang, Wei

    2017-02-02

    Chronic osteomyelitis is a prolonged persistent disease accompanied by bone destruction and sequestrum formation, it is very difficult to treat. Antibiotic loaded polymethyl methacrylate (PMMA) has been used in clinical. However, when PMMA was implanted in the body, the deficiencies is that it is non-biodegradable and a second operation is needed. Here, we synthesize a novel levofloxacin loaded mesoporous silica microspheres/nano-hydroxyapatite/polyurethane composite scaffolds, and evaluated the therapeutic effect in treating chronic osteomyelitis with bone defects in rabbit model compared with bulk PMMA. X-ray, Micro CT, gross pathology as well as immunohistochemical staining were performed at predesignated time points (1, 3, 6 and 12 weeks). Our results demonstrated that the efficiency of mesoporous silica microspheres/nano-hydroxyapatite/polyurethane composite scaffolds loaded with 5 mg levofloxacin was much better at treating bone defects than the other groups. This novel synthetic scaffold may provide a solution for the treatment of chronic osteomyelitis.

  10. Levofloxacin loaded mesoporous silica microspheres/nano-hydroxyapatite/polyurethane composite scaffold for the treatment of chronic osteomyelitis with bone defects

    PubMed Central

    Wang, Qi; Chen, Cheng; Liu, Wen; He, Xiaoqiang; Zhou, Nian; Zhang, Dongli; Gu, Hongchen; Li, Jidong; Jiang, Jiaxing; Huang, Wei

    2017-01-01

    Chronic osteomyelitis is a prolonged persistent disease accompanied by bone destruction and sequestrum formation, it is very difficult to treat. Antibiotic loaded polymethyl methacrylate (PMMA) has been used in clinical. However, when PMMA was implanted in the body, the deficiencies is that it is non-biodegradable and a second operation is needed. Here, we synthesize a novel levofloxacin loaded mesoporous silica microspheres/nano-hydroxyapatite/polyurethane composite scaffolds, and evaluated the therapeutic effect in treating chronic osteomyelitis with bone defects in rabbit model compared with bulk PMMA. X-ray, Micro CT, gross pathology as well as immunohistochemical staining were performed at predesignated time points (1, 3, 6 and 12 weeks). Our results demonstrated that the efficiency of mesoporous silica microspheres/nano-hydroxyapatite/polyurethane composite scaffolds loaded with 5 mg levofloxacin was much better at treating bone defects than the other groups. This novel synthetic scaffold may provide a solution for the treatment of chronic osteomyelitis. PMID:28150731

  11. Development of gelatin-chitosan-hydroxyapatite based bioactive bone scaffold with controlled pore size and mechanical strength.

    PubMed

    Maji, Kanchan; Dasgupta, Sudip; Kundu, Biswanath; Bissoyi, Akalabya

    2015-01-01

    Hydroxyapatite-chitosan/gelatin (HA:Chi:Gel) nanocomposite scaffold has potential to serve as a template matrix to regenerate extra cellular matrix of human bone. Scaffolds with varying composition of hydroxyapatite, chitosan, and gelatin were prepared using lyophilization technique where glutaraldehyde (GTA) acted as a cross-linking agent for biopolymers. First, phase pure hydroxyapatite-chitosan nanocrystals were in situ synthesized by coprecipitation method using a solution of 2% acetic acid dissolved chitosan and aqueous solution of calcium nitrate tetrahydrate [Ca(NO3)2,4H2O] and diammonium hydrogen phosphate [(NH4)2H PO4]. Keeping solid loading constant at 30 wt% and changing the composition of the original slurry of gelatin, HA-chitosan allowed control of the pore size, its distribution, and mechanical properties of the scaffolds. Microstructural investigation by scanning electron microscopy revealed the formation of a well interconnected porous scaffold with a pore size in the range of 35-150 μm. The HA granules were uniformly dispersed in the gelatin-chitosan network. An optimal composition in terms of pore size and mechanical properties was obtained from the scaffold with an HA:Chi:Gel ratio of 21:49:30. The composite scaffold having 70% porosity with pore size distribution of 35-150 μm exhibited a compressive strength of 3.3-3.5 MPa, which is within the range of that exhibited by cancellous bone. The bioactivity of the scaffold was evaluated after conducting mesenchymal stem cell (MSC) - materials interaction and MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay using MSCs. The scaffold found to be conducive to MSC's adhesion as evident from lamellipodia, filopodia extensions from cell cytoskeleton, proliferation, and differentiation up to 14 days of cell culture.

  12. Fabrication of pillared PLGA microvessel scaffold using femtosecond laser ablation

    PubMed Central

    Wang, Hsiao-Wei; Cheng, Chung-Wei; Li, Ching-Wen; Chang, Han-Wei; Wu, Ping-Han; Wang, Gou-Jen

    2012-01-01

    One of the persistent challenges confronting tissue engineering is the lack of intrinsic microvessels for the transportation of nutrients and metabolites. An artificial microvascular system could be a feasible solution to this problem. In this study, the femtosecond laser ablation technique was implemented for the fabrication of pillared microvessel scaffolds of polylactic-co-glycolic acid (PLGA). This novel scaffold facilitates implementation of the conventional cell seeding process. The progress of cell growth can be observed in vitro by optical microscopy. The problems of becoming milky or completely opaque with the conventional PLGA scaffold after cell seeding can be resolved. In this study, PLGA microvessel scaffolds consisting of 47 μm × 80 μm pillared branches were produced. Results of cell culturing of bovine endothelial cells demonstrate that the cells adhere well and grow to surround each branch of the proposed pillared microvessel networks. PMID:22605935

  13. Fabrication, Properties and Applications of Dense Hydroxyapatite: A Review

    PubMed Central

    Prakasam, Mythili; Locs, Janis; Salma-Ancane, Kristine; Loca, Dagnija; Largeteau, Alain; Berzina-Cimdina, Liga

    2015-01-01

    In the last five decades, there have been vast advances in the field of biomaterials, including ceramics, glasses, glass-ceramics and metal alloys. Dense and porous ceramics have been widely used for various biomedical applications. Current applications of bioceramics include bone grafts, spinal fusion, bone repairs, bone fillers, maxillofacial reconstruction, etc. Amongst the various calcium phosphate compositions, hydroxyapatite, which has a composition similar to human bone, has attracted wide interest. Much emphasis is given to tissue engineering, both in porous and dense ceramic forms. The current review focusses on the various applications of dense hydroxyapatite and other dense biomaterials on the aspects of transparency and the mechanical and electrical behavior. Prospective future applications, established along the aforesaid applications of hydroxyapatite, appear to be promising regarding bone bonding, advanced medical treatment methods, improvement of the mechanical strength of artificial bone grafts and better in vitro/in vivo methodologies to afford more particular outcomes. PMID:26703750

  14. Nanoparticle fabrication of hydroxyapatite by laser ablation in water

    SciTech Connect

    Musaev, O. R.; Wieliczka, D. M.; Wrobel, J. M.; Kruger, M. B.; Dusevich, V.

    2008-10-15

    Synthetic polycrystalline hydroxyapatite was ablated in water with 337 nm radiation from a UV nitrogen pulsed laser. According to transmission electron microscopy micrographs, the ablated particles were approximately spherical and had a size of {approx}80 nm. Raman spectroscopic analysis demonstrated that particles had the same structure as the original crystal. X-ray photoelectron spectroscopy showed that the surface chemical composition was close to that of the original material. The characteristics of the ablated particles and estimations of the temperature rise of the hydroxyapatite surface under laser irradiation are consistent with the mechanism of explosive boiling being responsible for ablation. The experimental observations offer the basis for preparation of hydroxyapatite nanoparticles by laser ablation in water.

  15. Fabrication and application of nanofibrous scaffolds in tissue engineering.

    PubMed

    Li, Wan-Ju; Tuan, Rocky S

    2009-03-01

    Nanofibers fabricated by electrospinning are morphological mimics of fibrous components of the native extracellular matrix, making nanofibrous scaffolds ideal for three-dimensional cell culture and tissue engineering applications. Although electrospinning is not a conventional technique in cell biology, the experimental setup may be constructed in a relatively straightforward manner, and the procedure can be carried out by individuals with limited engineering experience. Here, we detail a protocol for electrospinning of nanofibers and provide relevant specific details concerning the optimization of fiber formation (Basic Protocol 1). The protocol also includes conditions required for preparing biodegradable polymer solutions for the fabrication of nonwoven and aligned nanofibrous scaffolds suitable for various cell/tissue applications. In addition, information on effective cell loading into nanofibrous scaffolds and cellular constructs grown in a bioreactor is provided (Basic Protocol 2). Instructions for building the electrospinning apparatus are also included (see the Support Protocol).

  16. Porous scaffolds of polycaprolactone reinforced with in situ generated hydroxyapatite for bone tissue engineering.

    PubMed

    Fabbri, Paola; Bondioli, Federica; Messori, Massimo; Bartoli, Cristina; Dinucci, Dinuccio; Chiellini, Federica

    2010-01-01

    Polycaprolactone/hydroxyapatite (PCL/HA) composites were prepared by in situ generation of HA in the polymer solution starting from the precursors calcium nitrate tetrahydrate and ammonium dihydrogen phosphate via sol-gel process. Highly interconnected porosity was achieved by means of the salt-leaching technique using a mixture of sodium chloride and sodium bicarbonate as porogens. Structure and morphology of the PCL/HA composites were investigated by scanning electron microscopy, and mechanical properties were determined by means of tensile and compression tests. The possibility to employ the developed composites as scaffolds for bone tissue regeneration was assessed by cytotoxicity test of the PCL/HA composites extracts and cell adhesion and proliferation in vitro studies.

  17. Bone regeneration in critical bone defects using three-dimensionally printed β-tricalcium phosphate/hydroxyapatite scaffolds is enhanced by coating scaffolds with either dipyridamole or BMP-2.

    PubMed

    Ishack, Stephanie; Mediero, Aranzazu; Wilder, Tuere; Ricci, John L; Cronstein, Bruce N

    2017-02-01

    Bone defects resulting from trauma or infection need timely and effective treatments to restore damaged bone. Using specialized three-dimensional (3D) printing technology we have created custom 3D scaffolds of hydroxyapatite (HA)/beta-tri-calcium phosphate (β-TCP) to promote bone repair. To further enhance bone regeneration we have coated the scaffolds with dipyridamole, an agent that increases local adenosine levels by blocking cellular uptake of adenosine. Nearly 15% HA:85% β-TCP scaffolds were designed using Robocad software, fabricated using a 3D Robocasting system, and sintered at 1100°C for 4 h. Scaffolds were coated with BMP-2 (200 ng mL(-1) ), dypiridamole 100 µM or saline and implanted in C57B6 and adenosine A2A receptor knockout (A2AKO) mice with 3 mm cranial critical bone defects for 2-8 weeks. Dipyridamole release from scaffold was assayed spectrophotometrically. MicroCT and histological analysis were performed. Micro-computed tomography (microCT) showed significant bone formation and remodeling in HA/β-TCP-dipyridamole and HA/β-TCP-BMP-2 scaffolds when compared to scaffolds immersed in vehicle at 2, 4, and 8 weeks (n = 5 per group; p ≤ 0.05, p ≤ 0.05, and p ≤ 0.01, respectively). Histological analysis showed increased bone formation and a trend toward increased remodeling in HA/β-TCP- dipyridamole and HA/β-TCP-BMP-2 scaffolds. Coating scaffolds with dipyridamole did not enhance bone regeneration in A2AKO mice. In conclusion, scaffolds printed with HA/β-TCP promote bone regeneration in critical bone defects and coating these scaffolds with agents that stimulate A2A receptors and growth factors can further enhance bone regeneration. These coated scaffolds may be very useful for treating critical bone defects due to trauma, infection or other causes. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 366-375, 2017.

  18. The influence of plasma technology coupled to chemical grafting on the cell growth compliance of 3D hydroxyapatite scaffolds.

    PubMed

    Russo, Laura; Zanini, Stefano; Giannoni, Paolo; Landi, Elena; Villa, Anna; Sandri, Monica; Riccardi, Claudia; Quarto, Rodolfo; Doglia, Silvia M; Nicotra, Francesco; Cipolla, Laura

    2012-11-01

    The development of advanced materials with biomimetic features in order to elicit desired biological responses and to guarantee tissue biocompatibility is recently gaining attention for tissue engineering applications. Bioceramics, such as hydroxyapatite-based biomaterials are now used in a number of different applications throughout the body, covering all areas of the skeleton, due to their biological and chemical similarity to the inorganic phases of bones. When bioactive sintered hydroxyapatite (HA) is desired, biomolecular modification of these materials is needed. In the present work, we investigated the influence of plasma surface modification coupled to chemical grafting on the cell growth compliance of HA 3D scaffolds.

  19. Design, fabrication, and characterization of laminated hydroxyapatite-polysulfone composites

    NASA Astrophysics Data System (ADS)

    Wilson, Clifford Adams, II

    There exists a need to develop devices that can be used to replace hard tissues, such as bone, in load-bearing areas of the body. An ideal hard tissue replacement device is one that stimulates growth of natural tissues, and is slowly resorbed by the body. The implant is also required to have elastic modulus, strength, and toughness values similar to the tissues being replaced. Hydroxyapatite (HA) is the primary mineral phase of bone and has the potential for use in biomedical applications because it stimulates cell growth and is resorbable. Unfortunately, HA is a relatively low strength, low toughness material, which limits its application to only low load-bearing regions of the body. In order to apply HA to greater load-bearing areas of the body, strength and toughness must be improved through the formation of a composite structure. The goal of this study to show that a composite structure formed from HA and a biocompatible polymer can be fabricated with strength and toughness values that are within the range necessary for load-bearing biomedical applications. Therefore, Polysulfone-HA composites were developed and tested. Polysulfone (PSu) is a hard, glassy polymer that has been shown to be biocompatible. Composites were fabricated through a combination of tape casting, solvent casting, and lamination. Monolithic HA and laminate specimens were tested in biaxial flexure. A unique laminate theory solution was developed to characterize stress distributions for laminates. Failure loads, failure stress, work of fracture, and apparent toughness were compared for the laminates against monolithic HA specimens. Initial testing results showed that laminates had a failure stress of 60 +/- 10, which is a 170% improvement over the 22 +/- 2 MPa failure stress for monolithic HA. The work of fracture was improved by 5500% from 11 +/- 2 for the monolithic HA to 612 +/- 240 for the laminates. Work of fracture values gave the laminates an apparent fracture toughness of 7.2 MPa•m1

  20. Fabrication of cancellous biomimetic chitosan-based nanocomposite scaffolds applying a combinational method for bone tissue engineering.

    PubMed

    Jamalpoor, Zahra; Mirzadeh, Hamid; Joghataei, Mohammad Taghi; Zeini, Darya; Bagheri-Khoulenjani, Shadab; Nourani, Mohammad Reza

    2015-05-01

    The aim of this study was to mimic the specific structure of bone and fabricate a biomimetic nano-hydroxyapatite (HA)/chitosan (Cs)/gelatin scaffolds using combination of particle leaching and freeze drying methods eliminating mold effects. To achieve an optimum structure, scaffolds with different gelatin/Cs weight ratio were fabricated. Morphological characterization of scaffolds by scanning electron microscopy method showed highly interconnected porous structures similar to cancellous bone with mean pore size ranging from 140 to 190 μm. Nano-HA crystals were dispersed homogeneously in the polymer matrix according to the energy-dispersive X-ray spectroscopy and transmission electron microscopy images. Fourier transform infrared and X-ray diffraction results disclosed that chemical interactions were formed between nano-HA, Cs, gelatin and crystallinity of each material decreased with blending. It was found that increasing the gelatin content significantly improved water uptake, degradation rate as well as attachment, infiltration and proliferation of Saos2 cells to the scaffolds. The presented results confirm that the designed biomimetic nano-HA /Cs/gelatin scaffolds can be used as promising substitutes for bone tissue engineering.

  1. Repair of segmental long bone defect in a rabbit radius nonunion model: comparison of cylindrical porous titanium and hydroxyapatite scaffolds.

    PubMed

    Zhang, Ming; Wang, Guang-lin; Zhang, Hong-fang; Hu, Xu-dong; Shi, Xiao-yuan; Li, Sen; Lin, Wei

    2014-06-01

    A segmental long bone defect in a rabbit radius nonunion model was repaired using cylindrical porous titanium (Ti) and hydroxyapatite (HA) scaffolds. Each scaffold was produced using the same method, namely, a slurry foaming method. Repairing ability was characterized using x-radiographic score 12 and 24 weeks postprocedure; failure load of the radius-ulna construct, under three-point bending, 12 weeks postprocedure; and the percentage of newly formed bone within the implant, 12 and 24 weeks after postprocedure. For each of these parameters, the difference in the results when porous Ti scaffold was used compared with when HA scaffolds were used was not significant; both porous scaffolds showed excellent repairing ability. Because the trabecular bone is a porous tissue, the interconnected porous scaffolds have the advantages of natural bone, and vasculature can grow into the porous structure to accelerate the osteoconduction and osteointegration between the implant and bone. The porous Ti scaffold not only enhanced the bone repair process, similar to porous HA scaffolds, but also has superior biomechanical properties. The present results suggest that porous Ti scaffolds may have promise for use in the clinical setting.

  2. Protein Corona of Magnetic Hydroxyapatite Scaffold Improves Cell Proliferation via Activation of Mitogen-Activated Protein Kinase Signaling Pathway.

    PubMed

    Zhu, Yue; Yang, Qi; Yang, Minggang; Zhan, Xiaohui; Lan, Fang; He, Jing; Gu, Zhongwei; Wu, Yao

    2017-03-21

    The beneficial effect of magnetic scaffolds on the improvement of cell proliferation has been well documented. Nevertheless, the underlying mechanisms about the magnetic scaffolds stimulating cell proliferation remain largely unknown. Once the scaffold enters into the biological fluids, a protein corona forms and directly influences the biological function of scaffold. This study aimed at investigating the formation of protein coronas on hydroxyapatite (HA) and magnetic hydroxyapatite (MHA) scaffolds in vitro and in vivo, and consequently its effect on regulating cell proliferation. The results demonstrated that magnetic nanoparticles (MNP)-infiltrated HA scaffolds altered the composition of protein coronas and ultimately contributed to increased concentration of proteins related to calcium ions, G-protein coupled receptors (GPCRs), and MAPK/ERK cascades as compared with pristine HA scaffolds. Noticeably, the enriched functional proteins on MHA samples could efficiently activate of the MAPK/ERK signaling pathway, resulting in promoting MC3T3-E1 cell proliferation, as evidenced by the higher expression levels of the key proteins in the MAPK/ERK signaling pathway, including mitogen-activated protein kinase kinases1/2 (MEK1/2) and extracellular signal regulated kinase 1/2 (ERK1/2). Artificial down-regulation of MEK expression can significantly down-regulate the MAPK/ERK signaling and consequently suppress the cell proliferation on MHA samples. These findings not only provide a critical insight into the molecular mechanism underlying cellular proliferation on magnetic scaffolds, but also have important implications in the design of magnetic scaffolds for bone tissue engineering.

  3. Hydroxyapatite nanoparticle-containing scaffolds for the study of breast cancer bone metastasis.

    PubMed

    Pathi, Siddharth P; Lin, Debra D W; Dorvee, Jason R; Estroff, Lara A; Fischbach, Claudia

    2011-08-01

    Breast cancer frequently metastasizes to bone, where it leads to secondary tumor growth, osteolytic bone degradation, and poor clinical prognosis. Hydroxyapatite Ca(10)(PO(4))(6)(OH)(2) (HA), a mineral closely related to the inorganic component of bone, may be implicated in these processes. However, it is currently unclear how the nanoscale materials properties of bone mineral, such as particle size and crystallinity, which change as a result of osteolytic bone remodeling, affect metastatic breast cancer. We have developed a two-step hydrothermal synthesis method to obtain HA nanoparticles with narrow size distributions and varying crystallinity. These nanoparticles were incorporated into gas-foamed/particulate leached poly(lactide-co-glycolide) scaffolds, which were seeded with metastatic breast cancer cells to create mineral-containing scaffolds for the study of breast cancer bone metastasis. Our results suggest that smaller, poorly-crystalline HA nanoparticles promote greater adsorption of adhesive serum proteins and enhance breast tumor cell adhesion and growth relative to larger, more crystalline nanoparticles. Conversely, the larger, more crystalline HA nanoparticles stimulate enhanced expression of the osteolytic factor interleukin-8 (IL-8). Our data suggest an important role for nanoscale HA properties in the vicious cycle of bone metastasis and indicate that mineral-containing tumor models may be excellent tools to study cancer biology and to define design parameters for non-tumorigenic mineral-containing or mineralized matrices for bone regeneration.

  4. Extraction and characterization of biocompatible hydroxyapatite from fresh water fish scales for tissue engineering scaffold.

    PubMed

    Panda, Niladri Nath; Pramanik, Krishna; Sukla, Lala Behari

    2014-03-01

    In bone tissue engineering, porous hydroxyapatite (HAp) is used as filling material for bone defects, augmentation, artificial bone graft and scaffold material. The present paper compares the preparation and characterization of HAp from fish scale (FS) and synthetic body fluid (SBF) solution. Thermo gravimetric analysis, differential thermal analysis, Fourier transform infrared spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM) and particle size analysis of the samples have been performed. The analysis indicates that synthesized HAp consists of sub-micron HAp particle with Ca/P ratio corresponding to FS and SBF 1.62 and 1.71, respectively. MTT assay and quantitative DNA analysis show growth and proliferation of cells over the HA scaffold with the increase in time. The shape and size (morphology) of mesenchymal stem cells after 3 days show a transition from rounded shape to elongated and flattened shape expressing its spreading behavior. These results confirm that HAp bio-materials from fish scale are physico-chemically and biologically equivalent to the chemically synthesized HAp from SBF. Biological HAp, thus, possesses a great potential for conversion of industrial by-product into highly valuable compounds using simple effective and novel processes.

  5. Freeze casting of porous hydroxyapatite scaffolds. I. Processing and general microstructure.

    PubMed

    Fu, Qiang; Rahaman, Mohamed N; Dogan, Fatih; Bal, B Sonny

    2008-07-01

    Freeze casting of aqueous suspensions on a cold substrate was investigated as a method for preparing hydroxyapatite (HA) scaffolds with unidirectional porosity. In the present paper, we report on the ability to manipulate the microstructure of freeze-cast constructs by controlling the processing parameters. Constructs prepared from aqueous suspensions (5-20 volume percent particles) on a steel substrate at -20 degrees C had a lamellar-type microstructure, consisting of plate-like HA and unidirectional pores oriented in the direction of freezing. Sintering for 3 h at 1350 degrees C produced constructs with dense HA lamellas, porosity of approximately 50%, and inter-lamellar pore widths of 5-30 microm. The thickness of the HA lamellas decreased but the width of the pores increased with decreasing particle concentration. Decreasing the substrate temperature from -20 degrees C to -196 degrees C produced a finer lamellar microstructure. The use of water-glycerol mixtures (20 wt % glycerol) as the solvent in the suspension resulted in the production of finer pores (1-10 microm) and a larger number of dendritic growth connecting the HA lamellas. On the other hand, the use of water-dioxane mixtures (60 wt % dioxane) produced a cellular-type microstructure with larger pores (90-110 microm). The ability to produce a uniaxial microstructure and its manipulation by controlling the processing parameters indicate the potential of the present freeze casting route for the production of scaffolds for bone tissue engineering applications.

  6. Preparation of collagen/hydroxyapatite/alendronate hybrid hydrogels as potential scaffolds for bone regeneration.

    PubMed

    Ma, Xin; He, Zhiwei; Han, Fengxuan; Zhong, Zhiyuan; Chen, Liang; Li, Bin

    2016-07-01

    Development of biomimetic scaffolds represents a promising direction in bone tissue engineering. In this study, we designed a two-step process to prepare a type of biomimetic hybrid hydrogels that were composed of collagen, hydroxyapatite (HAP) and alendronate (ALN), an anti-osteoporosis drug. First, water-soluble ALN-conjugated HAP (HAP-ALN) containing 4.0wt.% of ALN was synthesized by treating HAP particles with ALN. Hydrogels were then formed from HAP-ALN conjugate and collagen under physiological conditions using genipin (GNP) as the crosslinker. Depending on the ALN/collagen molar ratio and GNP concentration, the gelation time of hydrogels ranged from 5 to 37min. Notably, these hybrid hydrogels exhibited markedly improved mechanical property (storage modulus G'=38-187kPa), higher gel contents, and lower swelling ratios compared to the hydrogels prepared from collagen alone under similar conditions. Moreover, they showed tunable degradation behaviors against collagenase. The collagen/HAP-ALN hybrid hydrogels supported the adhesion and growth of murine MC3T3-E1 osteoblastic cells well. Such tough yet enzymatically degradable hybrid hydrogels hold potential as scaffolds for bone tissue engineering. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. A novel two-step sintering for nano-hydroxyapatite scaffolds for bone tissue engineering

    PubMed Central

    Feng, Pei; Niu, Man; Gao, Chengde; Peng, Shuping; Shuai, Cijun

    2014-01-01

    In this study, nano-hydroxyapatite scaffolds with high mechanical strength and an interconnected porous structure were prepared using NTSS for the first time. The first step was performed using a laser characterized by the rapid heating to skip the surface diffusion and to obtain the driving force for grain boundary diffusion. Additionally, the interconnected porous structure was achieved by SLS. The second step consisted of isothermal heating in a furnace at a lower temperature (T2) than that of the laser beam to further increase the density and to suppress grain growth by exploiting the difference in kinetics between grain-boundary diffusion and grain-boundary migration. The results indicated that the mechanical properties first increased and then decreased as T2 was increased from 1050 to 1250°C. The optimal fracture toughness, compressive strength and stiffness were 1.69 MPam1/2, 18.68 MPa and 245.79 MPa, respectively. At the optimal point, the T2 was 1100°C, the grain size was 60 nm and the relative density was 97.6%. The decrease in mechanical properties was due to the growth of grains and the decomposition of HAP. The cytocompatibility test results indicated that cells adhered and spread well on the scaffolds. A bone-like apatite layer formed, indicating good bioactivity. PMID:24998362

  8. A novel two-step sintering for nano-hydroxyapatite scaffolds for bone tissue engineering.

    PubMed

    Feng, Pei; Niu, Man; Gao, Chengde; Peng, Shuping; Shuai, Cijun

    2014-07-07

    In this study, nano-hydroxyapatite scaffolds with high mechanical strength and an interconnected porous structure were prepared using NTSS for the first time. The first step was performed using a laser characterized by the rapid heating to skip the surface diffusion and to obtain the driving force for grain boundary diffusion. Additionally, the interconnected porous structure was achieved by SLS. The second step consisted of isothermal heating in a furnace at a lower temperature (T2) than that of the laser beam to further increase the density and to suppress grain growth by exploiting the difference in kinetics between grain-boundary diffusion and grain-boundary migration. The results indicated that the mechanical properties first increased and then decreased as T2 was increased from 1050 to 1250°C. The optimal fracture toughness, compressive strength and stiffness were 1.69 MPam(1/2), 18.68 MPa and 245.79 MPa, respectively. At the optimal point, the T2 was 1100°C, the grain size was 60 nm and the relative density was 97.6%. The decrease in mechanical properties was due to the growth of grains and the decomposition of HAP. The cytocompatibility test results indicated that cells adhered and spread well on the scaffolds. A bone-like apatite layer formed, indicating good bioactivity.

  9. Fabrication and Microstructure of Hydroxyapatite Coatings on Zirconia by Room Temperature Spray Process.

    PubMed

    Seo, Dong Seok; Chae, Hak Cheol; Lee, Jong Kook

    2015-08-01

    Hydroxyapatite coatings were fabricated on zirconia substrates by a room temperature spray process and were investigated with regards to their microstructure, composition and dissolution in water. An initial hydroxyapatite powder was prepared by heat treatment of bovine-bone derived powder at 1100 °C for 2 h, while dense zirconia substrates were fabricated by pressing 3Y-TZP powder and sintering it at 1350 °C for 2 h. Room temperature spray coating was performed using a slit nozzle in a low pressure-chamber with a controlled coating time. The phase composition of the resultant hydroxyapatite coatings was similar to that of the starting powder, however, the grain size of the hydroxyapatite particles was reduced to about 100 nm due to their formation by particle impaction and fracture. All areas of the coating had a similar morphology, consisting of reticulated structure with a high surface roughness. The hydroxyapatite coating layer exhibited biostability in a stimulated body fluid, with no severe dissolution being observed during in vitro experimentation.

  10. Regeneration of a goat femoral head using a tissue-specific, biphasic scaffold fabricated with CAD/CAM technology.

    PubMed

    Ding, Chunming; Qiao, Zhiguang; Jiang, Wenbo; Li, Haowei; Wei, Jianhe; Zhou, Guangdong; Dai, Kerong

    2013-09-01

    Tissue engineering is considered as a promising approach for the regeneration of biological joint theoretically and thus provides a potential treatment option for advanced osteoarthritis. However, no significant progresses so far have been made in regenerating biological joint. In this study, a biphasic scaffold, which was consisted of polylactic acid-coated polyglycolic acid (PGA/PLA) scaffold and poly-ε-caprolactone/hydroxyapatite (PCL/HA) scaffold, was designed and used for regeneration of goat femoral head. The content of PLA and HA was optimized to a proper ratio, thus the scaffolds could achieve appropriate stiffness which was more conducive to articular cartilage and bone regeneration respectively. Furthermore, computer-aided design and manufacturing (CAD/CAM) technology was employed to fabricate the biphasic scaffolds into the desired shape and structure. The biphasic scaffolds with fine cell biocompatibility matched perfectly. Chondrocytes and bone marrow stromal cells (BMSCs) were seeded into the scaffolds for cartilage and bone regeneration respectively. After 10 weeks of implantation in nude mice subcutaneously, the cell-scaffold constructs successfully regenerated goat femoral heads. The regenerated femoral heads presented a precise appearance in shape and size similar to that of native goat femoral heads with a smooth, continuous, avascular, and homogeneous cartilage layer on the surface and stiff bone-like tissue in the microchannels of PCL/HA scaffold. Additionally, histological examination of the regenerated cartilage and bone showed typical histological structures and biophysical properties similar to that of native ones with specific matrix deposition and a well-integrated osteochondral interface. The strategy established in the study provides a promising approach for regenerating a biological joint which could be used to reconstruct the impaired joint.

  11. Mag-seeding of rat bone marrow stromal cells into porous hydroxyapatite scaffolds for bone tissue engineering.

    PubMed

    Shimizu, Kazunori; Ito, Akira; Honda, Hiroyuki

    2007-09-01

    Bone tissue engineering has been investigated as an alternative strategy for autograft transplantation. In the process of tissue engineering, cell seeding into three-dimensional (3-D) scaffolds is the first step for constructing 3-D tissues. We have proposed a methodology of cell seeding into 3-D porous scaffolds using magnetic force and magnetite nanoparticles, which we term Mag-seeding. In this study, we applied this Mag-seeding technique to bone tissue engineering using bone marrow stromal cells (BMSCs) and 3-D hydroxyapatite (HA) scaffolds. BMSCs were magnetically labeled with our original magnetite cationic liposomes (MCLs) having a positive surface charge to improve adsorption to cell surface. Magnetically labeled BMSCs were seeded onto a scaffold, and a 1-T magnet was placed under the scaffold. By using Mag-seeding, the cells were successfully seeded into the internal space of scaffolds with a high cell density. The cell seeding efficiency into HA scaffolds by Mag-seeding was approximately threefold larger than that by static-seeding (conventional method, without a magnet). After a 14-d cultivation period using the osteogenic induction medium by Mag-seeding, the level of two representative osteogenic markers (alkaline phosphatase and osteocalcin) were significantly higher than those by static-seeding. These results indicated that Mag-seeding of BMSCs into HA scaffolds is an effective approach to bone tissue engineering.

  12. Design of a multiphase osteochondral scaffold. II. Fabrication of a mineralized collagen-glycosaminoglycan scaffold.

    PubMed

    Harley, Brendan A; Lynn, Andrew K; Wissner-Gross, Zachary; Bonfield, William; Yannas, Ioannis V; Gibson, Lorna J

    2010-03-01

    This paper is the second in a series of papers describing the design and development of an osteochondral scaffold using collagen-glycosaminoglycan and calcium phosphate technologies engineered for the regenerative repair of articular cartilage defects. The previous paper described a technology (concurrent mapping) for systematic variation and control of the chemical composition of triple coprecipitated collagen, glycosaminoglycan, and calcium phosphate (CGCaP) nanocomposites without using titrants. This paper describes (1) fabricating porous, three-dimensional scaffolds from the CGCaP suspensions, (2) characterizing the microstructure and mechanical properties of such scaffolds, and (3) modifying the calcium phosphate mineral phase. The methods build on the previously demonstrated ability to vary the composition of a CGCaP suspension (calcium phosphate mass fraction between 0 and 80 wt %) and enable the production of scaffolds whose pore architecture (mean pore size: 50-1000 microm), CaP phase chemistry (brushite, octacalcium phosphate, apatite) and crosslinking density (therefore mechanical properties and degradation rate) can be independently controlled. The scaffolds described in this paper combine the desirable biochemical properties and pore architecture of porous collagen-glycosaminoglycan scaffolds with the strength and direct bone-bonding properties of calcium phosphate biomaterials in a manner that can be tailored to meet the demands of a range of applications in orthopedics and regenerative medicine.

  13. Structure design and fabrication of porous hydroxyapatite microspheres for cell delivery

    NASA Astrophysics Data System (ADS)

    Li, Ruijing; Chen, Kexin; Li, Geng; Han, Guoxiang; Yu, Sheng; Yao, Juming; Cai, Yurong

    2016-09-01

    Porous microspheres fabricated from bioceramics have great potential for cell delivery in injectable tissue engineering application. The size and structure of pores in the microspheres are important for the effective protection and transportation of cells. In this study, porous hydroxyapatite microspheres are fabricated through the water-in-oil emulsion method followed by a calcination treatment at the high temperature. Both self-made resorcinol-formaldehyde (RF) composite spheres and camphene are used as pore-forming agents to produce big pores corresponding to the size of RF spheres and connected channel among big pores in hydroxyapatite matrix. The properties of the microspheres are characterized using X-ray diffraction, thermogravimetry analysis, universal material machine, field emission scanning electron microscopy. Cell assays are carried out to evaluate the cellular compatibility of the microspheres. The results showed that the hydroxyapatite microspheres with controllable pore structure and high porosity could be fabricated by this method, which have better strength to resist the compressive force. The microspheres are conducive to support adhesion, proliferation and differentiation of MC3T3-E1 cells. The results indicate that the obtained porous hydroxyapatite microspheres can be a permeable microenvironment for cell delivery in injectable tissue engineering.

  14. Chondrogenic regeneration using bone marrow clots and a porous polycaprolactone-hydroxyapatite scaffold by three-dimensional printing.

