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Sample records for porous polycaprolactone scaffold

  1. Polycaprolactone coated porous tricalcium phosphate scaffolds for controlled release of protein for tissue engineering

    PubMed Central

    Xue, Weichang; Bandyopadhyay, Amit; Bose, Susmita

    2010-01-01

    Polycaprolactone (PCL) was coated on porous tricalcium phosphate (TCP) scaffolds to achieve controlled protein delivery. Porous TCP scaffolds were fabricated using reticulated polyurethane foam as sacrificial scaffold with a porosity of 70–90 vol %. PCL was coated on sintered porous TCP scaffolds by dipping-drying process. The compressive strength of TCP scaffolds increased significantly after PCL coating. The highest strength of 2.41 MPa at a porosity of 70% was obtained for the TCP scaffold coated with 5% PCL solution. Model protein bovine serum albumin (BSA) was encapsulated efficiently within the PCL coating. The amount of BSA encapsulation was controlled by varying proteins’ composition in the PCL coating. The FTIR analysis confirmed that BSA retained its structural conformation and did not show significant denaturization during PCL coating. The release kinetics in phosphate buffer solution indicated that the protein release was controlled and sustained, and primarily dependant on protein concentration encapsulated in the PCL coating. PMID:19572301

  2. The interaction between bone marrow stromal cells and RGD modified three dimensional porous polycaprolactone scaffolds

    PubMed Central

    Zhang, Huina; Lin, Chia-Ying; Hollister, Scott J

    2015-01-01

    We previously established a simple method to immobilize the Arg-Gly-Asp (RGD) peptide on polycaprolactone (PCL) two-dimensional film surfaces that significantly improved bone marrow stromal cell (BMSC) adhesion to these films. The current work extends this modification strategy to three-dimensional (3D) PCL scaffolds to investigate BMSCs attachment, cellular distribution and cellularity, signal transduction and survival on the modified PCL scaffold compared to those on the untreated ones. The results demonstrated that treatment of 3D PCL scaffold surfaces with 1,6-hexanediamine introduced the amino functional groups onto the porous PCL scaffold homogenously as detected by a ninhydrin staining method. Followed by the cross-linking reaction, RGDC peptide was successfully immobilized on the surface of PCL scaffold. Although the static seeding method used in this study caused heterogeneous cell distribution, the RGD modified PCL scaffold still demonstrated the improved BMSC attachment and cellular distribution in the scaffold. More importantly, the integrin-mediated signal transduction FAK-PI3K-Akt pathway was significantly up-regulated by RGD modification and a subsequent increase in cell survival and growth was found in the modified scaffold. The present study introduces an easy method to immobilize RGD peptide on the 3D porous PCL scaffold and provides further evidence that modification of 3D PCL scaffolds with RGD peptides elicits specific cellular responses and improves the final cell-biomaterial interaction. PMID:19487019

  3. Polycaprolactone- and polycaprolactone/ceramic-based 3D-bioplotted porous scaffolds for bone regeneration: A comparative study.

    PubMed

    Gómez-Lizárraga, K K; Flores-Morales, C; Del Prado-Audelo, M L; Álvarez-Pérez, M A; Piña-Barba, M C; Escobedo, C

    2017-10-01

    One of the critical challenges that scaffolding faces in the organ and tissue regeneration field lies in mimicking the structure, and the chemical and biological properties of natural tissue. A high-level control over the architecture, mechanical properties and composition of the materials in contact with cells is essential to overcome such challenge. Therefore, definition of the method, materials and parameters for the production of scaffolds during the fabrication stage is critical. With the recent emergence of rapid prototyping (RP), it is now possible to create three-dimensional (3D) scaffolds with the essential characteristics for the proliferation and regeneration of tissues, such as porosity, mechanical strength, pore size and pore interconnectivity, and biocompatibility. In this study, we employed 3D bioplotting, a RP technology, to fabricate scaffolds made from (i) pure polycaprolactone (PCL) and (ii) a composite based on PCL and ceramic micro-powder. The ceramics used for the composite were bovine bone filling Nukbone® (NKB), and hydroxyapatite (HA) with 5%, 10% or 20% wt. The scaffolds were fabricated in a cellular lattice structure (i.e. meshing mode) using a 0/90° lay down pattern with a continuous contour filament in order to achieve interconnected porous reticular structures. We varied the temperature, as well as injection speed and pressure during the bioplotting process to achieve scaffolds with pore size ranging between 200 and 400μm and adequate mechanical stability. The resulting scaffolds had an average pore size of 323μm and an average porosity of 32%. Characterization through ATR-FTIR revealed the presence of the characteristic bands of hydroxyapatite in the PCL matrix, and presented an increase of the intensity of the phosphate and carbonyl bands as the ceramic content increased. The bioplotted 3D scaffolds have a Young's modulus (E) in the range between 0.121 and 0.171GPa, which is compatible with the modulus of natural bone. PCL

  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. The interaction between bone marrow stromal cells and RGD-modified three-dimensional porous polycaprolactone scaffolds.

    PubMed

    Zhang, Huina; Lin, Chia-Ying; Hollister, Scott J

    2009-09-01

    We previously established a simple method to immobilize the Arg-Gly-Asp (RGD) peptide on polycaprolactone (PCL) two-dimensional film surfaces that significantly improved bone marrow stromal cell (BMSC) adhesion to these films. The current work extends this modification strategy to three-dimensional (3D) PCL scaffolds to investigate BMSC attachment, cellular distribution and cellularity, signal transduction and survival on the modified PCL scaffold compared to those on the untreated ones. The results demonstrated that treatment of 3D PCL scaffold surfaces with 1,6-hexanediamine introduced the amino functional groups onto the porous PCL scaffold homogenously as detected by a ninhydrin staining method. Followed by the cross-linking reaction, RGDC peptide was successfully immobilized on the surface of PCL scaffold. Although the static seeding method used in this study caused heterogeneous cell distribution, the RGD-modified PCL scaffold still demonstrated the improved BMSC attachment and cellular distribution in the scaffold. More importantly, the integrin-mediated signal transduction FAK-PI3K-Akt pathway was significantly up-regulated by RGD modification and a subsequent increase in cell survival and growth was found in the modified scaffold. The present study introduces an easy method to immobilize RGD peptide on the 3D porous PCL scaffold and provides further evidence that modification of 3D PCL scaffolds with RGD peptides elicits specific cellular responses and improves the final cell-biomaterial interaction.

  6. 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.

  7. 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.

  8. 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.

  9. 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

  10. 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.

  11. 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.

  12. Ligament Tissue Engineering Using a Novel Porous Polycaprolactone Fumarate Scaffold and Adipose Tissue-Derived Mesenchymal Stem Cells Grown in Platelet Lysate

    PubMed Central

    Wagner, Eric R.; Bravo, Dalibel; Dadsetan, Mahrokh; Riester, Scott M.; Chase, Steven; Westendorf, Jennifer J.; Dietz, Allan B.; van Wijnen, Andre J.; Yaszemski, Michael J.

    2015-01-01

    Purpose: Surgical reconstruction of intra-articular ligament injuries is hampered by the poor regenerative potential of the tissue. We hypothesized that a novel composite polymer “neoligament” seeded with progenitor cells and growth factors would be effective in regenerating native ligamentous tissue. Methods: We synthesized a fumarate-derivative of polycaprolactone fumarate (PCLF) to create macro-porous scaffolds to allow cell–cell communication and nutrient flow. Clinical grade human adipose tissue-derived human mesenchymal stem cells (AMSCs) were cultured in 5% human platelet lysate (PL) and seeded on scaffolds using a dynamic bioreactor. Cell growth, viability, and differentiation were examined using metabolic assays and immunostaining for ligament-related markers (e.g., glycosaminoglycans [GAGs], alkaline phosphatase [ALP], collagens, and tenascin-C). Results: AMSCs seeded on three-dimensional (3D) PCLF scaffolds remain viable for at least 2 weeks with proliferating cells filling the pores. AMSC proliferation rates increased in PL compared to fetal bovine serum (FBS) (p < 0.05). Cells had a low baseline expression of ALP and GAG, but increased expression of total collagen when induced by the ligament and tenogenic growth factor fibroblast growth factor 2 (FGF-2), especially when cultured in the presence of PL (p < 0.01) instead of FBS (p < 0.05). FGF-2 and PL also significantly increased immunostaining of tenascin-C and collagen at 2 and 4 weeks compared with human fibroblasts. Summary: Our results demonstrate that AMSCs proliferate and eventually produce a collagen-rich extracellular matrix on porous PCLF scaffolds. This novel scaffold has potential in stem cell engineering and ligament regeneration. PMID:26413793

  13. Ligament Tissue Engineering Using a Novel Porous Polycaprolactone Fumarate Scaffold and Adipose Tissue-Derived Mesenchymal Stem Cells Grown in Platelet Lysate.

    PubMed

    Wagner, Eric R; Bravo, Dalibel; Dadsetan, Mahrokh; Riester, Scott M; Chase, Steven; Westendorf, Jennifer J; Dietz, Allan B; van Wijnen, Andre J; Yaszemski, Michael J; Kakar, Sanjeev

    2015-11-01

    Surgical reconstruction of intra-articular ligament injuries is hampered by the poor regenerative potential of the tissue. We hypothesized that a novel composite polymer "neoligament" seeded with progenitor cells and growth factors would be effective in regenerating native ligamentous tissue. We synthesized a fumarate-derivative of polycaprolactone fumarate (PCLF) to create macro-porous scaffolds to allow cell-cell communication and nutrient flow. Clinical grade human adipose tissue-derived human mesenchymal stem cells (AMSCs) were cultured in 5% human platelet lysate (PL) and seeded on scaffolds using a dynamic bioreactor. Cell growth, viability, and differentiation were examined using metabolic assays and immunostaining for ligament-related markers (e.g., glycosaminoglycans [GAGs], alkaline phosphatase [ALP], collagens, and tenascin-C). AMSCs seeded on three-dimensional (3D) PCLF scaffolds remain viable for at least 2 weeks with proliferating cells filling the pores. AMSC proliferation rates increased in PL compared to fetal bovine serum (FBS) (p < 0.05). Cells had a low baseline expression of ALP and GAG, but increased expression of total collagen when induced by the ligament and tenogenic growth factor fibroblast growth factor 2 (FGF-2), especially when cultured in the presence of PL (p < 0.01) instead of FBS (p < 0.05). FGF-2 and PL also significantly increased immunostaining of tenascin-C and collagen at 2 and 4 weeks compared with human fibroblasts. Our results demonstrate that AMSCs proliferate and eventually produce a collagen-rich extracellular matrix on porous PCLF scaffolds. This novel scaffold has potential in stem cell engineering and ligament regeneration.

  14. A novel nano-structured porous polycaprolactone scaffold improves hyaline cartilage repair in a rabbit model compared to a collagen type I/III scaffold: in vitro and in vivo studies.

    PubMed

    Christensen, Bjørn Borsøe; Foldager, Casper Bindzus; Hansen, Ole Møller; Kristiansen, Asger Albæk; Le, Dang Quang Svend; Nielsen, Agnete Desirée; Nygaard, Jens Vinge; Bünger, Cody Erik; Lind, Martin

    2012-06-01

    To develop a nano-structured porous polycaprolactone (NSP-PCL) scaffold and compare the articular cartilage repair potential with that of a commercially available collagen type I/III (Chondro-Gide) scaffold. By combining rapid prototyping and thermally induced phase separation, the NSP-PCL scaffold was produced for matrix-assisted autologous chondrocyte implantation. Lyophilizing a water-dioxane-PCL solution created micro and nano-pores. In vitro: The scaffolds were seeded with rabbit chondrocytes and cultured in hypoxia for 6 days. qRT-PCR was performed using primers for sox9, aggrecan, collagen type 1 and 2. In vivo: 15 New Zealand White Rabbits received bilateral osteochondral defects in the femoral intercondylar grooves. Autologous chondrocytes were harvested 4 weeks prior to surgery. There were 3 treatment groups: (1) NSP-PCL scaffold without cells. (2) The Chondro-Gide scaffold with autologous chondrocytes and (3) NSP-PCL scaffold with autologous chondrocytes. Observation period was 13 weeks. Histological evaluation was made using the O'Driscoll score. In vitro: The expressions of sox9 and aggrecan were higher in the NSP-PCL scaffold, while expression of collagen 1 was lower compared to the Chondro-Gide scaffold. In vivo: Both NSP-PCL scaffolds with and without cells scored significantly higher than the Chondro-Gide scaffold when looking at the structural integrity and the surface regularity of the repair tissue. No differences were found between the NSP-PCL scaffold with and without cells. The NSP-PCL scaffold demonstrated higher in vitro expression of chondrogenic markers and had higher in vivo histological scores compared to the Chondro-Gide scaffold. The improved chondrocytic differentiation can potentially produce more hyaline cartilage during clinical cartilage repair. It appears to be a suitable cell-free implant for hyaline cartilage repair and could provide a less costly and more effective treatment option than the Chondro-Gide scaffold with cells.

  15. Mechanical and microstructural properties of polycaprolactone scaffolds with one-dimensional, two-dimensional, and three-dimensional orthogonally oriented porous architectures produced by selective laser sintering.

    PubMed

    Eshraghi, Shaun; Das, Suman

    2010-07-01

    This article reports on the experimental determination and finite element modeling of tensile and compressive mechanical properties of solid polycaprolactone (PCL) and of porous PCL scaffolds with one-dimensional, two-dimensional and three-dimensional orthogonal, periodic porous architectures produced by selective laser sintering (SLS). PCL scaffolds were built using optimum processing parameters, ensuring scaffolds with nearly full density (>95%) in the designed solid regions and with excellent geometric and dimensional control (within 3-8% of design). The tensile strength of bulk PCL ranged from 10.5 to 16.1 MPa, its modulus ranged from 343.9 to 364.3 MPa, and the tensile yield strength ranged from 8.2 to 10.1 MPa. These values are consistent with reported literature values for PCL processed through various manufacturing methods. Across porosity ranged from 56.87% to 83.3%, the tensile strength ranged from 4.5 to 1.1 MPa, the tensile modulus ranged from 140.5 to 35.5 MPa, and the yield strength ranged from 3.2 to 0.76 MPa. The compressive strength of bulk PCL was 38.7 MPa, the compressive modulus ranged from 297.8 to 317.1 MPa, and the compressive yield strength ranged from 10.3 to 12.5 MPa. Across porosity ranged from 51.1% to 80.9%, the compressive strength ranged from 10.0 to 0.6 MPa, the compressive modulus ranged from 14.9 to 12.1 MPa, and the compressive yield strength ranged from 4.25 to 0.42 MPa. These values, while being in the lower range of reported values for trabecular bone, are the highest reported for PCL scaffolds produced by SLS and are among the highest reported for similar PCL scaffolds produced through other layered manufacturing techniques. Finite element analysis showed good agreement between experimental and computed effective tensile and compressive moduli. Thus, the construction of bone tissue engineering scaffolds endowed with oriented porous architectures and with predictable mechanical properties through SLS is demonstrated. Copyright

  16. Nanohydroxyapatite incorporated electrospun polycaprolactone/polycaprolactone-polyethyleneglycol-polycaprolactone blend scaffold for bone tissue engineering applications.

    PubMed

    Remya, K R; Joseph, Jasmin; Mani, Susan; John, Annie; Varma, H K; Ramesh, P

    2013-09-01

    The present work is a comparative evaluation of physical and biological properties of electrospun biodegradable fibrous scaffolds based on polycaprolactone (PCL) and its blend with polycaprolactone-polyethyleneglycol-polycaprolactone (CEC) with and without nanohydroxyapatite (nHAP) particles. The fiber morphology, porosity, surface wettability, and mechanical properties of electrospun PCL were distinctly influenced by the presence of both copolymer CEC and nHAP. The degradation in hydrolytic media affected both morphological and mechanical properties of the scaffolds and the tensile strength decreased by 58% for PCL, 83% for PCL/CEC, 36% for PCL/nHAP and 75% for PCL/CEC/nHAP in 90 days of PBS ageing. MTT assay using mouse fibroblast L929 cells proved all the scaffolds to be non-cytotoxic. An overall enhanced performance was shown by PCL/CEC/nHAP scaffold in cell viability (LPH) and proliferation (Picogreen). Simultaneously, ELF assay of ALP activity (bone marker) confirmed the presence of osteogenic-induced Rabbit adipose-derived mesenchymal stem cells (ADMSCs) on all the scaffolds. In comparison, the results reveal the potential of the cytocompatible PCL/CEC/nHAP scaffold for the fabrication of living bony constructs for tissue engineering applications.

  17. 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.

  18. Composite tissue engineering on polycaprolactone nanofiber scaffolds.

    PubMed

    Reed, Courtney R; Han, Li; Andrady, Anthony; Caballero, Montserrat; Jack, Megan C; Collins, James B; Saba, Salim C; Loboa, Elizabeth G; Cairns, Bruce A; van Aalst, John A

    2009-05-01

    Tissue engineering has largely focused on single tissue-type reconstruction (such as bone); however, the basic unit of healing in any clinically relevant scenario is a compound tissue type (such as bone, periosteum, and skin). Nanofibers are submicron fibrils that mimic the extracellular matrix, promoting cellular adhesion, proliferation, and migration. Stem cell manipulation on nanofiber scaffolds holds significant promise for future tissue engineering. This work represents our initial efforts to create the building blocks for composite tissue reflecting the basic unit of healing. Polycaprolactone (PCL) nanofibers were electrospun using standard techniques. Human foreskin fibroblasts, murine keratinocytes, and periosteal cells (4-mm punch biopsy) harvested from children undergoing palate repair were grown in appropriate media on PCL nanofibers. Human fat-derived mesenchymal stem cells were osteoinduced on PCL nanofibers. Cell growth was assessed with fluorescent viability staining; cocultured cells were differentiated using antibodies to fibroblast- and keratinocyte-specific surface markers. Osteoinduction was assessed with Alizarin red S. PCL nanofiber scaffolds supported robust growth of fibroblasts, keratinocytes, and periosteal cells. Cocultured periosteal cells (with fibroblasts) and keratinocytes showed improved longevity of the keratinocytes, though growth of these cell types was randomly distributed throughout the scaffold. Robust osteoinduction was noted on PCL nanofibers. Composite tissue engineering using PCL nanofiber scaffolds is possible, though the major obstacles to the trilaminar construct are maintaining an appropriate interface between the tissue types and neovascularization of the composite structure.