    PubMed

    Yao, Qingqiang; Wei, Bo; Liu, Nancy; Li, Chenshuang; Guo, Yang; Shamie, Arya Nick; Chen, James; Tang, Cheng; Jin, Chengzhe; Xu, Yan; Bian, Xiuwu; Zhang, Xinli; Wang, Liming

    2015-04-01

    Scaffolds play an important role in directing three-dimensional (3D) cartilage regeneration. Our recent study reported the potential advantages of bone marrow clots (MC) in promoting extracellular matrix (ECM) scaffold chondrogenic regeneration. The aim of this study is to build a new scaffold for MC, with improved characteristics in mechanics, shaping, and biodegradability, compared to our previous study. To address this issue, this study prepared a 3D porous polycaprolactone (PCL)-hydroxyapatite (HA) scaffold combined with MC (Group A), while the control group (Group B) utilized a bone marrow stem cell seeded PCL-HA scaffold. The results of in vitro cultures and in vivo implantation demonstrated that although an initial obstruction of nutrient exchange caused by large amounts of fibrin and erythrocytes led to a decrease in the ratio of live cells in Group A, these scaffolds also showed significant improvements in cell adhesion, proliferation, and chondrogenic differentiation with porous recanalization in the later culture, compared to Group B. After 4 weeks of in vivo implantation, Group A scaffolds have a superior performance in DNA content, Sox9 and RunX2 expression, cartilage lacuna-like cell and ECM accumulation, when compared to Group B. Furthermore, Group A scaffold size and mechanics were stable during in vitro and in vivo experiments, unlike the scaffolds in our previous study. Our results suggest that the combination with MC proved to be a highly efficient, reliable, and simple new method that improves the biological performance of 3D PCL-HA scaffold. The MC-PCL-HA scaffold is a candidate for future cartilage regeneration studies.

  15. Chondrogenic Regeneration Using Bone Marrow Clots and a Porous Polycaprolactone-Hydroxyapatite Scaffold by Three-Dimensional Printing

    PubMed Central

    Yao, Qingqiang; Wei, Bo; Liu, Nancy; Li, Chenshuang; Guo, Yang; Shamie, Arya Nick; Chen, James; Tang, Cheng; Jin, Chengzhe; Xu, Yan

    2015-01-01

    Scaffolds play an important role in directing three-dimensional (3D) cartilage regeneration. Our recent study reported the potential advantages of bone marrow clots (MC) in promoting extracellular matrix (ECM) scaffold chondrogenic regeneration. The aim of this study is to build a new scaffold for MC, with improved characteristics in mechanics, shaping, and biodegradability, compared to our previous study. To address this issue, this study prepared a 3D porous polycaprolactone (PCL)-hydroxyapatite (HA) scaffold combined with MC (Group A), while the control group (Group B) utilized a bone marrow stem cell seeded PCL-HA scaffold. The results of in vitro cultures and in vivo implantation demonstrated that although an initial obstruction of nutrient exchange caused by large amounts of fibrin and erythrocytes led to a decrease in the ratio of live cells in Group A, these scaffolds also showed significant improvements in cell adhesion, proliferation, and chondrogenic differentiation with porous recanalization in the later culture, compared to Group B. After 4 weeks of in vivo implantation, Group A scaffolds have a superior performance in DNA content, Sox9 and RunX2 expression, cartilage lacuna-like cell and ECM accumulation, when compared to Group B. Furthermore, Group A scaffold size and mechanics were stable during in vitro and in vivo experiments, unlike the scaffolds in our previous study. Our results suggest that the combination with MC proved to be a highly efficient, reliable, and simple new method that improves the biological performance of 3D PCL-HA scaffold. The MC-PCL-HA scaffold is a candidate for future cartilage regeneration studies. PMID:25530453

  16. Biomimetic scaffolds based on hydroxyapatite nanorod/poly(D,L) lactic acid with their corresponding apatite-forming capability and biocompatibility for bone-tissue engineering.

    PubMed

    Nga, Nguyen Kim; Hoai, Tran Thanh; Viet, Pham Hung

    2015-04-01

    This study presents a facile synthesis of biomimetic hydroxyapatite nanorod/poly(D,L) lactic acid (HAp/PDLLA) scaffolds with the use of solvent casting combined with a salt-leaching technique for bone-tissue engineering. Field emission scanning electron microscopy, Fourier transform infrared spectroscopy, and energy-dispersive X-ray spectroscopy were used to observe the morphologies, pore structures of synthesized scaffolds, interactions between hydroxyapatite nanorods and poly(D,L) lactic acid, as well as the compositions of the scaffolds, respectively. Porosity of the scaffolds was determined using the liquid substitution method. Moreover, the apatite-forming capability of the scaffolds was evaluated through simulated body fluid (SBF) incubation tests, whereas the viability, attachment, and distribution of human osteoblast cells (MG 63 cell line) on the scaffolds were determined through alamarBlue assay and confocal laser microscopy after nuclear staining with 4',6-diamidino-2-phenylindole and actin filaments of a cytoskeleton with Oregon Green 488 phalloidin. Results showed that hydroxyapatite nanorod/poly(D,L) lactic acid scaffolds that mimic the structure of natural bone were successfully produced. These scaffolds possessed macropore networks with high porosity (80-84%) and mean pore sizes ranging 117-183 μm. These scaffolds demonstrated excellent apatite-forming capabilities. The rapid formation of bone-like apatites with flower-like morphology was observed after 7 days of incubation in SBFs. The scaffolds that had a high percentage (30 wt.%) of hydroxyapatite demonstrated better cell adhesion, proliferation, and distribution than those with low percentages of hydroxyapatite as the days of culture increased. This work presented an efficient route for developing biomimetic composite scaffolds, which have potential applications in bone-tissue engineering. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Hollow carbonated hydroxyapatite microspheres with mesoporous structure: hydrothermal fabrication and drug delivery property.

    PubMed

    Guo, Ya-Jun; Wang, Ying-Ying; Chen, Ting; Wei, Yi-Ting; Chu, Lian-Feng; Guo, Ya-Ping

    2013-08-01

    Hollow carbonated hydroxyapatite microspheres with mesoporous structure (HCHAs) have been fabricated by using calcium carbonated microspheres as sacrificial templates according to the following routes: (i) the in situ deposit of carbonated hydroxyapatite on the surfaces of CaCO3 microspheres by hydrothermal method and (ii) the removal of CaCO3 by chemical etching. The HCHAs consist of a hollow core and a mesoporous shell. Interestingly, the shell of the microspheres is constructed by carbonated hydroxyapatite nanoplates as building blocks. Moreover, these nanoplates are composed of many smaller nanoparticles with different crystal orientations, and the mesopores exist among these nanoparticles. The HCHAs exhibit the high drug-loading capacity and sustained drug release property, suggesting that the hierarchically porous microspheres have great potentials for bone-implantable drug-delivery applications. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. A new injectable biphasic hydrogel based on partially hydrolyzed polyacrylamide and nano hydroxyapatite, crosslinked with chromium acetate, as scaffold for cartilage regeneration

    NASA Astrophysics Data System (ADS)

    Koushki, N.; Tavassoli, H.; Katbab, A. A.; Katbab, P.; Bonakdar, S.

    2015-05-01

    Polymer scaffolds are applied in the field of tissue engineering as three dimensional structures to organize cells and present stimuli to direct generation of a desired damaged tissue. In situ gelling scaffolds have attracted great attentions, as they are structurally similar to the extra cellular matrix (ECM). In the present work, attempts have been made to design and fabricate a new injectable and crosslinkable biphasic hydrogel based on partially hydrolyzed polyacrylamide (HPAM), chromium acetate as crosslink agent and nanocrystalline hydroxyapatite (nHAp) as reinforcing and bioactive agent for repair and regeneration of damaged cartilage. The distinct characteristic of HPAM is the presence of carboxylate anion groups on its backbone which allows to engineer the structure of the hydrogel for the desired bioactivity with appropriate cells differentiation towards both soft and hard (bone) tissues. The synthesized hydrogel exhibited bifunctional behavior which was derived by its biphasic structure in which one phase was loaded with nano hydroxyapatite to provide integration capability by subchondral bones and fix the hydrogel at cartilage defect without a need for suturing. The other phase differentiates the rabbit adipogenic mesenchymal stem cells (MSCs) towards soft tissue. Rheomechanical spectrometry (RMS) was employed to study the kinetic of the gelation including induction time and rate, as well as to measure the ultimate elastic modulus of the optimum crosslinked hydrogel. Surface tension measurement was also performed to tailor the surface characteristics of the gels. In vitro culturing of the cells inside the crosslinked hydrogel revealed high viability and high differentiation of the encapsulated rabbit stem cells, providing that the chromium acetate level was kept below 0.2 wt%. Based on the obtained results, the designed and fabricated biphasic hydrogel exhibited high potential as carrier for the stem cells for cartilage tissue engineering application

  19. Proliferation and osteogenic differentiation of human bone marrow stromal cells on alginate-gelatine-hydroxyapatite scaffolds with anisotropic pore structure.

    PubMed

    Bernhardt, A; Despang, F; Lode, A; Demmler, A; Hanke, T; Gelinsky, M

    2009-01-01

    Porous mineralized scaffolds are required for various applications in bone engineering. In particular, tube-like pores with controlled orientation inside the scaffold may support homogeneous cell seeding as well as sufficient nutrient supply and may facilitate blood vessel ingrowth. Scaffolds with parallely orientated tube-like pores were generated by diffusion-controlled ionotropic gelation of alginate. Incorporation of hydroxyapatite (HA) during the gelation process yielded stable scaffolds with an average pore diameter of approximately 90 microm. To evaluate the potential use of alginate-gelatine-HA scaffolds for bone tissue engineering, in vitro tests with human bone marrow stromal cells (hBMSCs) were carried out. We analysed biocompatibility and cell penetration into the capillary pores by microscopic methods. hBMSCs were also cultivated on alginate-gelatine-HA scaffolds for 3 weeks in the presence and absence of osteogenic supplements. We studied proliferation and osteogenic differentiation in terms of total lactate dehydrogenase (LDH) activity, DNA content and alkaline phosphatase (ALP) activity and found a 10-14-fold increase of cell number after 2 weeks of cultivation, as well as an increase of specific ALP activity for osteogenic-induced hBMSCs. Furthermore, the expression of bone-related genes [ALP, bone sialoprotein II (BSPII)] was analysed. We found an increase of ALP as well as BSPII expression for osteogenic-induced hBMSCs on alginate-gelatin-HA scaffolds. 2008 John Wiley & Sons, Ltd

  20. Ectopic osteogenesis and angiogenesis regulated by porous architecture of hydroxyapatite scaffolds with similar interconnecting structure in vivo.

    PubMed

    Li, Jinyu; Zhi, Wei; Xu, Taotao; Shi, Feng; Duan, Ke; Wang, Jianxin; Mu, Yandong; Weng, Jie

    2016-10-01

    The macro-pore sizes of porous scaffold play a key role for regulating ectopic osteogenesis and angiogenesis but many researches ignored the influence of interconnection between macro-pores with different sizes. In order to accurately reveal the relationship between ectopic osteogenesis and macro-pore sizes in dorsal muscle and abdominal cavities of dogs, hydroxyapatite (HA) scaffolds with three different macro-pore sizes of 500-650, 750-900 and 1100-1250 µm were prepared via sugar spheres-leaching process, which also had similar interconnecting structure determined by keeping the d/s ratio of interconnecting window diameter to macro-pore size constant. The permeability test showed that the seepage flow of fluid through the porous scaffolds increased with the increase of macro-pore sizes. The cell growth in three scaffolds was not affected by the macro-pore sizes. The in vivo ectopic implantation results indicated that the macro-pore sizes of HA scaffolds with the similar interconnecting structure have impact not only the speed of osteogenesis and angiogenesis but also the space distribution of newly formed bone. The scaffold with macro-pore sizes of 750-900 µm exhibited much faster angiogenesis and osteogenesis, and much more uniformly distribution of new bone than those with other macro-pore sizes. This work illustrates the importance of a suitable macro-pore sizes in HA scaffolds with the similar interconnecting structure which provides the environment for ectopic osteogenesis and angiogenesis.

  1. Ectopic osteogenesis and angiogenesis regulated by porous architecture of hydroxyapatite scaffolds with similar interconnecting structure in vivo

    PubMed Central

    Li, Jinyu; Zhi, Wei; Xu, Taotao; Shi, Feng; Duan, Ke; Wang, Jianxin; Mu, Yandong; Weng, Jie

    2016-01-01

    The macro-pore sizes of porous scaffold play a key role for regulating ectopic osteogenesis and angiogenesis but many researches ignored the influence of interconnection between macro-pores with different sizes. In order to accurately reveal the relationship between ectopic osteogenesis and macro-pore sizes in dorsal muscle and abdominal cavities of dogs, hydroxyapatite (HA) scaffolds with three different macro-pore sizes of 500–650, 750–900 and 1100–1250 µm were prepared via sugar spheres-leaching process, which also had similar interconnecting structure determined by keeping the d/s ratio of interconnecting window diameter to macro-pore size constant. The permeability test showed that the seepage flow of fluid through the porous scaffolds increased with the increase of macro-pore sizes. The cell growth in three scaffolds was not affected by the macro-pore sizes. The in vivo ectopic implantation results indicated that the macro-pore sizes of HA scaffolds with the similar interconnecting structure have impact not only the speed of osteogenesis and angiogenesis but also the space distribution of newly formed bone. The scaffold with macro-pore sizes of 750–900 µm exhibited much faster angiogenesis and osteogenesis, and much more uniformly distribution of new bone than those with other macro-pore sizes. This work illustrates the importance of a suitable macro-pore sizes in HA scaffolds with the similar interconnecting structure which provides the environment for ectopic osteogenesis and angiogenesis. PMID:27699059

  2. FABRICATION AND MODELING OF DYNAMIC MULTI-POLYMER NANOFIBROUS SCAFFOLDS

    PubMed Central

    Baker, Brendon M.; Nerurkar, Nandan L.; Burdick, Jason A.; Elliott, Dawn M.; Mauck, Robert L.

    2010-01-01

    Aligned nanofibrous scaffolds hold tremendous potential for the engineering of dense connective tissues. These biomimetic micro-patterns direct organized, cell-mediated matrix deposition, and can be tuned to possess nonlinear and anisotropic mechanical properties. For these scaffolds to function in vivo, however, they must either recapitulate the full dynamic mechanical range of the native tissue upon implantation, or must foster cell infiltration and matrix deposition so as to enable construct maturation to meet these criteria. In our recent studies, we noted that cell infiltration into dense aligned structures is limited, but could be expedited via the inclusion of a distinct, rapidly eroding sacrificial component. In the present study, we sought to further the fabrication of dynamic nanofibrous constructs by combining multiple fiber populations, each with distinct mechanical characteristics, into a single composite nanofibrous scaffold. Towards this goal, we developed a novel method for the generation of aligned electrospun composites containing rapidly eroding (PEO), moderately degradable (PLGA and PCL/PLGA), and slowly degrading (PCL) fiber populations. We evaluated the mechanical properties of these composites upon formation and with degradation in a physiologic environment. Further, we employed a hyperelastic constrained mixture model to capture the nonlinear and time-dependent properties of these scaffolds when formed as single-fiber populations or in multi-polymer composites. After validating this model, we demonstrated that by carefully selecting fiber populations with differing mechanical properties, and altering the relative fraction of each, a wide range of mechanical properties (and degradation characteristics) can be achieved. This advance allows for the rational design of nanofibrous scaffolds to match native tissue properties, and will significantly enhance our ability to fabricate replacements for load bearing tissues of the musculoskeletal system

  3. Fabrication and modeling of dynamic multipolymer nanofibrous scaffolds.

    PubMed

    Baker, Brendon M; Nerurkar, Nandan L; Burdick, Jason A; Elliott, Dawn M; Mauck, Robert L

    2009-10-01

    Aligned nanofibrous scaffolds hold tremendous potential for the engineering of dense connective tissues. These biomimetic micropatterns direct organized cell-mediated matrix deposition and can be tuned to possess nonlinear and anisotropic mechanical properties. For these scaffolds to function in vivo, however, they must either recapitulate the full dynamic mechanical range of the native tissue upon implantation or must foster cell infiltration and matrix deposition so as to enable construct maturation to meet these criteria. In our recent studies, we noted that cell infiltration into dense aligned structures is limited but could be expedited via the inclusion of a distinct rapidly eroding sacrificial component. In the present study, we sought to further the fabrication of dynamic nanofibrous constructs by combining multiple-fiber populations, each with distinct mechanical characteristics, into a single composite nanofibrous scaffold. Toward this goal, we developed a novel method for the generation of aligned electrospun composites containing rapidly eroding (PEO), moderately degradable (PLGA and PCL/PLGA), and slowly degrading (PCL) fiber populations. We evaluated the mechanical properties of these composites upon formation and with degradation in a physiologic environment. Furthermore, we employed a hyperelastic constrained-mixture model to capture the nonlinear and time-dependent properties of these scaffolds when formed as single-fiber populations or in multipolymer composites. After validating this model, we demonstrated that by carefully selecting fiber populations with differing mechanical properties and altering the relative fraction of each, a wide range of mechanical properties (and degradation characteristics) can be achieved. This advance allows for the rational design of nanofibrous scaffolds to match native tissue properties and will significantly enhance our ability to fabricate replacements for load-bearing tissues of the musculoskeletal system.

  4. New method for the fabrication of highly osteoconductive β-1,3-glucan/HA scaffold for bone tissue engineering: Structural, mechanical, and biological characterization.

    PubMed

    Klimek, Katarzyna; Przekora, Agata; Pałka, Krzysztof; Ginalska, Grażyna

    2016-10-01

    Recent studies have shown that thermal method for β-1,3-glucan (curdlan) gelation performed at temperature above 80°C enables fabrication of biocompatible bone scaffolds. The aim of this study was to establish new method for fabrication of β-1,3-glucan/hydroxyapatite (glu/HA) scaffold using ion-exchanging dialysis for curdlan gelation that allows for the modifications of the glu/HA material with thermo-sensitive agents like growth factors or adhesive proteins. Obtained results reveal that fabricated scaffold appears to be highly osteoconductive as it is nontoxic, promotes osteoblast growth and proliferation as well as increases bone alkaline phosphatase level thereby enhancing cell differentiation. It was demonstrated that developed new method for the glu/HA scaffold fabrication allows to obtain material that not only can be modified with thermo-sensitive agents at the stage of production process but also is a promising candidate for bone tissue engineering applications to act as a framework for osteoblasts to spread and form new bone. It should be noted that dialysis method for curdlan gelation has never been used before to fabricate bone scaffold. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2528-2536, 2016. © 2016 Wiley Periodicals, Inc.

  5. Bone Marrow Stem Cells Added to a Hydroxyapatite Scaffold Result in Better Outcomes after Surgical Treatment of Intertrochanteric Hip Fractures

    PubMed Central

    Gutierres, Manuel; Lopes, M. Ascenção; Santos, J. Domingos; Cabral, A. T.; Pinto, R.

    2014-01-01

    Introduction. Intertrochanteric hip fractures occur in the proximal femur. They are very common in the elderly and are responsible for high rates of morbidity and mortality. The authors hypothesized that adding an autologous bone marrow stem cells concentrate (ABMC) to a hydroxyapatite scaffold and placing it in the fracture site would improve the outcome after surgical fixation of intertrochanteric hip fractures. Material and Methods. 30 patients were randomly selected and divided into 2 groups of 15 patients, to receive either the scaffold enriched with the ABMC (Group A) during the surgical procedure, or fracture fixation alone (Group B). Results. There was a statistically significant difference in favor of group A at days 30, 60, and 90 for Harris Hip Scores (HHS), at days 30 and 60 for VAS pain scales, for bedridden period and time taken to start partial and total weight bearing (P < 0.05). Discussion. These results show a significant benefit of adding a bone marrow enriched scaffold to surgical fixation in intertrochanteric hip fractures, which can significantly reduce the associated morbidity and mortality rates. Conclusion. Bone marrow stem cells added to a hydroxyapatite scaffold result in better outcomes after surgical treatment of intertrochanteric hip fractures. PMID:24955356

  6. Biomimetic fiber mesh scaffolds based on gelatin and hydroxyapatite nano-rods: Designing intrinsic skills to attain bone reparation abilities.

    PubMed

    Sartuqui, Javier; Gravina, A Noel; Rial, Ramón; Benedini, Luciano A; Yahia, L'Hocine; Ruso, Juan M; Messina, Paula V

    2016-09-01

    Intrinsic material skills have a deep effect on the mechanical and biological performance of bone substitutes, as well as on its associated biodegradation properties. In this work we have manipulated the preparation of collagenous derived fiber mesh frameworks to display a specific composition, morphology, open macroporosity, surface roughness and permeability characteristics. Next, the effect of the induced physicochemical attributes on the scaffold's mechanical behavior, bone bonding potential and biodegradability were evaluated. It was found that the scaffold microstructure, their inherent surface roughness, and the compression strength of the gelatin scaffolds can be modulated by the effect of the cross-linking agent and, essentially, by mimicking the nano-scale size of hydroxyapatite in natural bone. A clear effect of bioactive hydroxyapatite nano-rods on the scaffolds skills can be appreciated and it is greater than the effect of the cross-linking agent, offering a huge perspective for the upcoming progress of bone implant technology. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Perfusion conditioning of hydroxyapatite-chitosan-gelatin scaffolds for bone tissue regeneration from human mesenchymal stem cells.

    PubMed

    Sellgren, Katelyn L; Ma, Teng

    2012-01-01

    Tissue-engineered bone grafts require an osteogenic cell source and a scaffold capable of supporting tissue regeneration. Hydroxyapatite (H), chitosan (C), and gelatin (G), when combined, produce a biomimetic scaffold with a chemical similarity to the main structural components of natural bone tissue. In this study a phase-separation technique was used to produce a porous 3D HCG scaffold, containing a network of cross-linked chitosan and gelatin fibrils coated in hydroxyapatite, with pore size readily controlled by freezing temperature. The HCG scaffolds were then seeded with human mesenchymal stem cells (hMSCs), using a depth filtration system after preconditioning with serum-containing medium for 7 days under either static or perfusion conditions. The effects of static and perfusion media preconditioning on protein adsorption, surface morphology, hMSC attachment, proliferation and osteogenic differentiation were examined. Perfusion preconditioning, as opposed to static preconditioning, enhances adsorption of ECM proteins, which in turn promotes hMSC proliferation and osteogenic differentiation. The results demonstrate the importance of convective flow in modulating the 3D HCG microenvironment and highlight its profound influence on 3D construct development. Copyright © 2011 John Wiley & Sons, Ltd.

  8. Stochastic modeling, virtual refinement and deterministic fabrication of tissue engineering scaffolds

    NASA Astrophysics Data System (ADS)

    Rajagopalan, Srinivasan; Robb, Richard A.

    2006-03-01

    Tissue engineering attempts to address the ever widening gap between the demand and supply of organ and tissue transplants using natural and biomimetic scaffolds. The current scaffold fabrication techniques can be broadly classified into (a) conventional, irreproducible, stochastic techniques producing biomorphic "secundam naturam" but sub optimal scaffold architecture and (b) rapidly emerging, repeatable, computer-controlled Solid Freeform Fabrication (SFF) producing, "contra naturam" scaffold architecture. This paper presents an image-based scaffold optimization strategy based on microCT images of the conventional scaffolds. This approach, attempted and perfected for the first time, synergistically exploits the orthogonal techniques to create repeatable, biomorphic scaffolds with optimal scaffold geometry. The ramifications of this image based computer assisted intervention to improve the status quo of scaffold fabrication might contribute to the previously elusive deployment of promising benchside tissue analogs to the clinical bedside.

  9. Hydroxyapatite and gelatin composite foams processed via novel freeze-drying and crosslinking for use as temporary hard tissue scaffolds.

    PubMed

    Kim, Hae-Won; Knowles, Jonathan C; Kim, Hyoun-Ee

    2005-02-01

    Hydroxyapatite (HA) and gelatin composites were fabricated in a foam type via a novel freeze-drying and crosslinking technique. The morphological and mechanical properties of and in vitro cellular responses to the foams were investigated. The HA powder was added at up to 30 wt % into the gelatin solution, and the mixtures were freeze-dried and further crosslinked. The pure gelatin foam had a well-developed pore configuration with porosity and pore size of approximately 90% and 400-500 microm, respectively. With HA addition, the porosity decreased and pore shape became more irregular. The HA particulates, in sizes of about 2-5 microm, were distributed within the gelatin network homogeneously and made the framework surface rougher. All the foams had high water absorption capacities, showing typical hydrogel characteristics, even though the HA addition decreased the degree of water absorption. The HA addition made the foam much stronger and stiffer (i.e., with increasing HA amount the foams sustained higher compressive stress and had higher elastic modulus in both dry and wet states). The osteoblast-like human osteosarcoma cells spread and grew actively on all the foams. The cell proliferation rate, quantified indirectly on the cells cultured on Ti discs coated with gelatin and gelatin-HA composites using MTT assay, exhibited an up-regulation with gelatin coating compared with bare Ti substrate, but a slight decrease on the composite coatings. However, the alkaline phosphatase activities expressed by the cells cultured on composites foams as well as their coatings on Ti discs were significantly enhanced compared with those on pure gelatin foam and coating. These findings suggest that the gelatin-HA composite foams have great potential for use as hard tissue regeneration scaffolds. Copyright 2004 Wiley Periodicals, Inc.

  10. Porous nano-hydroxyapatite/collagen scaffold containing drug-loaded ADM-PLGA microspheres for bone cancer treatment.

    PubMed

    Rong, Zi-Jie; Yang, Lian-Jun; Cai, Bao-Ta; Zhu, Li-Xin; Cao, Yan-Lin; Wu, Guo-Feng; Zhang, Zan-Jie

    2016-05-01

    To develop adriamycin (ADM)-encapsulated poly(lactic-co-glycolic acid) (PLGA) nanoparticles in a porous nano-hydroxyapatite/collagen scaffold (ADM-PLGA-NHAC). To provide novel strategies for future treatment of osteosarcoma, the properties of the scaffold, including its in vitro extended-release properties, the inhibition effects of ADM-PLGA-NHAC on the osteosarcoma MG63 cells, and its bone repair capacity, were investigated in vivo and in vitro. The PLGA copolymer was utilized as a drug carrier to deliver ADM-PLGA nanoparticles (ADM-PLGA-NP). Porous nano-hydroxyapatite and collagen were used to materials to produce the porous nano-hydroxyapatite/collagen scaffold (NHAC), into which the ADM-PLGA-NP was loaded. The performance of the drug-carrying scaffold was assessed using multiple techniques, including scanning electron microscopy and in vitro extended release. The antineoplastic activities of scaffold extracts on the human osteosarcoma MG63 cell line were evaluated in vitro using the cell counting kit-8 (CCK8) method and live-dead cell staining. The bone repair ability of the scaffold was assessed based on the establishment of a femoral condyle defect model in rabbits. ADM-PLGA-NHAC and NHAC were implanted into the rat muscle bag for immune response experiments. A tumor-bearing nude mice model was created, and the TUNEL and HE staining results were observed under optical microscopy to evaluate the antineoplastic activity and toxic side effects of the scaffold. The composite scaffold demonstrated extraordinary extended-release properties, and its extracts also exhibited significant inhibition of the growth of osteosarcoma MG63 cells. In the bone repair experiment, no significant difference was observed between ADM-PLGA-NHAC and NHAC by itself. In the immune response experiments, ADM-PLGA-NHAC exhibited remarkable biocompatibility. The in vivo antitumor experiment revealed that the implantation of ADM-PLGA-NHAC in the tumor resulted in a improved antineoplastic

  11. Hybrid 3D-2D printing for bone scaffolds fabrication

    NASA Astrophysics Data System (ADS)

    Seleznev, V. A.; Prinz, V. Ya

    2017-02-01

    It is a well-known fact that bone scaffold topography on micro- and nanometer scale influences the cellular behavior. Nano-scale surface modification of scaffolds allows the modulation of biological activity for enhanced cell differentiation. To date, there has been only a limited success in printing scaffolds with micro- and nano-scale features exposed on the surface. To improve on the currently available imperfect technologies, in our paper we introduce new hybrid technologies based on a combination of 2D (nano imprint) and 3D printing methods. The first method is based on using light projection 3D printing and simultaneous 2D nanostructuring of each of the layers during the formation of the 3D structure. The second method is based on the sequential integration of preliminarily created 2D nanostructured films into a 3D printed structure. The capabilities of the developed hybrid technologies are demonstrated with the example of forming 3D bone scaffolds. The proposed technologies can be used to fabricate complex 3D micro- and nanostructured products for various fields.

  12. Enhancement of osteoinduction by continual simvastatin release from poly(lactic-co-glycolic acid)-hydroxyapatite-simvastatin nano-fibrous scaffold.

    PubMed

    Jiang, Liming; Sun, Haizhu; Yuan, Anliang; Zhang, Kai; Li, Daowei; Li, Chen; Shi, Ce; Li, Xiangwei; Gao, Kai; Zheng, Changyu; Yang, Bai; Sun, Hongchen

    2013-11-01

    Simvastatin is considered as a stimulator for bone formation. However, the half-life for simvastatin is generally 2 hours, which means, it is difficult to maintain biologically active simvastatin in vivo. To overcome this limitation, we created a system to slowly release simvastatin in vitro and in vivo. We constructed a poly(lactic-co-glycolic acid)/hydroxyapatite nano-fibrous scaffold to carry simvastatin. Releasing assays showed that simvastatin was released from poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin quickly within - 15 days, and small amounts continued to be released through day 56 (experiments terminated). MTT assays demonstrated that both poly(lactic-co-glycolic acid)/hydroxyapatite and poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin promoted MC3T3-E1 cell proliferation. However, Alkaline phosphatase assays showed that only poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin scaffold significantly promoted the osteogenic differentiation of MC3T3-E1 cells in vitro on day 14. To further test in vivo, we created calvaria bone defect models and implanted either poly(lactic-co-glycolic acid)/hydroxyapatite or poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin. After 4 or 8 weeks post-implantation, the results indicated that poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin scaffold induced bone formation more efficiently than poly(lactic-co-glycolic acid)/hydroxyapatite alone. Our data demonstrates that poly(lactic-co-glycolic acid)/hydroxyapatite/simvastatin has the potential to aid in healing bone defects and promoting bone regeneration in the future although we still need to optimize this complex to efficiently promote bone regeneration.

  13. Effect of internal structure of collagen/hydroxyapatite scaffold on the osteogenic differentiation of mesenchymal stem cells.

    PubMed

    Chen, Guobao; Lv, Yonggang; Dong, Chanjuan; Yang, Li

    2015-01-01

    Consisting of seed cells and scaffold, regenerative medicine provides a new way for the repair and regeneration of tissue and organ. Collagen/hydroxyapatite (HA) biocomposite scaffold is highlighted due to its advantageous features of two major components of bone matrix: collagen and HA. The aim of this study is to investigate the effect of internal structure of collagen/HA scaffold on the fate of rat mesenchymal stem cells (MSCs). The internal structure of collagen/HA scaffold was characterized by micro-CT. It is found that the porosity decreased while average compressive modulus increased with the increase of collagen proportion. Within the collagen proportion of 0.35%, 0.5% and 0.7%, the porosities were 89.08%, 78.37% and 75.36%, the pore sizes were 140.6±75.5 μm, 133.9±48.4 μm and 160.7±119.6 μm, and the average compressive moduli were 6.74±1.16 kPa, 8.82±2.12 kPa and 23.61±8.06 kPa, respectively. Among these three kinds of scaffolds, MSCs on the Col 0.35/HA 22 scaffold have the highest viability and the best cell proliferation. On the contrary, the Col 0.7/HA 22 scaffold has the best ability to stimulate MSCs to differentiate into osteoblasts in a relatively short period of time. In vivo research also demonstrated that the internal structure of collagen/HA scaffold has significant effect on the cell infiltration. Therefore, precise control of the internal structure of collagen/HA scaffold can provide a more efficient carrier to the repair of bone defects.

  14. Fabrication of naturel pumice/hydroxyapatite composite for biomedical engineering.

    PubMed

    Komur, Baran; Lohse, Tim; Can, Hatice Merve; Khalilova, Gulnar; Geçimli, Zeynep Nur; Aydoğdu, Mehmet Onur; Kalkandelen, Cevriye; Stan, George E; Sahin, Yesim Muge; Sengil, Ahmed Zeki; Suleymanoglu, Mediha; Kuruca, Serap Erdem; Oktar, Faik Nuzhet; Salman, Serdar; Ekren, Nazmi; Ficai, Anton; Gunduz, Oguzhan

    2016-07-07

    We evaluated the Bovine hydroxyapatite (BHA) structure. BHA powder was admixed with 5 and 10 wt% natural pumice (NP). Compression strength, Vickers micro hardness, Fourier transform infrared spectroscopy, scanning electron microscopy (SEM) and X-ray diffraction studies were performed on the final NP-BHA composite products. The cells proliferation was investigated by MTT assay and SEM. Furthermore, the antimicrobial activity of NP-BHA samples was interrogated. Variances in the sintering temperature (for 5 wt% NP composites) between 1000 and 1300 °C, reveal about 700 % increase in the microhardness (~100 and 775 HV, respectively). Composites prepared at 1300 °C demonstrate the greatest compression strength with comparable result for 5 wt% NP content (87 MPa), which are significantly better than those for 10 wt% and those that do not include any NP (below 60 MPa, respectively). The results suggested the optimal parameters for the preparation of NP-BHA composites with increased mechanical properties and biocompatibility. Changes in micro-hardness and compression strength can be tailored by the tuning the NP concentration and sintering temperature. NP-BHA composites have demonstrated a remarkable potential for biomedical engineering applications such as bone graft and implant.

  15. Preparation of chitosan/silk fibroin/hydroxyapatite porous scaffold and its characteristics in comparison to bi-component scaffolds.

    PubMed

    Qi, Xiao-Ni; Mou, Zhao-Li; Zhang, Jing; Zhang, Zhi-Qi

    2014-02-01

    Composite porous scaffolds have attracted extensive attention in the biomedical material field. The aim of this research was to prepare a novel tri-component composite porous scaffold and to evaluate its relevant properties. The porous scaffold was composed of chitosan (CS), silk fibroin (SF), and nanohydroxyapatite particles (nHA), which we named CS/SF/nHA scaffold and prepared via salt fractionation method combined with lyophilization. The porous structure was achieved using a porogen (salt), and the pore size was controlled by the size of porogen. To evaluate the characteristics of the tri-component scaffold, three bi-component scaffolds, CS/SF, CS/nHA, and SF/nHA, were simultaneously prepared for comparison. The scaffolds were subjected to morphological, micro-structural, and biodegradation analyses. Results demonstrated that all of the scaffolds had pore sizes of 100-300 μm and a porosity of 90.5-96.1%. The biodegradation characteristics of all scaffolds meet the requirements of good biomedical materials. The investigation of the mechanical properties showed that the tri-component scaffold has better properties than the bi-component scaffolds. The in vitro biocompatibility with osteoblast-like MG-63 cells showed that all the scaffolds are suitable for cell attachment and proliferation; however, the CS/SF/nHA composite porous scaffold is much more effective than the others. © 2013 Wiley Periodicals, Inc.

  16. Scaffold-Free Fabrication of Osteoinductive Cellular Constructs Using Mouse Gingiva-Derived Induced Pluripotent Stem Cells

    PubMed Central

    Okawa, Hiroko; Kayashima, Hiroki; Sasaki, Jun-Ichi; Miura, Jiro; Kamano, Yuya; Kosaka, Yukihiro; Imazato, Satoshi; Yatani, Hirofumi; Matsumoto, Takuya; Egusa, Hiroshi

    2016-01-01

    Three-dimensional (3D) cell constructs are expected to provide osteoinductive materials to develop cell-based therapies for bone regeneration. The proliferation and spontaneous aggregation capability of induced pluripotent stem cells (iPSCs) thus prompted us to fabricate a scaffold-free iPSC construct as a transplantation vehicle. Embryoid bodies of mouse gingival fibroblast-derived iPSCs (GF-iPSCs) were seeded in a cell chamber with a round-bottom well made of a thermoresponsive hydrogel. Collected ball-like cell constructs were cultured in osteogenic induction medium for 30 days with gentle shaking, resulting in significant upregulation of osteogenic marker genes. The constructs consisted of an inner region of unstructured cell mass and an outer osseous tissue region that was surrounded by osteoblast progenitor-like cells. The outer osseous tissue was robustly calcified with elemental calcium and phosphorous as well as hydroxyapatite. Subcutaneous transplantation of the GF-iPSC constructs into immunodeficient mice contributed to extensive ectopic bone formation surrounded by teratoma tissue. These results suggest that mouse GF-iPSCs could facilitate the fabrication of osteoinductive scaffold-free 3D cell constructs, in which the calcified regions and surrounding osteoblasts may function as scaffolds and drivers of osteoinduction, respectively. With incorporation of technologies to inhibit teratoma formation, this system could provide a promising strategy for bone regenerative therapies. PMID:27110251

  17. Analyzing the hydrodynamic and crowding evolution of aqueous hydroxyapatite-gelatin networks: Digging deeper into bone scaffold design variables.