  19. [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.

  20. Tubular open-porous β-tricalcium phosphate polycaprolactone scaffolds as guiding structure for segmental bone defect regeneration in a novel sheep model.

    PubMed

    Pobloth, Anne-Marie; Schell, Hanna; Petersen, Ansgar; Beierlein, Katleen; Kleber, Christian; Schmidt-Bleek, Katharina; Duda, Georg N

    2017-05-08

    Large segmental bone defect reconstruction with sufficient functional restoration is one of the most demanding challenges in orthopaedic surgery. Available regenerative treatment options, as the vascularized bone graft transfer, the Masquelet technique or the Ilizarov distraction osteogenesis, are associated with specific indications and distinct limitations. As an alternative, a hollow cylindrical ceramic-polymer composite scaffold (β-tricalcium phosphate and poly-lactid co-ε- caprolactone), facilitating a strong surface guiding effect for tissue ingrowth (group 1; n = 6) was investigated here. In combination with an additional autologous, cancellous bone graft filling, the scaffold's ability to work as an open-porous membrane to improve the defect healing process was analysed (group 2; n = 6). A novel model of a critical size (40 mm) tibia osteotomy defect stabilized with an external hybrid-ring fixator, was established in sheep. Segmental defect regeneration and tissue organization in relation to the scaffold were analysed radiologically, (immune-) histologically, and with second-harmonic generation imaging 12 weeks after surgery. The scaffold's tubular shape and open-porous structure controlled the collagen fibre orientation within the bone defect and guided the following mineralization process along the scaffold surface. In combination with the osteoinductive stimulus, a unilateral bony bridging of the critically sized defect was achieved in one third of the animals. The external hybrid-ring fixator was appropriate for large segmental defect stabilization in sheep. Copyright © 2017 John Wiley & Sons, Ltd.

  1. Fluorescent composite scaffolds made of nanodiamonds/polycaprolactone

    NASA Astrophysics Data System (ADS)

    Cao, Li; Hou, Yanwen; Lafdi, Khalid; Urmey, Kirk

    2015-11-01

    Polycaprolactone (PCL) has been widely studied for biological applications. Biodegradable PCL fibrous scaffold can work as an appropriate substrate for tissue regeneration. In this letter, fluorescent nanodiamonds (FNDs) were prepared after surface passivation with octadecylamine. The FNDs were then mixed with PCL polymer and subsequently electrospun into FNDs/PCL fibrous scaffolds. The obtained scaffolds not only exhibited photoluminescence, but also showed reinforced mechanical strength. Toxicity study indicated FNDs/PCL scaffolds were nontoxic. This biocompatible fluorescent composite fibrous scaffold can support in vitro cell growth and also has the potential to act as an optical probe for tissue engineering application in vitro and in vivo.

  2. Manufacture of layered collagen/chitosan-polycaprolactone scaffolds with biomimetic microarchitecture.

    PubMed

    Zhu, Youjia; Wan, Ying; Zhang, Jun; Yin, Dengke; Cheng, Wenze

    2014-01-01

    Chitosan-polycaprolactone (CH-PCL) copolymers with various PCL percentages less than 45 wt% were synthesized. Different CH-PCLs were respectively blended with Type-II collagen at prescribed ratios to fabricate a type of layered porous scaffolds with some biomimetic features while using sodium tripolyphosphate as a crosslinker. The compositions of different layers inside scaffolds were designed in a way so that from the top layer to the bottom layer collagen content changed in a degressive trend contrary to that of chitosan. A combinatorial processing technique involving adjustable temperature gradients, collimated photothermal heating and freeze-drying was used to construct desired microstructures of scaffolds. The resultant scaffolds had highly interconnected porous layers with a layer thickness of around 1mm and porous interface zones without visual clefts. Results obtained from SEM observations and measurements of pore parameters and swelling properties as well as mechanical examinations confirmed that graded average pore-size and porosity, gradient swelling index and oriented compressive modulus for certain scaffolds were synchronously achieved. In addition, certain evaluations of cell-scaffold constructs indicated that the achieved scaffolds were able to well support the growth of seeded chondrocytes. The optimized collagen/CH-PCL scaffolds are partially similar to articular cartilage extracellular matrix in composition, porous microarchitecture, water content and compressive mechanical properties, suggesting that they have promising potential for applications in articular cartilage repair.

  3. Three-dimensional polycaprolactone scaffold via needleless electrospinning promotes cell proliferation and infiltration.

    PubMed

    Li, Dawei; Wu, Tong; He, Nanfei; Wang, Jing; Chen, Weiming; He, Liping; Huang, Chen; Ei-Hamshary, Hany A; Al-Deyab, Salem S; Ke, Qinfei; Mo, Xiumei

    2014-09-01

    Electrospinning has been widely used in fabrication of tissue engineering scaffolds. Currently, most of the electrospun nanofibers performed like a conventional two-dimensional (2D) membrane, which hindered their further applications. Moreover, the low production rate of the traditional needle-electrospinning (NE) also limited the commercialization. In this article, disc-electrospinning (DE) was utilized to fabricate a three-dimensional (3D) scaffold consisting of porous macro/nanoscale fibers. The morphology of the porous structure was investigated by scanning electron microscopy images, which showed irregular pores of nanoscale spreading on the surface of DE polycaprolactone (PCL) fibers. Protein adsorption assessment illustrated the porous structure could significantly enhance proteins pickup, which was 55% higher than that of solid fiber scaffolds. Fibroblasts were cultured on the scaffold. The results demonstrated that DE fiber scaffold could enhance initial cell attachment. In the 7 days of culture, fibroblasts grew faster on DE fiber scaffold in comparison with solid fiber, solvent cast (SC) film and TCP. Fibroblasts on DE fibers showed a stretched shape and integrated with the porous surface tightly. Cells were also found to migrate into the DE scaffold up to 800μm. Results supported the use of DE PCL fibers as a 3D tissue engineering scaffold in soft tissue regeneration.

  4. Polycaprolactone nanofiber interspersed collagen type-I scaffold for bone regeneration: a unique injectable osteogenic scaffold.

    PubMed

    Baylan, Nuray; Bhat, Samerna; Ditto, Maggie; Lawrence, Joseph G; Lecka-Czernik, Beata; Yildirim-Ayan, Eda

    2013-08-01

    There is an increasing demand for an injectable cell coupled three-dimensional (3D) scaffold to be used as bone fracture augmentation material. To address this demand, a novel injectable osteogenic scaffold called PN-COL was developed using cells, a natural polymer (collagen type-I), and a synthetic polymer (polycaprolactone (PCL)). The injectable nanofibrous PN-COL is created by interspersing PCL nanofibers within pre-osteoblast cell embedded collagen type-I. This simple yet novel and powerful approach provides a great benefit as an injectable bone scaffold over other non-living bone fracture stabilization polymers, such as polymethylmethacrylate and calcium content resin-based materials. The advantages of injectability and the biomimicry of collagen was coupled with the structural support of PCL nanofibers, to create cell encapsulated injectable 3D bone scaffolds with intricate porous internal architecture and high osteoconductivity. The effects of PCL nanofiber inclusion within the cell encapsulated collagen matrix has been evaluated for scaffold size retention and osteocompatibility, as well as for MC3T3-E1 cells osteogenic activity. The structural analysis of novel bioactive material proved that the material is chemically stable enough in an aqueous solution for an extended period of time without using crosslinking reagents, but it is also viscous enough to be injected through a syringe needle. Data from long-term in vitro proliferation and differentiation data suggests that novel PN-COL scaffolds promote the osteoblast proliferation, phenotype expression, and formation of mineralized matrix. This study demonstrates for the first time the feasibility of creating a structurally competent, injectable, cell embedded bone tissue scaffold. Furthermore, the results demonstrate the advantages of mimicking the hierarchical architecture of native bone with nano- and micro-size formation through introducing PCL nanofibers within macron-size collagen fibers and in

  5. Preparation of bioactive porous HA/PCL composite scaffolds

    NASA Astrophysics Data System (ADS)

    Zhao, J.; Guo, L. Y.; Yang, X. B.; Weng, J.

    2008-12-01

    Porous hydroxyapatite (HA) bioceramic scaffold has been widely attracted the attention to act as a three-dimensional (3D) template for cell adhesion, proliferation, differentiation and thus promoting bone and cartilage regeneration because of its osteoinduction. However, the porous bioceramic scaffold is fragile so that it is not suitable to be applied in clinic for bone repair or replacement. Therefore, it is significant to improve the mechanical property of porous HA bioceramics while the interconnected structure is maintained for tissue ingrowth in vivo. In the present research, a porous composite scaffold composed of HA scaffold and polycaprolactone (PCL) lining was fabricated by the method of polymer impregnating to produce HA scaffold coated with PCL lining. Subsequently, the composite scaffolds were deposited with biomimetic coating for improving the bioactivity. The HA/PCL composite scaffolds with improved mechanical property and bioactivity is expected to be a promising bone substitute in tissue engineering applications.

  6. 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.

  7. Embroidered and surface coated polycaprolactone-co-lactide scaffolds

    PubMed Central

    Rentsch, Barbe; Bernhardt, Ricardo; Scharnweber, Dieter; Schneiders, Wolfgang; Rammelt, Stefan; Rentsch, Claudia

    2012-01-01

    Tissue engineering and regenerative techniques targeting bone include a broad range of strategies and approaches to repair, augment, replace or regenerate bone tissue. Investigations that are aimed at optimization of these strategies until clinical translation require control of systemic factors as well as modification of a broad range of key parameters. This article reviews a possible strategy using a tissue engineering approach and systematically describes a series of experiments evaluating the properties of an embroidered and surface coated polycaprolactone-co-lactide scaffold being considered as bone graft substitute for large bone defects. The scaffold design and fabrication, the scaffolds properties, as well as its surface modification and their influence in vitro are evaluated, followed by in vivo analysis of the scaffolds using orthotopic implantation models in small and large animals. PMID:23507867

  8. Electrospun polycaprolactone scaffolds under strain and their application in cartilage tissue engineering

    NASA Astrophysics Data System (ADS)

    Nam, Jin

    Electrospinning is a promising fabrication method for three dimensional tissue engineering scaffolds due to its ability to produce a nano-/micro-sized non-woven fibrous structure which resembles the natural extracellular matrix. We investigated the mechanical behavior of two different electrospun microstructures. Polycaprolactone (PCL) fibers with or without "point-bonding" exhibited different deformation behaviors having significant biomedical consequences. While fibers with point-bonded structure failed due to the generation of voids by the fracture of fiber interconnections under strain, fibers without point-bonds produced a 'bamboo' structure with fiber joining visible at higher levels of strain. In addition, gelatin and PCL were electrospun and the residual solvent contents were systematically investigated. A simple and effective means of reducing residual solvent content was developed. The interaction between these electrospun matrices and chondrocytic cells were compared to other topographies having the same chemistry. Electrospun polycaprolactone fibers supported better proliferation and extracellular matrix production than the corresponding semi-porous and dense surfaces and even, at some time points, glass surfaces. The intrinsic capability of electrospinning to produce high porosity appears to offset the relative hydrophobicity of polycaprolactone resulting in a more uniform cell seeding. Electrospun fibers induced a higher level of glycosaminoglycans (GAG) production by providing a 'dynamic scaffold' in which chondrocytes are able to maintain a morphology associated with the appropriate phenotype. Finally, based on this study, a method producing macro-pores within an electrospun scaffold was developed. With this method, not only can cellular infiltration into a thick electrospun scaffold be facilitated, but scaffolds having designed, anisotropic structures can be produced that better approximate the final tissue.

  9. Poly(caprolactone) based magnetic scaffolds for bone tissue engineering

    NASA Astrophysics Data System (ADS)

    Bañobre-López, M.; Piñeiro-Redondo, Y.; De Santis, R.; Gloria, A.; Ambrosio, L.; Tampieri, A.; Dediu, V.; Rivas, J.

    2011-04-01

    Synthetic scaffolds for tissue engineering coupled to stem cells represent a promising approach aiming to promote the regeneration of large defects of damaged tissues or organs. Magnetic nanocomposites formed by a biodegradable poly(caprolactone) (PCL) matrix and superparamagnetic iron doped hydroxyapatite (FeHA) nanoparticles at different PCL/FeHA compositions have been successfully prototyped, layer on layer, through 3D bioplotting. Magnetic measurements, mechanical testing, and imaging were carried out to calibrate both model and technological processing in the magnetized scaffold prototyping. An amount of 10% w/w of magnetic FeHA nanoparticles represents a reinforcement for PCL matrix, however, a reduction of strain at failure is also observed. Energy loss (absorption) measurements under a radio-frequency applied magnetic field were performed in the resulting magnetic scaffolds and very promising heating properties were observed, making them very useful for potential biomedical applications.

  10. New porous polycaprolactone-silica composites for bone regeneration.

    PubMed

    Plazas Bonilla, Clara E; Trujillo, Sara; Demirdögen, Bermali; Perilla, Jairo E; Murat Elcin, Y; Gómez Ribelles, José L

    2014-07-01

    Polycaprolactone porous membranes were obtained by freeze extraction of dioxane from polycaprolactone-dioxane solid solutions. Porosities as high as 90% with interconnected structures were obtained by this technique. A silica phase was synthesized inside the pores of the polymer membrane by sol-gel reaction using tetraethylorthosilicate (TEOS) as a silica precursor and catalyzed in acidic and basic conditions. Two different morphologies of the inorganic phase were obtained depending on the type of catalyst. In acid catalyzed sol-gel reaction, a homogeneous layer of silica was deposited on the pores, and discrete microspheres were synthesized on the pore walls when a basic catalyst was used. The morphology of the inorganic phase influenced the mechanical and thermal behavior, as well as the hydrophilic character of the composites. Bioactivity of the porous materials was tested in vitro by measuring the deposition of hydroxyapatite on the surfaces of the porous composite membranes. Polycaprolactone/silica composites revealed a superior bioactivity performance compared with that of the pure polymer; evidenced by the characteristic cauliflower structures on the material surface, increase in weight and Ca/P ratio of the hydroxyapatite layer. Also, the acid catalyzed composites presented better bioactivity than the base catalyzed composites, evidencing the importance in the morphology of the silica phase. Copyright © 2014. Published by Elsevier B.V.

  11. 3D-Printing Composite Polycaprolactone-Decellularized Bone Matrix Scaffolds for Bone Tissue Engineering Applications.

    PubMed

    Rindone, Alexandra N; Nyberg, Ethan; Grayson, Warren L

    2017-05-11

    Millions of patients worldwide require bone grafts for treatment of large, critically sized bone defects from conditions such as trauma, cancer, and congenital defects. Tissue engineered (TE) bone grafts have the potential to provide a more effective treatment than current bone grafts since they would restore fully functional bone tissue in large defects. Most bone TE approaches involve a combination of stem cells with porous, biodegradable scaffolds that provide mechanical support and degrade gradually as bone tissue is regenerated by stem cells. 3D-printing is a key technique in bone TE that can be used to fabricate functionalized scaffolds with patient-specific geometry. Using 3D-printing, composite polycaprolactone (PCL) and decellularized bone matrix (DCB) scaffolds can be produced to have the desired mechanical properties, geometry, and osteoinductivity needed for a TE bone graft. This book chapter will describe the protocols for fabricating and characterizing 3D-printed PCL:DCB scaffolds. Moreover, procedures for culturing adipose-derived stem cells (ASCs) in these scaffolds in vitro will be described to demonstrate the osteoinductivity of the scaffolds.

  12. Porous silicon confers bioactivity to polycaprolactone composites in vitro.

    PubMed

    Henstock, J R; Ruktanonchai, U R; Canham, L T; Anderson, S I

    2014-04-01

    Silicon is an essential element for healthy bone development and supplementation with its bioavailable form (silicic acid) leads to enhancement of osteogenesis both in vivo and in vitro. Porous silicon (pSi) is a novel material with emerging applications in opto-electronics and drug delivery which dissolves to yield silicic acid as the sole degradation product, allowing the specific importance of soluble silicates for biomaterials to be investigated in isolation without the elution of other ionic species. Using polycaprolactone as a bioresorbable carrier for porous silicon microparticles, we found that composites containing pSi yielded more than twice the amount of bioavailable silicic acid than composites containing the same mass of 45S5 Bioglass. When incubated in a simulated body fluid, the addition of pSi to polycaprolactone significantly increased the deposition of calcium phosphate. Interestingly, the apatites formed had a Ca:P ratio directly proportional to the silicic acid concentration, indicating that silicon-substituted hydroxyapatites were being spontaneously formed as a first order reaction. Primary human osteoblasts cultured on the surface of the composite exhibited peak alkaline phosphatase activity at day 14, with a proportional relationship between pSi content and both osteoblast proliferation and collagen production over 4 weeks. Culturing the composite with J744A.1 murine macrophages demonstrated that porous silicon does not elicit an immune response and may even inhibit it. Porous silicon may therefore be an important next generation biomaterial with unique properties for applications in orthopaedic tissue engineering.

  13. The significance of grafting collagen on polycaprolactone composite scaffolds: processing-structure-functional property relationship.

    PubMed

    Kiran, S; Nune, K C; Misra, R D K

    2015-09-01

    The study concerns processing-structure-functional property relationship in organic-inorganic hybrid scaffolds based on grafted collagen for bone tissue engineering. Biodegradable polyester, polycaprolactone (PCL) and nanohydroxyapatite were used to fabricate three-dimensional porous scaffolds by adopting a combination of solvent casting, particulate leaching, and polymer leaching approaches. The PCL scaffold was subsequently surface modified by chemical bonding of 1,6-hexanediamine to the ester groups of PCL to introduce free NH2 groups. The introduction of NH2 groups as active sites enabled immobilization of biocompatible macromolecule, collagen, on the aminolyzed PCL via a cross-linking agent, glutaraldehyde. The osteoblasts' functions, notably cell adhesion, proliferation, and mineralization, were favorably modulated because of the chemical interaction between Arg-Gly-Asp domains in collagen molecule and integrin receptor in the cell membrane. The study underscores the significance of grafting collagen on PCL-nHA scaffold in modulating cellular activity and biological functions expanding its current use in soft tissue engineering to hard tissue regeneration. © 2015 Wiley Periodicals, Inc.