    PubMed

    Sartuqui, Javier; D' Elía, Noelia; Gravina, A Noel; Messina, Paula V

    2015-07-01

    The hydration of the polypeptide network is a determinant factor to be controlled on behalf of the design of precise functional tissue scaffolding. Here we present an exhaustive study of the hydrodynamic and crowding evolution of aqueous gelatin-hydroxyapatite systems with the aim of increasing the knowledge about the biomimesis of collagen mineralization; and how it can be manipulated for the preparation of collagenous derived frameworks with specific morphological characteristics. The solution's density and viscosity evaluation measurements in combination with spectroscopic techniques revealed that there is a progressive association of protein chain that can be influenced by the amount of hydroxyapatite nanorods. Gelatin and additives' concentration effect on the morphology of the gelatin scaffolds was investigated. Transverse and longitudinal sections of the obtained scaffolds were taken and analyzed using optical microscopy. It can be seen that the porous size and shape of gelatin assemblies can be easily adjusted by controlling the gelatin/HAp ratio in the solution used as template in agreement with our statement.

  18. Characterization of hydroxyapatite whisker reinforced composites and scaffolds for mechanical and biological function in orthopaedic and spinal implants

    NASA Astrophysics Data System (ADS)

    Conrad, Timothy L.

    The overall objective of this study was to investigate the mechanical and biological properties of HA whisker reinforced polyaryletherketone (PAEK) composites and scaffolds which are key to clinical translation for orthopedic and spinal implants. The fatigue behavior of polyetherketoneketone (PEKK) reinforced with 0, 20, and 40 vol% hydroxyapatite (HA) was investigated in four-point bending fatigue. The fatigue life decreased with increasing HA reinforcement. However, PEKK reinforced with 40 vol% HA whiskers exhibited a fatigue life greater than 2.106 cycles at 40 MPa. Moreover, HA whisker reinforcement resulted in decreased creep deformation and minimal modulus degradation. The effects of the mold temperature and polyetheretherketone (PEEK) powder were investigated on the mechanical properties and crystallinity of HA whisker reinforced PEEK scaffolds prepared using compression molding and porogen leaching. The mechanical properties of the scaffolds increased while the PEEK crystallinity decreased, with increasing mold temperature and suggested an optimal mold temperature of 370--375°C for PEEK scaffolds comprising of 75% porosity and 20 vol% HA whisker reinforcement, regardless of the PEEK powder size. The effects of the porogen morphology on the architecture, mechanical properties, and permeability of HA whisker reinforced PEEK scaffolds were investigated in 75--90% porous scaffolds. HA whisker reinforced PEEK scaffolds prepared with an ellipsoidal porogen exhibited a greater permeability than scaffolds prepared with a cubic porogen. The compressive modulus, yield strength, and yield strain were not affected by the porogen morphology. The effects of HA reinforcement morphology and content was investigated on the behavior of primary osteoblasts on dense HA reinforced PEEK substrates in vitro. At day 7, the number of osteoblasts attached to PEEK substrate surfaces increased with increasing HA content and for HA whiskers compared to equiaxed HA powder reinforcement

  19. Injectable alginate/hydroxyapatite gel scaffold combined with gelatin microspheres for drug delivery and bone tissue engineering.

    PubMed

    Yan, Jingxuan; Miao, Yuting; Tan, Huaping; Zhou, Tianle; Ling, Zhonghua; Chen, Yong; Xing, Xiaodong; Hu, Xiaohong

    2016-06-01

    Injectable and biodegradable alginate-based composite gel scaffolds doubly integrated with hydroxyapatite (HAp) and gelatin microspheres (GMs) were cross-linked via in situ release of calcium cations. As triggers of calcium cations, CaCO3 and glucono-D-lactone (GDL) were fixed as a mass ratio of 1:1 to control pH value ranging from 6.8 to 7.2 during gelation. Synchronously, tetracycline hydrochloride (TH) was encapsulated into GMs to enhance bioactivity of composite gel scaffolds. The effects of HAp and GMs on characteristics of gel scaffolds, including pH value, gelation time, mechanical properties, swelling ratio, degradation behavior and drug release, were investigated. The results showed that HAp and GMs successfully improved mechanical properties of gel scaffolds at strain from 0.1 to 0.5, which stabilized the gel network and decreased weight loss, as well as swelling ratio and gelation time. TH could be released from this composite gel scaffold into the local microenvironment in a controlled fashion by the organic/inorganic hybrid of hydrogel network. Our results demonstrate that the HAp and GMs doubly integrated alginate-based gel scaffolds, especially the one with 6% (w/v) HAp and 5% (w/v) GMs, have suitable physical performance and bioactive properties, thus provide a potential opportunity to be used for bone tissue engineering. The potential application of this gel scaffold in bone tissue engineering was confirmed by encapsulation behavior of osteoblasts. In combination with TH, the gel scaffold exhibited beneficial effects on osteoblast activity, which suggested a promising future for local treatment of pathologies involving bone loss.

  20. Evaluate the growth and adhesion of osteoblast cells on nanocomposite scaffold of hydroxyapatite/titania coated with poly hydroxybutyrate

    PubMed Central

    Pourmollaabbassi, Babak; Karbasi, Saeed; Hashemibeni, Batool

    2016-01-01

    Background: The generation of bioartificial bone tissues may help to overcome the problems related to donor site morbidity and size limitations. Materials and Methods: In this paper, hydroxyapatite (HA) powder was made out of bovine bone by thermal analysis at 900°C and first, and then, porous HA (50 weight percentage) was produced by polyurethane sponge replication method. In order to improve the scaffold mechanical properties, they have been coated with poly hydroxybutyrate. In terms of phase studies, morphology, and specifying agent groups, the specific characterization devices such as X-ray diffraction and Fourier transform infrared, were employed. To compare the behavior of cellular scaffolds, they were divided into four groups of scaffolds. The osteoblast cells were cultured. To perform phase studies, analysis of Methylthiazole tetrazolium (MTT) and Trypan blue were carried out for the viability and attachment on the surface of the scaffold, and the specification of Scanning electron microscopy was employed for the morphology of the cells. Results: The results of MTT analysis performed on four groups of scaffolds have shown that Titanium oxide (Tio2) had no effect on cell growth alone and HA was the main factor of growth and cell osteoblast adhesion on the scaffold. Moreover, the results showed that the use of coating with poly-3-hydroxybutyrate saved the factors and placed the osteoblasts within the pore. Since the main part of bone consists of HA, the TiO2 accelerates the formation of apatite crystals at the scaffold surface which is the evidence for bone tissue regeneration. Conclusions: It is likely that the relation between HA and TiO2 leads to an increase in osteoblast adhesion and growth of cells on the scaffold surface. PMID:27761431

  1. Supercritical CO2 assisted process for the production of high-purity and sterile nano-hydroxyapatite/chitosan hybrid scaffolds.

    PubMed

    Ruphuy, G; Souto-Lopes, M; Paiva, D; Costa, P; Rodrigues, A E; Monteiro, F J; Salgado, C L; Fernandes, M H; Lopes, J C; Dias, M M; Barreiro, M F

    2017-05-04

    Hybrid scaffolds composed of hydroxyapatite (HAp), in particular in its nanometric form (n-HAp), and chitosan (CS) are promising materials for non-load-bearing bone graft applications. The main constraints of their production concern the successful implementation of the final purification/neutralization and sterilization steps. Often, the used purification strategies can compromise scaffold structural features, and conventional sterilization techniques can result in material's thermal degradation and/or contamination with toxic residues. In this context, this work presents a process to produce n-HAp/CS scaffolds mimicking bone composition and structure, where an innovative single step based on supercritical CO2 extraction was used for both purification and sterilization. A removal of 80% of the residual acetic acid was obtained (T = 75°C, p = 8.0 MPa, 2 extraction cycles of 2 h) giving rise to scaffolds exhibiting adequate interconnected porous structure, fast swelling and storage modulus compatible with non-load-bearing applications. Moreover, the obtained scaffolds showed cytocompatibility and osteoconductivity without further need of disinfection/sterilization procedures. Among the main advantages, the proposed process comprises only three steps (n-HAp/CS dispersion preparation; freeze-drying; and supercritical CO2 extraction), and the supercritical CO2 extraction show clear advantages over currently used procedures based on neutralization steps. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2017. © 2017 Wiley Periodicals, Inc.

  2. Guided tissue fabrication from periosteum using preformed biodegradable polymer scaffolds.

    PubMed

    Thomson, R C; Mikos, A G; Beahm, E; Lemon, J C; Satterfield, W C; Aufdemorte, T B; Miller, M J

    1999-11-01

    A successful tissue engineering method for bone replacement would imitate natural bone graft by providing the essential elements for new bone formation using synthetic scaffolds, osteogenic cell populations, and bone induction factors. This is a study of the suitability of various formulations of poly(DL-lactic-co-glycolic acid) (PLGA) foams to provide a tissue conducting scaffold in an ovine model for bone flap fabrication. Three formulations were used of different copolymer ratio and molecular weight. Porous wafers of PLGA were stacked into rectangular chambers (volume 4 cm3) enclosed on five sides. Some chambers also contained autologous morcellized bone graft (MBG). The chambers were inserted with the open face adjacent to the cambium layer of the periosteum in rib beds of seven sheep and harvested after 8 weeks in vivo. Gross and histologic examination of the resulting tissue specimens demonstrated molded units of vascularized tissue generally conforming to the shape of the chambers and firmly attached to the periosteum. Polymer degradation appeared to occur by varying degrees based on polymer formulation. New bone formation was observed only in areas containing MBG. There was no evidence of significant inflammatory reaction or local tissue damage at 8 weeks. We conclude that a PLGA foam scaffold is (1) an efficient conductor of new tissue growth but not osteoinductive, (2) contributes to the shape of molded tissue, and (3) biocompatible when used in this model. Further studies are warranted to develop practical methods to deliver bone induction factors to the system to promote osseous tissue generation throughout the synthetic scaffold.

  3. [Current progress of fabricating tissue engineering scaffold using rapid prototyping techniques].

    PubMed

    Li, Xiang; Wang, Chengtao

    2008-08-01

    As one of the key factors for tissue engineering, scaffolds affect the spread and proliferation of seeded cells and the formation of new tissue. Although conventional methods can produce porous scaffolds with different porosities, they are lack controls the porous structures of the scaffolds. In recent years, rapid prototyping (RP) techniques have been developed and have successfully applied to fabricate TE scaffolds. RP techniques can provide accurate control over internal pore architectures and complex-shapes. As a result of these techniques, ideal tissue-engineered constructs could be prepared. This paper reviewed the advantages, potential and future directions of RP techniques in the design and fabrication of TE scaffolds.

  4. Fabrication of hydroxyapatite ceramics with controlled pore characteristics by slip casting.

    PubMed

    Yao, Xiumin; Tan, Shouhong; Jiang, Dongliang

    2005-02-01

    Porous hydroxyapatite (HAp) ceramics with controlled pore characteristics were fabricated using slip casting method by mixing PMMA with HAp powder. The optimum conditions of HAp slip for slip casting was achieved by employing various experimental techniques, zeta potential and sedimentation, as a function of pH of the slips in the pH range of 4-12. HAp suspensions displayed an absolute maximum in zeta potential values and a minimum in sedimentation height at pH 11.5. The optimal amount of dispersant for the HAp suspensions was found at 1.0 wt% according to the viscosity of 25 vol% HAp slurry. The rheological behaviour of HAp slurry displays a shear-thinning behavior without thixotropy, which is needed in slip casting processing. The pore characteristics of sintered porous hydroxyapatite bioceramics can be controlled by added PMMA particle size and volume. The obtained ceramics exhibit higher strength than those obtained by dry pressing.

  5. Fabrication of three-dimensional nano, micro and micro/nano scaffolds of porous poly(lactic acid) by electrospinning and comparison of cell infiltration by Z-stacking/three-dimensional projection technique.

    PubMed

    Shalumon, K T; Chennazhi, K P; Tamura, H; Kawahara, K; Nair, S V; Jayakumar, R

    2012-03-01

    The use of electrospun extracellular matrix (ECM)-mimicking nanofibrous scaffolds for tissue engineering is limited by poor cellular infiltration. The authors hypothesised that cell penetration could be enhanced in scaffolds by using a hierarchical structure where nano fibres are combined with micron-scale fibres while preserving the overall scaffold architecture. To assess this, we fabricated electrospun porous poly(lactic acid) (PLA) scaffolds having nanoscale, microscale and combined micro/nano architecture and evaluated the structural characteristics and biological response in detail. Although the bioactivity was intermediate to that for nanofibre and microfibre scaffold, a unique result of this study was that the micro/nano combined fibrous scaffold showed improved cell infiltration and distribution than the nanofibrous scaffold. Although the cells were found to be lining the scaffold periphery in the case of nanofibrous scaffold, micro/nano scaffolds had cells dispersed throughout the scaffold. Further, as expected, the addition of nanoparticles of hydroxyapatite (nHAp) improved the bioactivity, although it did not play a significant role in cell penetration. Thus, this strategy of creating a three-dimensional (3D) micro/nano architecture that would increase the porosity of the fibrous scaffold and thereby improving the cell penetration, can be utilised for the generation of functional tissue engineered constructs in vitro.

  6. Three-dimensional plotter technology for fabricating polymeric scaffolds with micro-grooved surfaces.

    PubMed

    Son, JoonGon; Kim, GeunHyung

    2009-01-01

    Various mechanical techniques have been used to fabricate biomedical scaffolds, including rapid prototyping (RP) devices that operate from CAD files of the target feature information. The three-dimensional (3-D) bio-plotter is one RP system that can produce design-based scaffolds with good mechanical properties for mimicking cartilage and bones. However, the scaffolds fabricated by RP have very smooth surfaces, which tend to discourage initial cell attachment. Initial cell attachment, migration, differentiation and proliferation are strongly dependent on the chemical and physical characteristics of the scaffold surface. In this study, we propose a new 3-D plotting method supplemented with a piezoelectric system for fabricating surface-modified scaffolds. The effects of the physically-modified surface on the mechanical and hydrophilic properties were investigated, and the results of cell culturing of chondrocytes indicate that this technique is a feasible new method for fabricating high-quality 3-D polymeric scaffolds.

  7. Evaluation of bone regeneration, angiogenesis, and hydroxyapatite conversion in critical-sized rat calvarial defects implanted with bioactive glass scaffolds.

    PubMed

    Bi, Lianxiang; Jung, Steve; Day, Delbert; Neidig, Katie; Dusevich, Vladimir; Eick, David; Bonewald, Lynda

    2012-12-01

    Bioactive glasses are biocompatible materials that convert to hydroxyapatite in vivo, and potentially support bone formation, but have mainly been available in particulate and not scaffold form. In this study, borosilicate and borate bioactive glass scaffolds were evaluated in critical-sized rat calvarial defects. Twelve-week-old rats were implanted with 45S5 silicate glass particles and scaffolds of 1393 silicate, 1393B1 borosilicate, and 1393B3 borate glass. After 12 weeks, the defects were harvested, stained with hematoxylin and eosin to evaluate bone regeneration, Periodic Acid Schiff to quantitate blood vessel area, and von Kossa and backscatter SEM to estimate newly mineralized bone and hydroxyapatite conversion of bioactive glasses. The amount of new bone was 12.4% for 45S5, 8.5% for 1393, 9.7% for 1393B1, and 14.9% for 1393B3 (*p = 0.04; cf. 1393 and 1393B1). Blood vessel area was significantly higher (p = 0.009) with 45S5 (3.8%), with no differences among 1393 (2.0%), 1393B1 (2.4%), or 1393B3 (2.2%). Percent von Kossa-positive area was 18.7% for 45S5, 25.4% for 1393, 29.5% for 1393B1, and 30.1% for 1393B3, significantly higher (p = 0.014) in 1393B1 and 1393B3 glasses than in 45S5. 45S5 and 1393B3 converted completely to HA in vivo. The 1393B3 glass provided greater bone formation and may be more promising for bone defect repair due to its capacity to be molded into scaffolds. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A 100A:3267-3275, 2012.

  8. Micromechanical finite-element modeling and experimental characterization of the compressive mechanical properties of polycaprolactone-hydroxyapatite composite scaffolds prepared by selective laser sintering for bone tissue engineering.

    PubMed

    Eshraghi, Shaun; Das, Suman

    2012-08-01

    Bioresorbable scaffolds with mechanical properties suitable for bone tissue engineering were fabricated from polycaprolactone (PCL) and hydroxyapatite (HA) by selective laser sintering (SLS) and modeled by finite-element analysis (FEA). Both solid gage parts and scaffolds having 1-D, 2-D and 3-D orthogonal, periodic porous architectures were made with 0, 10, 20 and 30 vol.% HA. PCL:HA scaffolds manufactured by SLS had nearly full density (99%) in the designed solid regions and had excellent geometric and dimensional control. Through optimization of the SLS process, the compressive moduli for our solid gage parts and scaffolds are the highest reported in the literature for additive manufacturing. The compressive moduli of solid gage parts were 299.3, 311.2, 415.5 and 498.3 MPa for PCL:HA loading at 100:0, 90:10, 80:20 and 70:30, respectively. The compressive effective stiffness tended to increase as the loading of HA was increased and the designed porosity was lowered. In the case of the most 3-D porous scaffold, the compressive modulus more than doubled from 14.9 to 36.2 MPa when changing the material from 100:0 to 70:30 PCL:HA. A micromechanical FEA model was developed to investigate the reinforcement effect of HA loading on the compressive modulus of the bulk material. Using a first-principles based approach, the random distribution of HA particles in a solidified PCL matrix was modeled for any HA loading to predict the bulk mechanical properties of the composites. The bulk mechanical properties were also used for FEA of the scaffold geometries. The results of the FEA were found to be in good agreement with experimental mechanical testing. The development of patient- and site-specific composite tissue-engineering constructs with tailored properties can be seen as a direct extension of this work on computational design, a priori modeling of mechanical properties and direct digital manufacturing. Copyright © 2012 Acta Materialia Inc. Published by Elsevier Ltd. All

  9. Micromechanical finite element modeling and experimental characterization of the compressive mechanical properties of polycaprolactone:hydroxyapatite composite scaffolds prepared by selective laser sintering for bone tissue engineering

    PubMed Central

    Eshraghi, Shaun; Das, Suman

    2012-01-01

    Bioresorbable scaffolds with mechanical properties suitable for bone tissue engineering were fabricated from polycaprolactone (PCL) and hydroxyapatite (HA) by selective laser sintering (SLS) and modeled by finite element analysis (FEA). Both solid gage parts and scaffolds having 1-D, 2-D and 3-D orthogonal, periodic porous architectures were made with 0, 10, 20 and 30% HA by volume. PCL:HA scaffolds manufactured by SLS had nearly full density (99%) in the designed solid regions and had excellent geometric and dimensional control. Through optimization of the SLS process, the compressive moduli for our solid gage parts and scaffolds are the highest reported in the literature for additive manufacturing. The compressive moduli of solid gage parts were 299.3, 311.2, 415.5 and 498.3 MPa for PCL:HA loading at 100:0, 90:10, 80:20 and 70:30 respectively. The compressive effective stiffness tended to increase as the loading of HA was increased and the designed porosity was lowered. In the case of the most 3-D porous scaffold, the compressive modulus more than doubled from 14.9 MPa to 36.2 MPa when changing the material from 100:0 to 70:30 PCL:HA. A micromechanical finite element analysis (FEA) model was developed to investigate the reinforcement effect of HA loading on the compressive modulus of the bulk material. Using a first-principles based approach, the random distribution of HA particles in a solidified PCL matrix was modeled for any loading of HA to predict the bulk mechanical properties of the composites. The bulk mechanical properties were also used for FEA of the scaffold geometries. Results of the FEA were found to be in good agreement with experimental mechanical testing. The development of patient and site-specific composite tissue engineering constructs with tailored properties can be seen as a direct extension of this work on computational design, a priori modeling of mechanical properties and direct digital manufacturing. PMID:22522129

  10. Comparison between PCL/hydroxyapatite (HA) and PCL/halloysite nanotube (HNT) composite scaffolds prepared by co-extrusion and gas foaming.

    PubMed

    Jing, Xin; Mi, Hao-Yang; Turng, Lih-Sheng

    2017-03-01

    In this work, three-dimensional poly(caprolactone) (PCL) tissue engineering scaffolds were prepared by co-extrusion and gas foaming. Biocompatible hydroxyapatite (HA) and halloysite nanotubes (HNT) were added to the polymer matrix to enhance the mechanical properties and bioactivity of the composite scaffolds. The effects of HA and HNT on the rheological behavior, microstructure, and mechanical properties of the composite scaffolds were systematically compared. It was found that the HNT improved viscosity more significantly than HA, and reduced the pore size of scaffolds, while the mechanical performance of PCL/HNT scaffolds was higher than PCL/HA scaffolds with the same filler content. Human mesenchymal stem cells (hMSCs) were used as the cell model to compare the biological properties of two composite scaffolds. The results demonstrated that cells could survive on all scaffolds, and showed a more flourishing living state on the composite scaffolds. The cell differentiation for 5% HA and 1% HNT scaffolds were significantly higher than other scaffolds, while the differentiation of 5% HNT scaffolds was lower than that of 1% HNT scaffolds mainly because of the reduced pore size and pore interconnectivity. Therefore, this study suggested that, with proper filler content and control of microstructure through processing, HNT could be a suitable substitute for HA for bone tissue engineering to reduce the cost and improve mechanical performance.

  11. 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.

  12. Biodegradable polymer scaffold fabrication and the creation of tissue-engineered bone

    NASA Astrophysics Data System (ADS)

    Thomson, Robert Craig

    It is estimated that over one million surgeries to restore lost bone function are performed each year in the U.S. Although existing therapies generally result in adequate restoration of mechanical or aesthetic function, they are by no means ideal. An alternative method for bone regeneration has been proposed which utilizes autologous cell transplantation within porous biodegradable polymer foam scaffolds. The work presented in this thesis was aimed at creating a porous biodegradable scaffold with appropriate mechanical properties, which in concert with an appropriate vascular environment, would be sufficient to create tissue engineered bone. A method was developed to fabric ate three-dimensional poly(DL-lactic-co-glycolic acid) (PLGA) foams which involves the production of PLGA/gelatin composites followed by leaching of the embedded gelatin microspheres. These PLGA foams had insufficient yield strength in compression. However, the results served as a basis for the design of foams with mechanical properties suitable for bone regeneration. As a means of enhancing foam compressive strength, this processing technique was modified to produce composite foams of PLGA and hydroxyapatite short fibers. Low porosity composite foams with enhanced compressive yield strengths were produced. We were able to manufacture: high porosity foams suitable for cell seeding but which have minimal compressive yield strength; or low porosity foams with enhanced compressive yield strength but which may not suitable for cell seeding. Using bone chambers filled with morcellized bone graft and implanted against ovine rib periosteum, we were able to form a clinically significant mass of vascularized bone with substantial compressive strength which was comparable to normal bone. Appropriately designed PLGA foams implanted against the periosteum allowed vascularized tissue ingrowth. We believe this tissue may have a large number of osteoprogenitor cells of periosteal origin and may therefore

  13. Polymer-ceramic spiral structured scaffolds for bone tissue engineering: effect of hydroxyapatite composition on human fetal osteoblasts.

    PubMed

    Zhang, Xiaojun; Chang, Wei; Lee, Paul; Wang, Yuhao; Yang, Min; Li, Jun; Kumbar, Sangamesh G; Yu, Xiaojun

    2014-01-01

    For successful bone tissue engineering, a scaffold needs to be osteoconductive, porous, and biodegradable, thus able to support attachment and proliferation of bone cells and guide bone formation. Recently, hydroxyapatites (HA), a major inorganic component of natural bone, and biodegrade polymers have drawn much attention as bone scaffolds. The present study was designed to investigate whether the bone regenerative properties of nano-HA/polycaprolactone (PCL) spiral scaffolds are augmented in an HA dose dependent manner, thereby establishing a suitable composition as a bone formation material. Nano-HA/PCL spiral scaffolds were prepared with different weight ratios of HA and PCL, while porosity was introduced by a modified salt leaching technique. Human fetal osteoblasts (hFOBs) were cultured on the nano-HA/PCL spiral scaffolds up to 14 days. Cellular responses in terms of cell adhesion, viability, proliferation, differentiation, and the expression of bone-related genes were investigated. These scaffolds supported hFOBs adhesion, viability and proliferation. Cell proliferation trend was quite similar on polymer-ceramic and neat polymer spiral scaffolds on days 1, 7, and 14. However, the significantly increased amount of alkaline phosphatase (ALP) activity and mineralized matrix synthesis was evident on the nano-HA/PCL spiral scaffolds. The HA composition in the scaffolds showed a significant effect on ALP and mineralization. Bone phenotypic markers such as bone sialoprotein (BSP), osteonectin (ON), osteocalcin (OC), and type I collagen (Col-1) were semi-quantitatively estimated by reverse transcriptase polymerase chain reaction analysis. All of these results suggested the osteoconductive characteristics of HA/PCL nanocomposite and cell maturation were HA dose dependent. For instance, HA∶PCL = 1∶4 group showed significantly higher ALP mineralization and elevated levels of BSP, ON, OC and Col-I expression as compared other lower or higher ceramic ratios

  14. Polymer-Ceramic Spiral Structured Scaffolds for Bone Tissue Engineering: Effect of Hydroxyapatite Composition on Human Fetal Osteoblasts

    PubMed Central

    Zhang, Xiaojun; Chang, Wei; Lee, Paul; Wang, Yuhao; Yang, Min; Li, Jun; Kumbar, Sangamesh G.; Yu, Xiaojun

    2014-01-01

    For successful bone tissue engineering, a scaffold needs to be osteoconductive, porous, and biodegradable, thus able to support attachment and proliferation of bone cells and guide bone formation. Recently, hydroxyapatites (HA), a major inorganic component of natural bone, and biodegrade polymers have drawn much attention as bone scaffolds. The present study was designed to investigate whether the bone regenerative properties of nano-HA/polycaprolactone (PCL) spiral scaffolds are augmented in an HA dose dependent manner, thereby establishing a suitable composition as a bone formation material. Nano-HA/PCL spiral scaffolds were prepared with different weight ratios of HA and PCL, while porosity was introduced by a modified salt leaching technique. Human fetal osteoblasts (hFOBs) were cultured on the nano-HA/PCL spiral scaffolds up to 14 days. Cellular responses in terms of cell adhesion, viability, proliferation, differentiation, and the expression of bone-related genes were investigated. These scaffolds supported hFOBs adhesion, viability and proliferation. Cell proliferation trend was quite similar on polymer-ceramic and neat polymer spiral scaffolds on days 1, 7, and 14. However, the significantly increased amount of alkaline phosphatase (ALP) activity and mineralized matrix synthesis was evident on the nano-HA/PCL spiral scaffolds. The HA composition in the scaffolds showed a significant effect on ALP and mineralization. Bone phenotypic markers such as bone sialoprotein (BSP), osteonectin (ON), osteocalcin (OC), and type I collagen (Col-1) were semi-quantitatively estimated by reverse transcriptase polymerase chain reaction analysis. All of these results suggested the osteoconductive characteristics of HA/PCL nanocomposite and cell maturation were HA dose dependent. For instance, HA∶PCL = 1∶4 group showed significantly higher ALP mineralization and elevated levels of BSP, ON, OC and Col-I expression as compared other lower or higher ceramic ratios

  15. Osteogenesis and Trophic Factor Secretion are Influenced by the Composition of Hydroxyapatite/Poly(Lactide-Co-Glycolide) Composite Scaffolds

    PubMed Central

    He, Jiawei; Genetos, Damian C.

    2010-01-01

    The use of composite biomaterials for bone repair capitalizes on the beneficial aspects of individual materials while tailoring the mechanical properties of the composite. We hypothesized that substrate composition would modulate the osteogenic response and secretion of potent trophic factors by human mesenchymal stem cells (hMSCs). Composite scaffolds were prepared by combining nanosized hydroxyapatite (HA) and microspheres formed of poly(lactic-co-glycolic acid) (PLG) at varying ratios between 0:1 and 5:1. Scaffolds were seeded with hMSCs for culture in osteogenic conditions or subcutaneous implantation into nude rats. Compressive moduli increased with HA content in a near-linear fashion. The osteogenic differentiation of hMSCs increased in a dose-dependent manner as determined by alkaline phosphatase activity and osteopontin secretion after 4 weeks of culture. Further, endogenous secretion of vascular endothelial growth factor was sustained at significantly higher levels over 28 days for hMSCs seeded in 2.5:1 and 5:1 HA:PLG scaffolds. Eight weeks after implantation, scaffolds with higher HA:PLG ratios exhibited greater vascularization and more mineralized tissue. These data demonstrate that the mechanical properties, osteogenic differentiation, as well as the timing and duration of trophic factor secretion by hMSCs can be tailored through controlling the composition of the polymer–bioceramic composite. PMID:19642853

  16. Templated repair of long bone defects in rats with bioactive spiral-wrapped electrospun amphiphilic polymer/hydroxyapatite scaffolds.

    PubMed

    Kutikov, Artem B; Skelly, Jordan D; Ayers, David C; Song, Jie

    2015-03-04

    Effective repair of critical-size long bone defects presents a significant clinical challenge. Electrospun scaffolds can be exploited to deliver protein therapeutics and progenitor cells, but their standalone application for long bone repair has not been explored. We have previously shown that electrospun composites of amphiphilic poly(d,l-lactic acid)-co-poly(ethylene glycol)-co-poly(d,l-lactic acid) (PELA) and hydroxyapatite (HA) guide the osteogenic differentiation of bone marrow stromal cells (MSCs), making these scaffolds uniquely suited for evaluating cell-based bone regeneration approaches. Here we examine whether the in vitro bioactivity of these electrospun scaffolds can be exploited for long bone defect repair, either through the participation of exogenous MSCs or through the activation of endogenous cells by a low dose of recombinant human bone morphogenetic protein-2 (rhBMP-2). In critical-size rat femoral segmental defects, spiral-wrapped electrospun HA-PELA with preseeded MSCs resulted in laminated endochondral ossification templated by the scaffold across the longitudinal span of the defect. Using GFP labeling, we confirmed that the exogenous MSCs adhered to HA-PELA survived at least 7 days postimplantation, suggesting direct participation of these exogenous cells in templated bone formation. When loaded with 500 ng of rhBMP-2, HA-PELA spirals led to more robust but less clearly templated bone formation than MSC-bearing scaffolds. Both treatment groups resulted in new bone bridging over the majority of the defect by 12 weeks. This study is the first demonstration of a standalone bioactive electrospun scaffold for templated bone formation in critical-size long bone defects.

  17. Induction and quantification of collagen fiber alignment in a three-dimensional hydroxyapatite-collagen composite scaffold.

    PubMed

    Banglmaier, Richard F; Sander, Edward A; VandeVord, Pamela J

    2015-04-01

    Hydroxyapatite-collagen composite scaffolds are designed to serve as a regenerative load bearing replacement that mimics bone. However, the material properties of these scaffolds are at least an order of magnitude less than that of bone and subject to fail under physiological loading conditions. These scaffolds compositionally resemble bone but they do not possess important structural attributes such as an ordered arrangement of collagen fibers, which is a correlate to the mechanical properties in bone. Furthermore, it is unclear how much ordering of structure is satisfactory to mimic bone. Therefore, quantitative methods are needed to characterize collagen fiber alignment in these scaffolds for better correlation between the scaffold structure and the mechanical properties. A combination of extrusion and compaction was used to induce collagen fiber alignment in composite scaffolds. Collagen fiber alignment, due to extrusion and compaction, was quantified from polarized light microscopy images with a Fourier transform image processing algorithm. The Fourier transform method was capable of resolving the degree of collagen alignment from polarized light images. Anisotropy indices of the image planes ranged from 0.08 to 0.45. Increases in the degree of fiber alignment induced solely by extrusion (0.08-0.25) or compaction (0.25-0.44) were not as great as those by the combination of extrusion and compaction (0.35-0.45). Additional measures of randomness and fiber direction corroborate these anisotropy findings. This increased degree of collagen fiber alignment was induced in a preferred direction that is consistent with the extrusion direction and parallel with the compacted plane.

  18. Osteogenic/Odontogenic Bioengineering with co-Administration of Simvastatin and Hydroxyapatite on Poly Caprolactone Based Nanofibrous Scaffold

    PubMed Central

    Samiei, Mohammad; Aghazadeh, Marziyeh; Alizadeh, Effat; Aslaminabadi, Naser; Davaran, Soodabeh; Shirazi, Sajjad; Ashrafi, Farhad; Salehi, Roya

    2016-01-01

    Purpose: Statin is an effective factor for promoting osteogenesis. The aim of the present study was to evaluate the effect of simvastatin (SIM) and/or HA addition on changes in osteogenesis levels by human DPSCs transferred onto three-dimensional (3D) nanofibrous Poly (ε-caprolactone) (PCL)/Poly lactic acide (PLLA) polymeric scaffolds. Methods: For this purpose, a 3D nanofibrous composite scaffold of PCL/PLLA/HA was prepared by electrospinning method. SIM was added to scaffolds during DPSCs culturing step. Cell proliferation and osteogenic activity levels were assessed by using MTT assay and Alizarin Red assay methods. In addition, the expression of genes responsible for osteogenesis, including BMP2, Osteocalcin, DSPP and RUNX2, were determined before and 2 weeks after incorporation of SIM. Results: The MTT assay showed that PCL/PLLA/HA scaffolds seeded with DPSCs has significant (p<0.05) more proliferative effect than PCL/PLLA or DMEM cultured cells, additionally SIM administration improved this result over the PCL/PLLA/HA scaffolds without SIM treatment. SEM imaging revealed improved adhesion and probably osteogenic differentiation of DPSCs on PCL/PLLA/HA nanofibers treated with SIM, moreover the alizarin red assay ensured significant (p<0.05) higher mineralization of this group. Finally, real time PCR confirmed the positive regulation (P<0.05) of the expression of osteo/odontogenesis markers BMP2, Osteocalcin, DSPP and RUNX2 genes in PLLA-PCL-HA (0.1)-SIM group. Conclusion: As a result, addition of simvastatin with incorporation of hydroxyapatite in PCL-PLLA scaffolds might increase the expression of osteogenesis markers in the DPSCs, with a possible increase in cell differentiation and bone formation. PMID:27766219

  19. Fabrication and characterization of modified-hydroxyapatite/polyetheretherketone coating materials

    NASA Astrophysics Data System (ADS)

    Ma, Rui; Fang, Lin; Luo, Zhongkuan; Zheng, Ruisheng; Song, Shenhua; Weng, Luqian; Lei, JinPing

    2014-09-01

    45 wt%-Hydroxyaptite/polyetheretherketone (HA/PEEK) coating materials modified by silane coupling agent (KH560) on PEEK substrate were successfully fabricated by solution casting method and characterized by Fourier transform infrared spectroscopy (FTIR), X-Ray Diffraction (XRD), scanning electron microscopy (SEM), and tensile testing. The modified HA fillers were obtained to be uniformly distributed in the HA/PEEK coating, which has better properties of tensile strength and fracture toughness than those of the unmodified specimen. A good bonding between the composite coating and the PEEK substrate was achieved by solution casting method, resulting in integral-fracture without falling apart or delaminating during tensile loading. The modified specimens dipped into simulated body fluid (SBF) were characterized by SEM, XRD and FTIR, indicating that the bioactivity of the dipped materials was demonstrated more apparent with extending the dipping time. Therefore, the coating materials may become the substitutes for the hard tissues of the human body in the future, which could realize the balance between the mechanical properties and the bioactivity by modifying the structural design of the coating.