  14. Reinforcing bioceramic scaffolds with in situ synthesized ε-polycaprolactone coatings.

    PubMed

    Martínez-Vázquez, Francisco J; Miranda, Pedro; Guiberteau, Fernando; Pajares, Antonia

    2013-12-01

    In situ ring-opening polymerization of ε-caprolactone (ε-CL) was performed to coat β-tricalcium phosphate (β-TCP) scaffolds fabricated by robocasting in order to enhance their mechanical performance while preserving the predesigned macropore architecture. Concentrated colloidal inks prepared from β-TCP commercial powders were used to fabricate porous structures consisting of a three-dimensional mesh of interpenetrating rods. Then, ε-CL was in situ polymerized within the ceramic structure using a lipase as catalyst and toluene as solvent, to obtain a highly homogeneous coating and full impregnation of in-rod microporosity. The strength and toughness of scaffolds coated by ε-polycaprolactone (ε-PCL) were significantly increased (twofold and fivefold increase, respectively) over those of the bare structures. Enhancement of both properties is associated to the healing of preexisting microdefects in the bioceramic rods. These enhancements are compared to results from previous work on fully impregnated structures. The implications of the results for the optimization of the mechanical and biological performance of scaffolds for bone tissue engineering applications are discussed. Copyright © 2013 Wiley Periodicals, Inc., a Wiley Company.

  15. Design and manufacture of neural tissue engineering scaffolds using hyaluronic acid and polycaprolactone nanofibers with controlled porosity.

    PubMed

    Entekhabi, Elahe; Haghbin Nazarpak, Masoumeh; Moztarzadeh, Fathollah; Sadeghi, Ali

    2016-12-01

    Given the large differences in nervous tissue and other tissues of the human body and its unique features, such as poor and/or lack of repair, there are many challenges in the repair process of this tissue. Tissue engineering is one of the most effective approaches to repair neural damages. Scaffolds made from electrospun fibers have special potential in cell adhesion, function and cell proliferation. This research attempted to design a high porous nanofibrous scaffold using hyaluronic acid and polycaprolactone to provide ideal conditions for nerve regeneration by applying proper physicochemical and mechanical signals. Chemical and mechanical properties of pure PCL and PCL/HA nanofibrous scaffolds were measured by FTIR and tensile test. Morphology, swelling behavior, and biodegradability of the scaffolds were evaluated too. Porosity of various layers of scaffolds was measured by image analysis method. To assess the cell-scaffold interaction, SH-SY5Y human neuroblastoma cell line were cultured on the electrospun scaffolds. Taken together, these results suggest that the blended nanofibrous scaffolds PCL/HA 95:5 exhibit the most balanced properties to meet all of the required specifications for neural cells and have potential application in neural tissue engineering. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. 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.

  17. Modeling of porous scaffold deformation induced by medium perfusion.

    PubMed

    Podichetty, Jagdeep T; Madihally, Sundararajan V

    2014-05-01

    In this study, we tested the possibility of calculating permeability of porous scaffolds utilized in soft tissue engineering using pore size and shape. We validated the results using experimental measured pressure drop and simulations with the inclusion of structural deformation. We prepared Polycaprolactone (PCL) and Chitosan-Gelatin (CG) scaffolds by salt leaching and freeze drying technique, respectively. Micrographs were assessed for pore characteristics and mechanical properties. Porosity for both scaffolds was nearly same but the permeability varied 10-fold. Elastic moduli were 600 and 9 kPa for PCL and CG scaffolds, respectively, while Poisson's ratio was 0.3 for PCL scaffolds and ∼1.0 for CG scaffolds. A flow-through bioreactor accommodating a 10 cm diameter and 0.2 cm thick scaffold was used to determine the pressure-drop at various flow rates. Additionally, computational fluid dynamic (CFD) simulations were performed by coupling fluid flow, described by Brinkman equation, with structural mechanics using a dynamic mesh. The experimentally obtained pressure drop matched the simulation results of PCL scaffolds. Simulations were extended to a broad range of permeabilities (10(-10) m(2) to 10(-14) m(2) ), elastic moduli (10-100,000 kPa) and Poisson's ratio (0.1-0.49). The results showed significant deviation in pressure drop due to scaffold deformation compared to rigid scaffold at permeabilities near healthy tissues. Also, considering the scaffold as a nonrigid structure altered the shear stress profile. In summary, scaffold permeability can be calculated using scaffold pore characteristics and deformation could be predicted using CFD simulation. These relationships could potentially be used in monitoring tissue regeneration noninvasively via pressure drop. Copyright © 2013 Wiley Periodicals, Inc.

  18. Establishment of a three-dimensional culture system of gastric stem cells supporting mucous cell differentiation using microfibrous polycaprolactone scaffolds.

    PubMed

    Pulikkot, S; Greish, Y E; Mourad, A-H I; Karam, S M

    2014-12-01

    To generate various polycaprolactone (PCL) scaffolds and test their suitability for growth and differentiation of immortalized mouse gastric stem (mGS) cells. Non-porous, microporous and three-dimensional electrospun microfibrous PCL scaffolds were prepared and characterized for culture of mGS cells. First, growth of mGS cells was compared on these different scaffolds after 3 days culture, using viability assay and microscopy. Secondly, growth pattern of the cells on microfibrous scaffolds was studied after 3, 6, 9 and 12 days culture using DNA PicoGreen assay and scanning electron microscopy. Thirdly, differentiation of the cells grown on microfibrous scaffolds for 3 and 9 days was analysed using lectin/immunohistochemistry. The mGS cells grew preferentially on microfibrous scaffolds. From 3 to 6 days, there was increase in cell number, followed by reduction by days 9 and 12. To test whether the reduction in cell number was associated with cell differentiation, cryosections of cell-containing scaffolds cultured for 3 and 9 days were probed with gastric epithelial cell differentiation markers. On day 3, none of the markers examined bound to the cells. However by day 9, approximately, 50% of them bound to N-acetyl-d-glucosamine-specific lectin and anti-trefoil factor 2 antibodies, indicating their differentiation into glandular mucus-secreting cells. Microfibrous PCL scaffolds supported growth and differentiation of mGS cells into mucus-secreting cells. These data will help lay groundwork for future experiments to explore use of gastric stem cells and PCL scaffolds in stomach tissue engineering. © 2014 John Wiley & Sons Ltd.

  19. Fibronectin adsorption on functionalized electrospun polycaprolactone scaffolds: experimental and molecular dynamics studies.

    PubMed

    Regis, Shawn; Youssefian, Sina; Jassal, Manisha; Phaneuf, Matthew D; Rahbar, Nima; Bhowmick, Sankha

    2014-06-01

    Designing scaffolds to modulate protein adsorption is a key to building advanced scaffolds for tissue regeneration. Protein adsorption to tissue engineering scaffolds is critical in early cell attachment, survival, and eventual proliferation. The goal of this study is to examine the effect of functionalization on fibronectin adsorption to electrospun polycaprolactone (PCL) scaffolds through experimentation using fluorescently labeled fibronectin and to couple this experimental data with analysis of interaction energies obtained through molecular dynamics (MD) simulations to develop a better understanding of the adsorption process. This study is the first to analyze and compare experimental and MD simulation results of fibronectin adsorption on functionalized electrospun PCL scaffolds. Electrospun nanofiber PCL scaffolds were treated with either 1 N NaOH (hydrolyzed) or 46% hexamethylenediamine (HMD) (aminated) solution to be compared with untreated (control) scaffolds. We found that aminated PCL scaffolds experimentally adsorbed more fibronectin than control scaffolds, whereas hydrolyzed scaffolds showed decreased adsorption. MD simulations carried out with NVT ensemble at a temperature of 310 K indicated a higher work of adhesion for both functionalized scaffolds over control. Also, the simulations revealed different conformations of fibronectin on each scaffold type after adsorption, with the arginine-glycine-aspartic acid sequence appearing most accessible on the aminated scaffolds. This suggests that functionalization affects not only the quantity of protein that will adsorb on a scaffold but how it attaches as well, which could affect subsequent cell attachment.

  20. The Proliferation Study of Hips Cell-Derived Neuronal Progenitors on Poly-Caprolactone Scaffold

    PubMed Central

    Havasi, Parvaneh; Soleimani, Masoud; Morovvati, Hassan; Bakhshandeh, Behnaz; Nabiuni, Mohammad

    2014-01-01

    Introduction The native inability of nervous system to regenerate, encourage researchers to consider neural tissue engineering as a potential treatment for spinal cord injuries. Considering the suitable characteristics of induced pluripotent stem cells (iPSCs) for tissue regeneration applications, in this study we investigated the adhesion, viability and proliferation of neural progenitors (derived from human iPSCs) on aligned poly-caprolactone (PCL) nanofibers. Methods Aligned poly-caprolactone nanofibrous scaffold was fabricated by electrospinning and characterized by scanning electron microscopy (SEM). Through neural induction, neural progenitor cells were derived from induced pluripotent stem cells. After cell seeding on the scaffolds, their proliferation was investigated on different days of culture. Results According to the SEM micrographs, the electrospun PCL scaffolds were aligned along with uniformed morphology. Evaluation of adhesion and viability of neural progenitor cells on plate (control) and PCL scaffold illustrated increasing trends in proliferation but this rate was higher in scaffold group. The statistical analyses confirmed significant differences between groups on 36h and 48h. Discussion Evaluation of cell proliferation along with morphological assessments, staining and SEM finding suggested biocompatibility of the PCL scaffolds and suitability of the combination of the mentioned scaffold and human iPS cells for neural regeneration. PMID:25337369

  1. Three-dimensional polycaprolactone hierarchical scaffolds supplemented with natural biomaterials to enhance mesenchymal stem cell proliferation.

    PubMed

    Yoon, Hyeon; Ahn, Seunghyun; Kim, Geunhyung

    2009-10-01

    A hybrid technology that combines a three-dimensional (3-D) dispensing system with an electrospinning process was used to produce a hierarchical 3-D scaffold consisting of micro-sized polycaprolactone (PCL) strands and micro/nano-sized fibres. The micro/nanofibre biocomposites electrospun with PCL/small intestine submucosa (SIS) and PCL/Silk fibroin were layered between melt-plotted micro-strands. The scaffold containing SIS exhibited a stronger hydrophilic property than other scaffolds due to the various hydrophilic components in SIS. The 3-D hierarchical scaffold having biocomposites exhibited an incredibly enhanced initial cell attachment and proliferation of bone marrow-derived mesenchymal stem cells relative to the normally designed 3-D scaffold.

  2. Mechanical properties and cell-culture characteristics of a polycaprolactone kagome-structure scaffold fabricated by a precision extruding deposition system.

    PubMed

    Lee, Se-Hwan; Cho, Yong Sang; Hong, Myoung Wha; Lee, Bu-Kyu; Park, Yongdoo; Park, Sang-Hyug; Kim, Young Yul; Cho, Young-Sam

    2017-09-13

    To enhance the mechanical properties of three-dimensional (3D) scaffolds used for bone regeneration in tissue engineering, many researchers have studied their structure and chemistry. In the structural engineering field, the kagome structure has been known to have an excellent relative strength. In this study, to enhance the mechanical properties of a synthetic polymer scaffold used for tissue engineering, we applied the 3D kagome structure to a porous scaffold for bone regeneration. Prior to fabricating the biocompatible-polymer scaffold, the ideal kagome structure, which was manufactured by a 3D printer of the digital light processing type, was compared with a grid-structure, which was used as the control group, using a compressive experiment. A polycaprolactone (PCL) kagome-structure scaffold was successfully fabricated by additive manufacturing using a 3D printer with a precision extruding deposition head. To assess the physical characteristics of the fabricated PCL-kagome-structure scaffold, we analyzed its porosity, pore size, morphological structure, surface roughness, compressive stiffness, and mechanical bending properties. The results showed that, the mechanical properties of proposed kagome-structure scaffold were superior to those of a grid-structure scaffold. Moreover, Sarcoma osteogenic (Saos-2) cells were used to evaluate the characteristics of in vitro cell proliferation. We carried out cell counting kit-8 (CCK-8) and DNA contents assays. Consequently, the cell proliferation of the kagome-structure scaffold was increased; this could be because the surface roughness of the kagome-structure scaffold enhances initial cell attachment.

  3. Preparation and characterization of bioactive mesoporous wollastonite - Polycaprolactone composite scaffold.

    PubMed

    Wei, Jie; Chen, Fangping; Shin, Jung-Woog; Hong, Hua; Dai, Chenglong; Su, Jiancan; Liu, Changsheng

    2009-02-01

    A well-defined mesoporous structure of wollastonite with high specific surface area was synthesized using surfactant P123 (triblock copolymer) as template, and its composite scaffolds with poly(epsilon-caprolactone) (PCL) were fabricated by a simple method of solvent casting-particulate leaching. The measurements of the water contact angles suggest that the incorporation of either mesoporous wollastonite (m-WS) or conventional wollastonite (c-WS) into PCL could improve the hydrophilicity of the composites, and the former was more effective than the later. The bioactivity of the composite scaffold was evaluated by soaking the scaffolds in a simulated body fluid (SBF) and the results show that the m-WS/PCL composite (m-WPC) scaffolds can induce a dense and continuous layer of apatite after soaking for 1 week, as compared with the scattered and discrete apatite particles on the c-WS/PCL composite (c-WPC) scaffolds. The m-WPC had a significantly enhanced apatite-forming bioactivity compared with the c-WPC owing to the high specific surface area and pore volume of m-WS. In addition, attachment and proliferation of MG(63) cells on m-WPC scaffolds were significantly higher than that of c-WPC, revealing that m-WPC scaffolds had excellent biocompatibility. Such improved properties of m-WPC should be helpful for developing new biomaterials and may have potential use in hard tissue repair.

  4. 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.

  5. Antimicrobial effects of nanofiber poly(caprolactone) tissue scaffolds releasing rifampicin.

    PubMed

    Ruckh, Timothy T; Oldinski, Rachael A; Carroll, Derek A; Mikhova, Krasimira; Bryers, James D; Popat, Ketul C

    2012-06-01

    This study quantified the antibiotic release kinetics and subsequent bactericidal efficacy of rifampicin (RIF) against Gram-positive and Gram-negative bacteria under in vitro static conditions. Antibiotic-loaded scaffolds were fabricated by electrospinning poly(caprolactone) (PCL) with 10% or 20% (w/w) RIF. Scaffold fiber diameter and RIF loading were characterized, and RIF release kinetics were measured. RIF-releasing and RIF-free scaffolds were inoculated with Pseudomonas aeruginosa and Staphylococcus epidermidis, and the suspended concentration live and dead bacteria were determined by fluorescent microscopy. Adherent bacteria and biofilm formation were examined using scanning electron microscopy. Mean fiber diameters were 557 ± 399 nm for RIF-free, 402 ± 225 nm for 10% RIF, and 665 ± 402 nm for 20% RIF scaffolds. RIF release kinetics exhibited a short-burst release during the first hour, followed by a 7 h, zero-order release during which both RIF scaffolds released ~50% of their initial RIF mass loading. P. aeruginosa and S. epidermidis suspended cell populations proliferated in accordance with logarithmic growth models when exposed to control scaffolds; however both RIF-containing scaffolds completely inhibited bacterial growth in suspension and, subsequently, prevented biofilm formation within the scaffolds through the first 6 h.

  6. The effects of Biodentine/polycaprolactone three-dimensional-scaffold with odontogenesis properties on human dental pulp cells.

    PubMed

    Ho, C-C; Fang, H-Y; Wang, B; Huang, T-H; Shie, M-Y

    2017-06-20

    To determine the feasibility of using three-dimensional printed Biodentine/polycaprolactone composite scaffolds for orthopaedic and dental applications. The physicochemical properties and the odontogenic differentiation of human dental pulp cells (hDPCs) were investigated. Biodentine was well-suspended in ethanol and dropped slowly into molten polycaprolactone with vigorous stirring. The Biodentine/polycaprolactone composite scaffolds were then fabricated into controlled macropore sizes and structures using an extrusion-based three-dimensional (3D) printer. The mechanical properties, bioactivity, and the proliferation and odontogenic differentiation of human dental pulp cells (hDPCs) cultured on the scaffolds were evaluated. Biodentine/polycaprolactone scaffolds had uniform macropores 550 μm in size with established interconnections and a compressive strength of 6.5 MPa. In addition, the composite scaffolds exhibited a good apatite-forming ability and were capable of supporting the proliferation and differentiation of hDPCs. The composite scaffolds fabricated by an extrusion-based 3D printing technique had similar characteristics to Biodentine cement, including bioactivity and the ability to promote the differentiation of hDPCs. These results indicate that the composite scaffold would be a candidate for dental and bone regeneration. © 2017 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  7. Evaluation of nanofibrous scaffolds obtained from blends of chitosan, gelatin and polycaprolactone for skin tissue engineering.

    PubMed

    Gomes, Susana; Rodrigues, Gabriela; Martins, Gabriel; Henriques, Célia; Silva, Jorge Carvalho

    2017-09-01

    Polymer blending is a strategy commonly used to obtain hybrid materials possessing properties better than those of the individual constituents regarding their use in scaffolds for Tissue Engineering. In the present work, the scaffolds produced by electrospinning solutions of polymeric blends obtained using a polyester (polycaprolactone, PCL), a polysaccharide (chitosan, CS) and a protein (gelatin extracted from cold water fish skin, GEL), were investigated. Solutions conductivity, shear viscosity and surface tension were determined. GEL-containing scaffolds were crosslinked with vapour phase glutaraldehyde (GTA). The scaffolds were characterized physico-chemically regarding fibre morphology, porosity, water contact angle, mechanical properties, chemical bonds and fibre and dimensional stability upon immersion in water and cell culture medium. The scaffolds were further tested in vitro for cell adhesion, growth and morphology of human foetal fibroblasts (cell line HFFF2). Results show that the nanofibrous scaffolds are hydrophilic and display the typical porosity of non-woven fibre mats. The CS/PCL and CS/PCL/GEL scaffolds have the highest elastic modulus (48MPa). Dimensional stability is best for the CS/PCL/GEL scaffolds. FTIR spectra confirm the occurrence of cross-linking reactions of GTA with both GEL and CS. Cell adhesion ratio ranked from excellent (close to 100%) to satisfactory (around 50%) in the order PCL/GEL>CS/GEL>CS/PCL/GEL>CS/PCL. Cell populations show an extended lag phase in comparison with the controls but cell proliferation occurs on all scaffolds until confluence is reached. In conclusion, all scaffolds studied possess characteristics that enable them to be used in skin tissue engineering but the CS/PCL/GEL scaffolds have better physical properties whereas the PCL/GEL scaffolds support a higher cell adhesion. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Mechanical properties and cell cultural response of polycaprolactone scaffolds designed and fabricated via fused deposition modeling.

    PubMed

    Hutmacher, D W; Schantz, T; Zein, I; Ng, K W; Teoh, S H; Tan, K C

    2001-05-01

    A number of different processing techniques have been developed to design and fabricate three-dimensional (3D) scaffolds for tissue-engineering applications. The imperfection of the current techniques has encouraged the use of a rapid prototyping technology known as fused deposition modeling (FDM). Our results show that FDM allows the design and fabrication of highly reproducible bioresorbable 3D scaffolds with a fully interconnected pore network. The mechanical properties and in vitro biocompatibility of polycaprolactone scaffolds with a porosity of 61 +/- 1% and two matrix architectures were studied. The honeycomb-like pores had a size falling within the range of 360 x 430 x 620 microm. The scaffolds with a 0/60/120 degrees lay-down pattern had a compressive stiffness and a 1% offset yield strength in air of 41.9 +/- 3.5 and 3.1 +/- 0.1 MPa, respectively, and a compressive stiffness and a 1% offset yield strength in simulated physiological conditions (a saline solution at 37 degrees C) of 29.4 +/- 4.0 and 2.3 +/- 0.2 MPa, respectively. In comparison, the scaffolds with a 0/72/144/36/108 degrees lay-down pattern had a compressive stiffness and a 1% offset yield strength in air of 20.2 +/- 1.7 and 2.4 +/- 0.1 MPa, respectively, and a compressive stiffness and a 1% offset yield strength in simulated physiological conditions (a saline solution at 37 degrees C) of 21.5 +/- 2.9 and 2.0 +/- 0.2 MPa, respectively. Statistical analysis confirmed that the five-angle scaffolds had significantly lower stiffness and 1% offset yield strengths under compression loading than those with a three-angle pattern under both testing conditions (p < or = 0.05). The obtained stress-strain curves for both scaffold architectures demonstrate the typical behavior of a honeycomb structure undergoing deformation. In vitro studies were conducted with primary human fibroblasts and periosteal cells. Light, environmental scanning electron, and confocal laser microscopy as well as

  9. Fabrication of chitosan-polycaprolactone composite nanofibrous scaffold for simultaneous delivery of ferulic acid and resveratrol.

    PubMed

    Poornima, Balan; Korrapati, Purna Sai

    2017-02-10

    Wound healing is a complex cellular process involving various mechanisms making it intricate for designing a scaffold for therapeutic use. This study deals with the designing and development of coaxial electrospun Chitosan-Polycaprolactone nanofiber wound dressings for efficient simultaneous drug delivery. Ferulic acid and resveratrol were chosen for incorporation into core-shell nanofibers for their efficacy in anti-inflammatory and pro-angiogenic activities respectively. Structural and physico-chemical characterization of the scaffold confirmed the encapsulation of both the molecules. The in vitro release studies showed sustained release of both resveratrol and ferulic acid up to 48% and 55% respectively till 120h. Functional characterization of the nanofibrous wound dressing exhibited good in vitro and in vivo biocompatibility. The scaffold treated rats exhibited complete healing in 15days in comparison to the controls that healed in 20days. The study therefore, opens up venues for designing sequential and sustained drug delivery systems in wound therapeutics.

  10. 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.

  11. Formation of porous HPCL/LPCL/HA scaffolds with supercritical CO2 gas foaming method.

    PubMed

    Moghadam, M Zahedi; Hassanajili, Sh; Esmaeilzadeh, F; Ayatollahi, M; Ahmadi, M

    2017-05-01

    Scaffold is a 3D porous structure that is made of different materials, such as synthetic and natural polymers. It plays the role of a synthetic extracellular matrix and permits adhesion, proliferation and differentiation of the cells. Porosity and pore size are the important factors for any 3D scaffold used in bone tissue engineering. In this study, porous scaffolds were prepared by adding hydroxyapatite (HA) nanoparticles as filler to the polymeric matrix of polycaprolactone (PCL) blends with two different molecular weight by using supercritical CO2 (ScCO2) foaming method. The effect of different parameters such as CO2 pressure, ratios of the polymers and amount of the filler on the scaffold properties was investigated. The results showed that porosity increased with increment of pressure and decreased with increasing the ratio of the high molecular weight PCL to the low molecular weight PCL in the scaffolds and also HA content. Optimum condition for obtaining adequate porous scaffold of HPCL/LPCL/HA occurred at 140bar and 45°C. The physical and mechanical properties of the prepared scaffolds were characterized using DSC, XRD, FTIR, SEM, contact angle and compression test. By analyzing the results of these tests, optimum sample for cell culture was selected. The biocompatibility of the selected HPCL/LPCL/HA scaffold (HPCL/LPCL 60/40 containing 2.5% HA) was assessed in vitro by using human mesenchymal stem cells (hMSCs). Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Exploring the Potential of Starch/Polycaprolactone Aligned Magnetic Responsive Scaffolds for Tendon Regeneration.

    PubMed

    Gonçalves, Ana I; Rodrigues, Márcia T; Carvalho, Pedro P; Bañobre-López, Manuel; Paz, Elvira; Freitas, Paulo; Gomes, Manuela E

    2016-01-21

    The application of magnetic nanoparticles (MNPs) in tissue engineering (TE) approaches opens several new research possibilities in this field, enabling a new generation of multifunctional constructs for tissue regeneration. This study describes the development of sophisticated magnetic polymer scaffolds with aligned structural features aimed at applications in tendon tissue engineering (TTE). Tissue engineering magnetic scaffolds are prepared by incorporating iron oxide MNPs into a 3D structure of aligned SPCL (starch and polycaprolactone) fibers fabricated by rapid prototyping (RP) technology. The 3D architecture, composition, and magnetic properties are characterized. Furthermore, the effect of an externally applied magnetic field is investigated on the tenogenic differentiation of adipose stem cells (ASCs) cultured onto the developed magnetic scaffolds, demonstrating that ASCs undergo tenogenic differentiation synthesizing a Tenascin C and Collagen type I rich matrix under magneto-stimulation conditions. Finally, the developed magnetic scaffolds were implanted in an ectopic rat model, evidencing good biocompatibility and integration within the surrounding tissues. Together, these results suggest that the effect of the magnetic aligned scaffolds structure combined with magnetic stimulation has a significant potential to impact the field of tendon tissue engineering toward the development of more efficient regeneration therapies.

  13. Artificial neural network for modeling the elastic modulus of electrospun polycaprolactone/gelatin scaffolds.

    PubMed

    Vatankhah, Elham; Semnani, Dariush; Prabhakaran, Molamma P; Tadayon, Mahdi; Razavi, Shahnaz; Ramakrishna, Seeram

    2014-02-01

    Scaffolds for tissue engineering (TE) require the consideration of multiple aspects, including polymeric composition and the structure and mechanical properties of the scaffolds, in order to mimic the native extracellular matrix of the tissue. Electrospun fibers are frequently utilized in TE due to their tunable physical, chemical, and mechanical properties and porosity. The mechanical properties of electrospun scaffolds made from specific polymers are highly dependent on the processing parameters, which can therefore be tuned for particular applications. Fiber diameter and orientation along with polymeric composition are the major factors that determine the elastic modulus of electrospun nano- and microfibers. Here we have developed a neural network model to investigate the simultaneous effects of composition, fiber diameter and fiber orientation of electrospun polycaprolactone/gelatin mats on the elastic modulus of the scaffolds under ambient and simulated physiological conditions. The model generated might assist bioengineers to fabricate electrospun scaffolds with defined fiber diameters, orientations and constituents, thereby replicating the mechanical properties of the native target tissue.

  14. Plasma Surface Modification for Immobilization of Bone Morphogenic Protein-2 on Polycaprolactone Scaffolds

    NASA Astrophysics Data System (ADS)

    Kim, Byung Hoon; Myung, Sung Woon; Jung, Sang Chul; Ko, Yeong Mu

    2013-11-01

    The immobilization of recombinant human bone formation protein-2 (rhBMP-2) on polycaprolactone (PCL) scaffolds was performed by plasma polymerization. RhBMP-2, which induces osteoblast differentiation in various cell types, is a growth factor that plays an important role in bone formation and repair. The surface of the PCL scaffold was functionalized with the carboxyl groups of plasma-polymerized acrylic acid (PPAA) thin films. Plasma polymerization was carried out at a discharge power of 60 W at an acrylic acid flow rate of 7 sccm for 5 min. The PPAA thin film exhibited moderate hydrophilic properties and possessed a high density of carboxyl groups. Carboxyl groups and rhBMP-2 on the PCL scaffolds surface were identified by attenuated total reflection Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy, respectively. The alkaline phosphatase activity assay showed that the rhBMP-2 immobilized PCL scaffold increased the level of MG-63 cell differentiation. Plasma surface modification for the preparation of biomaterials, such as biofunctionalized polymer scaffolds, can be used for the binding of bioactive molecules in tissue engineering.

  15. Osteogenesis of adipose-derived stem cells on polycaprolactone-β-tricalcium phosphate scaffold fabricated via selective laser sintering and surface coating with collagen type I.

    PubMed

    Liao, Han-Tsung; Lee, Ming-Yih; Tsai, Wen-Wei; Wang, Hsiu-Chen; Lu, Wei-Chieh

    2016-10-01

    The current study aimed to fabricate three-dimensional (3D) polycaprolactone (PCL), polycaprolactone and β-tricalcium phosphate (PCL-TCP) scaffolds via a selective laser-sintering technique (SLS). Collagen type I was further coated onto PCL-TCP scaffolds to form PCL-TCP-COL scaffolds. The physical characters of these three scaffolds were analysed. The osteogenic potential of porcine adipose-derived stem cells (pASCs) was compared among these three scaffolds in order to find an optimal scaffold for bone tissue engineering. The experimental results showed no significant differences in pore size and porosity among the three scaffolds; the porosity was ca. 75-77% and the pore size was ca. 300-500 µm in all three. The compressive modulus was increased from 6.77 ± 0.19 to 13.66 ± 0.19 MPa by adding 30% β-TCP into a 70% PCL scaffold. No significant increase of mechanical strength was found by surface-coating with collagen type I. Hydrophilicity and swelling ratios showed statistical elevation (p < 0.05) after collagen type I was coated onto the PCL-TCP scaffolds. The in vitro study demonstrated that pASCs had the best osteogenic differentiation on PCL-TCP-COL group scaffolds, due to the highest ALP activity, osteocalcin mRNA expression and mineralization. A nude mice experiment showed better woven bone and vascular tissue formation in the PCL-TCP-COL group than in the PCL group. In conclusion, the study demonstrated the ability to fabricate 3D, porous PCL-TCP composite scaffolds (PCL:TCP = 70:30 by weight) via an in-house-built SLS technique. In addition, the osteogenic ability of pASCs was found to be enhanced by coating COL onto the PCL-TCP scaffolds, both in vitro and in vivo. Copyright © 2013 John Wiley & Sons, Ltd.

  16. Three-dimensional electrospun polycaprolactone (PCL)/alginate hybrid composite scaffolds.

    PubMed

    Kim, Min Seong; Kim, GeunHyung

    2014-12-19

    Micro/nanofibrous scaffolds have been used widely in biomedical applications because the micro/nano-scale fibres resemble natural extracellular matrix and the high surface-to-volume ratio encourages cellular activities (attachment and proliferation). However, poor mechanical properties, low controllability of various shapes and difficulties in obtaining controllable pore structure have been obstacles to their use in hard-tissue regeneration. To overcome these shortcomings, we suggest a new composite system, which uses a combination method of wet electrospinning, rapid prototyping and a physical punching process. Using the process, we obtained polycaprolactone (PCL)/alginate composite scaffolds, consisting of electrospun PCL/alginate fibres and micro-sized PCL struts, with mean pore sizes of 821 ± 55 μm. To show the feasibility of the scaffolds for hard-tissue regeneration, the scaffolds were assessed not only for physical properties, including hydrophilicity, water absorption, and tensile and compressive strength, but also in vitro cellular responses (cell viability and proliferation) and osteogenic differentiation (alkaline phosphatase (ALP) activity, and mineralisation) by culturing with pre-osteoblasts (MC3T3-E1 cells). With the reinforcing micro-sized PCL struts, the elastic modulus of the PCL/alginate scaffold was significantly improved versus a pure PCL scaffold. Additionally, due to the alginate component in the fibrous scaffold, they showed significantly enhanced hydrophilic behaviour, water absorption (∼8-fold) and significant biological activities (∼1.6-fold for cell viability at 7 days, ∼2.3-fold for ALP activity at 14 days and ∼6.4-fold for calcium mineralisation at 14 days) compared with those of a pure PCL fibrous scaffold.

  17. 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.

  18. Polycaprolactone-Coated 3D Printed Tricalcium Phosphate Scaffolds for Bone Tissue Engineering: In Vitro Alendronate Release Behavior and Local Delivery Effect on In Vivo Osteogenesis

    PubMed Central

    2015-01-01

    The aim of this work was to evaluate the effect of in vitro alendronate (AD) release behavior through polycaprolactone (PCL) coating on in vivo bone formation using PCL-coated 3D printed interconnected porous tricalcium phosphate (TCP) scaffolds. Higher AD and Ca2+ ion release was observed at lower pH (5.0) than that at higher pH (7.4). AD and Ca2+ release, surface morphology, and phase analysis after release indicated a matrix degradation dominated AD release caused by TCP dissolution. PCL coating showed its effectiveness for controlled and sustained AD release. Six different scaffold compositions, namely, (i) TCP (bare TCP), (ii) TCP + AD (AD-coated TCP), (iii) TCP + PCL (PCL-coated TCP), (iv) TCP + PCL + AD, (v) TCP + AD + PCL, and (vi) TCP + AD + PCL + AD were tested in the distal femoral defect of Sprague–Dawley rats for 6 and 10 weeks. An excellent bone formation inside the micro and macro pores of the scaffolds was observed from histomorphology. Histomorphometric analysis revealed maximum new bone formation in TCP + AD + PCL scaffolds after 6 weeks. No adverse effect of PCL on bioactivity of TCP and in vivo bone formation was observed. All scaffolds with AD showed higher bone formation and reduced TRAP (tartrate resistant acid phosphatase) positive cells activity compared to bare TCP and TCP coated with only PCL. Bare TCP scaffolds showed the highest TRAP positive cells activity followed by TCP + PCL scaffolds, whereas TCP + AD scaffolds showed the lowest TRAP activity. A higher TRAP positive cells activity was observed in TCP + AD + PCL compared to TCP + AD scaffolds after 6 weeks. Our results show that in vivo local AD delivery from PCL-coated 3DP TCP scaffolds could further induce increased early bone formation. PMID:24826838

  19. Bioinspired Strong and Highly Porous Glass Scaffolds.

    PubMed

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

    2011-03-22

    The quest for more efficient energy-related technologies is driving the development of porous and high-performance structural materials with exceptional mechanical strength. Natural materials achieve their strength through complex hierarchical designs and anisotropic structures that are extremely difficult to replicate synthetically. We emulate nature's design by direct-ink-write assembling of glass scaffolds with a periodic pattern, and controlled sintering of the filaments into anisotropic constructs similar to biological materials. The final product is a porous glass scaffold with a compressive strength (136 MPa) comparable to that of cortical bone and a porosity (60%) comparable to that of trabecular bone. The strength of this porous glass scaffold is ~100 times that of polymer scaffolds and 4-5 times that of ceramic and glass scaffolds with comparable porosities reported elsewhere. The ability to create both porous and strong structures opens a new avenue for fabricating scaffolds for a broad array of applications, including tissue engineering, filtration, lightweight composites, and catalyst support.

  20. 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.

  1. 3D Printed Polycaprolactone Carbon Nanotube Composite Scaffolds for Cardiac Tissue Engineering.

    PubMed

    Ho, Chee Meng Benjamin; Mishra, Abhinay; Lin, Pearlyn Teo Pei; Ng, Sum Huan; Yeong, Wai Yee; Kim, Young-Jin; Yoon, Yong-Jin

    2017-04-01

    Fabrication of tissue engineering scaffolds with the use of novel 3D printing has gained lot of attention, however systematic investigation of biomaterials for 3D printing have not been widely explored. In this report, well-defined structures of polycaprolactone (PCL) and PCL- carbon nanotube (PCL-CNT) composite scaffolds have been designed and fabricated using a 3D printer. Conditions for 3D printing has been optimized while the effects of varying CNT percentages with PCL matrix on the thermal, mechanical and biological properties of the printed scaffolds are studied. Raman spectroscopy is used to characterise the functionalized CNTs and its interactions with PCL matrix. Mechanical properties of the composites are characterised using nanoindentation. Maximum peak load, elastic modulus and hardness increases with increasing CNT content. Differential scanning calorimetry (DSC) studies reveal the thermal and crystalline behaviour of PCL and its CNT composites. Biodegradation studies are performed in Pseudomonas Lipase enzymatic media, showing its specificity and effect on degradation rate. Cell imaging and viability studies of H9c2 cells from rat origin on the scaffolds are performed using fluorescence imaging and MTT assay, respectively. PCL and its CNT composites are able to show cell proliferation and have the potential to be used in cardiac tissue engineering.

  2. Surface plasma treatment of poly(caprolactone) micro, nano, and multiscale fibrous scaffolds for enhanced osteoconductivity.