  20. Janus emulsion mediated porous scaffold bio-fabrication.

    PubMed

    Kovach, Ildiko; Rumschöttel, Jens; Friberg, Stig E; Koetz, Joachim

    2016-09-01

    A three dimensional biopolymer network structure with incorporated nano-porous calcium phosphate (CaP) balls was fabricated by using gelatin-chitosan (GC) polymer blend and GC stabilized olive/silicone oil Janus emulsions, respectively. The emulsions were freeze-dried, and the oil droplets were washed out in order to prepare porous scaffolds with larger surface area. The morphology, pore size, chemical composition, thermal and swelling behavior was studied by Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FTIR) and micro-Differential Scanning Calorimetry (micro-DSC). Microscopic analysis confirmed that the pore size of the GC based sponges after freeze-drying may be drastically reduced by using Janus emulsions. Besides, the incorporation of nanoporous calcium phosphate balls is also lowering the pore size and enhancing thermal stability. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Alternate Calcification in Microcapillaries for the Fabrication of Hydroxyapatite Films without Light Exposure, Calcination, or Applied Voltage.

    PubMed

    Watanabe, Satoshi; Iida, Munetoyo; Matsumoto, Mutsuyoshi

    2016-10-01

    We discuss an alternate method for calcification in microcapillaries for fabricating calcium phosphate films using silicone molds and calcifying solutions. Calcium phosphate films with a line/space of 5-50 µm were fabricated by controlling the concentrations of calcium chloride and sodium phosphate solutions. Plate-type crystals of hydroxyapatite were grown when the calcium phosphate films were immersed in hydroxyapatite precursors. In the initial stage of hydroxyapatite crystal growth, the c-plane of the crystals was grown parallel to the substrates, and subsequently the growth followed with the c-plane growing perpendicular to the substrates. In narrow capillaries, dendritic structures were formed with a tendency to grow in a direction parallel to the direction of the microcapillaries. This technique is useful in the micropatterning of biocompatible ceramics with a minimized material consumption and a short fabrication time.

  2. Solvent-free Fabrication of Tissue Engineering Scaffolds with Immiscible Polymer Blends

    PubMed Central

    Ma, Liang; Jiang, Wei; Li, Wei

    2014-01-01

    A completely organic solvent-free fabrication method is developed for tissue engineering scaffolds by gas foaming of immiscible polylactic acid (PLA) and sucrose blends, followed by water leaching. PLA scaffolds with above 90% porosity and 25–200 μm pore size were fabricated. The pore size and porosity was controlled with process parameters including extrusion temperature and foaming process parameters. Dynamic mechanical analysis showed that the extrusion temperature could be used to control the scaffold strength. Both unfoamed and foamed scaffolds were used to culture glioblastoma (GBM) cells M059K. The results showed that the cells grew better in the foamed PLA scaffolds. The method presented in the paper is versatile and can be used to fabricate tissue engineering scaffolds without any residual organic solvents. PMID:24764605

  3. In Vivo Performance of Bilayer Hydroxyapatite Scaffolds for Bone Tissue Regeneration in the Rabbit Radius

    DTIC Science & Technology

    2011-02-02

    compared to the mineral density of contralateral limbs for animals receiving scaffolds or autologous bone grafts treatments (Fig. 7a). In agreement...significantly lower flex- ural toughness when compared to the contralateral limbs of animals receiving scaffolds or autologous bone graft treatments. Similarly... comparable to empty defects and flexural toughness being comparable to defects implanted with autologous bone grafts . The scaffolds

  4. Strategic design and recent fabrication techniques for bioengineered tissue scaffolds to improve peripheral nerve regeneration.

    PubMed

    Rajaram, Ajay; Chen, Xiong-Biao; Schreyer, David J

    2012-12-01

    Bioengineered tissue scaffolds are a potential tool for improving regenerative repair of damaged peripheral nerves. Novel modes of fabrication coupled with scaffold design strategies that are based on an understanding of the biology of nerve injury offer the prospect of intervention at a more sophisticated level. We review the etiology and incidence of peripheral nerve injury and the biological events that unfold during nerve regeneration after an injury. Newly available tissue scaffold fabrication technologies using bioplotting and laser-based techniques are described. Scaffold design strategies are also discussed, including the incorporation of living cells during scaffold fabrication, inclusion of neurotrophic gradients, use of electric stimulation, inclusion of antioxidant compounds to counteract neural apotosis, and promotion of angiogenesis. Use of these advanced fabrication techniques and incorporation of one or more of these active biological strategies may eventually lead to a greater success in peripheral nerve tissue engineering.

  5. A Versatile Method for Fabricating Tissue Engineering Scaffolds with a Three-Dimensional Channel for Prevasculature Networks.

    PubMed

    Li, Shuai; Liu, Yuan-Yuan; Liu, Li-Jun; Hu, Qing-Xi

    2016-09-28

    Despite considerable advances in tissue engineering over the past two decades, solutions to some crucial problems remain elusive. Vascularization is one of the most important factors that greatly influence the function of scaffolds. Many research studies have focused on the construction of a vascular-like network with prevascularization structure. Sacrificial materials are widely used to build perfusable vascular-like architectures, but most of these fabricated scaffolds only have a 2D plane-connected network. The fabrication of three-dimensional perfusable branched networks remains an urgent issue. In this work, we developed a novel sacrificial molding technique for fabricating biocompatible scaffolds with a three-dimensional perfusable branched network. Here, 3D-printed poly(vinyl alcohol) (PVA) filament was used as the sacrificial material. The fused PVA was deposited on the surface of a cylinder to create the 3D branched solid network. Gelatin was used to embed the solid network. Then, the PVA mold was dissolved after curing the hydrogel. The obtained architecture shows good perfusability. Cell experiment results indicated that human umbilical vein endothelial cells (HUVECs) successfully attached to the surface of the branched channel and maintained high viability after a few days in culture. In order to prevent deformation of the channel, paraffin was coated on the surface of the printed structure, and hydroxyapatite (HA) was added to gelatin. In conclusion, we demonstrate a novel strategy toward the engineering of prevasculature thick tissues through the integration of the fused PVA filament deposit. This approach has great potential in solving the issue of three-dimensional perfusable branched networks and opens the way to clinical applications.

  6. Surface modification of polycaprolactone scaffolds fabricated via selective laser sintering for cartilage tissue engineering.

    PubMed

    Chen, Chih-Hao; Lee, Ming-Yih; Shyu, Victor Bong-Hang; Chen, Yi-Chieh; Chen, Chien-Tzung; Chen, Jyh-Ping

    2014-07-01

    Surface modified porous polycaprolactone scaffolds fabricated via rapid prototyping techniques were evaluated for cartilage tissue engineering purposes. Polycaprolactone scaffolds manufactured by selective laser sintering (SLS) were surface modified through immersion coating with either gelatin or collagen. Three groups of scaffolds were created and compared for both mechanical and biological properties. Surface modification with collagen or gelatin improved the hydrophilicity, water uptake and mechanical strength of the pristine scaffold. From microscopic observations and biochemical analysis, collagen-modified scaffold was the best for cartilage tissue engineering in terms of cell proliferation and extracellular matrix production. Chondrocytes/collagen-modified scaffold constructs were implanted subdermally in the dorsal spaces of female nude mice. Histological and immunohistochemical staining of the retrieved implants after 8 weeks revealed enhanced cartilage tissue formation. We conclude that collagen surface modification through immersion coating on SLS-manufactured scaffolds is a feasible scaffold for cartilage tissue engineering in craniofacial reconstruction. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Engineered electrospun poly(caprolactone)/polycaprolactone-g-hydroxyapatite nano-fibrous scaffold promotes human fibroblasts adhesion and proliferation.

    PubMed

    Keivani, F; Shokrollahi, P; Zandi, M; Irani, S; F Shokrolahi; Khorasani, S C

    2016-11-01

    Polycaprolactone (PCL)/hydroxyapatite nano-composites are among the best candidates for tissue engineering. However, interactions between nHAp and PCL are difficult to control leading to inhomogeneous dispersion of the bio-ceramic particles. Grafting of polymer chains at high density/chain length while promotes the phase compatibility may result in reduced HAp exposed surface area and therefore, bioactivity is compromised. This issue is addressed here by grafting PCL chains onto HAp nano-particles through ring opening polymerization of ε-caprolactone (PCL-g-HAp). FTIR and TGA analysis showed that PCL (6.9wt%), was successfully grafted on the HAp. PCL/PCL-g-HAp nano-fibrous scaffold showed up to 10 and 33% enhancement in tensile strength and modulus, respectively, compared to those of PCL/HAp. The effects of HAp on the in vitro HAp formation were investigated for both the PCL/HAp and PCL/PCL-g-HAp scaffolds. Precipitation of HAp on the nano-composite scaffolds observed after 15days incubation in simulated body fluid (SBF), as confirmed by scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDX). Human fibroblasts were seeded on PCL, PCL/HAp and PCL/PCL-g-HAp scaffolds. According to MTT assay, the highest cell proliferation was recorded for PCL/PCL-g-HAp nano-composite, at all time intervals (1-21days, P<0.001). Fluorescent microscopy (of DAPI stained samples) and electron microscopy images showed that all nano-fibrous scaffolds (PCL, PCL/HAp, and PCL/PCL-g-HAp), were non-toxic against cells, while more cell adhesion, and the most uniform cell distribution observed on the PCL/PCL-g-HAp. Overall, grafting of relatively short chains of PCL on the surface of HAp nano-particles stimulates fibroblasts adhesion and proliferation on the PCL/PCL-g-HAp nano-composite.

  8. Fabrication and mechanical evaluation of hydroxyapatite/oxide nano-composite materials.

    PubMed

    Mohamed, Khaled R; Beherei, Hanan H; El Bassyouni, Gehan T; El Mahallawy, Nahed

    2013-10-01

    In the current study, the semiconducting metal oxides such as nano-ZnO and SiO2 powders were prepared via sol-gel technique and conducted on nano-hydroxyapatite (nHA) which was synthesized by chemical precipitation. The properties of fabricated nano-structured composites containing different ratios of HA, ZnO and SiO2 were examined using X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscope (SEM) and transmission electron microscope (TEM) techniques. The effect of the variation of ratios between the three components on mechanical, microstructure and in-vitro properties was assessed to explore the possibility of enhancing these properties. The results proved that the mechanical properties exhibited an increment with increasing the ZnO content at the extent of HA. In-vitro study proved the formation and nucleation of apatite onto the surface of the fabricated composites after one week of immersion. It is concluded that HA composites containing SiO2 or SiO2/ZnO content had a suitable mechanical properties and ability to form apatite particles onto the composite surface. Based on bioactivity behavior, Si-HA is more bioactive than pure hydroxyapatite and nano-arrangements will provide an interface for better bone formation. Therefore, these nano-composites will be promising as bone substitutes especially in load bearing sites.

  9. Improvement of the compressive strength of a cuttlefish bone-derived porous hydroxyapatite scaffold via polycaprolactone coating.

    PubMed

    Kim, Beom-Su; Kang, Hyo Jin; Lee, Jun

    2013-10-01

    Cuttlefish bones (CBs) have emerged as attractive biomaterials because of their porous structure and components that can be converted into hydroxyapatite (HAp) via a hydrothermal reaction. However, their brittleness and low strength restrict their application in bone tissue engineering. Therefore, to improve the compressive strength of the scaffold following hydrothermal conversion to a HAp form of CB (CB-HAp), the scaffold was coated using a polycaprolactone (PCL) polymer at various concentrations. In this study, raw CB was successfully converted into HAp via a hydrothermal reaction. We then evaluated their surface properties and composition by scanning electron microscopy and X-ray diffraction analysis. The CB-HAp coated with PCL showed improved compressive performance and retained a microporous structure. The compressive strength was significantly increased upon coating with 5 and 10% PCL, by 2.09- and 3.30-fold, respectively, as compared with uncoated CB-HAp. However, coating with 10% PCL resulted in a reduction in porosity. Furthermore, an in vitro biological evaluation demonstrated that MG-63 cells adhered well, proliferated and were able to be differentiated on the PCL-coated CB-HAp scaffold, which was noncytotoxic. These results suggest that a simple coating method is useful to improve the compressive strength of CB-HAp for bone tissue engineering applications. Copyright © 2013 Wiley Periodicals, Inc.

  10. Bone augmentation after ectopic implantation of a cell-free collagen-hydroxyapatite scaffold in the mouse

    PubMed Central

    Calabrese, Giovanna; Giuffrida, Raffaella; Forte, Stefano; Salvatorelli, Lucia; Fabbi, Claudia; Figallo, Elisa; Gulisano, Massimo; Parenti, Rosalba; Magro, Gaetano; Colarossi, Cristina; Memeo, Lorenzo; Gulino, Rosario

    2016-01-01

    The bone grafting is the classical way to treat large bone defects. Among the available techniques, autologous bone grafting is still the most used but, however, it can cause complications such as infection and donor site morbidity. Alternative and innovative methods rely on the development of biomaterials mimicking the structure and properties of natural bone. In this study, we characterized a cell-free scaffold, which was subcutaneously implanted in mice and then analyzed both in vivo and ex vivo after 1, 2, 4, 8 and 16 weeks, respectively. Two types of biomaterials, made of either collagen alone or collagen plus magnesium-enriched hydroxyapatite have been used. The results indicate that bone augmentation and angiogenesis could spontaneously occur into the biomaterial, probably by the recruitment of host cells, and that the composition of the scaffolds is crucial. In particular, the biomaterial more closely mimicking the native bone drives the process of bone augmentation more efficiently. Gene expression analysis and immunohistochemistry demonstrate the expression of typical markers of osteogenesis by the host cells populating the scaffold. Our data suggest that this biomaterial could represent a promising tool for the reconstruction of large bone defects, without using exogenous living cells or growth factors. PMID:27821853

  11. Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications.

    PubMed

    Jung, Hyun-Do; Lee, Hyun; Kim, Hyoun-Ee; Koh, Young-Hag; Song, Juha

    2015-12-08

    Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness and low bioactivity of metals. In this study, we have developed a new method towards fabricating a new type of bioactive and mechanically reliable porous metal scaffolds-densified porous Ti scaffolds. The method consists of two fabrication processes, 1) the fabrication of porous Ti scaffolds by dynamic freeze casting, and 2) coating and densification of the porous scaffolds. The dynamic freeze casting method to fabricate porous Ti scaffolds allowed the densification of porous scaffolds by minimizing the chemical contamination and structural defects. The densification process is distinctive for three reasons. First, the densification process is simple, because it requires a control of only one parameter (degree of densification). Second, it is effective, as it achieves mechanical enhancement and sustainable release of biomolecules from porous scaffolds. Third, it has broad applications, as it is also applicable to the fabrication of functionally graded porous scaffolds by spatially varied strain during densification.

  12. Novel biodegradable star-shaped polylactide scaffolds for bone regeneration fabricated by two-photon polymerization.

    PubMed

    Timashev, Peter; Kuznetsova, Daria; Koroleva, Anastasia; Prodanets, Natalia; Deiwick, Andrea; Piskun, Yuri; Bardakova, Ksenia; Dzhoyashvili, Nina; Kostjuk, Sergei; Zagaynova, Elena; Rochev, Yuri; Chichkov, Boris; Bagratashvili, Viktor

    2016-05-01

    To assess the properties of 3D biodegradable scaffolds fabricated from novel star-shaped poly(D,L-lactide) (SSL) materials for bone tissue regeneration. The SSL polymer was synthesized using an optimized synthetic procedure and applied for scaffold fabrication by the two-photon polymerization technique. The osteogenic differentiation was controlled using human adipose-derived stem cells cultured for 28 days. The SSL scaffolds with or without murine MSCs were implanted into the cranial bone of C57/Bl6 mice. The SSL scaffolds supported differentiation of human adipose-derived stem cells toward the osteogenic lineage in vitro. The SSL scaffolds with murine MSCs enhanced the mineralized tissue formation. The SSL scaffolds provide a beneficial microenvironment for the osteogenic MSCs' differentiation in vitro and support de novo bone formation in vivo.

  13. Preparation, characterization and antimicrobial activity of a bio-composite scaffold containing chitosan/nano-hydroxyapatite/nano-silver for bone tissue engineering.

    PubMed

    Saravanan, Sekaran; Nethala, Sricharan; Pattnaik, Soumitri; Tripathi, Anjali; Moorthi, Ambigapathi; Selvamurugan, Nagarajan

    2011-08-01

    In this study, a bio-composite scaffold containing chitosan/nano-hydroxyapatite/nano-silver particles (CS/nHAp/nAg) was developed by freeze drying technique, followed by introduction of silver ions in controlled amount through reduction phenomenon by functional groups of chitosan. The scaffolds were characterized using SEM, FT-IR, XRD, swelling, and biodegradation studies. The testing of the prepared scaffolds with Gram-positive and Gram-negative bacterial strains showed antibacterial activity. The scaffold materials were also found to be non-toxic to rat osteoprogenitor cells and human osteosarcoma cell line. Thus, these results suggested that CS/nHAp/nAg bio-composite scaffolds have the potential in controlling implant associated bacterial infection during reconstructive surgery of bone.

  14. Comparison of glutaraldehyde and carbodiimides to crosslink tissue engineering scaffolds fabricated by decellularized porcine menisci.

    PubMed

    Gao, Shuang; Yuan, Zhiguo; Guo, Weimin; Chen, Mingxue; Liu, Shuyun; Xi, Tingfei; Guo, Quanyi

    2017-02-01

    The objectives of this study were to fabricate porous scaffolds using decellularized meniscus, and to explore a preferable crosslinking condition to enhance mechanical properties of scaffolds. Moreover, the microstructure, porosity, biodegradation and cytotoxicity were also evaluated. EDAC or GTA in different concentration was used to crosslink scaffolds. FTIR demonstrated functional groups change in crosslinking process. SEM photography showed that crosslinked scaffolds had blurry edges, which resulted scaffolds crosslinked by 1.2mol/l EDAC had smaller porosity than other groups. The structure change enhanced antidegradation property. After immersing in enzyme solution for 96h, scaffolds crosslinked by GTA and EDAC could maintain their mass >70% and 80%. Most importantly, mechanical properties of crosslinked scaffolds were also improved. Uncrosslinked Scaffolds had only 0.49kPa in compression modulus and 12.81kPa in tensile modulus. The compression and tensile modulus of scaffolds crosslinked by 1.0% GTA were 1.42 and 567.44kPa respectively. The same value of scaffolds crosslinked by 1.2mol/l EDAC were 1.49 and 532.50kPa. Scaffolds crosslinked by 1.0% and 2.5% GTA were toxic to cells, while EDAC groups showed no cytotoxicity. Chondrocytes could proliferate and infiltrate within scaffolds after seeding. Overall, 1.2mol/l EDAC was a preferable crosslinking condition.

  15. Fabrication and Mechanical Characterization of Hydrogel Infused Network Silk Scaffolds

    PubMed Central

    Kundanati, Lakshminath; Singh, Saket K.; Mandal, Biman B.; Murthy, Tejas G.; Gundiah, Namrata; Pugno, Nicola M.

    2016-01-01

    Development and characterization of porous scaffolds for tissue engineering and regenerative medicine is of great importance. In recent times, silk scaffolds were developed and successfully tested in tissue engineering and drug release applications. We developed a novel composite scaffold by mechanical infusion of silk hydrogel matrix into a highly porous network silk scaffold. The mechanical behaviour of these scaffolds was thoroughly examined for their possible use in load bearing applications. Firstly, unconfined compression experiments show that the denser composite scaffolds displayed significant enhancement in the elastic modulus as compared to either of the components. This effect was examined and further explained with the help of foam mechanics principles. Secondly, results from confined compression experiments that resemble loading of cartilage in confinement, showed nonlinear material responses for all scaffolds. Finally, the confined creep experiments were performed to calculate the hydraulic permeability of the scaffolds using soil mechanics principles. Our results show that composite scaffolds with some modifications can be a potential candidate for use of cartilage like applications. We hope such approaches help in developing novel scaffolds for tissue engineering by providing an understanding of the mechanics and can further be used to develop graded scaffolds by targeted infusion in specific regions. PMID:27681725

  16. Bone tissue engineering using polycaprolactone scaffolds fabricated via selective laser sintering.

    PubMed

    Williams, Jessica M; Adewunmi, Adebisi; Schek, Rachel M; Flanagan, Colleen L; Krebsbach, Paul H; Feinberg, Stephen E; Hollister, Scott J; Das, Suman

    2005-08-01

    Polycaprolactone (PCL) is a bioresorbable polymer with potential applications for bone and cartilage repair. In this work, porous PCL scaffolds were computationally designed and then fabricated via selective laser sintering (SLS), a rapid prototyping technique. The microstructure and mechanical properties of the fabricated scaffolds were assessed and compared to the designed porous architectures and computationally predicted properties. Scaffolds were then seeded with bone morphogenetic protein-7 (BMP-7) transduced fibroblasts and implanted subcutaneously to evaluate biological properties and to demonstrate tissue in-growth. The work done illustrates the ability to design and fabricate PCL scaffolds with porous architecture that have sufficient mechanical properties for bone tissue engineering applications using SLS. Compressive modulus and yield strength values ranged from 52 to 67 MPa and 2.0 to 3.2 Mpa, respectively, lying within the lower range of properties reported for human trabecular bone. Finite element analysis (FEA) results showed that mechanical properties of scaffold designs and of fabricated scaffolds can be computationally predicted. Histological evaluation and micro-computed tomography (microCT) analysis of implanted scaffolds showed that bone can be generated in vivo. Finally, to demonstrate the clinical application of this technology, we designed and fabricated a prototype mandibular condyle scaffold based on an actual pig condyle. The integration of scaffold computational design and free-form fabrication techniques presented here could prove highly useful for the construction of scaffolds that have anatomy specific exterior architecture derived from patient CT or MRI data and an interior porous architecture derived from computational design optimization.

  17. Porogen-based solid freeform fabrication of polycaprolactone-calcium phosphate scaffolds for tissue engineering.

    PubMed

    Mondrinos, Mark J; Dembzynski, Robert; Lu, Lin; Byrapogu, Venkata K C; Wootton, David M; Lelkes, Peter I; Zhou, Jack

    2006-09-01

    Drop on demand printing (DDP) is a solid freeform fabrication (SFF) technique capable of generating microscale physical features required for tissue engineering scaffolds. Here, we report results toward the development of a reproducible manufacturing process for tissue engineering scaffolds based on injectable porogens fabricated by DDP. Thermoplastic porogens were designed using Pro/Engineer and fabricated with a commercially available DDP machine. Scaffolds composed of either pure polycaprolactone (PCL) or homogeneous composites of PCL and calcium phosphate (CaP, 10% or 20% w/w) were subsequently fabricated by injection molding of molten polymer-ceramic composites, followed by porogen dissolution with ethanol. Scaffold pore sizes, as small as 200 microm, were attainable using the indirect (porogen-based) method. Scaffold structure and porosity were analyzed by scanning electron microscopy (SEM) and microcomputed tomography, respectively. We characterized the compressive strength of 90:10 and 80:20 PCL-CaP composite materials (19.5+/-1.4 and 24.8+/-1.3 Mpa, respectively) according to ASTM standards, as well as pure PCL scaffolds (2.77+/-0.26 MPa) fabricated using our process. Human embryonic palatal mesenchymal (HEPM) cells attached and proliferated on all scaffolds, as evidenced by fluorescent nuclear staining with Hoechst 33258 and the Alamar Blue assay, with increased proliferation observed on 80:20 PCL-CaP scaffolds. SEM revealed multilayer assembly of HEPM cells on 80:20 PCL-CaP composite, but not pure PCL, scaffolds. In summary, we have developed an SFF-based injection molding process for the fabrication of PCL and PCL-CaP scaffolds that display in vitro cytocompatibility and suitable mechanical properties for hard tissue repair.

  18. Fabrication of functional PLGA-based electrospun scaffolds and their applications in biomedical engineering.

    PubMed

    Zhao, Wen; Li, Jiaojiao; Jin, Kaixiang; Liu, Wenlong; Qiu, Xuefeng; Li, Chenrui

    2016-02-01

    Electrospun PLGA-based scaffolds have been applied extensively in biomedical engineering, such as tissue engineering and drug delivery system. Due to lack of the recognition sites on cells, hydropholicity and single-function, the applications of PLGA fibrous scaffolds are limited. In order to tackle these issues, many works have been done to obtain functional PLGA-based scaffolds, including surface modifications, the fabrication of PLGA-based composite scaffolds and drug-loaded scaffolds. The functional PLGA-based scaffolds have significantly improved cell adhesion, attachment and proliferation. Moreover, the current study has summarized the applications of functional PLGA-based scaffolds in wound dressing, vascular and bone tissue engineering area as well as drug delivery system.

  19. Fabrication of Porous α-TCP/Gellan Gum Scaffold for Bone Tissue Engineering.

    PubMed

    Wen, Jian; Kim, Ill Yong; Kikuta, Koichi; Ohtsuki, Chikara

    2016-03-01

    α-tricalcium phosphate (α-TCP, α-Ca3(PO4)2) receives great attention for bone repairing due to its biodegradability and capability of transformation to human bone's main inorganic components, hydroxyapatite (HAp). α-TCP porous scaffold is easily procurable by sintering of the low-temperature polymorph of TCP, β-TCR Still, porous body of α-TCP is too brittle to being handled and shaped, limiting its clinical application as implant materials. To improve mechanical properties of α-TCP porous scaffold, the present study focused on coating of a type of polysaccharides on α-TCP scaffolds. Gellan gum was chosen as the polysaccharide for coating because of its biodegradability as well as the potential acting as substrate for HAp deposition during hydration of α-TCP after exposure to body fluid. After coating of gellan gum on α-TCP scaffolds with porosity of 75 vol%, the compressive strength increased from 0.45 MPa to around 2.00 MPa. Among the coated scaffold, the maximum compressive strength, 3.97 MPa, was obtained on the scaffold with porosity of 63 vol%. Improvement of mechanical properties of α-TCP/gellan gum composites was achieved to show easy handling performance for a bone substitute for tissue repairing. The dissolving rate of the coated scaffolds was also controlled by adjusting the concentration of GG solutions.

  20. Development of a novel alginate-polyvinyl alcohol-hydroxyapatite hydrogel for 3D bioprinting bone tissue engineered scaffolds.

    PubMed

    Bendtsen, Stephanie T; Quinnell, Sean P; Wei, Mei

    2017-05-01

    Three-dimensional printed biomaterials used as personalized tissue substitutes have the ability to promote and enhance regeneration in areas of defected tissue. The challenge with 3D printing for bone tissue engineering remains the selection of a material with optimal rheological properties for printing in addition to biocompatibility and capacity for uniform cell incorporation. Hydrogel biomaterials may provide sufficient printability to allow cell encapsulation and bioprinting of scaffolds with uniform cell distribution. In this study, a novel alginate-polyvinyl alcohol (PVA)-hydroxyapatite (HA) hydrogel formulation with optimal rheological properties for 3D bioprinting of mouse calvaria 3T3-E1 (MC3T3) cells into scaffolds of high shape fidelity has been developed. A systematic investigation was conducted to determine the effect of varying concentrations of alginate, phosphate, calcium, and the PVA-HA suspension in the formulation on the resulting viscosity and thus printability of the hydrogel. HA, the main mineral component in natural bone, was incorporated into the hydrogel formulation to create a favorable bone-forming environment due to its excellent osteoconductivity. Degradation studies in α-MEM cell culture media showed that the 3D printed alginate-PVA-HA scaffolds remained in-tact for 14 days. MC3T3 cells were well distributed and encapsulated throughout the optimal hydrogel formulation and expressed high viability through the completion of the 3D printing process. Thus, the development of this novel, osteoconductive, biodegradable, alginate-PVA-HA formulation and its ability to 3D bioprint tissue engineered scaffolds make it a promising candidate for treating personalized bone defects. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1457-1468, 2017.

  1. FEM modeling of the reinforcement mechanism of Hydroxyapatite in PLLA scaffolds produced by supercritical drying, for Tissue Engineering applications.

    PubMed

    Baldino, L; Naddeo, F; Cardea, S; Naddeo, A; Reverchon, E

    2015-11-01

    Scaffolds have been produced by supercritical CO2 drying of Poly-L-Lactid Acid (PLLA) gels loaded with micrometric fructose particles used as porogens. These structures show a microporous architecture generated by the voids left in the solid material by porogen leaching, while they maintain the nanostructure of the gel, consisting of a network of nanofilaments. These scaffolds have also been loaded with Hydroxyapatite (HA) nanoparticles, from 10 to 50% w/w with respect to the polymer, to improve the mechanical properties of the PLLA structure. Based on miscroscopic and mechanical considerations, we propose a parametric Finite Element Method (FEM) model of PLLA-HA composites that describes the microporous structure as a close-packing of equal spheres and the nanoscale structure as a space frame of isotropic curved fibers. The effect of HA on the mechanical properties of the scaffolds has been modeled on the basis of SEM images and by taking into consideration the formation of concentric cylinders of HA nanoparticles around PLLA nanofibers. Modeling analysis confirms that mechanical properties of these scaffolds depend on nanofibrous network connections and that bending is the major factor causing deformation of the network. The FEM model also takes into account the formation of HA multi-layer coating on some areas in the nanofiber network and its increase in thickness with HA percentage. The Young modulus tends to a plateau for HA percentages larger than 30% w/w and when the coverage of the nanofibers produced by HA nanoparticles reaches a loaded surface index of 0.14 in the FEM model.

  2. In vitro mineralization of MC3T3-E1 osteoblast-like cells on collagen/nano-hydroxyapatite scaffolds coated carbon/carbon composites.

    PubMed

    Cao, Sheng; Li, Hejun; Li, Kezhi; Lu, Jinhua; Zhang, Leilei

    2016-02-01

    Collagen/nano-hydroxyapatite (collagen/nHA) scaffolds were successfully prepared on carbon/carbon composites as bioactive films using the layer-by-layer coating method. Surface characterizations of collagen/nHA scaffolds were detected by scanning electron microscope (SEM), X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. Compressive strengths of the scaffolds were evaluated by a universal test machine. In vitro biological performances were determined using scaffolds seeded with MC3T3-E1 osteoblasts-like cells and cultured in mineralization medium for up to 21 days. In addition, cellular morphologies and several related gene expressions of MC3T3-E1 cells in the scaffolds were also evaluated. Chemical and morphological analysis showed that the scaffolds had uniform pore sizes and unified phase composition. Mechanical testing indicated that the collagen/nHA scaffolds had the highest compressive strength in 50% of strain condition when the proportion of collagen and nano-hydroxyapatite was 1:3. Cellular morphology observations and cytology tests indicated that MC3T3-E1 cells were adhered on these scaffolds and proliferated. SEM photographs and gene expressions showed that mineralized MC3T3-E1 cells and newly formed extra cellular matrix (ECM) filled up the pores of the scaffolds after the 3-week mineralization inducement. Nano-sized apatite particles were secreted from MC3T3-E1 cells and combined with the reconstructed ECM. Collectively, collagen/nHA scaffolds provided C/C composites with a biomimetic surface for cell adhesion, proliferation and mineralized extra cellular matrices formation.

  3. Biocompatibility and bone-repairing effects: comparison between porous poly-lactic-co-glycolic acid and nano-hydroxyapatite/poly(lactic acid) scaffolds.

    PubMed

    Zong, Chen; Qian, Xiaodan; Tang, Zihua; Hu, Qinghong; Chen, Jiarong; Gao, Changyou; Tang, Ruikang; Tong, Xiangmin; Wang, Jinfu

    2014-06-01

    Copolymer composite scaffolds and bioceramic/polymer composite scaffolds are two representative forms of composite scaffolds used for bone tissue engineering. Studies to compare biocompatibility and bone-repairing effects between these two scaffolds are significant for selecting or improving the scaffold for clinical application. We prepared two porous scaffolds comprising poly-lactic-acid/poly-glycolic-acid (PLGA) and poly-lactic-acid/nano-hydroxyapatite (nHAP/PLA) respectively, and examined their biocompatibility with human bone marrow-derived mesenchymal stem cells (hMSCs) through evaluating adhesion, proliferation and osteogenic differentiation potentials of hMSCs in the scaffold. Then, the PLGA scaffold with hMSCs (PM construct) and the nHAP/PLA scaffold with hMSCs (HPM construct) were transplanted into the rat calvarial defect areas to compare their effects on the bone reconstruction. The results showed that the nHAP/PLA scaffold was in favor of adhesion, matrix deposition and osteogenic differentiation of hMSCs. For in vivo transplantation, both HPM and PM constructs led to mineralization and osteogenesis in the defect area of rat. However, the area grafted with PM construct showed a better formation of mature bone than that with HPM construct. In addition, the evaluation of in vitro and in vivo degradation indicated that the degradation rate of nHAP/PLA scaffold was much lower than that of PLGA scaffold. It is inferred that the lower degradation of nHAP/PLA scaffold should result in its inferior bone reconstruction in rat calvaria. Therefore, the preparation of an ideal composite scaffold for bone tissue engineering should be taken into account of the balance between its biocompatibility, degradation rate, osteoconductivity and mechanical property.

  4. Woven silk fabric-reinforced silk nanofibrous scaffolds for regenerating load-bearing soft tissues.

    PubMed

    Han, F; Liu, S; Liu, X; Pei, Y; Bai, S; Zhao, H; Lu, Q; Ma, F; Kaplan, D L; Zhu, H

    2014-02-01

    Although three-dimensional (3-D) porous regenerated silk scaffolds with outstanding biocompatibility, biodegradability and low inflammatory reactions have promising application in different tissue regeneration, the mechanical properties of regenerated scaffolds, especially suture retention strength, must be further improved to satisfy the requirements of clinical applications. This study presents woven silk fabric-reinforced silk nanofibrous scaffolds aimed at dermal tissue engineering. To improve the mechanical properties, silk scaffolds prepared by lyophilization were reinforced with degummed woven silk fabrics. The ultimate tensile strength, elongation at break and suture retention strength of the scaffolds were significantly improved, providing suitable mechanical properties strong enough for clinical applications. The stiffness and degradation behaviors were then further regulated by different after-treatment processes, making the scaffolds more suitable for dermal tissue regeneration. The in vitro cell culture results indicated that these scaffolds maintained their excellent biocompatibility after being reinforced with woven silk fabrics. Without sacrifice of porous structure and biocompatibility, the fabric-reinforced scaffolds with better mechanical properties could facilitate future clinical applications of silk as matrices in skin repair.

  5. 3D Printing Facilitated Scaffold-free Tissue Unit Fabrication

    PubMed Central

    Tan, Yu; Richards, Dylan J.; Trusk, Thomas C.; Visconti, Richard P.; Yost, Michael J.; Kindy, Mark S.; Drake, Christopher J.; Argraves, William Scott; Markwald, Roger R.; Mei, Ying

    2014-01-01

    Tissue spheroids hold great potential in tissue engineering as building blocks to assemble into functional tissues. To date, agarose molds have been extensively used to facilitate fusion process of tissue spheroids. As a molding material, agarose typically requires low temperature plates for gelation and/or heated dispenser units. Here, we proposed and developed an alginate-based, direct 3D mold-printing technology: 3D printing micro-droplets of alginate solution into biocompatible, bio-inert alginate hydrogel molds for the fabrication of scaffold-free tissue engineering constructs. Specifically, we developed a 3D printing technology to deposit micro-droplets of alginate solution on calcium containing substrates in a layer-by-layer fashion to prepare ring-shaped 3D hydrogel molds. Tissue spheroids composed of 50% endothelial cells and 50% smooth muscle cells were robotically placed into the 3D printed alginate molds using a 3D printer, and were found to rapidly fuse into toroid-shaped tissue units. Histological and immunofluorescence analysis indicated that the cells secreted collagen type I playing a critical role in promoting cell-cell adhesion, tissue formation and maturation. PMID:24717646

  6. 3D printing facilitated scaffold-free tissue unit fabrication.