    PubMed

    Sankar, Deepthi; Shalumon, K T; Chennazhi, K P; Menon, Deepthy; Jayakumar, R

    2014-06-01

    In this study, poly(caprolactone) (PCL) was electrospun to nano, micro, and multiscale (micro-nano) fibers, which were then subjected to low pressure argon and nitrogen plasma treatment. The electrospun fibers contain microfibers of diameter 8-10 μm and nanofibers of diameter 200-300 nm. Characterization of the plasma-treated fibers showed that treatment using less oxidizing gas like nitrogen and inert gas like argon functionalize the surface with polar groups that significantly modify the properties of the scaffold. Highly hydrophobic PCL fibrous scaffolds were rendered hydrophilic, with significantly improved biomineralization after the plasma treatment. While plasma treatment on micro and multiscale fibers enhanced their protein adsorption, cell attachment, spreading, elongation, and proliferation, nanofibers showed remarkably improved cell attachment. The applicability of plasma-treated electrospun fibers for differentiation of mesenchymal stem cell toward osteogenic lineage was also studied. Accelerated differentiation toward osteoblast lineage, with maximum alkaline phosphatase (ALP) activity in 14 days was achieved in plasma-treated fibers. Another remarkable outcome was the enhanced ALP activity of the microfibers after plasma treatment, compared with multiscale and nanofibers. Alizarin red staining further confirmed the mineralization of the plasma-treated scaffolds, indicative of maturation of the differentiated cells. This work thus concentrates on harnessing the potential of plasma treatment, for improving the osteoconductivity of fibrous scaffolds, which could be used for bone tissue engineering/regenerative medicine.

  3. 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.

  4. Functionalization of Polycaprolactone Scaffolds with Hyaluronic Acid and β-TCP Facilitates Migration and Osteogenic Differentiation of Human Dental Pulp Stem Cells In Vitro

    PubMed Central

    Kraft, David Christian Evar; Lysdahl, Helle; Foldager, Casper Bindzus; Chen, Muwan; Kristiansen, Asger Albæk; Rölfing, Jan Hendrik Duedal; Bünger, Cody Eric

    2015-01-01

    In this study, we sought to assess the osteogenic potential of human dental pulp stem cells (DPSCs) on three different polycaprolactone (PCL) scaffolds. The backbone structure of the scaffolds was manufactured by fused deposition modeling (PCL scaffold). The composition and morphology was functionalized in two of the scaffolds. The first underwent thermal induced phase separation of PCL infused into the pores of the PCL scaffold. This procedure resulted in a highly variable micro- and nanostructured porous (NSP), interconnected, and isotropic tubular morphology (NSP-PCL scaffold). The second scaffold type was functionalized by dip-coating the PCL scaffold with a mixture of hyaluronic acid and β-TCP (HT-PCL scaffold). The scaffolds were cylindrical and measured 5 mm in height and 10 mm in diameter. They were seeded with 1×106 human DPSCs, a cell type known to express bone-related markers, differentiate into osteoblasts-like cells, and to produce a mineralized bone-like extracellular matrix. DPSCs were phenotypically characterized by flow cytometry for CD90+, CD73+, CD105+, and CD14−. DNA, ALP, and Ca2+ assays and real-time quantitative polymerase chain reaction for genes involved in osteogenic differentiation were analyzed on day 1, 7, 14, and 21. Cell viability and distribution were assessed on day 1, 7, 14, and 21 by fluorescent-, scanning electron-, and confocal microscopy. The results revealed that the DPSCs expressed relevant gene expression consistent with osteogenic differentiation. The NSP-PCL and HT-PCL scaffolds promoted osteogenic differentiation and Ca2+ deposition after 21 days of cultivation. Different gene expressions associated with mature osteoblasts were upregulated in these two scaffold types, suggesting that the methods in which the scaffolds promote osteogenic differentiation, depends on functionalization approaches. However, only the HT-PCL scaffold was also able to support cell proliferation and cell migration resulting in even cell

  5. Development of Composite Porous Scaffolds Based on Collagen and Biodegradable Poly(ester urethane)urea

    PubMed Central

    Guan, Jianjun; Stankus, John J.; Wagner, William R.

    2010-01-01

    Our objective in this work was to develop a flexible, biodegradable scaffold for cell transplantation that would incorporate a synthetic component for strength and flexibility and type I collagen for enzymatic lability and cytocompatibility. A biodegradable poly(ester urethane)urea was synthesized from poly(caprolactone), 1,4-diisocyanatobutane, and putrescine. Using a thermally induced phase separation process, porous scaffolds were created from a mixture containing this polyurethane and 0%, 10%, 20%, or 30% type I collagen. The resulting scaffolds were found to have open, interconnected pores (from 7 to >100 um) and porosities from 58% to 86% depending on the polyurethane/collagen ratio. The scaffolds were also flexible with breaking strains of 82–443% and tensile strengths of 0.97–4.11 MPa depending on preparation conditions. Scaffold degradation was significantly increased when collagenase was introduced into an incubating buffer in a manner that was dependent on the mass fraction of collagen present in the scaffold. Mass losses could be varied from 15% to 59% over 8 weeks. When culturing umbilical artery smooth muscle cells on these scaffolds higher cell numbers were observed over a 4-week culture period in scaffolds containing collagen. In summary, a strong and flexible scaffold system has been developed that can degrade by both hydrolysis and collagenase degradation pathways, as well as support cell growth. This scaffold possesses properties that would make it attractive for future use in soft tissue applications where such mechanical and biological features would be advantageous. PMID:16826792

  6. An experimental fatigue study of a porous scaffold for the regeneration of articular cartilage.

    PubMed

    Vikingsson, L; Gómez-Tejedor, J A; Gallego Ferrer, G; Gómez Ribelles, J L

    2015-05-01

    The aim of this experimental study is to predict the long-term mechanical behavior of a porous scaffold implanted in a cartilage defect for tissue engineering purpose. Fatigue studies were performed by up to 100,000 unconfined compression cycles in a polycaprolactone (PCL) scaffold with highly interconnected pores architecture. The scaffold compliance, stress-strain response and hysteresis energy have been measured after different number of fatigue cycles, while the morphology has been observed by scanning electron microscopy at the same fatigue times. To simulate the growing tissue in the scaffold/tissue construct, the scaffold was filled with an aqueous solution of polyvinyl alcohol (PVA) and subjected to repeating cycles of freezing and thawing that increase the hydrogel stiffness. Fatigue studies show that the mechanical loading provokes failure of the dry scaffold at a smaller number of deformation cycles than when it is immersed in water, and also that 100,000 compressive dynamic cycles do not affect the scaffold/gel construct. This shows the stability of the scaffold implanted in a chondral defect and gives a realistic simulation of the mechanical performance from implantation of the empty scaffold to regeneration of the new tissue inside the scaffold's pores.

  7. Engineering calcium deposits on polycaprolactone scaffolds for intravascular applications using primary human osteoblasts.

    PubMed

    Zhu, Beili; Bailey, Steven R; Mauli Agrawal, C

    2011-04-01

    Total atherosclerotic occlusions often include significant calcium deposits. Current animal models do not mimic the pathology of gradual occlusion of arteries and lack cell-mediated calcium. The primary goal of this project was to establish an animal model incorporating these features into chronic total occlusions, using biodegradable scaffolds. As the first step, this study sought to determine the optimal dosage of TGF-β1 on polycaprolactone (PCL) scaffolds cultured with primary human osteoblasts (HOBs) to effectively induce in vitro calcification. HOBs were cultured in TGF-β1 and dexamethsaone (Dex)-supplemented medium in well plates. Calcium in the cultures was visualized using alizarin red. The highest calcification was observed in groups with both TGF-β1 (0.02 ng/ml) and Dex (10(-10) M) in the medium. Next, HOBs were cultured on PCL scaffolds with different loadings of TGF-β1: 0 (control), 5, 10, 50 and 100 ng. These cultures were performed with or without Dex (10(-10) M) in the medium. DNA content, ALP activity and the amount and distribution of calcium were examined at 7, 14, 21 and 28 days. TGF-β1 appeared to have an inhibitory effect on scaffold calcification when grown in Dex-supplemented medium. When cultured without Dex, the lower amount of TGF-β1 loading (5 ng) showed the most calcification, high DNA synthesis and high ALP activity on scaffolds. This study demonstrates the potential of implanting a PCL-HOB construct in an animal artery to establish a model of atherosclerotic occlusion with calcification. Copyright © 2010 John Wiley & Sons, Ltd.

  8. 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.

  9. Nano/macro porous bioactive glass scaffold

    NASA Astrophysics Data System (ADS)

    Wang, Shaojie

    Bioactive glass (BG) and ceramics have been widely studied and developed as implants to replace hard tissues of the musculo-skeletal system, such as bones and teeth. Recently, instead of using bulk materials, which usually do not degrade rapidly enough and may remain in the human body for a long time, the idea of bioscaffold for tissue regeneration has generated much interest. An ideal bioscaffold is a porous material that would not only provide a three-dimensional structure for the regeneration of natural tissue, but also degrade gradually and, eventually be replaced by the natural tissue completely. Among various material choices the nano-macro dual porous BG appears as the most promising candidate for bioscaffold applications. Here macropores facilitate tissue growth while nanopores control degradation and enhance cell response. The surface area, which controls the degradation of scaffold can also be tuned by changing the nanopore size. However, fabrication of such 3D structure with desirable nano and macro pores has remained challenging. In this dissertation, sol-gel process combined with spinodal decomposition or polymer sponge replication method has been developed to fabricate the nano-macro porous BG scaffolds. Macropores up to 100microm are created by freezing polymer induced spinodal structure through sol-gel transition, while larger macropores (>200um) of predetermined size are obtained by the polymer sponge replication technique. The size of nanopores, which are inherent to the sol-gel method of glass fabrication, has been tailored using several approaches: Before gel point, small nanopores are generated using acid catalyst that leads to weakly-branched polymer-like network. On the other hand, larger nanopores are created with the base-catalyzed gel with highly-branched cluster-like structure. After the gel point, the nanostructure can be further modified by manipulating the sintering temperature and/or the ammonia concentration used in the solvent

  10. Evaluation of 3D-Printed Polycaprolactone Scaffolds Coated with Freeze-Dried Platelet-Rich Plasma for Bone Regeneration

    PubMed Central

    Li, Junda; Chen, Meilin; Wei, Xiaoying; Hao, Yishan; Wang, Jinming

    2017-01-01

    Three-dimensional printing is one of the most promising techniques for the manufacturing of scaffolds for bone tissue engineering. However, a pure scaffold is limited by its biological properties. Platelet-rich plasma (PRP) has been shown to have the potential to improve the osteogenic effect. In this study, we improved the biological properties of scaffolds by coating 3D-printed polycaprolactone (PCL) scaffolds with freeze-dried and traditionally prepared PRP, and we evaluated these scaffolds through in vitro and in vivo experiments. In vitro, we evaluated the interaction between dental pulp stem cells (DPSCs) and the scaffolds by measuring cell proliferation, alkaline phosphatase (ALP) activity, and osteogenic differentiation. The results showed that freeze-dried PRP significantly enhanced ALP activity and the mRNA expression levels of osteogenic genes (ALP, RUNX2 (runt-related gene-2), OCN (osteocalcin), OPN (osteopontin)) of DPSCs (p < 0.05). In vivo, 5 mm calvarial defects were created, and the PRP-PCL scaffolds were implanted. The data showed that compared with traditional PRP-PCL scaffolds or bare PCL scaffolds, the freeze-dried PRP-PCL scaffolds induced significantly greater bone formation (p < 0.05). All these data suggest that coating 3D-printed PCL scaffolds with freeze-dried PRP can promote greater osteogenic differentiation of DPSCs and induce more bone formation, which may have great potential in future clinical applications. PMID:28773189

  11. In vitro small intestinal epithelial cell growth on a nanocomposite polycaprolactone scaffold

    PubMed Central

    Gupta, Ashish; Vara, Dina S.; Punshon, Geoffrey; Sales, Kevin M.; Winslet, Marc C.; Seifalian, Alexander M.

    2009-01-01

    Tissue engineering of the small intestine remains experimental despite worldwide attempts to develop a functional substitute for short bowel syndrome. Most published studies have reported predominant use of PLLA (poly-L-lactide acid)/PGA (polyglycolic acid) copolymer as the scaffold material, and studies have been limited by in vivo experiments. This lack of progress has inspired a fresh perspective and provoked further investigation and development in this field of tissue engineering. In the present paper, we exploit a relatively new nanocomposite of POSS (polyhedral oligomeric silsesquioxane) and PCL [poly(caprolactone-urea)urethane] as a material to develop porous scaffolds using a solvent casting/particulate leaching technique to fabricate porous scaffolds in different pore sizes and porosities. Scaffolds were characterized for pore morphology and porosity using scanning electron microscopy and micro-computed tomography. Rat intestinal epithelial cells were then seeded on to the polymer scaffolds for an in vitro study of cell compatibility and proliferation, which was assessed by Alamar Blue™ and lactate dehydrogenase assays performed for 21 days post-seeding. The results obtained demonstrate that POSS–PCL nanocomposite was produced as a macroporous scaffold with porosity over the range of 40–80% and pore size over the range of 150–250 μm. This scaffold was shown to support epithelial cell proliferation and growth. In conclusion, as a further step in investigating small intestinal tissue engineering, the nanocomposite employed in this study may prove to be a useful alternative to poly(lactic-co-glycolic acid) in the future. PMID:19860739

  12. Embedding of magnetic nanoparticles in polycaprolactone nanofiber scaffolds to facilitate bone healing and regeneration

    NASA Astrophysics Data System (ADS)

    Kannarkat, Jacob T.; Battogtokh, Jugdersuren; Philip, John; Wilson, Otto C.; Mehl, Patrick M.

    2010-05-01

    Scaffolds used for tissue engineering are made to mimic natural surroundings of tissues, the extracellular matrix (ECM). The ECM plays a large part in maintaining the structural integrity of the connective tissue. When producing a tissue in the laboratory, structural integrity of the cells is ensured only when a biomimetic ECM is present. Nanofibrous polymer fibers have been chosen for their resemblance to natural fibers of the ECM and their capability to provide the support necessary for cells to grow and differentiate into tissue. Polycaprolactone based nanofibrous scaffolds for tissue engineering have been fabricated through the electrospinning process. Electrospinning is a simple and cost-effective method for producing nanofibers which involves applying a high voltage to a falling polymer solution to form a fluid jet producing nanofibers. Magnetic nanoparticles (MNPs) have been incorporated within the nanofibers by addition of MNPs to the polymer solution to increase the rate of bone cell growth, proliferation, and differentiation. Studies by Nomura and Takano-Yamamoto, [Matrix Biol. 19, 91 (2000)] demonstrated an increase in the expression levels of multiple genes in bone tissue including growth factors when shear stress was applied at the cellular level. MNPs are around 1-100 nm and exhibit superparamagnetism. These properties of MNPs allow for high noninvasive control over them using an external magnetic field. While under an ac (15 Hz, 1-6 Gauss) or pulsed magnetic fields, MNPs will induce low level mechanical stresses within the scaffold causing shear stresses at the cellular level of the preosteoblast MC3T3-E1 cells to stimulate their growth, proliferation, and differentiation.

  13. Effect of the internal microstructure in rapid-prototyped polycaprolactone scaffolds on physical and cellular properties for bone tissue regeneration

    NASA Astrophysics Data System (ADS)

    Jeon, Hojun; Kim, Geun Hyung

    2012-09-01

    Biomedical scaffolds should be designed to optimize their inter-microstructure to enable cell infiltration and nutrient/waste transport. To acquire these properties, several structural parameters, such as pore size, pore shape, porosity, pore interconnectivity, permeability, and tortuosity are required. In this study, we explored the effect of tortuosity on the viable cell proliferation and mineralization of osteoblast-like-cells (MG63) in polycaprolactone scaffolds. For analysis, we designed four different scaffolds of various tortuosities ranging from 1.0 to 1.3 under the same porosity (56 %) and 100 % pore interconnectivity. The pore size of the scaffolds was set as 150 and 300 µm, and a mixture of these sizes. We found that despite the porosity being same, the elastic modulus was dependent on the pore size of the scaffolds due to the distributed stress concentration. In addition, the relative water movement within scaffolds was also related to the internal microstructure. Cell viability and Ca2+ deposition of the cell-seeded scaffolds showed that the proliferation of viable cells and mineralization in the scaffolds with appropriate tortuosity (1.2) was relatively high compared to those of the scaffolds displaying low (1.05 and 1.1) or high (1.3) tortuosity. Our findings indicated that the internal microstructure of the scaffolds may influence not only the physical properties, but in addition the cellular behavior.

  14. Porous ceramic scaffolds with complex architectures

    SciTech Connect

    Saiz, Eduardo; Munch, Etienne; Franco, Jaime; Deville, Sylvain; Hunger, Phillip; Saiz, Eduardo; Tomsia, Antoni P.

    2008-03-15

    This work compares two novel techniques for the fabrication of ceramic scaffolds for bone tissue engineering with complex porosity: robocasting and freeze casting. Both techniques are based on the preparation of concentrated ceramic suspensions with suitable properties for the process. In robocasting, the computer-guided deposition of the suspensions is used to build porous materials with designed three dimensional (3-D) geometries and microstructures. Freeze casting uses ice crystals as a template to form porous lamellar ceramic materials. Preliminary results on the compressive strengths of the materials are also reported.

  15. Examinations of a new long-term degradable electrospun polycaprolactone scaffold in three rat abdominal wall models.

    PubMed

    Jangö, Hanna; Gräs, Søren; Christensen, Lise; Lose, Gunnar

    2017-02-01

    Alternative approaches to reinforce native tissue in reconstructive surgery for pelvic organ prolapse are warranted. Tissue engineering combines the use of a scaffold with the regenerative potential of stem cells and is a promising new concept in urogynecology. Our objective was to evaluate whether a newly developed long-term degradable polycaprolactone scaffold could provide biomechanical reinforcement and function as a scaffold for autologous muscle fiber fragments. We performed a study with three different rat abdominal wall models where the scaffold with or without muscle fiber fragments was placed (1) subcutaneously (minimal load), (2) in a partial defect (partial load), and (3) in a full-thickness defect (heavy load). After 8 weeks, no animals had developed hernia, and the scaffold provided biomechanical reinforcement, even in the models where it was subjected to heavy load. The scaffold was not yet degraded but showed increased thickness in all groups. Histologically, we found a massive foreign body response with numerous large giant cells intermingled with the fibers of the scaffold. Cells from added muscle fiber fragments could not be traced by PKH26 fluorescence or desmin staining. Taken together, the long-term degradable polycaprolactone scaffold provided biomechanical reinforcement by inducing a marked foreign-body response and attracting numerous inflammatory cells to form a strong neo-tissue construct. However, cells from the muscle fiber fragments did not survive in this milieu. Properties of the new neo-tissue construct must be evaluated at the time of full degradation of the scaffold before its possible clinical value in pelvic organ prolapse surgery can be evaluated.