    PubMed

    Tan, Yu; Richards, Dylan J; Trusk, Thomas C; Visconti, Richard P; Yost, Michael J; Kindy, Mark S; Drake, Christopher J; Argraves, William Scott; Markwald, Roger R; Mei, Ying

    2014-06-01

    Tissue spheroids hold great potential in tissue engineering as building blocks to assemble into functional tissues. To date, agarose molds have been extensively used to facilitate fusion process of tissue spheroids. As a molding material, agarose typically requires low temperature plates for gelation and/or heated dispenser units. Here, we proposed and developed an alginate-based, direct 3D mold-printing technology: 3D printing microdroplets of alginate solution into biocompatible, bio-inert alginate hydrogel molds for the fabrication of scaffold-free tissue engineering constructs. Specifically, we developed a 3D printing technology to deposit microdroplets of alginate solution on calcium containing substrates in a layer-by-layer fashion to prepare ring-shaped 3D hydrogel molds. Tissue spheroids composed of 50% endothelial cells and 50% smooth muscle cells were robotically placed into the 3D printed alginate molds using a 3D printer, and were found to rapidly fuse into toroid-shaped tissue units. Histological and immunofluorescence analysis indicated that the cells secreted collagen type I playing a critical role in promoting cell-cell adhesion, tissue formation and maturation.

  7. Functional Scaffolds of Bicontinuous, Thermoresponsive L3-phase Silica/Hydroxyapatite Nanocomposites

    SciTech Connect

    Chang, Jeong H.; Park, Moo E.; Shin, Yongsoon; Exarhos, Gregory J.; Kim, Kyung J.; Lee, Sang Cheon; Oh, Kyung Sik

    2007-01-15

    The silicified L3-poly(N-sopropylacrylamide)/hydroxyapatite nanocomposites by the integration of thermosensitive polymer and heterogenious continuity in 3-D interconnected porous structure have been prepared for the highly controlled drug release without the burst release at the initial time during step-wised temperature changes, and demonstrated the in-vitro cytotoxicity test for biocompatibility.

  8. PCL/PHBV blended three dimensional scaffolds fabricated by fused deposition modeling and responses of chondrocytes to the scaffolds.

    PubMed

    Kosorn, Wasana; Sakulsumbat, Morakot; Uppanan, Paweena; Kaewkong, Pakkanun; Chantaweroad, Surapol; Jitsaard, Jaturong; Sitthiseripratip, Kriskrai; Janvikul, Wanida

    2017-07-01

    In this study, poly(ε-caprolactone)/poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PCL/PHBV) blended porous scaffolds were fabricated by fused deposition modeling (FDM). PCL/PHBV filaments, initially prepared at different weight ratios, that is, 100/0, 75/25, 50/50, and 25/75, were fabricated by the lay-down pattern of 0/90/45/135° to obtain scaffolds with dimension of 6.0 × 6.0 × 2.5 mm(3) and average filament diameters and channel sizes in the ranges of 370-390 µm and 190-210 µm, respectively. To enhance the surface hydrophilicity of the materials, the scaffolds were subsequently subjected to a low pressure oxygen plasma treatment. The untreated and plasma-treated scaffolds were comparatively characterized, in terms of surface properties, mechanical strength, and biological properties. From SEM, AFM, water contact angle, and XPS results, the surface roughness, wettability, and hydrophilicity of the blended scaffolds were found to be enhanced after plasma treatment, while the compressive strength of the scaffolds was scarcely changed. It was, however, found to increase with an increasing content of PHBV incorporated. The porcine chondrocytes exhibited higher proliferative capacity and chondrogenic potential when being cultured on the scaffolds with greater PHBV contents, especially when they were plasma-treated. The PCL/PHBV scaffolds were proven to possess good physical, mechanical, and biological properties that could be appropriately used in articular cartilage regeneration. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1141-1150, 2017. © 2016 Wiley Periodicals, Inc.

  9. Hydroxyapatite-anatase-carbon nanotube nanocomposite coatings fabricated by electrophoretic codeposition for biomedical applications.

    PubMed

    Zhang, Bokai; Kwok, Chi Tat

    2011-10-01

    In order to eliminate micro-cracks in the monolithic hydroxyapatite (HA) and composite hydroxyapatite/carbon nanotube (HA/CNT) coatings, novel HA/TiO(2)/CNT nanocomposite coatings on Ti6Al4V were attempted to fabricate by a single-step electrophoretic codeposition process for biomedical applications. The electrophoretically deposited layers with difference contents of HA, TiO(2) (anatase) and CNT nanoparticles were sintered at 800°C for densification with thickness of about 7-10 μm. A dense and crack-free coating was achieved with constituents of 85 wt% HA, 10 wt% TiO(2) and 5 wt% CNT. Open-circuit potential measurements and cyclic potentiodynamic polarization tests were used to investigate the electrochemical corrosion behavior of the coatings in vitro conditions (Hanks' solution at 37°C). The HA/TiO(2)/CNT coatings possess higher corrosion resistance than that of the Ti6Al4V substrate as reflected by nobler open circuit potential and lower corrosion current density. In addition, the surface hardness and adhesion strength of the HA/TiO(2)/CNT coatings are higher than that of the monolithic HA and HA/CNT coatings without compromising their apatite forming ability. The enhanced properties were attributed to the nanostructure of the coatings with the appropriate TiO(2) and CNT contents for eliminating micro-cracks and micro-pores.

  10. Biocompatibility and biomechanical characteristics of loofah based scaffolds combined with hydroxyapatite, cellulose, poly-l-lactic acid with chondrocyte-like cells.

    PubMed

    Cecen, Berivan; Kozaci, Leyla Didem; Yuksel, Mithat; Ustun, Ozcan; Ergur, Bekir Ugur; Havitcioglu, Hasan

    2016-12-01

    The current study reports the biocompatibility and biomechanical characteristics of loofah-based scaffolds combined with hydroxyapatite (HA), cellulose, poly-l-lactic acid (PLLA) with chondrocytes-like cells. Scanning electron microscope (SEM) micrographs of the scaffolds showed that the addition of PLLA usually resulted in an increase in cell's attachment on scaffolds. Mechanical and elemental analyzes were assessed using tensile test and Energy Dispersive X-ray spectrometry (EDS), respectively. In summary, we showed that the loofah+PLLA+HA scaffolds perform significantly better than other loofah-based scaffolds employed in terms of increasing a diversity of mechanical properties including tensile strength and Young's modulus. Based on the analysis of the differential scanning calorimetry (DSC) thermograms and EDS spectrums that give an idea about the calcium phosphate (CaP) ratios, the improvement in the mechanical properties could principally be recognized to the strong interaction formed between loofah, PLLA and HA. The viability of chondrocytes on loofah-based scaffolds was analyzed by XTT tests. However, none of the scaffolds have proved to be toxic in metabolic activity. The histological evaluation obtained by hematoxylin and eosin (H&E), Masson trichrome, toluidine blue and immunohistochemistry methods showed that cells in all scaffolds produced extracellular matrix that defined proteoglycan and type I-II collagens. The results of this study suggest that the loofah-based scaffold with desirable properties can be considered as an ideal candidate for cartilage tissue engineering applications. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Increasing the pore sizes of bone-mimetic electrospun scaffolds comprised of polycaprolactone, collagen I and hydroxyapatite to enhance cell infiltration

    PubMed Central

    Phipps, Matthew C.; Clem, William C.; Grunda, Jessica M.; Clines, Gregory A.; Bellis, Susan L.

    2012-01-01

    Bone-mimetic electrospun scaffolds consisting of polycaprolactone (PCL), collagen I and nanoparticulate hydroxyapatite (HA) have previously been shown to support the adhesion, integrin-related signaling and proliferation of mesenchymal stem cells (MSCs), suggesting these matrices serve as promising degradable substrates for osteoregeneration. However, the small pore sizes in electrospun scaffolds hinder cell infiltration in vitro and tissue-ingrowth into the scaffold in vivo, limiting their clinical potential. In this study, three separate techniques were evaluated for their capability to increase the pore size of the PCL/col I/nanoHA scaffolds: limited protease digestion, decreasing the fiber packing density during electro-spinning, and inclusion of sacrificial fibers of the water-soluble polymer PEO. The PEO sacrificial fiber approach was found to be the most effective in increasing scaffold pore size. Furthermore, the use of sacrificial fibers promoted increased MSC infiltration into the scaffolds, as well as greater infiltration of endogenous cells within bone upon placement of scaffolds within calvarial organ cultures. These collective findings support the use of sacrificial PEO fibers as a means to increase the porosity of complex, bone-mimicking electrospun scaffolds, thereby enhancing tissue regenerative processes that depend upon cell infiltration, such as vascularization and replacement of the scaffold with native bone tissue. PMID:22014462

  12. Preparation, in vitro degradability, cytotoxicity, and in vivo biocompatibility of porous hydroxyapatite whisker-reinforced poly(L-lactide) biocomposite scaffolds.

    PubMed

    Xie, Lu; Yu, Haiyang; Yang, Weizhong; Zhu, Zhuoli; Yue, Li

    2016-01-01

    Biodegradable and bioactive scaffolds with interconnected macroporous structures, suitable biodegradability, adequate mechanical property, and excellent biocompatibility have drawn increasing attention in bone tissue engineering. Hence, in this work, porous hydroxyapatite whisker-reinforced poly(L-lactide) (HA-w/PLLA) composite scaffolds with different ratios of HA and PLLA were successfully developed through compression molding and particle leaching. The microstructure, in vitro mineralization, cytocompatibility, hemocompatibility, and in vivo biocompatibility of the porous HA-w/PLLA were investigated for the first time. The SEM results revealed that these HA-w/PLLA scaffolds possessed interconnected pore structures. Compared with porous HA powder-reinforced PLLA (HA-p/PLLA) scaffolds, HA-w/PLLA scaffolds exhibited better mechanical property and in vitro bioactivity, as more formation of bone-like apatite layers were induced on these scaffolds after mineralization in SBF. Importantly, in vitro cytotoxicity displayed that porous HA-w/PLLA scaffold with HA/PLLA ratio of 1:1 (HA-w1/PLLA1) produced no deleterious effect on human mesenchymal stem cells (hMSCs), and cells performed elevated cell proliferation, indicating a good cytocompatibility. Simultaneously, well-behaved hemocompatibility and favorable in vivo biocompatibility determined from acute toxicity test and histological evaluation were also found in the porous HA-w1/PLLA1 scaffold. These findings may provide new prospects for utilizing the porous HA whisker-based biodegradable scaffolds in bone repair, replacement, and augmentation applications.

  13. Fabrication of polylactide nanocomposite scaffolds for bone tissue engineering applications

    NASA Astrophysics Data System (ADS)

    Mkhabela, Vuyiswa J.; Ray, Suprakas Sinha

    2015-05-01

    Highly porous three-dimensional polylactide (PLA) scaffolds were obtained from PLA incorporated with different amounts of chitosan-modified montmorillonite (CS-MMT), through solvent casting and particulate leaching method. The processed scaffolds were tested in vitro for their possible application in bone tissue engineering. Scaffolds were characterized by Focused Ion Beam Scanning Electron Microscopy (FIB SEM), Fourier Transform Infra-Red (FTIR), and X-Ray Diffraction (XRD) to study their structure and intermolecular interactions. Bioresorbability tests in simulated body fluid (pH 7.4) were conducted to assess the response of the scaffolds in a simulated physiological condition. The FIB SEM images of the scaffolds showed a porous architecture with gradual change in morphology with increasing CS-MMT concentration. FTIR analysis revealed the presence of both PLA and CS-MMT particles on the surface of the scaffolds. XRD showed that the crystalline unit cell type was the same for all the scaffolds, and crystallinity decreased with an increase in CS-MMT concentration. The scaffolds were found to be bioresorbable, with rapid bioresorbability on the scaffolds with a high CS-MMT concentration.

  14. Fabrication of polylactide nanocomposite scaffolds for bone tissue engineering applications

    SciTech Connect

    Mkhabela, Vuyiswa J.; Ray, Suprakas Sinha

    2015-05-22

    Highly porous three-dimensional polylactide (PLA) scaffolds were obtained from PLA incorporated with different amounts of chitosan-modified montmorillonite (CS-MMT), through solvent casting and particulate leaching method. The processed scaffolds were tested in vitro for their possible application in bone tissue engineering. Scaffolds were characterized by Focused Ion Beam Scanning Electron Microscopy (FIB SEM), Fourier Transform Infra-Red (FTIR), and X-Ray Diffraction (XRD) to study their structure and intermolecular interactions. Bioresorbability tests in simulated body fluid (pH 7.4) were conducted to assess the response of the scaffolds in a simulated physiological condition. The FIB SEM images of the scaffolds showed a porous architecture with gradual change in morphology with increasing CS-MMT concentration. FTIR analysis revealed the presence of both PLA and CS-MMT particles on the surface of the scaffolds. XRD showed that the crystalline unit cell type was the same for all the scaffolds, and crystallinity decreased with an increase in CS-MMT concentration. The scaffolds were found to be bioresorbable, with rapid bioresorbability on the scaffolds with a high CS-MMT concentration.

  15. Biomaterials and Fabrication to Optimise Scaffold Properties for Musculoskeletal Tissue Engineering.

    PubMed

    Wheelton, Andrew; Mace, James; Khan, Wasim S; Anand, Sanjay

    2016-01-01

    Tissue engineering has emerged as a promising scientific field potentially yielding in vitro developed tissue to replace degenerative or injured tissues in vivo, thus avoiding the donor site morbidity associated with reconstructive surgery. Integral to the process is the role of scaffolds and the biomaterials used to form them. This review explores the concept of scaffold based tissue engineering and design considerations. The scaffold needs to have certain mechanical and architectural properties, it needs to be biocompatible and biodegradable, and allow combination with bioactive molecules. We also discuss scaffolding techniques, different biomaterial options and fabrication technologies, and future areas of development.

  16. Fabrication of 3D porous SF/β-TCP hybrid scaffolds for bone tissue reconstruction.

    PubMed

    Park, Hyun Jung; Min, Kyung Dan; Lee, Min Chae; Kim, Soo Hyeon; Lee, Ok Joo; Ju, Hyung Woo; Moon, Bo Mi; Lee, Jung Min; Park, Ye Ri; Kim, Dong Wook; Jeong, Ju Yeon; Park, Chan Hum

    2016-07-01

    Bio-ceramic is a biomaterial actively studied in the field of bone tissue engineering. But, only certain ceramic materials can resolve the corrosion problem and possess the biological affinity of conventional metal biomaterials. Therefore, the recent development of composites of hybrid composites and polymers has been widely studied. In this study, we aimed to select the best scaffold of silk fibroin and β-TCP hybrid for bone tissue engineering. We fabricated three groups of scaffold such as SF (silk fibroin scaffold), GS (silk fibroin/small granule size of β-TCP scaffold) and GM (silk fibroin/medium granule size of β-TCP scaffold), and we compared the characteristics of each group. During characterization of the scaffold, we used scanning electron microscopy (SEM) and a Fourier transform infrared spectroscopy (FTIR) for structural analysis. We compared the physiological properties of the scaffold regarding the swelling ratio, water uptake and porosity. To evaluate the mechanical properties, we examined the compressive strength of the scaffold. During in vitro testing, we evaluated cell attachment and cell proliferation (CCK-8). Finally, we confirmed in vivo new bone regeneration from the implanted scaffolds using histological staining and micro-CT. From these evaluations, the fabricated scaffold demonstrated high porosity with good inter-pore connectivity, showed good biocompatibility and high compressive strength and modulus. In particular, the present study indicates that the GM scaffold using β-TCP accelerates new bone regeneration of implanted scaffolds. Accordingly, our scaffold is expected to act a useful application in the field of bone tissue engineering. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1779-1787, 2016.

  17. Replicable and shape-controllable fabrication of electrospun fibrous scaffolds for tissue engineering.

    PubMed

    Cho, Seong J; Nam, Hyoryung; An, Taechang; Lim, Geunbae

    2012-12-01

    Controlling the architecture of electrospun fibers is one of the key issues in tissue engineering. This report describes a rapid and reproducible patterning method for the fabrication of an electrospun fibrous scaffold. The electrospun fibers were deposited on a patterned electrode. The patterned scaffold was fabricated using a thin insulating film between layers of this electrode. For a tissue engineering application, poly(lactic-co-glycolic acid) (PLGA), a biocompatible and biodegradable material, was electrospun. Fibroblast cells were cultured on the fibrous PLGA scaffold and the viability, morphology, and distribution of the cells were studied.

  18. Porous titanium scaffolds fabricated using a rapid prototyping and powder metallurgy technique.

    PubMed

    Ryan, Garrett E; Pandit, Abhay S; Apatsidis, Dimitrios P

    2008-09-01

    One of the main issues in orthopaedic implant design is the fabrication of scaffolds that closely mimic the biomechanical properties of the surrounding bone. This research reports on a multi-stage rapid prototyping technique that was successfully developed to produce porous titanium scaffolds with fully interconnected pore networks and reproducible porosity and pore size. The scaffolds' porous characteristics were governed by a sacrificial wax template, fabricated using a commercial 3D-printer. Powder metallurgy processes were employed to generate the titanium scaffolds by filling around the wax template with titanium slurry. In the attempt to optimise the powder metallurgy technique, variations in slurry concentration, compaction pressure and sintering temperature were investigated. By altering the wax design template, pore sizes ranging from 200 to 400 microm were achieved. Scaffolds with porosities of 66.8 +/- 3.6% revealed compression strengths of 104.4+/-22.5 MPa in the axial direction and 23.5 +/- 9.6 MPa in the transverse direction demonstrating their anisotropic nature. Scaffold topography was characterised using scanning electron microscopy and microcomputed tomography. Three-dimensional reconstruction enabled the main architectural parameters such as pore size, interconnecting porosity, level of anisotropy and level of structural disorder to be determined. The titanium scaffolds were compared to their intended designs, as governed by their sacrificial wax templates. Although discrepancies in architectural parameters existed between the intended and the actual scaffolds, overall the results indicate that the porous titanium scaffolds have the properties to be potentially employed in orthopaedic applications.

  19. Ectopic Osteogenesis and Scaffold Biodegradation of Nano-Hydroxyapatite-Chitosan in a Rat Model

    PubMed Central

    He, Yiqun; Dong, Youhai; Cui, Fuzhai; Chen, Xujun; Lin, Rongqiang

    2015-01-01

    The bone-formation and scaffold-biodegradation processes have not been fully characterized. This study aimed to determine the osteogenic ability of nHA-CS osteo-induced bone marrow mesenchymal stem cell (BMSC) composites and to explore the relationship between bone formation and scaffold biodegradation. The nHA-CS osteo-induced BMSC composites (nHA-CS+cells group) and the nHA-CS scaffolds (nHA-CS group) were implanted into the femoral spatium intermusculare of SD rats. At 2, 4, 6, 8, and 12 weeks post-implantation, the rat femurs were scanned using computerized tomography (CT), and the CT values of the implants were measured and comparatively analyzed. The implants were then harvested and subjected to hematoxylin and eosin (HE) and Masson's trichrome staining, and the percentages of bone area, scaffold area and collagen area were compared between the two groups. The CT values of the implants were higher in the nHA-CS+cells group than the nHA-CS group at the same time points (P < 0.05). Histological analysis revealed that de novo bone and collagen formation in the pores of the scaffolds gradually increased from 2 weeks post-implantation in both groups and that the scaffold gradually degraded as bone formation proceeded. However, more de novo bone and collagen formation and scaffold degradation occurred in the nHA-CS+cells group than in the nHA-CS group at the same time points (P < 0.05). In conclusion, nHA-CS osteo-induced BMSC composites are promising bone tissue engineering substitutes, and osteo-induced BMSCs can significantly enhance the osteogenic ability and play an active role in the degradation of nHA-CS scaffolds on par with bone formation. PMID:26258851

  20. Sustained delivery of BMP-2 enhanced osteoblastic differentiation of BMSCs based on surface hydroxyapatite nanostructure in chitosan-HAp scaffold.

    PubMed

    Wang, Guancong; Qiu, Jichuan; Zheng, Lin; Ren, Na; Li, Jianhua; Liu, Hong; Miao, Junying

    2014-01-01

    The surface characteristics of biomaterials, especially regarding the sustained delivery of bone morphogenetic protein-2 (BMP-2), can possibly provide a novel and effective drug delivery system that can enhance osteogenesis. In this study, we evaluated the BMP-2 adsorption and release ability of the surface biomimetic hydroxyapatite (HAp) nanostructure on a new HAp-coated genipin-chitosan conjugation scaffold (HGCCS), and the resulting osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (BMSCs) in vitro. HGCCS exhibited a loading efficiency of 65% (1.30 μg), which is significantly higher than 28% (0.56 μg, p < 0.01) for the genipin cross-linked chitosan framework, as quantified by an enzyme-linked immunosorbent assay. More importantly, we found that the release of BMP-2 from HGGCS sustained for at least 14 days in simulated body fluid in vitro, which is much better than the burst release within 3 days for CGF. Moreover, the BMP-2 release from HGCCS induced an increase in alkaline phosphatase activity as an indicator of osteogenic differentiation of seeded BMSCs for 14 days in vitro. HGCCS also stimulated a high mRNA expression of osteogenic differentiation makers, runt-related transcription factor 2 for 14 days, osteopontin for 3 days, and osteocalcin for 14 days. The results of this study suggest that the surface biomimetic HAp nanostructure of HGCCS used as a delivery system for BMP-2 is capable of promoting osteogenic differentiation in vitro. These findings demonstrated that HAp nanostructure assembled on organic porous scaffold could work as both calcium source and absorption/release platform, which opened a new research avenue for cell growth factor release, and provided a promising strategy for design and preparation of bioactive scaffold for bone tissue engineering.

  1. Fabrication of orderly nanostructured PLGA scaffolds using anodic aluminum oxide templates.

    PubMed

    Wang, Gou-Jen; Lin, Yan-Cheng; Li, Ching-Wen; Hsueh, Cheng-Chih; Hsu, Shan-Hui; Hung, Huey-Shan

    2009-08-01

    In this research, two simple fabrication methods to fabricate orderly nanostructured PLGA scaffolds using anodic aluminum oxide (AAO) template were conducted. In the vacuum air-extraction approach, the PLGA solution was cast on an AAO template first. The vacuum air-extraction process was then applied to suck the semi-congealed PLGA into the nanopores of the AAO template to form a bamboo sprouts array of PLGA. The surface roughness of the nanostructured scaffolds, ranging from 20 nm to 76 nm, can be controlled by the sucking time of the vacuum air-extraction process. In the replica molding approach, the PLGA solution was cast on the orderly scraggy barrier-layer surface of an AAO membrane to fabricate a PLGA scaffold of concave nanostructure. Cell culture experiments using the bovine endothelial cells (BEC) demonstrated that the nanostructured PLGA membrane can increase the cell growing rate, especially for the bamboo sprouts array scaffolds with smaller surface roughness.

  2. Comparative evaluation of a biomimic collagen/hydroxyapatite/β-tricaleium phosphate scaffold in alveolar ridge preservation with Bio-Oss Collagen

    NASA Astrophysics Data System (ADS)

    Wang, Tong; Li, Qing; Zhang, Gui-feng; Zhou, Gang; Yu, Xin; Zhang, Jing; Wang, Xiu-mei; Tang, Zhi-hui

    2016-06-01

    Bone scaffolds are critical in current implant and periodontal regeneration approaches. In this study, we prepared a novel composite type-I collagen and hydroxyapatite (HA)/β-tricaleium phosphate (TCP) scaffold (CHTS) by incorporating type-I collagen and bovine calcined bone granules, prepared as a mixture of 50% HA and 50% TCP, by freeze drying. We then characterized the CHTS and determined its cytotoxic effects. Additionally, ridge preservation experiments were carried out to evaluate the clinical effects of the CHTS. The results demonstrated that the composite scaffolds had good surface morphology and no cytotoxicity. Additionally, an in vivo experiment in an animal model showed that the CHTS performed equally as well as Bio-Oss Collagen, a widely used bone graft in ridge preservation. These findings revealed that the CHTS, which contained natural constituents of bone, could be used as a scaffold for bone regeneration and clinical use.

  3. Preparation and biocompatibility of nanohybrid scaffolds by in situ homogeneous formation of nano hydroxyapatite from biopolymer polyelectrolyte complex for bone repair applications.

    PubMed

    Chen, Jingdi; Yu, Qifeng; Zhang, Guodong; Yang, Shen; Wu, Jiulin; Zhang, Qiqing

    2012-05-01

    The achievement of nano distribution for inorganic reinforced filler is a big challenge to three-dimensional porous composite scaffolds. In this paper, a homogeneous nano hydroxyapatite/polyelectrolyte complex (HAP/PEC) hybrid scaffold was developed and investigated. Based on the enhancing properties of the formation of PEC between chitosan and hyaluronic acid, the introduction of nano HAP via in situ crystallization from the PEC achieved nano distribution in the PEC matrix and supplied nano topographies of extracellular environments for the nanohybrid scaffold. The biocompatibility and bioactivity were evaluated by Human bone mesenchymal stem cells (hBMSCs) proliferation (MTT assay), maturation (alkaline phosphatase (ALP) activity) and histological analysis. The in vitro tests show the scaffold is excellent for cell penetration, growth, and proliferation and it is promising for bone repair application.

  4. Fabrication of fibrinogen/P(LLA-CL) hybrid nanofibrous scaffold for potential soft tissue engineering applications.

    PubMed

    He, Chuanglong; Xu, Xiaohong; Zhang, Fan; Cao, Lijun; Feng, Wei; Wang, Hongsheng; Mo, Xiumei

    2011-06-01

    Coelectrospinning of native proteins and elastic synthetic polymers is an attractive technique to fabricate hybrid fibrous scaffolds that combine the bioactivity and mechanical features of each material component. In this study, hybrid fibrous scaffolds composed of synthetic P(LLA-CL) elastomeric and naturally derived fibrinogen protein were fabricated and characterized for their bioactive and physiochemical properties. Fiber diameters of hybrid scaffolds increased with increasing P(LLA-CL) content, and the shape of fibers changed from cylindrical shape on pure polymer scaffolds to flat structure on hybrid scaffolds. Characterizations of ATR-FTIR, XRD, and thermal properties indicated that the hybrid scaffolds contain two different phases, one composed of pure fibrinogen and the other corresponding to a mixture of fibrinogen and P(LLA-CL), and no obvious chemical reaction takes place between two components. The hybrid fibrous scaffolds showed tailorable degradation rates than pure P(LLA-CL) and higher mechanical properties than pure fibrinogen, and both tensile strength and breaking strain increased with increasing P(LLA-CL) content. In Vitro studies revealed that L929 cells on hybrid scaffolds achieved relatively higher level of cell attachment after 12 h of culture and significant increased cell proliferation rate after 7 days of culture, when compared with pure fibrinogen and P(LLA-CL) scaffolds, and the cells exhibited a spreading polygonal shape on the hybrid fibrous surfaces compared to a round shape on surfaces of pure polymer scaffolds. Therefore, the fibrinogen/P(LLA-CL) hybrid fibrous scaffolds possess the combined benefits of each individual component, which make it capable as scaffolds for soft tissue reconstruction.

  5. Bio-scaffolds produced from irradiated squid pen and crab chitosan with hydroxyapatite/β-tricalcium phosphate for bone-tissue engineering.

    PubMed

    Shavandi, Amin; Bekhit, Alaa El-Din A; Sun, Zhifa; Ali, M Azam

    2016-12-01

    In this study, bio-scaffolds have been developed using irradiated chitosan from different sources - squid pen (RS) and crab shell (RC) - with hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) at a chitosan/HA/β-TCP ratio of 50/30/20. The bio-scaffolds were prepared at two different freezing temperature (-20°C and -80°C) followed by lyophilisation. To enhance the mechanical properties, the bio-scaffolds were cross-linked using sodium tripolyphosphate (TPP) followed by lyophilisation. The composition and morphology of the bio-scaffolds were characterized using XRD, SEM, TEM and μ-CT. The pore size of the porous scaffolds ranged from 90 to 220μm and the scaffolds had 70-80% porosity. The scaffolds had a water uptake ratio of more than 10, and a controlled biodegradation in the range of 30-40%. These results suggest that the physical and biological properties of chitosan-based bio-scaffolds can be a promising biomaterial for bone-tissue regeneration. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Net shape fabrication of calcium phosphate scaffolds with multiple material domains.

    PubMed

    Xie, Yangmin; Rustom, Laurence E; McDermott, Anna M; Boerckel, Joel D; Johnson, Amy J Wagoner; Alleyne, Andrew G; Hoelzle, David J

    2016-01-08

    Calcium phosphate (CaP) materials have been proven to be efficacious as bone scaffold materials, but are difficult to fabricate into complex architectures because of the high processing temperatures required. In contrast, polymeric materials are easily formed into scaffolds with near-net-shape forms of patient-specific defects and with domains of different materials; however, they have reduced load-bearing capacity compared to CaPs. To preserve the merits of CaP scaffolds and enable advanced scaffold manufacturing, this manuscript describes an additive manufacturing process that is coupled with a mold support for overhanging features; we demonstrate that this process enables the fabrication of CaP scaffolds that have both complex, near-net-shape contours and distinct domains with different microstructures. First, we use a set of canonical structures to study the manufacture of complex contours and distinct regions of different material domains within a mold. We then apply these capabilities to the fabrication of a scaffold that is designed for a 5 cm orbital socket defect. This scaffold has complex external contours, interconnected porosity on the order of 300 μm throughout, and two distinct domains of different material microstructures.

  7. Fabrication of Metallic Biomedical Scaffolds with the Space Holder Method: A Review

    PubMed Central

    Arifvianto, Budi; Zhou, Jie

    2014-01-01

    Bone tissue engineering has been increasingly studied as an alternative approach to bone defect reconstruction. In this approach, new bone cells are stimulated to grow and heal the defect with the aid of a scaffold that serves as a medium for bone cell formation and growth. Scaffolds made of metallic materials have preferably been chosen for bone tissue engineering applications where load-bearing capacities are required, considering the superior mechanical properties possessed by this type of materials to those of polymeric and ceramic materials. The space holder method has been recognized as one of the viable methods for the fabrication of metallic biomedical scaffolds. In this method, temporary powder particles, namely space holder, are devised as a pore former for scaffolds. In general, the whole scaffold fabrication process with the space holder method can be divided into four main steps: (i) mixing of metal matrix powder and space-holding particles; (ii) compaction of granular materials; (iii) removal of space-holding particles; (iv) sintering of porous scaffold preform. In this review, detailed procedures in each of these steps are presented. Technical challenges encountered during scaffold fabrication with this specific method are addressed. In conclusion, strategies are yet to be developed to address problematic issues raised, such as powder segregation, pore inhomogeneity, distortion of pore sizes and shape, uncontrolled shrinkage and contamination. PMID:28788638

  8. Freeform extrusion fabrication of titanium fiber reinforced 13-93 bioactive glass scaffolds.

    PubMed

    Thomas, Albin; Kolan, Krishna C R; Leu, Ming C; Hilmas, Gregory E

    2017-05-01

    Although implants made with bioactive glass have shown promising results for bone repair, their application in repairing load-bearing long bone is limited due to their poor mechanical properties in comparison to human bone. This work investigates the freeform extrusion fabrication of bioactive silicate 13-93 glass scaffolds reinforced with titanium (Ti) fibers. A composite paste prepared with 13-93 glass and Ti fibers (~16µm in diameter and lengths varying from ~200µm to ~2 mm) was extruded through a nozzle to fabricate scaffolds (0-90° filament orientation pattern) on a heated plate. The sintered scaffolds measured pore sizes ranging from 400 to 800µm and a porosity of ~50%. Scaffolds with 0.4vol% Ti fibers measured fracture toughness of ~0.8MPam(1/2) and a flexural strength of ~15MPa. 13-93 glass scaffolds without Ti fibers had a toughness of ~0.5MPam(1/2) and a strength of ~10MPa. The addition of Ti fibers increased the fracture toughness of the scaffolds by ~70% and flexural strength by ~40%. The scaffolds' biocompatibility and their degradation in mechanical properties in vitro were assessed by immersing the scaffolds in a simulated body fluid over a period of one to four weeks.

  9. Biomimetic composite scaffolds based on mineralization of hydroxyapatite on electrospun poly(ɛ-caprolactone)/nanocellulose fibers.

    PubMed

    Si, Junhui; Cui, Zhixiang; Wang, Qianting; Liu, Qiong; Liu, Chuntai

    2016-06-05

    A biomimetic nanocomposite scaffold with HA formation on the electrospun poly(ɛ-caprolactone) (PCL)/nanocellulose (NC) fibrous matrix was developed in this study. The electrospun PCL/NC fiber mat was built and then biomineralized by treatment in simulated body fluid (SBF). Using such a rapid and effective procedure, a continuous biomimetic crystalline HA layer could be successfully formed without the need of any additional chemical modification of the substrate surface. The results showed that the introduction of NC into composite fibers is an effective approach to induce the deposition of HA nucleus as well as to improve their distribution and growth of a crystalline HA layer on the fibrous scaffolds. The water contact angle (WCA) of the PCL/NC/HA scaffolds decreases with increasing NC content and mineralization time, resulting in the enhancement of their hydrophilicity. These results indicated that HA-mineralized on PCL/NC fiber can be prepared directly by simply using SBF immersion.

  10. Photoinitiator-free 3D scaffolds fabricated by excimer laser photocuring.

    PubMed

    Farkas, Balázs; Dante, Silvia; Brandi, Fernando

    2017-01-20

    Photoinitiator-free fabrication of poly(ethylene glycol) diacrylate (PEGDA) scaffolds is achieved using a novel three-dimensional (3D) printing method called mask projected excimer laser stereolithography (MPExSL). The spatial resolution of photoinitiator-free curing is suitable for 3D layer-by-layer fabrication with a single layer thickness well controllable at tens to hundreds of microns using 248 nm wavelength for the irradiation. The photoinitiator-free scaffolds are superior compared to their counterparts fabricated by using photoinitiator molecules, showing a higher level of biocompatibility. A release of toxic chemicals from the photoinitiator containing scaffolds is proven by cell proliferation tests. In contrast, no toxic release is found from the photoinitiator-free scaffolds, resulting in the very same level of cell proliferation as the control sample. The demonstration of photoinitiator-free PEGDA scaffolds enables the fabrication of 3D scaffolds with the highest level of biocompatibility for both in vitro and in vivo applications.

  11. Photoinitiator-free 3D scaffolds fabricated by excimer laser photocuring

    NASA Astrophysics Data System (ADS)

    Farkas, Balázs; Dante, Silvia; Brandi, Fernando

    2017-01-01

    Photoinitiator-free fabrication of poly(ethylene glycol) diacrylate (PEGDA) scaffolds is achieved using a novel three-dimensional (3D) printing method called mask projected excimer laser stereolithography (MPExSL). The spatial resolution of photoinitiator-free curing is suitable for 3D layer-by-layer fabrication with a single layer thickness well controllable at tens to hundreds of microns using 248 nm wavelength for the irradiation. The photoinitiator-free scaffolds are superior compared to their counterparts fabricated by using photoinitiator molecules, showing a higher level of biocompatibility. A release of toxic chemicals from the photoinitiator containing scaffolds is proven by cell proliferation tests. In contrast, no toxic release is found from the photoinitiator-free scaffolds, resulting in the very same level of cell proliferation as the control sample. The demonstration of photoinitiator-free PEGDA scaffolds enables the fabrication of 3D scaffolds with the highest level of biocompatibility for both in vitro and in vivo applications.

  12. Fabrication of Novel Film Containing Hydroxyapatite Nanoparticles/Cellulose Derivative and its Evaluation

    NASA Astrophysics Data System (ADS)

    Shimizu, Akinori; Umeda, Tomohiro; Isogai, Akira; Saito, Tsuguyuki; Koda, Seiichiro; Itatani, Kiyoshi

    2011-10-01

    The novel and flexible film for biomedical application could be fabricated by using hydroxyapatite (Ca10(PO4)6(OH)2; HAp) and cellulose derivatives. Firstly, 0.2 mass% 2,2,6,6-tetramethylpitiperidine-1-oxyl radical (TEMPO) oxidized cellulose (TOC) was agitated at 20,000 rpm for 3 min and then ultrasonically treated for 20 min at the frequency of 45,000 Hz to form the solution containing TOC nanofiber (TOCN). After the nano-sized HAp powder was put into TOCN solution (100 cm3; HAp/TOCN mass ratio of 0.10 or 0.50), the composite film was fabricated by drying it at 50°C for 3 d. The TOCN film with no HAp addition (the thickness: approximately 15 μm) possessed transparency, flexibility and high tensile strength (mean strength: 193 MPa). On the other hand, the tensile strength of HAp-TOCN composite film (thickness: 30 μm) was reduced from 60.1 to 50.2MPa with increasing HAp/TOCN mass ratio from 0.10 to 0.50. When the HAp/TOCN composite films were immersed in SBF at 37.0°C for 2 weeks, the spherical apatite particles were notably found to form on their surfaces.

  13. Novel chelate-setting calcium-phosphate cements fabricated with wet-synthesized hydroxyapatite powder.

    PubMed

    Konishi, Toshiisa; Horiguchi, Yukiko; Mizumoto, Minori; Honda, Michiyo; Oribe, Kazuya; Morisue, Hikaru; Ishii, Ken; Toyama, Yoshiaki; Matsumoto, Morio; Aizawa, Mamoru

    2013-03-01

    The hydroxyapatite (HAp) powder preparation process was optimized to fabricate inositol phosphate-HAp (IP6-HAp) cement with enhanced mechanical properties. Starting HAp powders were synthesized via a wet chemical process. The effect of the powder preparation process on the morphology, crystallinity, median particle size, and specific surface area (SSA) of the cement powders was examined, together with the mechanical properties of the resulting cement specimens. The smallest crystallite and median particle sizes, and the highest SSA were obtained from ball-milling of as-synthesized HAp powder under wet conditions and then freeze-drying. IP6-HAp cement fabricated with this powder had a maximum compressive strength of 23.1 ± 2.1 MPa. In vivo histological studies using rabbit models revealed that the IP6-HAp cements were directly in contact with newly formed and host bones. Thus, the present chelate-setting HAp cement is promising for application as a novel paste-like artificial bone.

  14. Bioactive composite gradient coatings of nano-hydroxyapatite/polyamide66 fabricated on polyamide66 substrates

    PubMed Central

    Huang, Di; Zuo, Yi; Li, Jidong; Zou, Qin; Zhang, Li; Gong, Mei; Wang, Li; Li, Limei; Li, Yubao

    2012-01-01

    Tightly bonding of bioactive coating is the first crucial need for orthopaedic implants. This study describes a novel and convenient technique to prepare bioactive coating with high adhesion on orthopaedic substitutes made of polymeric matrix. Here, a chemical corrosion method has been adopted to fabricate a coating on the surface of injection-moulded polyamide66 (PA66) substrates by corrosive nano-hydroxyapatite/polyamide66 (n-HA/PA66) composite slurry. Scanning electron microscopy observation shows that a porous chemical corrosion region presents between the coating and dense PA66 substrate. Energy-dispersive X-ray spectroscopy analysis indicates that the chemical corrosion region is mainly composed of PA66 matrix, and the coating layer is an n-HA-rich layer. Both the pore size and n-HA composition increase gradually from the polymeric substrate towards the coating surface. Mechanical testing shows the bonding strength can reach 13.7 ± 0.2 MPa, which is much higher than that fabricated on polymeric matrix by other coating methods. The gradual transition in coating structure and composition benefits for the interface bonding and for the surface bone-bonding bioactivity. Subsequent cell experiments corroborate n-HA-rich coating and a porous structure is benefitting for cell attachment and proliferation. The convenient coating method could be popularized and applied on similar polymer implants to produce a tightly and porous bioactive coating for bone tissue regeneration. PMID:22258549

  15. Physico-Chemical, In Vitro, and In Vivo Evaluation of a 3D Unidirectional Porous Hydroxyapatite Scaffold for Bone Regeneration

    PubMed Central

    Tanaka, Manabu; Haniu, Hisao; Kamanaka, Takayuki; Takizawa, Takashi; Sobajima, Atsushi; Yoshida, Kazushige; Aoki, Kaoru; Okamoto, Masanori; Kato, Hiroyuki; Saito, Naoto

    2017-01-01

    The unidirectional porous hydroxyapatite HAp (UDPHAp) is a scaffold with continuous communicated pore structure in the axial direction. We evaluated and compared the ability of the UDPHAp as a three-dimensional (3D) bone tissue engineering scaffold to the interconnected calcium porous HAp ceramic (IP-CHA). To achieve this, we evaluated in vitro the compressive strength, controlled rhBMP-2 release behavior, adherent cell morphology, cell adhesion manner, and cell attachment of UDPHAp. As a further in vivo experiment, UDPHAp and IP-CHA with rhBMP-2 were transplanted into mouse calvarial defects to evaluate their bone-forming ability. The Results demonstrated that the maximum compressive strengths of the UDPHAp was 7.89 ± 1.23 MPa and higher than that of IP-CHA (1.92 ± 0.53 MPa) (p = 0.0039). However, the breaking energies were similar (8.99 ± 2.72 vs. 13.95 ± 5.69 mJ, p = 0.055). The UDPHAp released rhBMP-2 more gradually in vivo. Cells on the UDPHAp adhered tightly to the surface, which had grown deeply into the scaffolds. A significant increase in cell number on the UDPHAp was observed compared to the IP-CHA on day 8 (102,479 ± 34,391 vs. 32,372 ± 29,061 estimated cells per scaffold, p = 0.0495). In a mouse calvarial defect model, the percentages of new bone area (mature bone + trabecular bone) in the 2x field were 2.514% ± 1.224% for the IP-CHA group and 7.045% ± 2.055% for the UDPHAp group, and the percentage was significantly higher in the UDPHAp group (p = 0.0209). While maintaining the same strength as the IP-CHA, the UDPHAp with 84% porosity showed a high cell number, high cell invasiveness, and excellent bone formation. We believe the UDPHAp is an excellent material that can be applied to bone regenerative medicine. PMID:28772390

  16. Sol-gel assisted fabrication of collagen hydrolysate composite scaffold: a novel therapeutic alternative to the traditional collagen scaffold.

    PubMed

    Ramadass, Satiesh Kumar; Perumal, Sathiamurthi; Gopinath, Arun; Nisal, Anuya; Subramanian, Saravanan; Madhan, Balaraman

    2014-09-10

    Collagen is one of the most widely used biomaterial for various biomedical applications. In this Research Article, we present a novel approach of using collagen hydrolysate, smaller fragments of collagen, as an alternative to traditionally used collagen scaffold. Collagen hydrolysate composite scaffold (CHCS) was fabricated with sol-gel transition procedure using tetraethoxysilane as the silica precursor. CHCS exhibits porous morphology with pore sizes varying between 380 and 780 μm. Incorporation of silica conferred CHCS with controlled biodegradation and better water uptake capacity. Notably, 3T3 fibroblast proliferation was seen to be significantly better under CHCS treatment when compared to treatment with collagen scaffold. Additionally, CHCS showed excellent antimicrobial activity against the wound pathogens Staphylococcus aureus, Bacillus subtilis, and Escherichia coli due to the inherited antimicrobial activity of collagen hydrolysate. In vivo wound healing experiments with full thickness excision wounds in rat model demonstrated that wounds treated with CHCS showed accelerated healing when compared to wounds treated with collagen scaffold. These findings indicate that the CHCS scaffold from collagen fragments would be an effective and affordable alternative to the traditionally used collagen structural biomaterials.

  17. A novel porous bioceramics scaffold by accumulating hydroxyapatite spherulites for large bone tissue engineering in vivo. II. Construct large volume of bone grafts.

    PubMed

    Zhi, Wei; Zhang, Cong; Duan, Ke; Li, Xiaohong; Qu, Shuxin; Wang, Jianxin; Zhu, Zhuoli; Huang, Peng; Xia, Tian; Liao, Ga; Weng, Jie

    2014-08-01

    In vivo engineering of bone autografts using bioceramic scaffolds with appropriate porous structures is a potential approach to prepare autologous bone grafts for the repair of critical-sized bone defects. This study investigated the evolutionary process of osteogenesis, angiogenesis, and compressive strength of bioceramic scaffolds implanted in two non-osseous sites of dogs: the abdominal cavity and the dorsal muscle. Hydroxyapatite (HA) sphere-accumulated scaffolds with controlled porous structures were prepared and placed in the two sites for up to 6 months. Analyses of retrieved scaffolds found that osteogenesis and angiogenesis were faster in scaffolds implanted in dorsal muscles compared with those placed in abdominal cavities. The abdominal cavity, however, can accommodate larger bone grafts with designed shape. Analyses of scaffolds implanted in abdominal cavities [an environment of a low mesenchymal stem cell (MSC) density] further demonstrated that angiogenesis play critical roles during osteogenesis in the scaffolds, presumably by supplying progenitor cells and/or MSCs as seed cells. This study also examined the relationship between the volume of bone grafts and the physiological environment of in vivo bioreactor. These results provide basic information for the selection of appropriate implanting sites and culture time required to engineer autologous bone grafts for the clinical bone defect repair. Based on these positive results, a pilot study has applied the grafts constructed in canine abdominal cavity to repair segmental bone defect in load-bearing sites (limbs).

  18. Enhancement of bone formation by BMP-7 transduced MSCs on biomimetic nano-hydroxyapatite/polyamide composite scaffolds in repair of mandibular defects.

    PubMed

    Li, Jihua; Li, Yubao; Ma, Sancheng; Gao, Ying; Zuo, Yi; Hu, Jing

    2010-12-15

    This study was to evaluate enhanced bone formation by bone morphogenetic protein-7 (BMP-7) transduced MSCs on nano-hydroxyapatite/polyamide (n-HA/PA) composite scaffolds for bone tissue engineering in repair of mandibular defect. n-HA/PA scaffolds were prepared and rabbit MSCs were separated and expanded; and then infected with adenoviral-mediated BMP-7 in vitro. The MSCs-BMP-7 and MSCs were seeded on the porous scaffolds. Scaffold/MSCs-BMP-7 constructs and scaffold/MSCs constructs were implanted in the defects of rabbits' mandible as the experimental groups A (n = 18) and groups B (n = 18), respectively, the pure scaffolds were implanted as controls (group C, n = 18). Six animals were sacrificed at 4-, 8-, and 16-week postimplantation, respectively. Their mandibles were removed and processed for radiographic, biomechanical tests, histological, and histomorphometric analysis. Group A animals showed greater bone formation and earlier mineralization than group B at 4- and 8-week postimplantation and similarly group B more than group C. However, no difference was found among three groups at 16-week postimplantation. The results of this study suggest that BMP-7 transduced MSCs-n-HA/PA composite could significantly accelerate bone formation in the implant at early stage. BMP-7 mediated ex vivo gene transfer based on MSCs as seed cells, combined with porous n-HA/PA as scaffolds for bone tissue engineering might be an alternative or supplemental approach to repair the mandibular defects. Copyright © 2010 Wiley Periodicals, Inc.

  19. Fabrication and Characterization of Three-Dimensional Macroscopic All-Carbon Scaffolds

    PubMed Central

    Lalwani, Gaurav; Kwaczala, Andrea Trinward; Kanakia, Shruti; Patel, Sunny C.; Judex, Stefan; Sitharaman, Balaji

    2012-01-01

    We report a simple method to fabricate macroscopic, 3-D, free standing, all-carbon scaffolds (porous structures) using multiwalled carbon nanotubes (MWCNTs) as the starting materials. The scaffolds prepared by radical initiated thermal crosslinking, and annealing of MWCNTs possess macroscale interconnected pores, robust structural integrity, stability, and conductivity. The porosity of the three-dimensional structure can be controlled by varying the amount of radical initiator, thereby allowing the design of porous scaffolds tailored towards specific potential applications. This method also allows the fabrication of 3-D scaffolds using other carbon nanomaterials such as single-walled carbon nanotubes, fullerenes, and graphene indicating that it could be used as a versatile method for 3-D assembly of carbon nanostructures with pi bond networks. PMID:23436939

  20. Tunable tissue scaffolds fabricated by in situ crosslink in phase separation system

    PubMed Central

    Liu, Xifeng; Chen, Wenjian; Gustafson, Carl T.; Miller, A. Lee; Waletzki, Brian E.; Yaszemski, Michael J.; Lu, Lichun

    2015-01-01

    Three-dimensional (3-D) scaffolds with intrinsic porous structures are desirable in various tissue regeneration applications. In this study, a unique method that combines thermally induced phase separation with a photocrosslinking process was developed for the fabrication of 3-D crosslinked polymer scaffolds with densely interconnected porous structures. Biodegradable poly(propylene fumarate)-co-poly(L-lactic acid) with crosslinkable fumarate bonds were used as the structural polymer material and a dioxane/water binary system was applied for the phase separation. By altering the polymer composition (9, 5 and 3 wt%), different types of scaffolds with distinct morphology, mechanical strength, degradation rate, cell growth and morphology, and extracellular matrix production were fabricated. These crosslinked 3-D porous scaffolds with tunable strength and biological responses show promise for potential applications in regenerative therapies, including bone and neural tissue engineering. PMID:26989479

  1. Fabrication, characterization, and in vitro degradation of composite scaffolds based on PHBV and bioactive glass.

    PubMed

    Li, Haiyan; Du, Ruilin; Chang, Jiang

    2005-10-01

    Composite scaffolds of polyhydroxybutyrate-polyhydroxyvalerate (PHBV) with sol-gel-derived bioactive glass (BG, 58S) are fabricated by compression molding, thermal processing, and salt particulate leaching method. Structure and mechanical properties of the scaffolds are determined. The bioactivity of the composites is evaluated by soaking the scaffolds in a simulated body fluid (SBF), and the formation of the apatite layer on the scaffolds is determined by scanning electron microscopy (SEM) and energy-dispersive spectrometry (EDS). The results show that the PHBV/BG composites are bioactive as they induce the formation of apatite on the composite scaffolds after soaking in SBF for 3 days. In addition, the measurements of the water contact angles suggest that incorporation of BG into PHBV can improve the hydrophilicity of the composites and the enhancement is dependent on the BG content. Furthermore, the degradation assessment of the scaffolds is performed in phosphate-buffered saline (PBS) solution at 37 C. Weight loss and water absorption of the scaffolds, pH of the incubation media, and molecular weight measurements of the PHBV in the scaffolds are used to monitor the degradation of the scaffolds during a nine-week incubation in PBS. It has been found that the incorporation of bioactive glass into the PHBV delayed the degradation of PHBV in the composite scaffolds for the period investigated. The present results show not only a useful method to prepare composite scaffolds with improved properties but also a way of adjusting the in vitro degradation behavior of composite scaffolds by tailoring the content of bioactive glass.

  2. A novel porous scaffold fabrication technique for epithelial and endothelial tissue engineering.

    PubMed

    McHugh, Kevin J; Tao, Sarah L; Saint-Geniez, Magali

    2013-07-01

    Porous scaffolds have the ability to minimize transport barriers for both two- (2D) and three-dimensional tissue engineering. However, current porous scaffolds may be non-ideal for 2D tissues such as epithelium due to inherent fabrication-based characteristics. While 2D tissues require porosity to support molecular transport, pores must be small enough to prevent cell migration into the scaffold in order to avoid non-epithelial tissue architecture and compromised function. Though electrospun meshes are the most popular porous scaffolds used today, their heterogeneous pore size and intense topography may be poorly-suited for epithelium. Porous scaffolds produced using other methods have similar unavoidable limitations, frequently involving insufficient pore resolution and control, which make them incompatible with 2D tissues. In addition, many of these techniques require an entirely new round of process development in order to change material or pore size. Herein we describe "pore casting," a fabrication method that produces flat scaffolds with deterministic pore shape, size, and location that can be easily altered to accommodate new materials or pore dimensions. As proof-of-concept, pore-cast poly(ε-caprolactone) (PCL) scaffolds were fabricated and compared to electrospun PCL in vitro using canine kidney epithelium, human colon epithelium, and human umbilical vein endothelium. All cell types demonstrated improved morphology and function on pore-cast scaffolds, likely due to reduced topography and universally small pore size. These results suggest that pore casting is an attractive option for creating 2D tissue engineering scaffolds, especially when the application may benefit from well-controlled pore size or architecture.

  3. Cartilage Tissue Engineering with Silk Fibroin Scaffolds Fabricated by Indirect Additive Manufacturing Technology.

    PubMed

    Chen, Chih-Hao; Liu, Jolene Mei-Jun; Chua, Chee-Kai; Chou, Siaw-Meng; Shyu, Victor Bong-Hang; Chen, Jyh-Ping

    2014-03-13

    Advanced tissue engineering (TE) technology based on additive manufacturing (AM) can fabricate scaffolds with a three-dimensional (3D) environment suitable for cartilage regeneration. Specifically, AM technology may allow the incorporation of complex architectural features. The present study involves the fabrication of 3D TE scaffolds by an indirect AM approach using silk fibroin (SF). From scanning electron microscopic observations, the presence of micro-pores and interconnected channels within the scaffold could be verified, resulting in a TE scaffold with both micro- and macro-structural features. The intrinsic properties, such as the chemical structure and thermal characteristics of SF, were preserved after the indirect AM manufacturing process. In vitro cell culture within the SF scaffold using porcine articular chondrocytes showed a steady increase in cell numbers up to Day 14. The specific production (per cell basis) of the cartilage-specific extracellular matrix component (collagen Type II) was enhanced with culture time up to 12 weeks, indicating the re-differentiation of chondrocytes within the scaffold. Subcutaneous implantation of the scaffold-chondrocyte constructs in nude mice also confirmed the formation of ectopic cartilage by histological examination and immunostaining.

  4. (*) Fabrication and Characterization of Biphasic Silk Fibroin Scaffolds for Tendon/Ligament-to-Bone Tissue Engineering.

    PubMed

    Font Tellado, Sònia; Bonani, Walter; Balmayor, Elizabeth R; Foehr, Peter; Motta, Antonella; Migliaresi, Claudio; van Griensven, Martijn

    2017-08-01

    Tissue engineering is an attractive strategy for tendon/ligament-to-bone interface repair. The structure and extracellular matrix composition of the interface are complex and allow for a gradual mechanical stress transfer between tendons/ligaments and bone. Thus, scaffolds mimicking the structural features of the native interface may be able to better support functional tissue regeneration. In this study, we fabricated biphasic silk fibroin scaffolds designed to mimic the gradient in collagen molecule alignment present at the interface. The scaffolds had two different pore alignments: anisotropic at the tendon/ligament side and isotropic at the bone side. Total porosity ranged from 50% to 80% and the majority of pores (80-90%) were <100-300 μm. Young's modulus varied from 689 to 1322 kPa depending on the type of construct. In addition, human adipose-derived mesenchymal stem cells were cultured on the scaffolds to evaluate the effect of pore morphology on cell proliferation and gene expression. Biphasic scaffolds supported cell attachment and influenced cytoskeleton organization depending on pore alignment. In addition, the gene expression of tendon/ligament, enthesis, and cartilage markers significantly changed depending on pore alignment in each region of the scaffolds. In conclusion, the biphasic scaffolds fabricated in this study show promising features for tendon/ligament-to-bone tissue engineering.

  5. Cartilage Tissue Engineering with Silk Fibroin Scaffolds Fabricated by Indirect Additive Manufacturing Technology

    PubMed Central

    Chen, Chih-Hao; Liu, Jolene Mei-Jun; Chua, Chee-Kai; Chou, Siaw-Meng; Shyu, Victor Bong-Hang; Chen, Jyh-Ping

    2014-01-01

    Advanced tissue engineering (TE) technology based on additive manufacturing (AM) can fabricate scaffolds with a three-dimensional (3D) environment suitable for cartilage regeneration. Specifically, AM technology may allow the incorporation of complex architectural features. The present study involves the fabrication of 3D TE scaffolds by an indirect AM approach using silk fibroin (SF). From scanning electron microscopic observations, the presence of micro-pores and interconnected channels within the scaffold could be verified, resulting in a TE scaffold with both micro- and macro-structural features. The intrinsic properties, such as the chemical structure and thermal characteristics of SF, were preserved after the indirect AM manufacturing process. In vitro cell culture within the SF scaffold using porcine articular chondrocytes showed a steady increase in cell numbers up to Day 14. The specific production (per cell basis) of the cartilage-specific extracellular matrix component (collagen Type II) was enhanced with culture time up to 12 weeks, indicating the re-differentiation of chondrocytes within the scaffold. Subcutaneous implantation of the scaffold-chondrocyte constructs in nude mice also confirmed the formation of ectopic cartilage by histological examination and immunostaining. PMID:28788558

  6. Techniques for fabrication and construction of three-dimensional scaffolds for tissue engineering.

    PubMed

    Lu, Tingli; Li, Yuhui; Chen, Tao

    2013-01-01

    Three-dimensional biomimetic scaffolds have widespread applications in biomedical tissue engineering because of their nanoscaled architecture, eg, nanofibers and nanopores, similar to the native extracellular matrix. In the conventional "top-down" approach, cells are seeded onto a biocompatible and biodegradable scaffold, in which cells are expected to populate in the scaffold and create their own extracellular matrix. The top-down approach based on these scaffolds has successfully engineered thin tissues, including skin, bladder, and cartilage in vitro. However, it is still a challenge to fabricate complex and functional tissues (eg, liver and kidney) due to the lack of vascularization systems and limited diffusion properties of these large biomimetic scaffolds. The emerging "bottom-up" method may hold great potential to address these challenges, and focuses on fabricating microscale tissue building blocks with a specific microarchitecture and assembling these units to engineer larger tissue constructs from the bottom up. In this review, state-of-the-art methods for fabrication of three-dimensional biomimetic scaffolds are presented, and their advantages and drawbacks are discussed. The bottom-up methods used to assemble microscale building blocks (eg, microscale hydrogels) for tissue engineering are also reviewed. Finally, perspectives on future development of the bottom-up approach for tissue engineering are addressed.

  7. Magnetic resonance imaging-three-dimensional printing technology fabricates customized scaffolds for brain tissue engineering.

    PubMed

    Fu, Feng; Qin, Zhe; Xu, Chao; Chen, Xu-Yi; Li, Rui-Xin; Wang, Li-Na; Peng, Ding-Wei; Sun, Hong-Tao; Tu, Yue; Chen, Chong; Zhang, Sai; Zhao, Ming-Liang; Li, Xiao-Hong

    2017-04-01

    Conventional fabrication methods lack the ability to control both macro- and micro-structures of generated scaffolds. Three-dimensional printing is a solid free-form fabrication method that provides novel ways to create customized scaffolds with high precision and accuracy. In this study, an electrically controlled cortical impactor was used to induce randomized brain tissue defects. The overall shape of scaffolds was designed using rat-specific anatomical data obtained from magnetic resonance imaging, and the internal structure was created by computer-aided design. As the result of limitations arising from insufficient resolution of the manufacturing process, we magnified the size of the cavity model prototype five-fold to successfully fabricate customized collagen-chitosan scaffolds using three-dimensional printing. Results demonstrated that scaffolds have three-dimensional porous structures, high porosity, highly specific surface areas, pore connectivity and good internal characteristics. Neural stem cells co-cultured with scaffolds showed good viability, indicating good biocompatibility and biodegradability. This technique may be a promising new strategy for regenerating complex damaged brain tissues, and helps pave the way toward personalized medicine.

  8. Techniques for fabrication and construction of three-dimensional scaffolds for tissue engineering

    PubMed Central

    Lu, Tingli; Li, Yuhui; Chen, Tao

    2013-01-01

    Three-dimensional biomimetic scaffolds have widespread applications in biomedical tissue engineering because of their nanoscaled architecture, eg, nanofibers and nanopores, similar to the native extracellular matrix. In the conventional “top-down” approach, cells are seeded onto a biocompatible and biodegradable scaffold, in which cells are expected to populate in the scaffold and create their own extracellular matrix. The top-down approach based on these scaffolds has successfully engineered thin tissues, including skin, bladder, and cartilage in vitro. However, it is still a challenge to fabricate complex and functional tissues (eg, liver and kidney) due to the lack of vascularization systems and limited diffusion properties of these large biomimetic scaffolds. The emerging “bottom-up” method may hold great potential to address these challenges, and focuses on fabricating microscale tissue building blocks with a specific microarchitecture and assembling these units to engineer larger tissue constructs from the bottom up. In this review, state-of-the-art methods for fabrication of three-dimensional biomimetic scaffolds are presented, and their advantages and drawbacks are discussed. The bottom-up methods used to assemble microscale building blocks (eg, microscale hydrogels) for tissue engineering are also reviewed. Finally, perspectives on future development of the bottom-up approach for tissue engineering are addressed. PMID:23345979

  9. Strategic Design and Fabrication of Engineered Scaffolds for Articular Cartilage Repair

    PubMed Central

    Izadifar, Zohreh; Chen, Xiongbiao; Kulyk, William

    2012-01-01

    Damage to articular cartilage can eventually lead to osteoarthritis (OA), a debilitating, degenerative joint disease that affects millions of people around the world. The limited natural healing ability of cartilage and the limitations of currently available therapies make treatment of cartilage defects a challenging clinical issue. Hopes have been raised for the repair of articular cartilage with the help of supportive structures, called scaffolds, created through tissue engineering (TE). Over the past two decades, different designs and fabrication techniques have been investigated for developing TE scaffolds suitable for the construction of transplantable artificial cartilage tissue substitutes. Advances in fabrication technologies now enable the strategic design of scaffolds with complex, biomimetic structures and properties. In particular, scaffolds with hybrid and/or biomimetic zonal designs have recently been developed for cartilage tissue engineering applications. This paper reviews critical aspects of the design of engineered scaffolds for articular cartilage repair as well as the available advanced fabrication techniques. In addition, recent studies on the design of hybrid and zonal scaffolds for use in cartilage tissue repair are highlighted. PMID:24955748

  10. Fabrication and characterization of porous PHBV scaffolds for tissue engineering

    NASA Astrophysics Data System (ADS)

    Ruiz, I.; Hermida, É. B.; Baldessari, A.

    2011-12-01

    Porous scaffolds of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) were elaborated by three different techniques: salt leaching (SL), emulsion solvent evaporation (ESE) and temperature induced phase separation (TIPS). For SL partially fused sieved grains of sodium chloride (106-355 μm) were used as porogen. Emulsions, prepared from a solution of PHBV in chloroform allow getting flexible films; the content of surfactant may be used to control the pore size. The pore size of the TIPS scaffolds decreased on increasing the cooling rate and the morphology of the interconnected structure could be controlled by changing the temperature gradient. Finally, chemical changes associated to the enhancement of hydrophilic behaviour of the scaffolds after alkaline and enzymatic hydrolysis as well as after sterilization by γ irradiation are presented.

  11. Fabricating gradient hydrogel scaffolds for 3D cell culture.

    PubMed

    Chatterjee, Kaushik; Young, Marian F; Simon, Carl G

    2011-05-01

    Optimizing cell-material interactions is critical for maximizing regeneration in tissue engineering. Combinatorial and high-throughput (CHT) methods can be used to systematically screen tissue scaffolds to identify optimal biomaterial properties. Previous CHT platforms in tissue engineering have involved a two-dimensional (2D) cell culture format where cells were cultured on material surfaces. However, these platforms are inadequate to predict cellular response in a three-dimensional (3D) tissue scaffold. We have developed a simple CHT platform to screen cell-material interactions in 3D culture format that can be applied to screen hydrogel scaffolds. Herein we provide detailed instructions on a method to prepare gradients in elastic modulus of photopolymerizable hydrogels.

  12. Hyper-elastic modeling and mechanical behavior investigation of porous poly-D-L-lactide/nano-hydroxyapatite scaffold material.

    PubMed

    Han, Quan Feng; Wang, Ze Wu; Tang, Chak Yin; Chen, Ling; Tsui, Chi Pong; Law, Wing Cheung

    2017-03-28

    Poly-D-L-lactide/nano-hydroxyapatite (PDLLA/nano-HA) can be used as the biological scaffold material in bone tissue engineering as it can be readily made into a porous composite material with excellent performance. However, constitutive modeling for the mechanical response of porous PDLLA/nano-HA under various stress conditions has been very limited so far. In this work, four types of fundamental compressible hyper-elastic constitutive models were introduced for constitutive modeling and investigation of mechanical behaviors of porous PDLLA/nano-HA. Moreover, the unitary expressions of Cauchy stress tensor have been derived for the PDLLA/nano-HA under uniaxial compression (or stretch), biaxial compression (or stretch), pure shear and simple shear load by using the theory of continuum mechanics. The theoretical results determined from the approach based on the Ogden compressible hyper-elastic constitutive model were in good agreement with the experimental data from the uniaxial compression tests. Furthermore, this approach can also be used to predict the mechanical behaviors of the porous PDLLA/nano-HA material under the biaxial compression (or stretch), pure shear and simple shear.

  13. A mild one-pot process for synthesising hydroxyapatite/biomolecule bone scaffolds for sustained and controlled antibiotic release.

    PubMed

    Hess, Ulrike; Hill, Sebastian; Treccani, Laura; Streckbein, Philipp; Heiss, Christian; Rezwan, Kurosch

    2015-01-16

    The release of active molecules or the control of nosocomial infections for improved osteoinduction is ideally addressed by a bone substitute material. For this purpose, the feasibility of a mild one-pot process is probed for incorporating directly active proteins and antibiotics in a hydroxyapatite (HAp) based scaffold. The effect of two serum model proteins, bovine serum albumin (BSA) and fibrinogen (FIB), on the microstructure, on selected mechanical properties as well as on degradation behaviour and on protein release are investigated. By protein incorporation, the porosity can be adjusted between 54 and 70% especially due to the foaming ability of BSA. The addition of 5 wt% FIB doubles the biaxial flexural strength up to 6 MPa in comparison to samples without proteins (3 MPa). Protein release experiments show that a rapid release takes place within the first days (between around 3% for FIB and 38% for BSA). As a possible application for osteomyelitis treatment, vancomycin and gentamicin were subsequently added instead of proteins to study their release behaviour and their antibacterial activity, respectively. A controlled antibiotic release was observed for a period of 18 d. By varying the protein type, mixture and quantity, the mechanical strength porosity as well as the protein release and calcium solubility can be controlled. Our studies underpin the suitability of this mild one-pot process as a promising simple-to-use platform for controlled local drug release and bone treatment.

  14. Fabrication of PLGA microvessel scaffolds with circular microchannels using soft lithography

    NASA Astrophysics Data System (ADS)

    Wang, Gou-Jen; Hsueh, Cheng-Chih; Hsu, Shan-hui; Hung, Huey-Shan

    2007-10-01

    A simple micromolding method for fabricating PLGA (poly(lactic-co-glycolic acid)) microstructures made up of microchannels with a circular cross-section is presented. The thermal reflow technique is adopted to fabricate the semi-cylindrical photoresist master. The PLGA solution is prepared by dissolving PLGA polymer in acetone and then casting the solution onto the semi-cylindrical photoresist master to produce PLGA microstructures. Two PLGA membranes are bonded together to form microstructures consisting of circular microchannels. A microvessel scaffold suitable for tissue engineering was fabricated using the proposed method, and bovine endothelial cells were cultured into the scaffold by semi-dynamic seeding. The cell stain Calcein-AM was used to overcome the problem of the PLGA scaffolds becoming opaque, which in the past had made it difficult to effectively monitor the progress of cell seeding.

  15. Modern materials in fabrication of scaffolds for bone defect replacement

    NASA Astrophysics Data System (ADS)

    Bazlov, V. A.; Mamuladze, T. Z.; Pavlov, V. V.; Kirilova, I. A.; Sadovoy, M. A.

    2016-08-01

    The article defines the requirements for modern scaffold-forming materials and describes the main advantages and disadvantages of various synthetic materials. Osseointegration of synthetic scaffolds approved for use in medical practice is evaluated. Nylon 618 (certification ISO9001 1093-1-2009) is described as the most promising synthetic material used in medical practice. The authors briefly highlight the issues of individual bone grafting with the use of 3D printing technology. An example of contouring pelvis defect after removal of a giant tumor with the use of 3D models is provided.

  16. New paradigms in internal architecture design and freeform fabrication of tissue engineering porous scaffolds.

    PubMed

    Yoo, Dongjin

    2012-07-01

    Advanced additive manufacture (AM) techniques are now being developed to fabricate scaffolds with controlled internal pore architectures in the field of tissue engineering. In general, these techniques use a hybrid method which combines computer-aided design (CAD) with computer-aided manufacturing (CAM) tools to design and fabricate complicated three-dimensional (3D) scaffold models. The mathematical descriptions of micro-architectures along with the macro-structures of the 3D scaffold models are limited by current CAD technologies as well as by the difficulty of transferring the designed digital models to standard formats for fabrication. To overcome these difficulties, we have developed an efficient internal pore architecture design system based on triply periodic minimal surface (TPMS) unit cell libraries and associated computational methods to assemble TPMS unit cells into an entire scaffold model. In addition, we have developed a process planning technique based on TPMS internal architecture pattern of unit cells to generate tool paths for freeform fabrication of tissue engineering porous scaffolds.

  17. High-precision flexible fabrication of tissue engineering scaffolds using distinct polymers

    SciTech Connect

    Wei, Chuang; Cai, Lei; Sonawane, Bhushan; Wang, Shanfeng; Dong, Jingyan

    2012-01-01

    Three-dimensional porous structures using biodegradable materials with excellent biocompatibility are critically important for tissue engineering applications. We present a multi-nozzle-based versatile deposition approach to flexibly construct porous tissue engineering scaffolds using distinct polymeric biomaterials such as thermoplastic and photo-crosslinkable polymers. We first describe the development of the deposition system and fabrication of scaffolds from two types of biodegradable polymers using this system. The thermoplastic sample is semi-crystalline poly({var_epsilon}-caprolactone) (PCL) that can be processed at a temperature higher than its melting point and solidifies at room temperature. The photo-crosslinkable one is polypropylene fumarate (PPF) that has to be dissolved in a reactive solvent as a resin for being cured into solid structures. Besides the direct fabrication of thermoplastic PCL scaffolds, we specifically develop a layer molding approach for the fabrication of crosslinkable polymers, which traditionally can only be fabricated by stereolithography. In this approach, a thermoplastic supporting material (paraffin wax) is first deposited to make a mold for each specific layer, and then PPF is deposited on demand to fill the mold and cured by the UV light. The supporting material can be removed to produce a porous scaffold of crosslinked PPF. Both PCL and crosslinked PPF scaffolds fabricated using the developed system have been characterized in terms of compressive mechanical properties, morphology, pore size and porosity. Mouse MC3T3-E1 pre-osteoblastic cell studies on the fabricated scaffolds have been performed to demonstrate their capability of supporting cell proliferation and ingrowth, aiming for bone tissue engineering applications.