  16. Electrospun polycaprolactone scaffolds with tailored porosity using two approaches for enhanced cellular infiltration.

    PubMed

    Zander, Nicole E; Orlicki, Joshua A; Rawlett, Adam M; Beebe, Thomas P

    2013-01-01

    The impact of mat porosity of polycaprolactone (PCL) electrospun fibers on the infiltration of neuron-like PC12 cells was evaluated using two different approaches. In the first method, bi-component aligned fiber mats were fabricated via the co-electrospinning of PCL with polyethylene oxide (PEO). Variation of the PEO flow rate, followed by selective removal of PEO from the PCL/PEO mesh, allowed for control of the porosity of the resulting scaffold. In the second method, aligned fiber mats were fabricated from various concentrations of PCL solutions to generate fibers with diameters between 0.13 ± 0.06 and 9.10 ± 4.1 μm. Of the approaches examined, the variation of PCL fiber diameter was found to be the better method for increasing the infiltration of PC12 cells, with the optimal infiltration into the ca. 1.5-mm-thick meshes observed for the mats with the largest fiber diameters, and hence largest pore sizes.

  17. Electrospun Polycaprolactone Scaffolds for Small-Diameter Tissue Engineered Blood Vessels

    NASA Astrophysics Data System (ADS)

    Lee, Carol Hsiu-Yueh

    Cardiovascular disease is the leading cause of death in the United States with many patients requiring coronary artery bypass grafting. The current standard is using autografts such as the saphenous vein or intimal mammary artery, however creating a synthetic graft could eliminate this painful and inconvenient procedure. Large diameter grafts have long been established with materials such as DacronRTM and TeflonRTM, however these materials have not proved successful in small-diameter (< 6 mm) grafts where thrombosis and intimal hyperplasia are common in graft failure. With the use of a synthetic biodegradable polymer (polycaprolactone) we utilize our expertise in electrospinning and femtosecond laser ablation to create a novel tri-layered tissue engineered blood vessel containing microchannels. The benefits of creating a tri-layer is to mimic native arteries that contain an endothelium to prevent thrombosis in the inner layer, aligned smooth muscle cells in the middle to control vasodilation and constriction, and a mechanically robust outer layer. The following work evaluates the mechanical properties of such a graft (tensile, fatigue, burst pressure, and suture retention strength), the ability to rapidly align cells in laser ablated microchannels in PCL scaffolds, and the biological integration (co-culture of endothelial and smooth muscle cells) with electrospun PCL scaffolds. The conclusions from this work establish that the electrospun tri-layers provide adequate mechanical strength as a tissue engineered blood vessel, that laser ablated microchannels are able to contain the smooth muscle cells, and that cells are able to adhere to PCL fibers. However, future work includes adjusting microchannel dimensions to properly align smooth muscle cells along with perfect co-cultures of endothelial and smooth muscle cells on the electrospun tri-layer.

  18. Material properties and electrical stimulation regimens through polycaprolactone fumarate-polypyrrole scaffolds as potential conductive nerve conduits

    PubMed Central

    Moroder, Philipp; Wang, Huan; Ruesink, Terry; Lu, Lichun; Windebank, Anthony J.; Yaszemski, Michael J.; Runge, M. Brett

    2010-01-01

    Mechanical and electrical properties of polycaprolactone fumarate-polypyrrole (PCLF-PPy) scaffolds were studied under physiological conditions to evaluate their ability to maintain material properties necessary for application as conductive nerve conduits. PC12 cells cultured on PCLF-PPy scaffolds were stimulated with regimens of 10 μA of constant or 20 Hz frequency current passed through the scaffolds for 1 h/day. PC12 cellular morphologies were analyzed by fluorescence microscopy after 48 h. PCLF-PPy scaffolds exhibited excellent mechanical properties at 37°C which would allow suturing and flexibility. The surface resistivity of the scaffolds was 2kΩ and the scaffolds were electrically stable during application of electrical stimulation (ES). In vitro studies showed significant increases in percentage of neurite bearing cells, number of neurites per cell and neurite length in the presence of ES compared to no ES. Additionally, extending neurites were observed to align in the direction of the applied current. This study shows that electrically conductive PCLF-PPy scaffolds possess material properties necessary for application as nerve conduits. Additionally, the capability to significantly enhance and direct neurite extension by passing electrical current through PCLF-PPy scaffolds renders them even more promising as future therapeutic treatments for severe nerve injuries. PMID:20965280

  19. In vivo efficacy of bone-marrow-coated polycaprolactone scaffolds for the reconstruction of orbital defects in the pig.

    PubMed

    Rohner, Dennis; Hutmacher, Dietmar W; Cheng, Tan Kim; Oberholzer, Martin; Hammer, Beat

    2003-08-15

    Alloplastic materials offer a number of advantages over bone autografts in the reconstruction of craniofacial defects. These include: lack of donor site morbidity, unlimited quantities of available material, and the possibility to conform exactly to the defect. An ideal bioresorbable material would degrade slowly, and have osteoconductive properties to allow replacement and remodeling by osseous tissue. This is seldom observed, the materials instead being replaced by fibrous tissue. Polycaprolactone (PCL), an FDA-approved bioresorbable polymer, has several properties that might make it suitable for reconstruction of craniofacial defects. The technique of fused deposition modeling (FDM) allows for the fabrication of highly reproducible bioresorbable 3D scaffolds. The nature of the fully interconnected pore network might enhance vascular ingrowth and osteoconductive properties. It was hypothesized that coating the scaffolds in bone marrow might enhance bone formation due to the osteoinductive nature of the bone-marrow mesenchymal cells. This study aimed to test these hypotheses in the pig model. Defects measuring 2 x 2 cm were surgically created in each orbit of eight Yorkshire pigs. The orbits were divided into three groups: Group 1 (n=4), no reconstruction (control); Group 2 (n=6), reconstruction with no coated PCL scaffolds; and Group 3 (n=6) reconstruction with bone-marrow-coated PCL scaffolds. The results were evaluated at 3 months by histological and histomorphometric analyses. The defects in Group 1 were covered with fibrous scar tissue. The shape of the reconstructed area was insufficient. The defects in Groups 2 and 3 were reconstructed correctly. In Group 2 the noncoated scaffolds showed 4.5% of new bone formation compared with 14.1% in Group 3, which is statistically significant (p<0.05). The entirely interconnected 3D polycaprolactone scaffold seems to be a promising material. It induces the bone ingrowth required for reconstructing craniofacial and

  20. Hybrid biomimetic scaffold composed of electrospun polycaprolactone nanofibers and self-assembled peptide amphiphile nanofibers

    PubMed Central

    Tambralli, Ajay; Blakeney, Bryan; Anderson, Joel; Kushwaha, Meenakshi; Andukuri, Adinarayana; Dean, Derrick; Jun, Ho-Wook

    2011-01-01

    Nanofibrous electrospun poly (ε-caprolactone) (ePCL) scaffolds have inherent structural advantages, but lack of bioactivity has limited their usefulness in biomedical applications. Thus, here we report the development of a hybrid, nanostructured, extracellular matrix (ECM) mimicking scaffold by a combination of ePCL nanofibers and self-assembled peptide amphiphile (PA) nanofibers. The PAs have ECM mimicking characteristics including a cell adhesive ligand (RGDS) and matrix metalloproteinase-2 (MMP-2) mediated degradable sites. TEM imaging verified successful PA self-assembly into nanofibers (diameters of 8 – 10 nm) using a solvent evaporation method. This evaporation coating method was then used to successfully coat PAs onto ePCL nanofibers (diameters of 300 – 400 nm), to develop the hybrid, bioactive scaffolds. SEM characterization showed that the PA coatings did not interfere with the porous ePCL nanofiber network. Human mesenchymal stem cells (hMSCs) were seeded onto the hybrid scaffolds to evaluate their bioactivity. Significantly greater attachment and spreading of hMSCs were observed on ePCL nanofibers coated with PA-RGDS as compared to ePCL nanofibers coated with PA-S (no cell adhesive ligand) and uncoated ePCL nanofibers. Overall, this novel strategy presents a new solution to overcome the current bioactivity challenges of electrospun scaffolds and combines the unique characteristics of ePCL nanofibers and self-assembled PA nanofibers to provide an ECM mimicking environment. This has great potential to be applied to many different electrospun scaffolds for various biomedical applications. PMID:20811101

  1. Surface modification of nanofibrous polycaprolactone/gelatin composite scaffold by collagen type I grafting for skin tissue engineering.

    PubMed

    Gautam, Sneh; Chou, Chia-Fu; Dinda, Amit K; Potdar, Pravin D; Mishra, Narayan C

    2014-01-01

    In the present study, a tri-polymer polycaprolactone (PCL)/gelatin/collagen type I composite nanofibrous scaffold has been fabricated by electrospinning for skin tissue engineering and wound healing applications. Firstly, PCL/gelatin nanofibrous scaffold was fabricated by electrospinning using a low cost solvent mixture [chloroform/methanol for PCL and acetic acid (80% v/v) for gelatin], and then the nanofibrous PCL/gelatin scaffold was modified by collagen type I (0.2-1.5wt.%) grafting. Morphology of the collagen type I-modified PCL/gelatin composite scaffold that was analyzed by field emission scanning electron microscopy (FE-SEM), showed that the fiber diameter was increased and pore size was decreased by increasing the concentration of collagen type I. Fourier transform infrared (FT-IR) spectroscopy and thermogravimetric (TG) analysis indicated the surface modification of PCL/gelatin scaffold by collagen type I immobilization on the surface of the scaffold. MTT assay demonstrated the viability and high proliferation rate of L929 mouse fibroblast cells on the collagen type I-modified composite scaffold. FE-SEM analysis of cell-scaffold construct illustrated the cell adhesion of L929 mouse fibroblasts on the surface of scaffold. Characteristic cell morphology of L929 was also observed on the nanofiber mesh of the collagen type I-modified scaffold. Above results suggest that the collagen type I-modified PCL/gelatin scaffold was successful in maintaining characteristic shape of fibroblasts, besides good cell proliferation. Therefore, the fibroblast seeded PCL/gelatin/collagen type I composite nanofibrous scaffold might be a potential candidate for wound healing and skin tissue engineering applications. © 2013.

  2. Evaluation of electrospinning parameters on the tensile strength and suture retention strength of polycaprolactone nanofibrous scaffolds through surface response methodology.

    PubMed

    Asvar, Zahra; Mirzaei, Esmaeil; Azarpira, Negar; Geramizadeh, Bita; Fadaie, Milad

    2017-11-01

    Scaffolds should provide sufficient biomechanical support during tissue regeneration for tissue engineering (TE) applications. Electrospun scaffolds are commonly applied in TE applications due to their tunable physical, chemical, and mechanical properties as well as their similarity to extracellular matrix. Although the mechanical properties of electrospun scaffolds are highly dependent on processing parameters, a limited number of studies have systematically investigated this subject. The present study has investigated the effects of the main electrospinning parameters on tensile and suture retention strength of polycaprolactone (PCL) scaffolds using response surface methodology. Scaffolds morphology and cell-scaffold interaction were also investigated in this study. According to the fitted model, polymer concentration and feed rate have the most significant positive effect on both the tensile and suture retention strength. Whereas applied voltage negatively affected both the tensile and suture retention strength. The effect of distance on tensile strength was not significant while its effect on suture retention was different depending on its values. Changes in biomechanical properties were associated with gross alterations in morphology of the fibers and cell-scaffold interaction. Scaffolds with lowest tensile strength presented a beaded morphology while scaffolds with higher tensile strength presented beadless morphology with worm-like fibers. The increase in tensile strength was correlated with the increase in average diameter of the fibers and pore size. The results of cell culture study showed that fibroblasts stretched and proliferated more on scaffolds with lower tensile strength. The generated model might be helpful when PCL scaffold with desirable tensile and suture retention strength are required. Furthermore, the results suggest that changes in morphology and subsequent cell-scaffold interaction should be considered when these biomechanical properties

  3. Nanofibrous yet injectable polycaprolactone-collagen bone tissue scaffold with osteoprogenitor cells and controlled release of bone morphogenetic protein-2.

    PubMed

    Subramanian, Gayathri; Bialorucki, Callan; Yildirim-Ayan, Eda

    2015-06-01

    In this work, we developed a nanofibrous, yet injectable orthobiologic tissue scaffold that is capable of hosting osteoprogenitor cells and controlling kinetic release profile of the encapsulated pro-osteogenic factor without diminishing its bioactivity over 21days. This innovative injectable scaffold was synthesized by incorporating electrospun and subsequently O2 plasma-functionalized polycaprolactone (PCL) nanofibers within the collagen type-I solution along with MC3T3-E1 cells (pre-osteoblasts) and bone morphogenetic protein-2 (BMP2). Through changing the PCL nanofiber concentration within the injectable scaffolds, we were able to tailor the mechanical strength, protein retention capacity, bioactivity preservation, and osteoinductive potential of the scaffolds. The nanofibrous internal structure of the scaffold allowed us to use a low dose of BMP2 (200ng/ml) to achieve osteoblastic differentiation in in vitro culture. The osteogenesis capacity of the injectable scaffolds were evaluated though measuring MC3T3-E1 cell proliferation, ALP activity, matrix mineralization, and early- and late-osteoblast specific gene expression profiles over 21days. The results demonstrated that the nanofibrous injectable scaffold provides not only an osteoinductive environment for osteoprogenitor cells to differentiate, but also a suitable biomechanical and biochemical environment to act as a reservoir for osteogenic factors with controlled release profile.

  4. A comparison study between electrospun polycaprolactone and piezoelectric poly(3-hydroxybutyrate-co-3-hydroxyvalerate) scaffolds for bone tissue engineering.

    PubMed

    Gorodzha, Svetlana N; Muslimov, Albert R; Syromotina, Dina S; Timin, Alexander S; Tcvetkov, Nikolai Y; Lepik, Kirill V; Petrova, Aleksandra V; Surmeneva, Maria A; Gorin, Dmitry A; Sukhorukov, Gleb B; Surmenev, Roman A

    2017-09-05

    In this study, bone scaffolds composed of polycaprolactone (PCL), piezoelectric poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and a combination of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and silicate containing hydroxyapatite (PHBV-SiHA) were successfully fabricated by a conventional electrospinning process. The morphological, chemical, wetting and biological properties of the scaffolds were examined. All fabricated scaffolds are composed of randomly oriented fibres with diameters from 800nm to 12μm. Fibre size increased with the addition of SiHA to PHBV scaffolds. Moreover, fibre surface roughness in the case of hybrid scaffolds was also increased. XRD, FTIR and Raman spectroscopy were used to analyse the chemical composition of the scaffolds, and contact angle measurements were performed to reveal the wetting behaviour of the synthesized materials. To determine the influence of the piezoelectric nature of PHBV in combination with SiHA nanoparticles on cell attachment and proliferation, PCL (non-piezoelectric), pure PHBV, and PHBV-SiHA scaffolds were seeded with human mesenchymal stem cells (hMSCs). In vitro study on hMSC adhesion, viability, spreading and osteogenic differentiation showed that the PHBV-SiHA scaffolds had the largest adhesion and differentiation abilities compared with other scaffolds. Moreover, the piezoelectric PHBV scaffolds have demonstrated better calcium deposition potential compared with non-piezoelectric PCL. The results of the study revealed pronounced advantages of hybrid PHBV-SiHA scaffolds to be used in bone tissue engineering. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Mechanical improvements to reinforced porous silk scaffolds.

    PubMed

    Gil, Eun Seok; Kluge, Jonathan A; Rockwood, Danielle N; Rajkhowa, Rangam; Wang, Lijing; Wang, Xungai; Kaplan, David L

    2011-10-01

    Load-bearing porous biodegradable scaffolds are required to engineer functional tissues such as bone. Mechanical improvements to porogen leached scaffolds prepared from silk proteins were systematically studied through the addition of silk particles in combination with silk solution concentration, exploiting interfacial compatibility between the two components. Solvent solutions of silk up to 32 w/v % were successfully prepared in hexafluoroisopropanol (HFIP) for the study. The mechanical properties of the reinforced silk scaffolds correlated to the material density and matched by a power law relationship, independent of the ratio of silk particles to matrix. These results were similar to the relationships previously shown for cancellous bone. From these data we conclude that the increased mechanical properties were due to a densification effect and not due to the inclusion of stiffer silk particles into the softer silk matrix. A continuous interface between the silk matrix and the silk particles, as well as homogeneous distribution of the silk particles within the matrix was observed. Furthermore, we note that the roughness of the pore walls was controllable by varying the ratio of the particles matrix, providing a route to control topography. The rate of proteolytic hydrolysis of the scaffolds decreased with increase in mass of silk used in the matrix and with increasing silk particle content.

  6. Hierarchical porous polymer scaffolds from block copolymers.

    PubMed

    Sai, Hiroaki; Tan, Kwan Wee; Hur, Kahyun; Asenath-Smith, Emily; Hovden, Robert; Jiang, Yi; Riccio, Mark; Muller, David A; Elser, Veit; Estroff, Lara A; Gruner, Sol M; Wiesner, Ulrich

    2013-08-02

    Hierarchical porous polymer materials are of increasing importance because of their potential application in catalysis, separation technology, or bioengineering. Examples for their synthesis exist, but there is a need for a facile yet versatile conceptual approach to such hierarchical scaffolds and quantitative characterization of their nonperiodic pore systems. Here, we introduce a synthesis method combining well-established concepts of macroscale spinodal decomposition and nanoscale block copolymer self-assembly with porosity formation on both length scales via rinsing with protic solvents. We used scanning electron microscopy, small-angle x-ray scattering, transmission electron tomography, and nanoscale x-ray computed tomography for quantitative pore-structure characterization. The method was demonstrated for AB- and ABC-type block copolymers, and resulting materials were used as scaffolds for calcite crystal growth.