  18. High-precision flexible fabrication of tissue engineering scaffolds using distinct polymers.

    PubMed

    Wei, Chuang; Cai, Lei; Sonawane, Bhushan; Wang, Shanfeng; Dong, Jingyan

    2012-06-01

    Three-dimensional porous structures using biodegradable materials with excellent biocompatibility are critically important for tissue engineering applications. We present a multi-nozzle-based versatile deposition approach to flexibly construct porous tissue engineering scaffolds using distinct polymeric biomaterials such as thermoplastic and photo-crosslinkable polymers. We first describe the development of the deposition system and fabrication of scaffolds from two types of biodegradable polymers using this system. The thermoplastic sample is semi-crystalline poly(ε-caprolactone) (PCL) that can be processed at a temperature higher than its melting point and solidifies at room temperature. The photo-crosslinkable one is polypropylene fumarate (PPF) that has to be dissolved in a reactive solvent as a resin for being cured into solid structures. Besides the direct fabrication of thermoplastic PCL scaffolds, we specifically develop a layer molding approach for the fabrication of crosslinkable polymers, which traditionally can only be fabricated by stereolithography. In this approach, a thermoplastic supporting material (paraffin wax) is first deposited to make a mold for each specific layer, and then PPF is deposited on demand to fill the mold and cured by the UV light. The supporting material can be removed to produce a porous scaffold of crosslinked PPF. Both PCL and crosslinked PPF scaffolds fabricated using the developed system have been characterized in terms of compressive mechanical properties, morphology, pore size and porosity. Mouse MC3T3-E1 pre-osteoblastic cell studies on the fabricated scaffolds have been performed to demonstrate their capability of supporting cell proliferation and ingrowth, aiming for bone tissue engineering applications.

  19. Friction of sodium alginate hydrogel scaffold fabricated by 3-D printing.

    PubMed

    Yang, Qian; Li, Jian; Xu, Heng; Long, Shijun; Li, Xuefeng

    2017-04-01

    A rapid prototyping technology, formed by three-dimensional (3-D) printing and then crosslinked by spraying Ca(2+) solution, is developed to fabricate a sodium alginate (SA) hydrogel scaffold. The porosity, swelling ratio, and compression modulus of the scaffold are investigated. A friction mechanism is developed by studying the reproducible friction behavior. Our results show that the scaffold can have 3-D structure with a porosity of 52%. The degree of swelling of the SA hydrogel scaffold is 8.5, which is nearly the same as bulk SA hydrogel. SA hydrogel exhibits better compressive resilience than bulk hydrogel despite its lower compressive modulus compared to bulk hydrogel. The SA hydrogel scaffold exhibits a higher frictional force at low sliding velocity (10(-6) to 10(-3) m/s) compared to bulk SA hydrogel, and they are equal at high sliding velocity (10(-2) to 1 m/s). For a small pressure (0.3 kPa), the SA hydrogel scaffold shows good friction reproducibility. In contrast, bulk SA hydrogel shows poor reproducibility with respect to friction behavior. The differences in friction behaviors between the SA hydrogel scaffold and bulk SA hydrogel are related to the structure of the scaffold, which can keep a stable hydrated lubrication layer.

  20. Dewetting based fabrication of fibrous micro-scaffolds as potential injectable cell carriers.

    PubMed

    Song, Hokyung; Yin, Liya; Chilian, William M; Zhang Newby, Bi-Min

    2015-03-01

    Although regenerative medicine utilizing tissue scaffolds has made enormous strides in recent years, many constraints still hamper their effectiveness. A limitation of many scaffolds is that they form surface patches, which are not particularly effective for some types of "wounds" that are deep within tissues, e.g., stroke and myocardial infarction. In this study, we reported the generation of fibrous micro-scaffolds feasible for delivering cells by injection into the tissue parenchyma. The micro-scaffolds (widths<100μm) were made by dewetting of poly(lactic-co-glycolic acid) thin films containing parallel strips, and cells were seeded to form cell/polymer micro-constructs during or post the micro-scaffold fabrication process. Five types of cells including rat induced vascular progenitor cells were assessed for the formation of the micro-constructs. Critical factors in forming fibrous micro-scaffolds via dewetting of polymer thin films were found to be properties of polymers and supporting substrates, temperature, and proteins in the culture medium. Also, the ability of cells to attach to the micro-scaffolds was essential in forming cell/polymer micro-constructs. Both in vitro and in vivo assessments of injecting these micro-scaffolding constructs showed, as compared to free cells, enhanced cell retention at the injected site, which could lead to improved tissue engineering and regeneration.

  1. Recent prospective of nanofiber scaffolds fabrication approaches for skin regeneration.

    PubMed

    Ahmadi-Aghkand, Fateme; Gholizadeh-Ghaleh Aziz, Shiva; Panahi, Yunes; Daraee, Hadis; Gorjikhah, Fateme; Gholizadeh-Ghaleh Aziz, Sara; Hsanzadeh, Arash; Akbarzadeh, Abolfazl

    2016-11-01

    The largest organ of human body is skin, which acting as a barrier with immunologic, sensorial and protective functions. It is always in exposure to the external environment, which can result many different types of damage and injury with loss of variable volumes of extracellular matrix (ECM). For the treatment of skin lesions and damages, several approaches are now accessible, such as the application of allografts, autografts, and tissue-engineered substitutes, wound dressings and nanofiber scaffolds approaches. Even though proven clinically effective, these methods are still characterized by main drawbacks such as patient inadequate vascularization, morbidity, the inability to reproduce skin appendages, low adherence to the wound bed and high manufacturing costs. Advanced approaches based on nanofiber scaffolds approaches offer a permanent, viable and effective substitute to explain the drawbacks of skin regeneration and repair by combining growth factors, cells, and biomaterials and advanced biomanufacturing methods. This review details recent advances of nanofiber scaffolds in skin regeneration and repair strategies, and describes a synthesis method of nanofiber scaffolds.

  2. Fabrication of biodegradable textile scaffold based on hydrophobized hyaluronic acid.

    PubMed

    Zapotocky, Vojtech; Pospisilova, Martina; Janouchova, Katerina; Svadlak, Daniel; Batova, Jana; Sogorkova, Jana; Cepa, Martin; Betak, Jiri; Stepankova, Veronika; Sulakova, Romana; Kulhanek, Jaromir; Pitucha, Tomas; Vranova, Jana; Duffy, Garry; Velebny, Vladimir

    2017-02-01

    In this work, we report on the preparation of a novel biodegradable textile scaffold made of palmitoyl-hyaluronan (palHA). Monofilament fibres of palHA with a diameter of 120μm were prepared by wet spinning. The wet-spun fibres were subsequently processed into a warp-knitted textile. To find a compromise between swelling in water and degradability of the final textile scaffold, a series of palHA derivatives with different degrees of substitution of the palmitoyl chain was synthesized. Freeze-drying not only provided shape fixation, but also speeded up scaffold degradation in vitro. Fibronectin, fibrinogen, laminin and collagen IV were physically adsorbed on the textile surface to enhance cell adhesion on the material. The highest amount of adsorbed cell-adhesive proteins was achieved with fibronectin (89%), followed by fibrinogen (81%). Finally, textiles modified with fibronectin or fibrinogen both supported the adhesion and proliferation of normal human fibroblasts in vitro, proving to be a useful cellular scaffold for tissue engineering.

  3. Metal nanodot arrays fabricated via seed-mediated electroless plating with block copolymer thin film scaffolding.

    PubMed

    Komiyama, Hideaki; Iyoda, Tomokazu; Sanji, Takanobu

    2015-10-02

    We present an alternative approach to fabricating hexagonally arranged nanodot arrays of various metals by seed-mediated electroless plating with a cylinder-forming block copolymer thin film, PEO-b-PMA(Az), as a scaffold. Metal ions were selectively incorporated into PEO cylinders, followed by their reduction to metal and the etching of the scaffold to obtain highly ordered seed arrays of Au, Pd, and Pt. Nanodot arrays of the target metals (Au, Ag, and Ni) were selectively grown on the seed with their highly ordered arrangement by electroless plating. We studied the fabrication processes' suitability for control of the nanodot array size, as well as the plasmonic properties thereof.

  4. Metal nanodot arrays fabricated via seed-mediated electroless plating with block copolymer thin film scaffolding

    NASA Astrophysics Data System (ADS)

    Komiyama, Hideaki; Iyoda, Tomokazu; Sanji, Takanobu

    2015-10-01

    We present an alternative approach to fabricating hexagonally arranged nanodot arrays of various metals by seed-mediated electroless plating with a cylinder-forming block copolymer thin film, PEO-b-PMA(Az), as a scaffold. Metal ions were selectively incorporated into PEO cylinders, followed by their reduction to metal and the etching of the scaffold to obtain highly ordered seed arrays of Au, Pd, and Pt. Nanodot arrays of the target metals (Au, Ag, and Ni) were selectively grown on the seed with their highly ordered arrangement by electroless plating. We studied the fabrication processes’ suitability for control of the nanodot array size, as well as the plasmonic properties thereof.

  5. Histological Comparison in Rats between Carbonate Apatite Fabricated from Gypsum and Sintered Hydroxyapatite on Bone Remodeling.

    PubMed

    Ayukawa, Yasunori; Suzuki, Yumiko; Tsuru, Kanji; Koyano, Kiyoshi; Ishikawa, Kunio

    2015-01-01

    Carbonate apatite (CO3Ap), the form of apatite found in bone, has recently attracted attention. The purpose of the present study was to histologically evaluate the tissue/cellular response toward the low-crystalline CO3Ap fabricated using a dissolution-precipitation reaction with set gypsum as a precursor. When set gypsum was immersed in a 100°C 1 mol/L Na3PO4 aqueous solution for 24 h, the set gypsum transformed into CO3Ap. Both CO3Ap and sintered hydroxyapatite (s-HAp), which was used as a control, were implanted into surgically created tibial bone defects of rats for histological evaluation. Two and 4 weeks after the implantation, histological sections were created and observed using light microscopy. The CO3Ap granules revealed both direct apposition of the bone matrix by osteoblasts and osteoclastic resorption. In contrast, the s-HAp granules maintained their contour even after 4 weeks following implantation which implied that there was a lack of replacement into the bone. The s-HAp granules were sometimes encapsulated with fibrous tissue, and macrophage polykaryon was occasionally observed directly apposed to the implanted granules. From the viewpoint of bone remodeling, the CO3Ap granules mimicked the bone matrix, suggesting that CO3Ap may be an appropriate bone substitute.

  6. Towards the Design of 3D Fiber-Deposited Poly(ε-caprolactone)/lron-Doped Hydroxyapatite Nanocomposite Magnetic Scaffolds for Bone Regeneration.

    PubMed

    De Santis, Roberta; Russo, Alessandro; Gloria, Antonio; D'Amora, Ugo; Russo, Teresa; Panseri, Silvia; Sandri, Monica; Tampieri, Anna; Marcacci, Maurilio; Dediu, Valentin A; Wilde, Colin J; Ambrosio, Luigi

    2015-07-01

    In the past few years, researchers have focused on the design and development of three-dimensional (3D) advanced scaffolds, which offer significant advantages in terms of cell performance. The introduction of magnetic features into scaffold technology could offer innovative opportunities to control cell populations within 3D microenvironments, with the potential to enhance their use in tissue regeneration or in cell-based analysis. In the present study, 3D fully biodegradable and magnetic nanocomposite scaffolds for bone tissue engineering, consisting of a poly(ε-caprolactone) (PCL) matrix reinforced with iron-doped hydroxyapatite (FeHA) nanoparticles, were designed and manufactured using a rapid prototyping technique. The performances of these novel 3D PCL/FeHA scaffolds were assessed through a combination of theoretical evaluation, experimental in vitro analyses and in vivo testing in a rabbit animal model. The results from mechanical com- pression tests were consistent with FEM simulations. The in vitro results showed that the cell growth in the magnetized scaffolds was 2.2-fold greater than that in non-magnetized ones. In vivo experiments further suggested that, after only 4 weeks, the PCL/FeHA scaffolds were completely filled with newly formed bone, proving a good level of histocompatibility. All of the results suggest that the introduction of magnetic features into biocompatible materials may confer significant advantages in terms of 3D cell assembly.

  7. In vitro and in vivo drug release and antibacterial properties of the novel vancomycin-loaded bone-like hydroxyapatite/poly amino acid scaffold

    PubMed Central

    Cao, Zhidong; Jiang, Dianming; Yan, Ling; Wu, Jun

    2017-01-01

    Antibiotic-loaded carriers were developed to fill cavities and locally deliver antibiotics following implantation. However, the most commonly used antibiotic carrier, polymethyl methacrylate (PMMA), has many disadvantages including that it does not promote bone regeneration or conduction. Vancomycin-loaded bone-like hydroxyapatite/poly amino acid (V-BHA/PAA) was successfully fabricated by a homogeneous method, certified as biosafe and known to promote osteogenesis. To evaluate its drug-release features, the quantity of the vancomycin in the elution was obtained every 2 days after in vitro simulated body fluid immersion. The drug concentration in the elution was determined to obtain the drug-release curve. The in vitro drug release was a three-phase process with two release peaks. Its antibacterial activity was evaluated in vitro using an antibacterial zone assay, antibacterial inhibition, and scanning electron microscopy (SEM) observation. Scaffolds of V-BHA/PAA were implanted into a rabbit model of chronic osteomyelitis. The antibacterial activity of the material was evaluated in vivo by gross observations, X-ray, and histological and ultrastructural observations. During the first 48 h, the vancomycin release was more rapid, followed by a period of sustained slow release. Use of V-BHA/PAA could achieve relatively long-term vancomycin delivery of 38 days in vitro and 42 days in vivo. V-BHA/PAA showed a significant and consistent bactericidal effect toward both Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) in vitro and in vivo. Moreover, the bactericidal effect was stronger than that of vancomycin-loaded polymethyl meth acrylate (V-PMMA). The duration of the antibacterial effect of V-BHA/PAA toward both S. aureus and MRSA exceeded 28 days in vitro, while that of V-PMMA lasted only 14 days. The curative rate for V-BHA/PAA in the chronic osteomyelitis model was 75% for regular S. aureus and 66.67% for MRSA infection, which significantly exceeded

  8. In vitro and in vivo drug release and antibacterial properties of the novel vancomycin-loaded bone-like hydroxyapatite/poly amino acid scaffold.

    PubMed

    Cao, Zhidong; Jiang, Dianming; Yan, Ling; Wu, Jun

    2017-01-01

    Antibiotic-loaded carriers were developed to fill cavities and locally deliver antibiotics following implantation. However, the most commonly used antibiotic carrier, polymethyl methacrylate (PMMA), has many disadvantages including that it does not promote bone regeneration or conduction. Vancomycin-loaded bone-like hydroxyapatite/poly amino acid (V-BHA/PAA) was successfully fabricated by a homogeneous method, certified as biosafe and known to promote osteogenesis. To evaluate its drug-release features, the quantity of the vancomycin in the elution was obtained every 2 days after in vitro simulated body fluid immersion. The drug concentration in the elution was determined to obtain the drug-release curve. The in vitro drug release was a three-phase process with two release peaks. Its antibacterial activity was evaluated in vitro using an antibacterial zone assay, antibacterial inhibition, and scanning electron microscopy (SEM) observation. Scaffolds of V-BHA/PAA were implanted into a rabbit model of chronic osteomyelitis. The antibacterial activity of the material was evaluated in vivo by gross observations, X-ray, and histological and ultrastructural observations. During the first 48 h, the vancomycin release was more rapid, followed by a period of sustained slow release. Use of V-BHA/PAA could achieve relatively long-term vancomycin delivery of 38 days in vitro and 42 days in vivo. V-BHA/PAA showed a significant and consistent bactericidal effect toward both Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) in vitro and in vivo. Moreover, the bactericidal effect was stronger than that of vancomycin-loaded polymethyl meth acrylate (V-PMMA). The duration of the antibacterial effect of V-BHA/PAA toward both S. aureus and MRSA exceeded 28 days in vitro, while that of V-PMMA lasted only 14 days. The curative rate for V-BHA/PAA in the chronic osteomyelitis model was 75% for regular S. aureus and 66.67% for MRSA infection, which significantly exceeded

  9. Collagen-gelatin-genipin-hydroxyapatite composite scaffolds colonized by human primary osteoblasts are suitable for bone tissue engineering applications: in vitro evidences.

    PubMed

    Vozzi, G; Corallo, C; Carta, S; Fortina, M; Gattazzo, F; Galletti, M; Giordano, N

    2014-05-01

    The application of porous hydroxyapatite (HAp)-collagen as a bone tissue engineering scaffold represents a new trend of mimicking the specific bone extracellular matrix (ECM). The use of HAp in reconstructive surgery has shown that it is slowly invaded by host tissue. Therefore, implant compatibility may be augmented by seeding cells before implantation. Human primary osteoblasts were seeded onto innovative collagen-gelatin-genipin (GP)-HAp scaffolds containing respectively 10%, 20%, and 30% HAp. Cellular adhesion, proliferation, alkaline phosphatase (ALP) activity, osteopontin (OPN), and osteocalcin (OC) expressions were evaluated after 3, 7, 15, and 21 days. The three types of scaffolds showed increased cellular proliferation over time in culture (maximum at 21 days) but the highest was recorded in 10% HAp scaffolds. ALP activity was the highest in 10% HAp scaffolds in all the times of evaluation. OC and OPN resulted in higher concentration in 10% HAp scaffolds compared to 20% and 30% HAp (maximum at 21 days). Finally, scanning electron microscopy analysis showed progressive scaffolds adhesion and colonization from the surface to the inside from day 3 to day 21. In vitro attachment, proliferation, and colonization of human primary osteoblasts on collagen-GP-HAp scaffolds with different percentages of HAp (10%, 20%, and 30%) all increased over time in culture, but comparing different percentages of HAp, they seem to increase with decreasing of HAp component. Therefore, the mechanical properties (such as the stiffness due to the HAp%) coupled with a good biomimetic component (collagen) are the parameters to set up in composite scaffolds design for bone tissue engineering.

  10. ELECTRICALLY CONDUCTIVE SURFACE MODIFICATIONS OF THREE-DIMENSIONAL POLYPROPYLENE FUMARATE SCAFFOLDS

    PubMed Central

    Runge, M. Brett; Dadsetan, Mahrokh; Baltrusaitis, Jonas; Yaszemski, Michael J.

    2014-01-01

    Summary Polypropylene fumarate (PPF) scaffolds fabricated by rapid prototyping technique were surface modified by solution deposition of electrically conductive polypyrrole coatings with or without hydroxyapatite. Scaffolds were electrically conductive with resistivity as low as 2Ω. Scaffold characterization by Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy and thermo gravimetric analysis shows both polypyrrole and hydroxyapatite are present. Cell viability, attachment, proliferation, and differentiation were analyzed using human fetal osteoblast cells. These studies show that surface modification using hydroxyapatite improved cell attachment and proliferation of osteoblasts onto the PPF scaffolds. Alkaline phosphatase activity as a marker for osteogenic differentiation of cell to mature osteoblasts was analyzed. Our data reveal that osteoblasts maintained their phenotype on PPF scaffolds with and without coatings. Thus, these scaffolds could be appropriate candidates for our future in vivo studies. PMID:22051167

  11. Fabrication and in vitro biocompatibility of biomorphic PLGA/nHA composite scaffolds for bone tissue engineering.

    PubMed

    Qian, Junmin; Xu, Weijun; Yong, Xueqing; Jin, Xinxia; Zhang, Wei

    2014-03-01

    In this study, biomorphic poly(dl-lactic-co-glycolic acid)/nano-hydroxyapatite (PLGA/nHA) composite scaffolds were successfully prepared using cane as a template. The porous morphology, phase, compression characteristics and in vitro biocompatibility of the PLGA/nHA composite scaffolds and biomorphic PLGA scaffolds as control were investigated. The results showed that the biomorphic scaffolds preserved the original honeycomb-like architecture of cane and exhibited a bimodal porous structure. The average channel diameter and micropore size of the PLGA/nHA composite scaffolds were 164 ± 52 μm and 13 ± 8 μm, respectively, with a porosity of 89.3 ± 1.4%. The incorporation of nHA into PLGA decreased the degree of crystallinity of PLGA, and significantly improved the compressive modulus of biomorphic scaffolds. The in vitro biocompatibility evaluation with MC3T3-E1 cells demonstrated that the biomorphic PLGA/nHA composite scaffolds could better support cell attachment, proliferation and differentiation than the biomorphic PLGA scaffolds. The localization depth of MC3T3-E1 cells within the channels of the biomorphic PLGA/nHA composite scaffolds could reach approximately 400 μm. The results suggested that the biomorphic PLGA/nHA composite scaffolds were promising candidates for bone tissue engineering.

  12. Hybrid 3D-2D printing of bone scaffolds Hybrid 3D-2D printing methods for bone scaffolds fabrication.

    PubMed

    Prinz, V Ya; Seleznev, Vladimir

    2016-12-13

    It is a well-known fact that bone scaffold topography on micro- and nanometer scale influences the cellular behavior. Nano-scale surface modification of scaffolds allows the modulation of biological activity for enhanced cell differentiation. To date, there has been only a limited success in printing scaffolds with micro- and nano-scale features exposed on the surface. To improve on the currently available imperfect technologies, in our paper we introduce new hybrid technologies based on a combination of 2D (nano imprint) and 3D printing methods. The first method is based on using light projection 3D printing and simultaneous 2D nanostructuring of each of the layers during the formation of the 3D structure. The second method is based on the sequential integration of preliminarily created 2D nanostructured films into a 3D printed structure. The capabilities of the developed hybrid technologies are demonstrated with the example of forming 3D bone scaffolds. The proposed technologies can be used to fabricate complex 3D micro- and nanostructured products for various fields. Copyright 2016 IOP Publishing Ltd.

  13. Optimized fabrication of Ca-P/PHBV nanocomposite scaffolds via selective laser sintering for bone tissue engineering.

    PubMed

    Duan, Bin; Cheung, Wai Lam; Wang, Min

    2011-03-01

    Biomaterials for scaffolds and scaffold fabrication techniques are two key elements in scaffold-based tissue engineering. Nanocomposites that consist of biodegradable polymers and osteoconductive bioceramic nanoparticles and advanced scaffold manufacturing techniques, such as rapid prototyping (RP) technologies, have attracted much attention for developing new bone tissue engineering strategies. In the current study, poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) microspheres and calcium phosphate (Ca-P)/PHBV nanocomposite microspheres were fabricated using the oil-in-water (O/W) and solid-in-oil-in-water (S/O/W) emulsion solvent evaporation methods. The microspheres with suitable sizes were then used as raw materials for scaffold fabrication via selective laser sintering (SLS), which is a mature RP technique. A three-factor three-level complete factorial design was applied to investigate the effects of the three factors (laser power, scan spacing, and layer thickness) in SLS and to optimize SLS parameters for producing good-quality PHBV polymer scaffolds and Ca-P/PHBV nanocomposite scaffolds. The plots of the main effects of these three factors and the three-dimensional response surface were constructed and discussed. Based on the regression equation, optimized PHBV scaffolds and Ca-P/PHBV scaffolds were fabricated using the optimized values of SLS parameters. Characterization of optimized PHBV scaffolds and Ca-P/PHBV scaffolds verified the optimization process. It has also been demonstrated that SLS has the capability of constructing good-quality, sophisticated porous structures of complex shape, which some tissue engineering applications may require.

  14. Effects of Nano-hydroxyapatite/Poly(DL-lactic-co-glycolic acid) Microsphere-Based Composite Scaffolds on Repair of Bone Defects: Evaluating the Role of Nano-hydroxyapatite Content.

    PubMed

    He, Shu; Lin, Kai-Feng; Sun, Zhen; Song, Yue; Zhao, Yi-Nan; Wang, Zheng; Bi, Long; Liu, Jian

    2016-07-01

    The aim of the current study was to prepare microsphere-based composite scaffolds made of nano-hydroxyapatite (nHA)/poly (DL-lactic-co-glycolic acid) (PLGA) at different ratios and evaluate the effects of nHA on the characteristics of scaffolds for tissue engineering application. First, microsphere-based composite scaffolds made of two ratios of nHA/PLGA (nHA/PLGA = 20/80 and nHA/PLGA = 50/50) were prepared. Then, the effects of nHA on the wettability, mechanical strength, and degradation of scaffolds were investigated. Second, the biocompatibility and osteoinductivity were evaluated and compared by co-culture of scaffolds with bone marrow stromal stem cells (BMSCs). The results showed that the adhesion, proliferation, and osteogenic differentiation of BMSCs with nHA/PLGA (50/50) were better than those with nHA/PLGA (20/80). Finally, we implanted the scaffolds into femur bone defects in a rabbit model, then the capacity of guiding bone regeneration as well as the in vivo degradation were observed by micro-CT and histological examinations. After 4 weeks' implantation, there was no significant difference on the repair of bone defects. However, after 8 and 12 weeks' implantation, the nHA/PLGA (20/80) exhibited better bone formation than nHA/PLGA (50/50). These results suggested that a proper concentration of nHA in the nHA/PLGA composite should be taken into account when the composite scaffolds were prepared, which plays an important role in the biocompatibility, degradation rate and osteoconductivity.

  15. Improved fabrication of melt electrospun tissue engineering scaffolds using direct writing and advanced electric field control.

    PubMed

    Ristovski, Nikola; Bock, Nathalie; Liao, Sam; Powell, Sean K; Ren, Jiongyu; Kirby, Giles T S; Blackwood, Keith A; Woodruff, Maria A

    2015-03-25

    Direct writing melt electrospinning is an additive manufacturing technique capable of the layer-by-layer fabrication of highly ordered 3d tissue engineering scaffolds from micron-diameter fibers. The utility of these scaffolds, however, is limited by the maximum achievable height of controlled fiber deposition, beyond which the structure becomes increasingly disordered. A source of this disorder is charge build-up on the deposited polymer producing unwanted coulombic forces. In this study, the authors introduce a novel melt electrospinning platform with dual voltage power supplies to reduce undesirable charge effects and improve fiber deposition control. The authors produced and characterized several 90° cross-hatched fiber scaffolds using a range of needle/collector plate voltages. Fiber thickness was found to be sensitive only to overall potential and invariant to specific tip/collector voltage. The authors also produced ordered scaffolds up to 200 layers thick (fiber spacing 1 mm and diameter 40 μm) and characterized structure in terms of three distinct zones: ordered, semiordered, and disordered. Our in vitro analysis indicates successful cell attachment and distribution throughout the scaffolds, with little evidence of cell death after seven days. This study demonstrates the importance of electrostatic control for reducing destabilizing polymer charge effects and enabling the fabrication of morphologically suitable scaffolds for tissue engineering.

  16. Living nano-micro fibrous woven fabric/hydrogel composite scaffolds for heart valve engineering.

    PubMed

    Wu, Shaohua; Duan, Bin; Qin, Xiaohong; Butcher, Jonathan T

    2017-03-15

    Regeneration and repair of injured or diseased heart valves remains a clinical challenge. Tissue engineering provides a promising treatment approach to facilitate living heart valve repair and regeneration. Three-dimensional (3D) biomimetic scaffolds that possess heterogeneous and anisotropic features that approximate those of native heart valve tissue are beneficial to the successful in vitro development of tissue engineered heart valves (TEHV). Here we report the development and characterization of a novel composite scaffold consisting of nano- and micro-scale fibrous woven fabrics and 3D hydrogels by using textile techniques combined with bioactive hydrogel formation. Embedded nano-micro fibrous scaffolds within hydrogel enhanced mechanical strength and physical structural anisotropy of the composite scaffold (similar to native aortic valve leaflets) and also reduced its compaction. We determined that the composite scaffolds supported the growth of human aortic valve interstitial cells (HAVIC), balanced the remodeling of heart valve ECM against shrinkage, and maintained better physiological fibroblastic phenotype in both normal and diseased HAVIC over single materials. These fabricated composite scaffolds enable the engineering of a living heart valve graft with improved anisotropic structure and tissue biomechanics important for maintaining valve cell phenotypes.

  17. Fabrication of co-continuous poly(ε-caprolactone)/polyglycolide blend scaffolds for tissue engineering

    SciTech Connect

    Allaf, Rula M.; Rivero, Iris V.; Ivanov, Ilia N.

    2015-06-06

    The apparent inability of a single biomaterial to meet all the requirements for tissue engineering scaffolds has led to continual research in novel engineered biomaterials. One method to provide new materials and fine-tune their properties is via mixing materials. Here in this study, a biodegradable powder blend of poly(ε-caprolactone) (PCL), polyglycolide (PGA), and poly(ethylene oxide) (PEO) was prepared and three-dimensional interconnected porous PCL/PGA scaffolds were fabricated by combining cryomilling and compression molding/polymer leaching techniques. The resultant porous scaffolds exhibited co-continuous morphologies with ~50% porosity. Mean pore sizes of 24 and 56 μm were achieved by varying milling time. The scaffolds displayed high mechanical properties and water uptake, in addition to a remarkably fast degradation rate. The results demonstrate the potential of this fabrication approach to obtain PCL/PGA blend scaffolds with interconnected porosity. In general, these results provide significant insight into an approach that will lead to the development of new composites and blends in scaffold manufacturing.

  18. Fabrication of scalable and structured tissue engineering scaffolds using water dissolvable sacrificial 3D printed moulds.

    PubMed

    Mohanty, Soumyaranjan; Larsen, Layla Bashir; Trifol, Jon; Szabo, Peter; Burri, Harsha Vardhan Reddy; Canali, Chiara; Dufva, Marin; Emnéus, Jenny; Wolff, Anders

    2015-10-01

    One of the major challenges in producing large scale engineered tissue is the lack of ability to create large highly perfused scaffolds in which cells can grow at a high cell density and viability. Here, we explore 3D printed polyvinyl alcohol (PVA) as a sacrificial mould in a polymer casting process. The PVA mould network defines the channels and is dissolved after curing the polymer casted around it. The printing parameters determined the PVA filament density in the sacrificial structure and this density resulted in different stiffness of the corresponding elastomer replica. It was possible to achieve 80% porosity corresponding to about 150 cm(2)/cm(3) surface to volume ratio. The process is easily scalable as demonstrated by fabricating a 75 cm(3) scaffold with about 16,000 interconnected channels (about 1m(2) surface area) and with a channel to channel distance of only 78 μm. To our knowledge this is the largest scaffold ever to be produced with such small feature sizes and with so many structured channels. The fabricated scaffolds were applied for in-vitro culturing of hepatocytes over a 12-day culture period. Smaller scaffolds (6×4 mm) were tested for cell culturing and could support homogeneous cell growth throughout the scaffold. Presumably, the diffusion of oxygen and nutrient throughout the channel network is rapid enough to support cell growth. In conclusion, the described process is scalable, compatible with cell culture, rapid, and inexpensive.

  19. Bicomponent electrospinning to fabricate three-dimensional hydrogel-hybrid nanofibrous scaffolds with spatial fiber tortuosity.

    PubMed

    Jin, Gyuhyung; Lee, Slgirim; Kim, Seung-Hyun; Kim, Minhee; Jang, Jae-Hyung

    2014-12-01

    Electrospun fibrous mats have emerged as powerful tissue engineering scaffolds capable of providing highly effective and versatile physical guidance, mimicking the extracellular environment. However, electrospinning typically produces a sheet-like structure, which is a major limitation associated with current electrospinning technologies. To address this challenge, highly porous, volumetric hydrogel-hybrid fibrous scaffolds were fabricated by one Taylor cone-based side-by-side dual electrospinning of poly (ε-caprolactone) (PCL) and poly (vinyl pyrrolidone) (PVP), which possess distinct properties (i.e., hydrophobic and hydrogel properties, respectively). Immersion of the resulting scaffolds in water induced spatial tortuosity of the hydrogel PVP fibers while maintaining their aligned fibrous structures in parallel with the PCL fibers. The resulting conformational changes in the entire bicomponent fibers upon immersion in water led to volumetric expansion of the fibrous scaffolds. The spatial fiber tortuosity significantly increased the pore volumes of electrospun fibrous mats and dramatically promoted cellular infiltration into the scaffold interior both in vitro and in vivo. Harmonizing the flexible PCL fibers with the soft PVP-hydrogel layers produced highly ductile fibrous structures that could mechanically resist cellular contractile forces upon in vivo implantation. This facile dual electrospinning followed by the spatial fiber tortuosity for fabricating three-dimensional hydrogel-hybrid fibrous scaffolds will extend the use of electrospun fibers toward various tissue engineering applications.

  20. Physico-chemical characterization and biological response of Labeo rohita-derived hydroxyapatite scaffold.

    PubMed

    Mondal, S; Mondal, A; Mandal, N; Mondal, B; Mukhopadhyay, S S; Dey, A; Singh, S

    2014-07-01

    The chemically treated Labeo rohita scale is used for synthesizing hydroxyapatite (HAp) biomaterials. Thermogravimetric and differential thermal analyses of fish scale materials reveal the different phase changes with temperature and find out the suitable calcination temperatures. The composition and structures of wet ball-milled calcined HAp powders are characterized by Fourier transform infrared spectroscopy, X-ray diffraction, field emission scanning electron microscopy, transmission electron microscopy, energy dispersive X-ray analysis (EDX). The EDX as well as chemical analysis of fish scale-derived apatite materials confirms that the Ca/P ratio is 1.71. The compressive stress, hardness and porosity have been evaluated on sintered HAp biomaterials. The cell attachment on HAp surfaces, cytotoxicity evaluation and MTT assay, which are carried out in RAW macrophage-like cell line media demonstrate good biocompatibility. The histological analysis also supports the bioaffinity of processed HAp biomaterials in Wistar rat model for investigating the contact reaction and stability at the artificial or natural prosthesis interface.

  1. 3D printing for the design and fabrication of polymer-based gradient scaffolds.

    PubMed

    Bracaglia, Laura G; Smith, Brandon T; Watson, Emma; Arumugasaamy, Navein; Mikos, Antonios G; Fisher, John P

    2017-03-22

    To accurately mimic the native tissue environment, tissue engineered scaffolds often need to have a highly controlled and varied display of three-dimensional (3D) architecture and geometrical cues. Additive manufacturing in tissue engineering has made possible the development of complex scaffolds that mimic the native tissue architectures. As such, architectural details that were previously unattainable or irreproducible can now be incorporated in an ordered and organized approach, further advancing the structural and chemical cues delivered to cells interacting with the scaffold. This control over the environment has given engineers the ability to unlock cellular machinery that is highly dependent upon the intricate heterogeneous environment of native tissue. Recent research into the incorporation of physical and chemical gradients within scaffolds indicates that integrating these features improves the function of a tissue engineered construct. This review covers recent advances on techniques to incorporate gradients into polymer scaffolds through additive manufacturing and evaluate the success of these techniques. As covered here, to best replicate different tissue types, one must be cognizant of the vastly different types of manufacturing techniques available to create these gradient scaffolds. We review the various types of additive manufacturing techniques that can be leveraged to fabricate scaffolds with heterogeneous properties and discuss methods to successfully characterize them.