  7. Polylactic acid fibre-reinforced polycaprolactone scaffolds for bone tissue engineering.

    PubMed

    Guarino, Vincenzo; Causa, Filippo; Taddei, Paola; di Foggia, Michele; Ciapetti, Gabriela; Martini, Desirèe; Fagnano, Concezio; Baldini, Nicola; Ambrosio, Luigi

    2008-09-01

    The employment of composite scaffolds with a well-organized architecture and multi-scale porosity certainly represents a valuable approach for achieving a tissue engineered construct to reproduce the middle and long-term behaviour of hierarchically complex tissues such as spongy bone. In this paper, fibre-reinforced composites scaffold for bone tissue engineering applications is described. These are composed of poly-L-lactide acid (PLLA) fibres embedded in a porous poly(epsilon-caprolactone) matrix, and were obtained by synergistic use of phase inversion/particulate leaching technique and filament winding technology. Porosity degree as high as 79.7% was achieved, the bimodal pore size distribution showing peaks at ca 10 and 200 microm diameter, respectively, accounting for 53.7% and 46.3% of the total porosity. In vitro degradation was carried out in PBS and SBF without significant degradation of the scaffold after 35 days, while in NaOH solution, a linear increase of weight lost was observed with preferential degradation of PLLA component. Subsequently, marrow stromal cells (MSC) and human osteoblasts (HOB) reached a plateau at 3 weeks, while at 5 weeks the number of cells was almost the same. Human marrow stromal cell and trabecular osteoblasts rapidly proliferate on the scaffold up to 3 weeks, promoting an oriented migration of bone cells along the fibre arrangement. Moreover, the role of seeded HOB and MSC on composite degradation mechanism was assessed by demonstrating a more relevant contribution to PLLA degradation of MSC when compared to HOB. The novel PCL/PLLA composite scaffolds thus showed promise whenever tuneable porosity, controlled degradability and guided cell-material interaction are simultaneously requested.

  8. Laser sintered porous polycaprolacone scaffolds loaded with hyaluronic acid and gelatin-grafted thermoresponsive hydrogel for cartilage tissue engineering.

    PubMed

    Lee, Ming-Yih; Tsai, Wen-Wei; Chen, His-Jung; Chen, Jyh-Ping; Chen, Chih-Hao; Yeh, Wen-Lin; An, Jia

    2013-01-01

    The aim of this study is to evaluate a soft/hard bi-phase scaffold for cartilage tissue engineering. Chondrocyte proliferation, glycoaminoglycan production and total collagen content are compared between laser-sintered porous polycaprolactone (PCL) scaffolds with and without a thermoresponsive hydrogel grafted with hyaluronic acid and gelatin. The in vitro results show that scaffolds loaded with hydrogel have a higher initial chondrocyte attachment than PCL scaffolds. At day 21 and 28, scaffolds loaded with hydrogel have a significantly higher glycosaminoglycan (GAG) production than PCL scaffolds alone, and total collagen content including collagen type II in the hydrogel-loaded group is three times higher than the group without hydrogel. It is concluded that the laser-sintered porous PCL scaffold has good cytocompatibility, and that the hydrogel phase is able to enhance initial chondrocytes attachment as well as GAG and collagen production of chondrocytes. This study suggests that a soft/hard bi-phase scaffold may be used for cartilage tissue engineering to enhance in vitro chondrogenesis.

  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

    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

  10. 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

  11. Porous allograft bone scaffolds: doping with strontium.

    PubMed

    Zhao, Yantao; Guo, Dagang; Hou, Shuxun; Zhong, Hongbin; Yan, Jun; Zhang, Chunli; Zhou, Ying

    2013-01-01

    Strontium (Sr) can promote the process of bone formation. To improve bioactivity, porous allograft bone scaffolds (ABS) were doped with Sr and the mechanical strength and bioactivity of the scaffolds were evaluated. Sr-doped ABS were prepared using the ion exchange method. The density and distribution of Sr in bone scaffolds were investigated by inductively coupled plasma optical emission spectrometry (ICP-OES), X-ray photoelectron spectroscopy (XPS), and energy-dispersive X-ray spectroscopy (EDS). Controlled release of strontium ions was measured and mechanical strength was evaluated by a compressive strength test. The bioactivity of Sr-doped ABS was investigated by a simulated body fluid (SBF) assay, cytotoxicity testing, and an in vivo implantation experiment. The Sr molar concentration [Sr/(Sr+Ca)] in ABS surpassed 5% and Sr was distributed nearly evenly. XPS analyses suggest that Sr combined with oxygen and carbonate radicals. Released Sr ions were detected in the immersion solution at higher concentration than calcium ions until day 30. The compressive strength of the Sr-doped ABS did not change significantly. The bioactivity of Sr-doped material, as measured by the in vitro SBF immersion method, was superior to that of the Sr-free freeze-dried bone and the Sr-doped material did not show cytotoxicity compared with Sr-free culture medium. The rate of bone mineral deposition for Sr-doped ABS was faster than that of the control at 4 weeks (3.28 ± 0.23 µm/day vs. 2.60 ± 0.20 µm/day; p<0.05). Sr can be evenly doped into porous ABS at relevant concentrations to create highly active bone substitutes.

  12. Porous Allograft Bone Scaffolds: Doping with Strontium

    PubMed Central

    Zhao, Yantao; Guo, Dagang; Hou, Shuxun; Zhong, Hongbin; Yan, Jun; Zhang, Chunli; Zhou, Ying

    2013-01-01

    Strontium (Sr) can promote the process of bone formation. To improve bioactivity, porous allograft bone scaffolds (ABS) were doped with Sr and the mechanical strength and bioactivity of the scaffolds were evaluated. Sr-doped ABS were prepared using the ion exchange method. The density and distribution of Sr in bone scaffolds were investigated by inductively coupled plasma optical emission spectrometry (ICP-OES), X-ray photoelectron spectroscopy (XPS), and energy-dispersive X-ray spectroscopy (EDS). Controlled release of strontium ions was measured and mechanical strength was evaluated by a compressive strength test. The bioactivity of Sr-doped ABS was investigated by a simulated body fluid (SBF) assay, cytotoxicity testing, and an in vivo implantation experiment. The Sr molar concentration [Sr/(Sr+Ca)] in ABS surpassed 5% and Sr was distributed nearly evenly. XPS analyses suggest that Sr combined with oxygen and carbonate radicals. Released Sr ions were detected in the immersion solution at higher concentration than calcium ions until day 30. The compressive strength of the Sr-doped ABS did not change significantly. The bioactivity of Sr-doped material, as measured by the in vitro SBF immersion method, was superior to that of the Sr-free freeze-dried bone and the Sr-doped material did not show cytotoxicity compared with Sr-free culture medium. The rate of bone mineral deposition for Sr-doped ABS was faster than that of the control at 4 weeks (3.28±0.23 µm/day vs. 2.60±0.20 µm/day; p<0.05). Sr can be evenly doped into porous ABS at relevant concentrations to create highly active bone substitutes. PMID:23922703

  13. Electrospun polycaprolactone/gelatin composites with enhanced cell-matrix interactions as blood vessel endothelial layer scaffolds.

    PubMed

    Jiang, Yong-Chao; Jiang, Lin; Huang, An; Wang, Xiao-Feng; Li, Qian; Turng, Lih-Sheng

    2017-02-01

    During the fabrication of tissue engineering scaffolds and subsequent tissue regeneration, surface bioactivity is vital for cell adhesion, spreading, and proliferation, especially for endothelium dysfunction repair. In this paper, synthetic polymer polycaprolactone (PCL) was blended with natural polymer gelatin at four different weight ratios followed by crosslinking (i.e., 100:0, 70:30, 50:50, 30:70, labeled as PCL-C, P7G3-C, P5G5-C, and P3G7-C) to impart enhanced bioactivity and tunable mechanical properties. The PCL/gelatin blends were first dissolved in 2,2,2-trifluroethanol (TFE) and supplementary acetic acid (1% relative to TFE) solvent, electrospun, and then cross-linked to produce PBS-proof fibrous scaffolds. Scanning electron micrographs (SEM) indicated that fibers of each sample were smooth and homogeneous, with the fiber diameters increasing from 1.01±0.51μm to 1.61±0.46μm as the content of gelatin increased. While thermal resistance and crystallization of the blends were affected by the presence of gelatin, as reflected by differential scanning calorimetry (DSC) results, water contact angle (WCA) tests confirmed that the scaffold surfaces became more hydrophilic. Tensile tests showed that PCL-C and P7G3-C scaffolds had mechanical properties comparable to those of human coronary arteries. As for cytocompatibility, skeleton staining images showed that human mesenchymal stem cells (hMSCs) had more favorable binding sites on PCL/gelatin scaffolds than those on PCL scaffolds. Cell proliferation assays revealed that P7G3-C scaffolds could support the most number of hMSCs. The results of this study demonstrated the enhanced cell-matrix interactions and potential use of electrospun PCL/gelatin scaffolds in the tissue engineering field, especially in wound dressings and endothelium regeneration. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Preparation and characterization of bioactive composite scaffolds from polycaprolactone nanofibers-chitosan-oxidized starch for bone regeneration.

    PubMed

    Nourmohammadi, Jhamak; Ghaee, Azadeh; Liavali, Samira Hosseini

    2016-03-15

    The objective of this study was to fabricate and investigate the characteristics of a suitable scaffold for bone regeneration. Therefore, chitosan was combined with various amounts of oxidized starch through reductive alkylation process. Afterwards, chopped CaP-coated PCL nanofibers were added into the chitosan-starch composite scaffolds in order to obtain bioactivity and mimic bone extracellular matrix structure. Scanning electron microscopy confirmed that all scaffolds had well-interconnected porous structure. The mean pore size, porosity, and water uptake of the composite scaffolds increased by incorporation of higher amounts of starch, while this trend was opposite for compressive modulus and strength. Osteoblast-like cells (MG63) culturing on the scaffolds demonstrated that higher starch content could improve cell viability. Moreover, the cells spread and anchored well on the scaffolds, on which the surface was covered with a monolayer of cells. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. A new method for the production of gelatin microparticles for controlled protein release from porous polymeric scaffolds.

    PubMed

    Ozkizilcik, Asya; Tuzlakoglu, Kadriye

    2014-03-01

    Tissue engineering using scaffolds and growth factors is a crucial approach in bone regeneration and repair. The combination of bioactive agents carrying microparticles with porous scaffolds can be an efficient solution when controlled release of bio-signalling molecules is required. The present study was based on a recent approach using a biodegradable scaffold and protein-loaded microparticles produced in an innovative manner in which protein loss is minimized during the loading process. Bovine serum albumin (BSA)-loaded gelatin microparticles were obtained by grinding freeze-dried membranes of gelatin and BSA. Porous scaffolds (250-355 µm pore size) produced from a polyactide (PLLA) and polycaprolactone (PCL) blend by salt leaching/supercritical CO₂ methods were used for the experiments. Gelatin microparticles containing three different BSA amounts were incorporated into the porous scaffolds by using a surfactant. In vitro release profiles showed up to 90% protein loading efficiency. This novel method appears to be an effective approach for producing particles that can minimize protein loss during the loading process.

  16. 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.

  17. Polymer powder processing of cryomilled polycaprolactone for solvent-free generation of homogeneous bioactive tissue engineering scaffolds.

    PubMed

    Lim, Jing; Chong, Mark Seow Khoon; Chan, Jerry Kok Yen; Teoh, Swee-Hin

    2014-06-25

    Synthetic polymers used in tissue engineering require functionalization with bioactive molecules to elicit specific physiological reactions. These additives must be homogeneously dispersed in order to achieve enhanced composite mechanical performance and uniform cellular response. This work demonstrates the use of a solvent-free powder processing technique to form osteoinductive scaffolds from cryomilled polycaprolactone (PCL) and tricalcium phosphate (TCP). Cryomilling is performed to achieve micrometer-sized distribution of PCL and reduce melt viscosity, thus improving TCP distribution and improving structural integrity. A breakthrough is achieved in the successful fabrication of 70 weight percentage of TCP into a continuous film structure. Following compaction and melting, PCL/TCP composite scaffolds are found to display uniform distribution of TCP throughout the PCL matrix regardless of composition. Homogeneous spatial distribution is also achieved in fabricated 3D scaffolds. When seeded onto powder-processed PCL/TCP films, mesenchymal stem cells are found to undergo robust and uniform osteogenic differentiation, indicating the potential application of this approach to biofunctionalize scaffolds for tissue engineering applications.

  18. Bio-functionalization of polycaprolactone infiltrated BCP scaffold with silicon and fibronectin enhances osteoblast activity in vitro

    NASA Astrophysics Data System (ADS)

    Kwak, Kyung-A.; Kim, Young-Hee; Kim, Minsung; Lee, Byong-Taek; Song, Ho-Yeon

    2013-08-01

    The surface property of a biomaterial plays a fundamental role in cell attachment, proliferation, differentiation, resorption, and biomolecular expression. In this study, the surface of a polycaprolactone-infiltrated biphasic calcium phosphate scaffold was biofunctionalized by silicon (Si) and fibronectin (FN) coating to evaluate the osteoblast-like cells activity in vitro. The surfaces of all scaffolds were characterized by scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS), whereas the presence of the functional group was determined by Fourier-transform infrared spectroscopy (FT-IR). Coomassie brilliant blue staining was applied to confirm the presence of FN on the scaffold surface. The in vitro bioactivity of the osteoblast-like cells was determined by one cell morphology and proliferation assay at 3, 7, and 14 days by SEM. Cell viability assay by MTT showed higher cell viability rate on coated scaffolds than in those coated with Si only or non-coated surfaces. The mRNA expressions of alkaline phosphatase (ALP) and osteocalcin (OC) were determined using RT-PCR and the expressions of osteopontin (OPN), type I collagen, and osteocalcin (OC) proteins were determined using Western blot. Thus the expression of genes and proteins further confirmed both early and intermediate phases of osteoblast-like cell activity which was found increased by Si-and Fn coating on PCL infiltrated BCP surfaces.

  19. Manufacturing of biodegradable polyurethane scaffolds based on polycaprolactone using a phase separation method: physical properties and in vitro assay

    PubMed Central

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

    2011-01-01

    Background Biodegradable polyurethanes have found widespread use in soft tissue engineering due to their suitable mechanical properties and biocompatibility. Methods In this study, polyurethane samples were synthesized from polycaprolactone, hexamethylene diisocyanate, and a copolymer of 1,4-butanediol as a chain extender. Polyurethane scaffolds were fabricated by a combination of liquid–liquid phase separation and salt leaching techniques. The effect of the NCO:OH ratio on porosity content and pore morphology was investigated. Results Scanning electron micrographs demonstrated that the scaffolds had a regular distribution of interconnected pores, with pore diameters of 50–300 μm, and porosities of 64%–83%. It was observed that, by increasing the NCO:OH ratio, the average pore size, compressive strength, and compressive modulus increased. L929 fibroblast and chondrocytes were cultured on the scaffolds, and all samples exhibited suitable cell attachment and growth, with a high level of biocompatibility. Conclusion These biodegradable polyurethane scaffolds demonstrate potential for soft tissue engineering applications. PMID:22072874

  20. Flow-Induced Stress Distribution in Porous Scaffolds

    NASA Astrophysics Data System (ADS)

    Papavassiliou, Dimitrios; Voronov, Roman; Vangordon, Samuel; Sikavitsas, Vassilios

    2010-11-01

    Flow-induced stresses help the differentiation and proliferation of mesenchymal cells cultured in porous scaffolds within perfusion bioreactors. The distribution of stresses in a scaffold is thus important for understanding the tissue growth process in such reactors. Computational results for flow through Poly-L-Lactic Acid porous scaffolds that have been produced with salt-leaching techniques, and for scaffolds that have been constructed with nonwoven fibers, indicate that the probability density function (pdf) of the wall stress, when normalized with the mean and the standard deviation of the pdf, appears to follow a single type of pdf. The scaffolds were imaged with micro-CT and the simulations were run with lattice Boltzmann methods. The parameters of the distribution can be obtained using Darcy's law and the Blake-Kozeny-Carman equation. Experimental results available in the literature appear to corroborate the computational findings, leading to the conclusion that stresses in high-porosity porous materials follow a single distribution.

  1. Porous Biodegradable Metals for Hard Tissue Scaffolds: A Review

    PubMed Central

    Yusop, A. H.; Bakir, A. A.; Shaharom, N. A.; Abdul Kadir, M. R.; Hermawan, H.

    2012-01-01

    Scaffolds have been utilized in tissue regeneration to facilitate the formation and maturation of new tissues or organs where a balance between temporary mechanical support and mass transport (degradation and cell growth) is ideally achieved. Polymers have been widely chosen as tissue scaffolding material having a good combination of biodegradability, biocompatibility, and porous structure. Metals that can degrade in physiological environment, namely, biodegradable metals, are proposed as potential materials for hard tissue scaffolding where biodegradable polymers are often considered as having poor mechanical properties. Biodegradable metal scaffolds have showed interesting mechanical property that was close to that of human bone with tailored degradation behaviour. The current promising fabrication technique for making scaffolds, such as computation-aided solid free-form method, can be easily applied to metals. With further optimization in topologically ordered porosity design exploiting material property and fabrication technique, porous biodegradable metals could be the potential materials for making hard tissue scaffolds. PMID:22919393

  2. 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.

  3. Integration of PCL and PLA in a monolithic porous scaffold for interface tissue engineering.

    PubMed

    Scaffaro, Roberto; Lopresti, Francesco; Botta, Luigi; Rigogliuso, Salvatrice; Ghersi, Giulio

    2016-10-01

    A novel bi-layered multiphasic scaffold (BLS) have been fabricated for the first time by combining melt mixing, compression molding and particulate leaching. One layer has been composed by polylactic acid (PLA) presenting pore size in the range of 90-110µm while the other layer has been made of polycaprolactone (PCL) with pores ranging from 5 to 40µm. The different chemo-physical properties of the two biopolymers combined with the tunable pore architecture permitted to realize monolithic functionally graded scaffolds engineered to be potentially used for interface tissues regenerations. BLS have been characterized from a morphological and a mechanical point of view. In particular, mechanical tests have been carried out both in air and immersing the specimens in phosphate buffered saline (PBS) solution at 37°C, in order to evaluate the elastic modulus and the interlayer adhesion strength. Fibroblasts and osteoblasts have been cultured and co-cultured in order to investigate the cells permeation trough the different layers. The results indicate that the presented method is appropriate for the preparation of multiphasic porous scaffolds with tunable morphological and mechanical characteristics. Furthermore, the cells seeded were found to grow with a different trend trough the different layers thus demonstrating that the presented device has good potential to be used in interface tissue regeneration applications.