  2. Fabrication and characterization of bioactive β-Ca2SiO4/PHBV composite scaffolds.

    PubMed

    Wang, Nana; Zhou, Zheng; Xia, Leilei; Dai, Yao; Liu, Hairong

    2013-05-01

    A key challenge in tissue engineering is the construction of a scaffold with adequate properties which would mimic extracellular matrix (ECM) to induce the cells' efficient adhesion, proliferation and proper differentiation. Novel β-Ca2SiO4/PHBV composite scaffolds were fabricated by integrating β-Ca2SiO4 nanoparticles with PHBV backbone via a modified solvent casting-particulates leaching method, which generates interconnected porous structure and the high porosity, about 87%, of these scaffolds. Compared with PHBV scaffolds, β-Ca2SiO4/PHBV composite scaffolds facilitate the adhesion of human osteoblast-like MG-63 cells due to their increased hydrophilicity. The β-Ca2SiO4/PHBV composite scaffolds containing 2.5 or 5% β-Ca2SiO4 nanoparticles significantly enhance the proliferation of MG-63 cells by stimulating the transcription of the transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-7 (BMP-7) genes. These scaffolds also induce early differentiation via promoting the transcription of alkaline phosphatase (ALP). The results suggest the potential application of β-Ca2SiO4/PHBV composites in bone tissue engineering.

  3. Laser fabrication of three-dimensional CAD scaffolds from photosensitive gelatin for applications in tissue engineering.

    PubMed

    Ovsianikov, Aleksandr; Deiwick, Andrea; Van Vlierberghe, Sandra; Dubruel, Peter; Möller, Lena; Dräger, Gerald; Chichkov, Boris

    2011-04-11

    In the present work, 3D CAD scaffolds for tissue engineering applications were developed starting from methacrylamide-modified gelatin (GelMOD) using two-photon polymerization (2PP). The scaffolds were cross-linked employing the biocompatible photoinitiator Irgacure 2959. Because gelatin is derived from collagen (i.e., the main constituent of the ECM), the developed materials mimic the cellular microenvironment from a chemical point of view. In addition, by applying the 2PP technique, structural properties of the cellular microenvironment can also be mimicked. Furthermore, in vitro degradation assays indicated that the enzymatic degradation capability of gelatin is preserved for the methacrylamide-modified derivative. An in depth morphological analysis of the 2PP-fabricated scaffolds demonstrated that the parameters of the CAD model are reproduced with great precision, including the ridge-like surface topography on the order of 1.5 μm. The developed scaffolds showed an excellent stability in culture medium. In a final part of the present work, the suitability of the developed scaffolds for tissue engineering applications was verified. The results indicated that the applied materials are suitable to support porcine mesenchymal stem cell adhesion and subsequent proliferation. Upon applying osteogenic stimulation, the seeded cells differentiated into the anticipated lineage. Energy dispersive X-ray (EDX) analysis showed the induced calcification of the scaffolds. The results clearly indicate that 2PP is capable of manufacturing precisely constructed 3D tissue engineering scaffolds using photosensitive polymers as starting material.

  4. Comparison of three-dimensional printing and vacuum freeze-dried techniques for fabricating composite scaffolds.

    PubMed

    Sun, Kai; Li, Ruixin; Jiang, Wenxue; Sun, Yufu; Li, Hui

    2016-09-02

    In this study, the performances of different preparation methods of the scaffolds were analyzed for chondrocyte tissue engineering. Silk fibroin/collagen (SF/C) was fabricated using a vacuum freeze-dried technique and by 3D printing. The porosity, water absorption expansion rates, mechanical properties, and pore sizes of the resulting materials were evaluated. The proliferation and metabolism of the cells was detected at different time points using an MTT assay. Cell morphologies and distributions were observed by histological analysis and scanning electron microscopy (SEM). The porosity, water absorption expansion rate, and Young's modulus of the material obtained via 3D printing were significantly higher than those obtained by the freeze-dried method, while the pore size did not differ significantly between the two methods. MTT assay results showed that the metabolism of cells seeded on the 3D printed scaffolds was more viable than the metabolism on the freeze-dried material. H&E staining of the scaffolds revealed that the number of cells in the 3D printed scaffold was higher in comparison to a similar measurement on the freeze-dried material. Consequently, stem cells grew well inside the 3D printed scaffolds, as measured by SEM, while the internal structure of the freeze-dried scaffold was disordered. Compared with the freeze-dried technique, the 3D printed scaffold exhibited better overall performance and was more suitable for cartilage tissue engineering.

  5. Fabrication and development of artificial osteochondral constructs based on cancellous bone/hydrogel hybrid scaffold.

    PubMed

    Song, Kedong; Li, Liying; Yan, Xinyu; Zhang, Yu; Li, Ruipeng; Wang, Yiwei; Wang, Ling; Wang, Hong; Liu, Tianqing

    2016-06-01

    Using tissue engineering techniques, an artificial osteochondral construct was successfully fabricated to treat large osteochondral defects. In this study, porcine cancellous bones and chitosan/gelatin hydrogel scaffolds were used as substitutes to mimic bone and cartilage, respectively. The porosity and distribution of pore size in porcine bone was measured and the degradation ratio and swelling ratio for chitosan/gelatin hydrogel scaffolds was also determined in vitro. Surface morphology was analyzed with the scanning electron microscope (SEM). The physicochemical properties and the composition were tested by using an infrared instrument. A double layer composite scaffold was constructed via seeding adipose-derived stem cells (ADSCs) induced to chondrocytes and osteoblasts, followed by inoculation in cancellous bones and hydrogel scaffolds. Cell proliferation was assessed through Dead/Live staining and cellular activity was analyzed with IpWin5 software. Cell growth, adhesion and formation of extracellular matrix in composite scaffolds blank cancellous bones or hydrogel scaffolds were also analyzed. SEM analysis revealed a super porous internal structure of cancellous bone scaffolds and pore size was measured at an average of 410 ± 59 μm while porosity was recorded at 70.6 ± 1.7 %. In the hydrogel scaffold, the average pore size was measured at 117 ± 21 μm and the porosity and swelling rate were recorded at 83.4 ± 0.8 % and 362.0 ± 2.4 %, respectively. Furthermore, the remaining hydrogel weighed 80.76 ± 1.6 % of the original dry weight after hydration in PBS for 6 weeks. In summary, the cancellous bone and hydrogel composite scaffold is a promising biomaterial which shows an essential physical performance and strength with excellent osteochondral tissue interaction in situ. ADSCs are a suitable cell source for osteochondral composite reconstruction. Moreover, the bi-layered scaffold significantly enhanced cell proliferation compared to the cells seeded on

  6. Partially degradable film/fabric composites: textile scaffolds for liver cell culture.

    PubMed

    Karamuk, E; Mayer, J; Wintermantel, E; Akaike, T

    1999-09-01

    In this study, a composite scaffold combining textile superstructures and biomimetic glycopolymers is introduced, which may allow engineering of organotypic liver tissue in vitro. Woven poly(ethylene therephtalat) (PET) fabrics were coated on one side with a thin biodegradable polymer film (poly[D-L-lactic-co-glycolic acid] PLGA), in order to obtain a polar structure. The composite structure ensured the stability of the membrane during in vitro degradation, independently of mesh size. Matrix porosity increased when a polymer blend matrix was used. For hepatocyte culturing studies, the scaffolds were additionally coated with an artificial glycopolymer (poly[N-p-vinylbenzyl-D-lactoamide], PVLA) in order to improve cell attachment. It was observed that formation of aggregates depends on the scaffold geometry as well as on the pretreatment and medium conditions. After 4 days in culture, the pores of the fabric were filled with aggregates illustrating the possibility of immobilizing hepatocyte aggregates in well-defined spatial configurations on textile structures.

  7. Comparison of three-dimensional printing and vacuum freeze-dried techniques for fabricating composite scaffolds

    SciTech Connect

    Sun, Kai; Li, Ruixin; Jiang, Wenxue; Sun, Yufu; Li, Hui

    2016-09-02

    In this study, the performances of different preparation methods of the scaffolds were analyzed for chondrocyte tissue engineering. Silk fibroin/collagen (SF/C) was fabricated using a vacuum freeze-dried technique and by 3D printing. The porosity, water absorption expansion rates, mechanical properties, and pore sizes of the resulting materials were evaluated. The proliferation and metabolism of the cells was detected at different time points using an MTT assay. Cell morphologies and distributions were observed by histological analysis and scanning electron microscopy (SEM). The porosity, water absorption expansion rate, and Young’s modulus of the material obtained via 3D printing were significantly higher than those obtained by the freeze-dried method, while the pore size did not differ significantly between the two methods. MTT assay results showed that the metabolism of cells seeded on the 3D printed scaffolds was more viable than the metabolism on the freeze-dried material. H&E staining of the scaffolds revealed that the number of cells in the 3D printed scaffold was higher in comparison to a similar measurement on the freeze-dried material. Consequently, stem cells grew well inside the 3D printed scaffolds, as measured by SEM, while the internal structure of the freeze-dried scaffold was disordered. Compared with the freeze-dried technique, the 3D printed scaffold exhibited better overall performance and was more suitable for cartilage tissue engineering. - Highlights: • Silk fibroin/collagen was fabricated using 3D printing. • Physical characterization and Cell compatibility were compared. • 3D printed scaffold exhibited better overall performance.

  8. Assessment of cellular viability on calcium sulphate/hydroxyapatite injectable scaffolds

    PubMed Central

    Naudi, Kurt; Dalby, Matthew J; Tanner, K Elizabeth; McMahon, Jeremy D; Ayoub, Ashraf

    2013-01-01

    Cements for maxillofacial reconstruction of jaw defects through calcification of rotated muscle have been tested. The objective of this study was to investigate the visibility of loading of two types of commercially available cements, Cerament™ Spine Support and Cerament Bone Void Filler with mesenchymal cells and cytokines (bone morphogenetic protein) to act as a biomimetic scaffolding for future clinical application. Determination of basic biocompatibility (cell viability) using methyl thiazolyl tetrazolium and live/dead assay was carried out using MG-63 cells at various time points. Next, in order to inform potential subsequent in vivo experiments, a collagen tissue mimic was used for characterization of rabbit mesenchymal stromal cells using immunofluorescent cytoskeleton staining, and simultaneous and then sequential injection of Cerament Spine Support cement and cells into collagen gels. Results indicated that Cerament Spine Support was more biocompatible and that sequential injection of cement and then rabbit mesenchymal stromal cells into the tissue mimics is an optimal approach for clinical applications. PMID:24555009

  9. A synergistic approach to the design, fabrication and evaluation of 3D printed micro and nano featured scaffolds for vascularized bone tissue repair

    NASA Astrophysics Data System (ADS)

    Holmes, Benjamin; Bulusu, Kartik; Plesniak, Michael; Zhang, Lijie Grace

    2016-02-01

    3D bioprinting has begun to show great promise in advancing the development of functional tissue/organ replacements. However, to realize the true potential of 3D bioprinted tissues for clinical use requires the fabrication of an interconnected and effective vascular network. Solving this challenge is critical, as human tissue relies on an adequate network of blood vessels to transport oxygen, nutrients, other chemicals, biological factors and waste, in and out of the tissue. Here, we have successfully designed and printed a series of novel 3D bone scaffolds with both bone formation supporting structures and highly interconnected 3D microvascular mimicking channels, for efficient and enhanced osteogenic bone regeneration as well as vascular cell growth. Using a chemical functionalization process, we have conjugated our samples with nano hydroxyapatite (nHA), for the creation of novel micro and nano featured devices for vascularized bone growth. We evaluated our scaffolds with mechanical testing, hydrodynamic measurements and in vitro human mesenchymal stem cell (hMSC) adhesion (4 h), proliferation (1, 3 and 5 d) and osteogenic differentiation (1, 2 and 3 weeks). These tests confirmed bone-like physical properties and vascular-like flow profiles, as well as demonstrated enhanced hMSC adhesion, proliferation and osteogenic differentiation. Additional in vitro experiments with human umbilical vein endothelial cells also demonstrated improved vascular cell growth, migration and organization on micro-nano featured scaffolds.

  10. A synergistic approach to the design, fabrication and evaluation of 3D printed micro and nano featured scaffolds for vascularized bone tissue repair.

    PubMed

    Holmes, Benjamin; Bulusu, Kartik; Plesniak, Michael; Zhang, Lijie Grace

    2016-02-12

    3D bioprinting has begun to show great promise in advancing the development of functional tissue/organ replacements. However, to realize the true potential of 3D bioprinted tissues for clinical use requires the fabrication of an interconnected and effective vascular network. Solving this challenge is critical, as human tissue relies on an adequate network of blood vessels to transport oxygen, nutrients, other chemicals, biological factors and waste, in and out of the tissue. Here, we have successfully designed and printed a series of novel 3D bone scaffolds with both bone formation supporting structures and highly interconnected 3D microvascular mimicking channels, for efficient and enhanced osteogenic bone regeneration as well as vascular cell growth. Using a chemical functionalization process, we have conjugated our samples with nano hydroxyapatite (nHA), for the creation of novel micro and nano featured devices for vascularized bone growth. We evaluated our scaffolds with mechanical testing, hydrodynamic measurements and in vitro human mesenchymal stem cell (hMSC) adhesion (4 h), proliferation (1, 3 and 5 d) and osteogenic differentiation (1, 2 and 3 weeks). These tests confirmed bone-like physical properties and vascular-like flow profiles, as well as demonstrated enhanced hMSC adhesion, proliferation and osteogenic differentiation. Additional in vitro experiments with human umbilical vein endothelial cells also demonstrated improved vascular cell growth, migration and organization on micro-nano featured scaffolds.

  11. A synergistic approach to the design, fabrication and evaluation of 3D printed micro and nano featured scaffolds for vascularized bone tissue repair

    PubMed Central

    Holmes, Benjamin; Bulusu, Kartik; Plesniak, Michael; Zhang, Lijie Grace

    2016-01-01

    3D bioprinting has begun to show great promise in advancing the development of functional tissue/organ replacements. However, to realize the true potential of 3D bioprinted tissues for clinical use requires the fabrication of an interconnected and effective vascular network. Solving this challenge is critical, as human tissue relies on an adequate network of blood vessels to transport oxygen, nutrients, other chemicals, biological factors and waste, in and out of the tissue. Here, we have successfully designed and printed a series of novel 3D bone scaffolds with both bone formation supporting structures and highly interconnected 3D microvascular mimicking channels, for efficient and enhanced osteogenic bone regeneration as well as vascular cell growth. Using a chemical functionalization process, we have conjugated our samples with nano hydroxyapatite (nHA), for the creation of novel micro and nano featured devices for vascularized bone growth. We evaluated our scaffolds with mechanical testing, hydrodynamic measurements and in vitro human mesenchymal stem cell (hMSC) adhesion (4 h), proliferation (1, 3 and 5 d) and osteogenic differentiation (1, 2 and 3 weeks). These tests confirmed bone-like physical properties and vascular-like flow profiles, as well as demonstrated enhanced hMSC adhesion, proliferation and osteogenic differentiation. Additional in vitro experiments with human umbilical vein endothelial cells also demonstrated improved vascular cell growth, migration and organization on micro-nano featured scaffolds. PMID:26758780

  12. Development of an Indirect Stereolithography Technology for Scaffold Fabrication with a Wide Range of Biomaterial Selectivity

    PubMed Central

    Kang, Hyun-Wook

    2012-01-01

    Tissue engineering, which is the study of generating biological substitutes to restore or replace tissues or organs, has the potential to meet current needs for organ transplantation and medical interventions. Various approaches have been attempted to apply three-dimensional (3D) solid freeform fabrication technologies to tissue engineering for scaffold fabrication. Among these, the stereolithography (SL) technology not only has the highest resolution, but also offers quick fabrication. However, a lack of suitable biomaterials is a barrier to applying the SL technology to tissue engineering. In this study, an indirect SL method that combines the SL technology and a sacrificial molding process was developed to address this challenge. A sacrificial mold with an inverse porous shape was fabricated from an alkali-soluble photopolymer by the SL technology. A sacrificial molding process was then developed for scaffold construction using a variety of biomaterials. The results indicated a wide range of biomaterial selectivity and a high resolution. Achievable minimum pore and strut sizes were as large as 50 and 65 μm, respectively. This technology can also be used to fabricate three-dimensional organ shapes, and combined with traditional fabrication methods to construct a new type of scaffold with a dual-pore size. Cytotoxicity tests, as well as nuclear magnetic resonance and gel permeation chromatography analyses, showed that this technology has great potential for tissue engineering applications. PMID:22443315

  13. [Evaluation of biocompatibility of Ti-6Al-4V scaffolds fabricated by electron beam melting].

    PubMed

    Wang, H; Zhao, B J; Yan, R Z; Wang, C; Luo, C C; Hu, M

    2016-11-09

    Objective: To investigate the biocompatibility of Ti-6Al-4V scaffolds fabricated by electron beam melting(EBM). Methods: Bone marrow mesenchymal stem cells(BMSC) co-cultured with Ti-6Al-4V specimens fabricated with EBM was prepared as experimental group and the regular cells culture was employed as control. The biocompatibility was detected using CCK-8 and cytoskeleton staining. The osteogenic differentiation ability was assessed using mineralization nodule formation. A 24 mm defect was created on the right mandibular body in 12 beagles. The mandibular defects were repaired with Ti-6Al-4V scaffolds mesh fabricated by EBM. General observation, CT and histology examination was carried out to evaluated the biocompatibility of Ti-6Al-4V scaffolds in vivo. Results: CCK-8 result showed the A values of the two groups had no significant difference(P >0.05). There was no significant difference between the two groups (P>0.05). Cytoskeletal staining showed that cells were fully stretched out and grew well on T-i6Al-4V specimen. The actin fibers were arranged in parallel and stained uniformly with fluorescent. After osteogenic culture, the quantity of the nodule formation of the experimental group and control group were 5.7±0.7 and 5.1 ± 0.6, respectively(P>0.05). All animals had tolerated the surgery and healed well. CT examination showed that Ti-6Al-4V scaffolds mesh had good retention with surrounding bone and the continuity of mandible was restored. Histological examination showed that no inflammation reaction or toxity was caused in the soft tissue surrounding the scaffolds and in the liver and kidney after implantation. Ti-6Al-4V scaffolds had good retention with surrounding bone. Conclusions: Ti-6Al-4V fabricated with electron beam melting has good biocompatibility.

  14. Fabrication of a customized bone scaffold using a homemade medical 3D printer for comminuted fractures

    NASA Astrophysics Data System (ADS)

    Yoon, Do-Kun; Jung, Joo-Young; Shin, Han-Back; Kim, Moo-Sub; Choe, Bo-Young; Kim, Sunmi; Suh, Tae Suk; Lee, Keum Sil; Xing, Lei

    2016-09-01

    The purpose of this study was to show a 3D printed reconstruction model of a bone destroyed by a comminuted fracture. After a thoracic limb of a cow with a comminuted fracture was scanned by using computed tomography, a scaffold was designed by using a 3D modeling tool for its reconstruction and fabricated by using a homemade medical 3D printer. The homemade medical 3D printer was designed for medical use. In order to reconstruct the geometry of the destroyed bone, we use the geometry of a similar section (reference geometry) of normal bone in the 3D modeling process. The missing part between the destroyed ridge and the reference geometry was filled with an effective space by using a manual interpolation. Inexpensive materials and free software were used to construct the medical 3D printer system. The fabrication of the scaffold progressed according to the design of reconstructed bone by using this medical 3D printer. The material of the scaffold was biodegradable material, and could be transplanted into the human body. The fabricated scaffold was correctly inserted into the fractured bone in place of the destroyed portion, with good agreement. According to physical stress test results, the performance of printing resolution was 0.1 mm. The average geometrical error of the scaffold was below 0.3 mm. The reconstructed bone by using the fabricated scaffold was able to support the weight of the human body. No process used to obtain the result was complex or required many resources. The methods and results in this study show several possible clinical applications in fields such as orthopedics or oncology without a need to purchase high-price instruments for 3D printing.

  15. Fabrication of Highly-Densified Hydroxyapatite Ceramic with Boron Oxide Addition and Its Superplastic Deformation

    NASA Astrophysics Data System (ADS)

    Itatani, Kiyoshi; Tsuchiya, Kenta; Sakka, Yoshio; Davies, Ian J.; Koda, Seiichiro

    2011-03-01

    Highly-densified and fine-grained hydroxyapatite (Ca10(PO4)6(OH)2: HAp) ceramics with and without 3.0 mol% B2O3 addition were fabricated by pulse-current pressure firing at 1000°C for 10 min under a pressure of 50 MPa. The tensile elongations of HAp ceramics with and without 3.0 mol% B2O3 addition were examined by changing the strain rate from 0.15 to 2.4 × 10-3 s-1 at the test temperature of 1000°C. The tensile elongation of HAp ceramic with no B2O3 addition showed a maximum (474%) at the strain rate of 0.59 × 10-3 s-1. On the other hand, the tensile elongation of HAp ceramic increased to 583% at the strain rate of 2.4 × 10-3 s-1 due to the addition of 3.0 mol% B2O3. Microscopic observation of fractured surfaces of HAp ceramic with 3.0 mol% B2O3 addition showed that the mean grain size decreased from 5.1 to 1.9 μm with increasing strain rate from 0.15 to 2.4 × 10-3 s-1 and that notable amounts of cavities were created among the grains. The time required to achieve the maximum tensile elongation (~580%) of the HAp ceramic could thus be significantly reduced (approximately 1/16, i.e., 12 h to 45 min), due to increasing the strain rate from 0.15 to 2.4 × 10-3 s-1.

  16. Fabrication of biodegradable elastomeric scaffolds with sub-micron morphologies

    PubMed Central

    Stankus, John J.; Guan, Jianjun; Wagner, William R.

    2010-01-01

    The native extracellular matrix (ECM) of elastic tissues is strong and flexible and supports cell adhesion and enzymatic matrix remodeling. In an attempt to convey these ECM properties to a synthetic scaffold appropriate for soft tissue engineering applications, a biodegradable, elastomeric poly(ester urethane)urea (PEUU) was combined with type I collagen at various ratios (2.5, 5, 10, 20, 50, 60, 70, 80, and 90 wt% collagen) and electrospun to construct elastic matrices. Randomly orientated fibers in the electrospun matrices ranged in diameter from 100–900 nm, dependent on initial polymer concentration. Picrosirius red staining of matrices and CD spectroscopy of released collagen confirmed collagen incorporation and preservation of collagen structure at the higher collagen mass fractions. Matrices were strong and distensible possessing strengths of 2–13 MPa with breaking strains of 160–280% even with low PEUU content. Collagen incorporation significantly enhanced smooth muscle cell adhesion onto electrospun scaffolds. An approach has been demonstrated that mimics elastic extracellular matrices by using a synthetic component to provide mechanical function together with a biomacromolecule, collagen. Such matrices may find application in engineering soft tissue. PMID:15307165

  17. Design and fabrication of 3D porous scaffolds to facilitate cell-based gene therapy.

    PubMed

    El-Ayoubi, Rouwayda; Eliopoulos, Nicoletta; Diraddo, Robert; Galipeau, Jacques; Yousefi, Azizeh-Mitra

    2008-06-01

    Biomaterials capable of efficient gene delivery by embedded cells provide a fundamental tool for the treatment of acquired or hereditary diseases. A major obstacle is maintaining adequate nutrient and oxygen diffusion to cells within the biomaterial. In this study, we combined the solid free-form fabrication and porogen leaching techniques to fabricate three-dimensional scaffolds, with bimodal pore size distribution, for cell-based gene delivery. The objective of this study was to design micro-/macroporous scaffolds to improve cell viability and drug delivery. Murine bone marrow-derived mesenchymal stromal cells (MSCs) genetically engineered to secrete erythropoietin (EPO) were seeded onto poly-L-lactide (PLLA) scaffolds with different microporosities. Over a period of 2 weeks in culture, an increase in cell proliferation and metabolic activity was observed with increasing scaffold microporosity. The concentration of EPO detected in supernatants also increased with increasing microporosity level. Our study shows that these constructs can promote cell viability and release of therapeutic proteins, and clearly demonstrates their capacity for a dual role as scaffolds for tissue regeneration and as delivery systems for soluble gene products.

  18. An Overview of Scaffold Design and Fabrication Technology for Engineered Knee Meniscus

    PubMed Central

    Sun, Jie; Vijayavenkataraman, Sanjairaj; Liu, Hang

    2017-01-01

    Current surgical treatments for meniscal tears suffer from subsequent degeneration of knee joints, limited donor organs and inconsistent post-treatment results. Three clinical scaffolds (Menaflex CMI, Actifit® scaffold and NUsurface® Meniscus Implant) are available on the market, but additional data are needed to properly evaluate their safety and effectiveness. Thus, many scaffold-based research activities have been done to develop new materials, structures and fabrication technologies to mimic native meniscus for cell attachment and subsequent tissue development, and restore functionalities of injured meniscus for long-term effects. This study begins with a synopsis of relevant structural features of meniscus and goes on to describe the critical considerations. Promising advances made in the field of meniscal scaffolding technology, in terms of biocompatible materials, fabrication methods, structure design and their impact on mechanical and biological properties are discussed in detail. Among all the scaffolding technologies, additive manufacturing (AM) is very promising because of its ability to precisely control fiber diameter, orientation, and pore network micro-architecture to mimic the native meniscus microenvironment. PMID:28772388

  19. Fabrication of Aligned Nanofiber Polymer Yarn Networks for Anisotropic Soft Tissue Scaffolds.

    PubMed

    Wu, Shaohua; Duan, Bin; Liu, Penghong; Zhang, Caidan; Qin, Xiaohong; Butcher, Jonathan T

    2016-07-06

    Nanofibrous scaffolds with defined architectures and anisotropic mechanical properties are attractive for many tissue engineering and regenerative medicine applications. Here, a novel electrospinning system is developed and implemented to fabricate continuous processable uniaxially aligned nanofiber yarns (UANY). UANY were processed into fibrous tissue scaffolds with defined anisotropic material properties using various textile-forming technologies, i.e., braiding, weaving, and knitting techniques. UANY braiding dramatically increased overall stiffness and strength compared to the same number of UANY unbraided. Human adipose derived stem cells (HADSC) cultured on UANY or woven and knitted 3D scaffolds aligned along local fiber direction and were >90% viable throughout 21 days. Importantly, UANY supported biochemical induction of HADSC differentiation toward smooth muscle and osteogenic lineages. Moreover, we integrated an anisotropic woven fiber mesh within a bioactive hydrogel to mimic the complex microstructure and mechanical behavior of valve tissues. Human aortic valve interstitial cells (HAVIC) and human aortic root smooth muscle cells (HASMC) were separately encapsulated within hydrogel/woven fabric composite scaffolds for generating scaffolds with anisotropic biomechanics and valve ECM like microenvironment for heart valve tissue engineering. UANY have great potential as building blocks for generating fiber-shaped tissues or tissue microstructures with complex architectures.

  20. Extrusion based rapid prototyping technique: an advanced platform for tissue engineering scaffold fabrication.

    PubMed

    Hoque, M Enamul; Chuan, Y Leng; Pashby, Ian

    2012-02-01

    Advances in scaffold design and fabrication technology have brought the tissue engineering field stepping into a new era. Conventional techniques used to develop scaffolds inherit limitations, such as lack of control over the pore morphology and architecture as well as reproducibility. Rapid prototyping (RP) technology, a layer-by-layer additive approach offers a unique opportunity to build complex 3D architectures overcoming those limitations that could ultimately be tailored to cater for patient-specific applications. Using RP methods, researchers have been able to customize scaffolds to mimic the biomechanical properties (in terms of structural integrity, strength, and microenvironment) of the organ or tissue to be repaired/replaced quite closely. This article provides intensive description on various extrusion based scaffold fabrication techniques and review their potential utility for TE applications. The extrusion-based technique extrudes the molten polymer as a thin filament through a nozzle onto a platform layer-by-layer and thus building 3D scaffold. The technique allows full control over pore architecture and dimension in the x- and y- planes. However, the pore height in z-direction is predetermined by the extruding nozzle diameter rather than the technique itself. This review attempts to assess the current state and future prospects of this technology.

  1. On-Demand Guided Bone Regeneration with Microbial Protection of Ornamented SPU Scaffold with Bismuth-Doped Single Crystalline Hydroxyapatite: Augmentation and Cartilage Formation.

    PubMed

    Selvakumar, M; Srivastava, Priyanka; Pawar, Harpreet Singh; Francis, Nimmy K; Das, Bodhisatwa; Sathishkumar, G; Subramanian, Bhuvaneshwaran; Jaganathan, Saravana Kumar; George, Gibin; Anandhan, S; Dhara, Santanu; Nando, Golok B; Chattopadhyay, Santanu

    2016-02-17

    Guided bone regeneration (GBR) scaffolds are futile in many clinical applications due to infection problems. In this work, we fabricated GBR with an anti-infective scaffold by ornamenting 2D single crystalline bismuth-doped nanohydroxyapatite (Bi-nHA) rods onto segmented polyurethane (SPU). Bi-nHA with high aspect ratio was prepared without any templates. Subsequently, it was introduced into an unprecedented synthesized SPU matrix based on dual soft segments (PCL-b-PDMS) of poly(ε-caprolactone) (PCL) and poly(dimethylsiloxane) (PDMS), by an in situ technique followed by electrospinning to fabricate scaffolds. For comparison, undoped pristine nHA rods were also ornamented into it. The enzymatic ring-opening polymerization technique was adapted to synthesize soft segments of PCL-b-PDMS copolymers of SPU. Structure elucidation of the synthesized polymers is done by nuclear magnetic resonance spectroscopy. Sparingly, Bi-nHA ornamented scaffolds exhibit tremendous improvement (155%) in the mechanical properties with excellent antimicrobial activity against various human pathogens. After confirmation of high osteoconductivity, improved biodegradation, and excellent biocompatibility against osteoblast cells (in vitro), the scaffolds were implanted in rabbits by subcutaneous and intraosseous (tibial) sites. Various histological sections reveal the signatures of early cartilage formation, endochondral ossification, and rapid bone healing at 4 weeks of the critical defects filled with ornamented scaffold compared to SPU scaffold. This implies osteogenic potential and ability to provide an adequate biomimetic microenvironment for mineralization for GBR of the scaffolds. Organ toxicity studies further confirm that no tissue architecture abnormalities were observed in hepatic, cardiac, and renal tissue sections. This finding manifests the feasibility of fabricating a mechanically adequate nanofibrous SPU scaffold by a biomimetic strategy and the advantages of Bi

  2. Fabrication of anatomically-shaped cartilage constructs using decellularized cartilage-derived matrix scaffolds.

    PubMed

    Rowland, Christopher R; Colucci, Lina A; Guilak, Farshid

    2016-06-01

    The native extracellular matrix of cartilage contains entrapped growth factors as well as tissue-specific epitopes for cell-matrix interactions, which make it a potentially attractive biomaterial for cartilage tissue engineering. A limitation to this approach is that the native cartilage extracellular matrix possesses a pore size of only a few nanometers, which inhibits cellular infiltration. Efforts to increase the pore size of cartilage-derived matrix (CDM) scaffolds dramatically attenuate their mechanical properties, which makes them susceptible to cell-mediated contraction. In previous studies, we have demonstrated that collagen crosslinking techniques are capable of preventing cell-mediated contraction in CDM disks. In the current study, we investigated the effects of CDM concentration and pore architecture on the ability of CDM scaffolds to resist cell-mediated contraction. Increasing CDM concentration significantly increased scaffold mechanical properties, which played an important role in preventing contraction, and only the highest CDM concentration (11% w/w) was able to retain the original scaffold dimensions. However, the increase in CDM concentration led to a concomitant decrease in porosity and pore size. Generating a temperature gradient during the freezing process resulted in unidirectional freezing, which aligned the formation of ice crystals during the freezing process and in turn produced aligned pores in CDM scaffolds. These aligned pores increased the pore size of CDM scaffolds at all CDM concentrations, and greatly facilitated infiltration by mesenchymal stem cells (MSCs). These methods were used to fabricate of anatomically-relevant CDM hemispheres. CDM hemispheres with aligned pores supported uniform MSC infiltration and matrix deposition. Furthermore, these CDM hemispheres retained their original architecture and did not contract, warp, curl, or splay throughout the entire 28-day culture period. These findings demonstrate that given the

  3. A new method of fabricating a blend scaffold using an indirect three-dimensional printing technique.

    PubMed

    Jung, Jin Woo; Lee, Hyungseok; Hong, Jung Min; Park, Jeong Hun; Shim, Jung Hee; Choi, Tae Hyun; Cho, Dong-Woo

    2015-11-03

    Due to its simplicity and effectiveness, the physical blending of polymers is considered to be a practical strategy for developing a versatile scaffold having desirable mechanical and biochemical properties. In the present work, an indirect three-dimensional (i3D) printing technique was proposed to fabricate a 3D free-form scaffold using a blend of immiscible materials, such as polycaprolactone (PCL) and gelatin. The i3D printing technique includes 3D printing of a mold and a sacrificial molding process. PCL/chloroform and gelatin/water were physically mixed to prepare the blend solution, which was subsequently injected into the cavity of a 3D printed mold. After solvent removal and gelatin cross-linking, the mold was dissolved to obtain a PCL-gelatin (PG) scaffold, with a specific 3D structure. Scanning electron microscopy and Fourier transform infrared spectroscopy analysis indicated that PCL masses and gelatin fibers in the PG scaffold homogenously coexisted without chemical bonding. Compression tests confirmed that gelatin incorporation into the PCL enhanced its mechanical flexibility and softness, to the point of being suitable for soft-tissue engineering, as opposed to pure PCL. Human adipose-derived stem cells, cultured on a PG scaffold, exhibited enhanced in vitro chondrogenic differentiation and tissue formation, compared with those on a PCL scaffold. The i3D printing technique can be used to blend a variety of materials, facilitating 3D scaffold fabrication for specific tissue regeneration. Furthermore, this convenient and versatile technique may lead to wider application of 3D printing in tissue engineering.

  4. Fabrication of poly-DL-lactide/polyethylene glycol scaffolds using the gas foaming technique.

    PubMed

    Ji, Chengdong; Annabi, Nasim; Hosseinkhani, Maryam; Sivaloganathan, Sobana; Dehghani, Fariba

    2012-02-01

    The aim of this study was to prepare poly-DL-lactide/polyethylene glycol (PDLLA/PEG) blends to improve medium absorption and cell proliferation in the three-dimensional (3-D) structure of their scaffolds. Carbon dioxide (CO2) was used as a foaming agent to create porosity in these blends. The results of Fourier transform infrared (FTIR) spectroscopy demonstrated that the blends were homogeneous mixtures of PDLLA and PEG. The peak shifts at 1092 and 1744 cm(-1) confirmed the presence of molecular interactions between these two compounds. Increasing the PEG weight ratio enhanced the relative crystallinity and hydrophilicity. The PDLLA/PEG blends (especially 80/20 and 70/30 weight ratios) exhibited linear degradation profiles over an incubation time of 8 weeks. The mechanical properties of PDLLA/PEG blends having less than 30 wt.% PEG were suitable for the fabrication of porous scaffolds. Increasing the concentration of PEG to above 50% resulted in blends that were brittle and had low mechanical integrity. Highly porous scaffolds with controllable pore size were produced for 30 wt.% PEG samples using the gas foaming technique at temperatures between 25 and 55 °C and pressures between 60 and 160 bar. The average pore diameters achieved by gas foaming process were between 15 and 150 μm, and had an average porosity of 84%. The medium uptake and degradation rate of fabricated PDLLA/PEG scaffolds were increased compared with neat PDLLA film due to the presence of PEG and porosity. The porous scaffolds also demonstrated a lower modulus of elasticity and a higher elongation at break compared to the non-porous film. The fabricated PDLLA/PEG scaffolds have high potential for various tissue-engineering applications.

  5. Structure-property relationship of regenerated spider silk protein nano/microfibrous scaffold fabricated by electrospinning.

    PubMed

    Yu, Qiaozhen; Xu, Shuiling; Zhang, Hong; Gu, Li; Xu, Yepei; Ko, Frank

    2014-11-01

    The regenerated Araneus ventricosus spider dragline silk protein fibrous scaffold with moderate strength and flexibility was fabricated by electrospinning and po