  4. Porous Collagen Scaffold Reinforced with Surfaced Activated PLLA Nanoparticles

    PubMed Central

    Xu, Cancan; Lu, Wei; Bian, Shaoquan; Liang, Jie; Fan, Yujiang; Zhang, Xingdong

    2012-01-01

    Porous collagen scaffold is integrated with surface activated PLLA nanoparticles fabricated by lyophilizing and crosslinking via EDC treatment. In order to prepare surface-modified PLLA nanoparticles, PLLA was firstly grafted with poly (acrylic acid) (PAA) through surface-initiated polymerization of acrylic acid. Nanoparticles of average diameter 316 nm and zeta potential −39.88 mV were obtained from the such-treated PLLA by dialysis method. Porous collagen scaffold were fabricated by mixing PLLA nanoparticles with collagen solution, freeze drying, and crosslinking with EDC. SEM observation revealed that nanoparticles were homogeneously dispersed in collagen matrix, forming interconnected porous structure with pore size ranging from 150 to 200 μm, irrespective of the amount of nanoparticles. The porosity of the scaffolds kept almost unchanged with the increment of the nanoparticles, whereas the mechanical property was obviously improved, and the degradation was effectively retarded. In vitro L929 mouse fibroblast cells seeding and culture studies revealed that cells infiltrated into the scaffolds and were distributed homogeneously. Compared with the pure collagen sponge, the number of cells in hybrid scaffolds greatly increased with the increment of incorporated nanoparticles. These results manifested that the surface-activated PLLA nanoparticles effectively reinforced the porous collagen scaffold and promoted the cells penetrating into the scaffold, and proliferation. PMID:22448137

  5. Novel Biodegradable Porous Scaffold Applied to Skin Regeneration

    PubMed Central

    Wang, Hui-Min; Chou, Yi-Ting; Wen, Zhi-Hong; Wang, Zhao-Ren; Chen, Chun-Hong; Ho, Mei-Ling

    2013-01-01

    Skin wound healing is an important lifesaving issue for massive lesions. A novel porous scaffold with collagen, hyaluronic acid and gelatin was developed for skin wound repair. The swelling ratio of this developed scaffold was assayed by water absorption capacity and showed a value of over 20 g water/g dried scaffold. The scaffold was then degraded in time- and dose-dependent manners by three enzymes: lysozyme, hyaluronidase and collagenase I. The average pore diameter of the scaffold was 132.5±8.4 µm measured from SEM images. With human skin cells growing for 7 days, the SEM images showed surface fractures on the scaffold due to enzymatic digestion, indicating the biodegradable properties of this scaffold. To simulate skin distribution, the human epidermal keratinocytes, melanocytes and dermal fibroblasts were seeded on the porous scaffold and the cross-section immunofluorescent staining demonstrated normal human skin layer distributions. The collagen amount was also quantified after skin cells seeding and presented an amount 50% higher than those seeded on culture wells. The in vivo histological results showed that the scaffold ameliorated wound healing, including decreasing neutrophil infiltrates and thickening newly generated skin compared to the group without treatments. PMID:23762223

  6. Novel biodegradable porous scaffold applied to skin regeneration.

    PubMed

    Wang, Hui-Min; Chou, Yi-Ting; Wen, Zhi-Hong; Wang, Chau-Zen; Wang, Zhao-Ren; Chen, Chun-Hong; Ho, Mei-Ling

    2013-01-01

    Skin wound healing is an important lifesaving issue for massive lesions. A novel porous scaffold with collagen, hyaluronic acid and gelatin was developed for skin wound repair. The swelling ratio of this developed scaffold was assayed by water absorption capacity and showed a value of over 20 g water/g dried scaffold. The scaffold was then degraded in time- and dose-dependent manners by three enzymes: lysozyme, hyaluronidase and collagenase I. The average pore diameter of the scaffold was 132.5±8.4 µm measured from SEM images. With human skin cells growing for 7 days, the SEM images showed surface fractures on the scaffold due to enzymatic digestion, indicating the biodegradable properties of this scaffold. To simulate skin distribution, the human epidermal keratinocytes, melanocytes and dermal fibroblasts were seeded on the porous scaffold and the cross-section immunofluorescent staining demonstrated normal human skin layer distributions. The collagen amount was also quantified after skin cells seeding and presented an amount 50% higher than those seeded on culture wells. The in vivo histological results showed that the scaffold ameliorated wound healing, including decreasing neutrophil infiltrates and thickening newly generated skin compared to the group without treatments.

  7. Porous magnesium-based scaffolds for tissue engineering.

    PubMed

    Yazdimamaghani, Mostafa; Razavi, Mehdi; Vashaee, Daryoosh; Moharamzadeh, Keyvan; Boccaccini, Aldo R; Tayebi, Lobat

    2017-02-01

    Significant amount of research efforts have been dedicated to the development of scaffolds for tissue engineering. Although at present most of the studies are focused on non-load bearing scaffolds, many scaffolds have also been investigated for hard tissue repair. In particular, metallic scaffolds are being studied for hard tissue engineering due to their suitable mechanical properties. Several biocompatible metallic materials such as stainless steels, cobalt alloys, titanium alloys, tantalum, nitinol and magnesium alloys have been commonly employed as implants in orthopedic and dental treatments. They are often used to replace and regenerate the damaged bones or to provide structural support for healing bone defects. Among the common metallic biomaterials, magnesium (Mg) and a number of its alloys are effective because of their mechanical properties close to those of human bone, their natural ionic content that may have important functional roles in physiological systems, and their in vivo biodegradation characteristics in body fluids. Due to such collective properties, Mg based alloys can be employed as biocompatible, bioactive, and biodegradable scaffolds for load-bearing applications. Recently, porous Mg and Mg alloys have been specially suggested as metallic scaffolds for bone tissue engineering. With further optimization of the fabrication techniques, porous Mg is expected to make a promising hard substitute scaffold. The present review covers research conducted on the fabrication techniques, surface modifications, properties and biological characteristics of Mg alloys based scaffolds. Furthermore, the potential applications, challenges and future trends of such degradable metallic scaffolds are discussed in detail.

  8. Porous shape memory alloy scaffolds for biomedical applications: a review

    NASA Astrophysics Data System (ADS)

    Wen, C. E.; Xiong, J. Y.; Li, Y. C.; Hodgson, P. D.

    2010-05-01

    The interest in using porous shape memory alloy (SMA) scaffolds as implant materials has been growing in recent years due to the combination of their unique mechanical and functional properties, i.e. shape memory effect and superelasticity, low elastic modulus combined with new bone tissue ingrowth ability and vascularization. These attractive properties are of great benefit to the healing process for implant applications. This paper reviews current state-of-the art on the processing, porous characteristics and mechanical properties of porous SMAs for biomedical applications, with special focus on the most widely used SMA nickel-titanium (NiTi), including (i) microstructural features, mechanical and functional properties of NiTi SMAs; (ii) main processing methods for the fabrication of porous NiTi SMAs and their mechanical properties and (iii) new-generation Ni-free, biocompatible porous SMA scaffolds.

  9. Multilayer porous UHMWPE scaffolds for bone defects replacement.

    PubMed

    Maksimkin, A V; Senatov, F S; Anisimova, N Yu; Kiselevskiy, M V; Zalepugin, D Yu; Chernyshova, I V; Tilkunova, N A; Kaloshkin, S D

    2017-04-01

    Reconstruction of the structural integrity of the damaged bone tissue is an urgent problem. UHMWPE may be potentially used for the manufacture of porous implants simulating as closely as possible the porous cancellous bone tissue. But the extremely high molecular weight of the polymer does not allow using traditional methods of foaming. Porous and multilayer UHMWPE scaffolds with nonporous bulk layer and porous layer that mimics cancellous bone architecture were obtained by solid-state mixing, thermopressing and washing in subcritical water. Structural and mechanical properties of the samples were studied. Porous UHMWPE samples were also studied in vitro and in vivo. The pores of UHMWPE scaffold are open and interconnected. Volume porosity of the obtained samples was 79±2%; the pore size range was 80-700μm. Strong connection of the two layers in multilayer UHMWPE scaffolds was observed with decreased number of fusion defects. Functionality of implants based on multilayer UHMWPE scaffolds is provided by the fixation of scaffolds in the bone defect through ingrowths of the connective tissue into the pores, which ensures the maintenance of the animals' mobility.

  10. Porous Three-Dimensional Carbon Nanotube Scaffolds for Tissue Engineering

    PubMed Central

    Lalwani, Gaurav; Gopalan, Anu; D’Agati, Michael; Sankaran, Jeyantt Srinivas; Judex, Stefan; Qin, Yi-Xian; Sitharaman, Balaji

    2015-01-01

    Assembly of carbon nanomaterials into three-dimensional (3D) architectures is necessary to harness their unique physiochemical properties for tissue engineering and regenerative medicine applications. Herein, we report the fabrication and comprehensive cytocompatibility assessment of 3D chemically crosslinked macro-sized (5–8 mm height and 4–6 mm diameter) porous carbon nanotube (CNT) scaffolds. Scaffolds prepared via radical initiated thermal crosslinking of single- or multi- walled CNTs (SWCNTs and MWCNTs) possess high porosity (>80%), and nano-, micro- and macro-scale interconnected pores. MC3T3 pre-osteoblast cells on MWCNT and SWCNT scaffolds showed good cell viability comparable to poly(lactic-co-glycolic) acid (PLGA) scaffolds after 5 days. Confocal live cell and immunofluorescence imaging showed that MC3T3 cells were metabolically active and could attach, proliferate and infiltrate MWCNT and SWCNT scaffolds. SEM imaging corroborated cell attachment and spreading and suggested that cell morphology is governed by scaffold surface roughness. MC3T3 cells were elongated on scaffolds with high surface roughness (MWCNTs) and rounded on scaffolds with low surface roughness (SWCNTs). The surface roughness of scaffolds may be exploited to control cellular morphology, and in turn govern cell fate. These results indicate that crosslinked MWCNTs and SWCNTs scaffolds are cytocompatible, and open avenues towards development of multifunctional all-carbon scaffolds for tissue engineering applications. PMID:25788440

  11. Image-based metrology of porous tissue engineering scaffolds

    NASA Astrophysics Data System (ADS)

    Rajagopalan, Srinivasan; Robb, Richard A.

    2006-03-01

    Tissue engineering is an interdisciplinary effort aimed at the repair and regeneration of biological tissues through the application and control of cells, porous scaffolds and growth factors. The regeneration of specific tissues guided by tissue analogous substrates is dependent on diverse scaffold architectural indices that can be derived quantitatively from the microCT and microMR images of the scaffolds. However, the randomness of pore-solid distributions in conventional stochastic scaffolds presents unique computational challenges. As a result, image-based characterization of scaffolds has been predominantly qualitative. In this paper, we discuss quantitative image-based techniques that can be used to compute the metrological indices of porous tissue engineering scaffolds. While bulk averaged quantities such as porosity and surface are derived directly from the optimal pore-solid delineations, the spatially distributed geometric indices are derived from the medial axis representations of the pore network. The computational framework proposed (to the best of our knowledge for the first time in tissue engineering) in this paper might have profound implications towards unraveling the symbiotic structure-function relationship of porous tissue engineering scaffolds.

  12. Surface-modified functionalized polycaprolactone scaffolds for bone repair: in vitro and in vivo experiments.

    PubMed

    Jensen, Jonas; Rölfing, Jan Hendrik Duedal; Le, Dang Quang Svend; Kristiansen, Asger Albaek; Nygaard, Jens Vinge; Hokland, Lea Bjerre; Bendtsen, Michael; Kassem, Moustapha; Lysdahl, Helle; Bünger, Cody Eric

    2014-09-01

    A porcine calvaria defect study was carried out to investigate the bone repair potential of three-dimensional (3D)-printed poly-ε-caprolactone (PCL) scaffolds embedded with nanoporous PCL. A microscopic grid network was created by rapid prototyping making a 3D-fused deposition model (FDM-PCL). Afterward, the FDM-PCL scaffolds were infused with a mixture of PCL, water, and 1,4-dioxane and underwent a thermal-induced phase separation (TIPS) followed by lyophilization. The TIPS process lead to a nanoporous structure shielded by the printed microstructure (NSP-PCL). Sixteen Landrace pigs were divided into two groups with 8 and 12 weeks follow-up, respectively. A total of six nonpenetrating holes were drilled in the calvaria of each animal. The size of the cylindrical defects was h 10 mm and Ø 10 mm. The defects were distributed randomly using following groups: (a) NSP-PCL scaffold, (b) FDM-PCL scaffold, (c) autograft, (d) empty defect, (a1) NSP-PCL scaffold + autologous mononuclear cells, and (a2) NSP-PCL scaffold + bone morphogenetic protein 2. Bone volume to total volume was analyzed using microcomputed tomography (µCT) and histomorphometry. The µCT and histological data showed significantly less bone formation in the NSP-PCL scaffolds in all three variations after both 8 and 12 weeks compared to all other groups. The positive autograft control had significantly higher new bone formation compared to all other groups except the FDM-PCL when analyzed using histomorphometry. The NSP-PCL scaffolds were heavily infiltrated with foreign body giant cells suggesting an inflammatory response and perhaps active resorption of the scaffold material. The unmodified FDM-PCL scaffold showed good osteoconductivity and osseointegration after both 8 and 12 weeks. © 2013 Wiley Periodicals, Inc.

  13. Development of porous scaffolds for bone tissue engineering

    NASA Astrophysics Data System (ADS)

    Ramay, Hassna Rehman

    In bone tissue engineering, biodegradable scaffolds are used as a temporary biological and mechanical support for new tissue growth. A scaffold must have good biocompatibility, controllable degradation rate, and enough mechanical strength to support bone cell attachment, differentiation, and proliferation as it gradually degrades and finally is completely replaced by new bone tissues. Biological studies and clinical practices have established that a three-dimensional interconnected porous structure is necessary to allow cell attachment, proliferation, and differentiation, and to provide pathways for biofluids. However, the mechanical strength of a material generally decreases as increasing porosity. The conflicting interests between biological and mechanical requirements thus pose a challenge in developing porous scaffolds for load-bearing bone tissue engineering. Two types of ceramic scaffolds, (1) Hydroxaypatite and (2) Hydroxaypatite/tricalcium phosphate, are prepared in this study utilizing a novel technique that combines the gel casting and polymer sponge methods. This technique provides better control over material microstructure and can produce scaffolds with enhanced mechanical toughness and strength. The hydroxyapatite scaffolds prepared by this technique have an open, uniform and interconnected porous structure (˜porosity = 76%) with compressive modulus of 7 GPa, comparable to that of cortical bone, and compressive strength of 5 MPa, comparable to that of cancellous bone. The second type of ceramic scaffold is a biphasic nano composite with tricalcium phosphate as the main matrix reinforced with hydroxyapatite (HA) nano-fibers. The porous scaffold attained a compressive strength of 9.6 MPa (˜porosity = 73%), comparable to the high-end value of cancellous bone. The toughness of the scaffold increased from 1.00 to 1.72 kN/m (˜porosity = 73%), as the addition of HA nano-fibers increased up to 5 wt.%. Polymer scaffolds are prepared using a solid

  14. The role of biodegradable engineered random polycaprolactone nanofiber scaffolds seeded with nestin-positive hair follicle stem cells for tissue engineering

    PubMed Central

    Yari, Abazar; Teimourian, Shahram; Amidi, Fardin; Bakhtiyari, Mehrdad; Heidari, Fatemeh; Sajedi, Nayereh; Veijouye, Sanaz Joulai; Dodel, Masumeh; Nobakht, Maliheh

    2016-01-01

    Background: Tissue engineering is a new approach to reconstruction and/or regeneration of lost or damaged tissue. The purpose of this study was to fabricate the polycaprolactone (PCL) random nanofiber scaffold as well as evaluation of the cell viability, adhesion, and proliferation of rat nestin-positive hair follicle stem cells (HFSCs) in the graft material using electrospun PCL nanofiber scaffold in regeneration medicine. Materials and Methods: The bulge HFSCs was isolated from rat whiskers and cultivated in Dulbecco's modified Eagle's medium/F12. To evaluate the biological nature of the bulge stem cells, flow cytometry using nestin, CD34 and K15 antibodies was performed. Electrospinning was used for the production of PCL nanofiber scaffolds. Furthermore, scanning electron microscopy (SEM) for HFSCs attachment, infiltration, and morphology, 3-(4, 5-di-methylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay for cell viability and cytotoxicity, tensile strength of the scaffolds mesh, and histology analysis were used. Results: Flow cytometry showed that HFSCs were nestin and CD34 positive but K15 negative. The results of the MTT assay showed cell viability and cell proliferation of the HFSCs on PCL nanofiber scaffolds. SEM microscopy photographs indicated that HFSCs are attached and spread on PCL nanofiber scaffolds. Furthermore, tensile strength of the scaffolds mesh was measured. Conclusion: The results of this study revealed that modified PCL nanofiber scaffolds are suitable for HFSCs seeding, attachment, and proliferation. Furthermore, HFSCs are attached and proliferated on PCL nanofiber scaffolds. PMID:26962524

  15. 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

  16. Multiwall carbon nanotubes/polycaprolactone scaffolds seeded with human dental pulp stem cells for bone tissue regeneration.

    PubMed

    Flores-Cedillo, M L; Alvarado-Estrada, K N; Pozos-Guillén, A J; Murguía-Ibarra, J S; Vidal, M A; Cervantes-Uc, J M; Rosales-Ibáñez, R; Cauich-Rodríguez, J V

    2016-02-01

    Conventional approaches to bone regeneration rarely use multiwall carbon nanotubes (MWCNTs) but instead use polymeric matrices filled with hydroxyapatite, calcium phosphates and bioactive glasses. In this study, we prepared composites of MWCNTs/polycaprolactone (PCL) for bone regeneration as follows: (a) MWCNTs randomly dispersed on PCL, (b) MWCNTs aligned with an electrical field to determine if the orientation favors the growing of human dental pulp stem cells (HDPSCs), and (c) MWCNTs modified with β-glycerol phosphate (BGP) to analyze its osteogenic potential. Raman spectroscopy confirmed the presence of MWCNTs and BGP on PCL, whereas the increase in crystallinity by the addition of MWCNTs to PCL was confirmed by X-ray diffraction and differential scanning calorimetry. A higher elastic modulus (608 ± 4.3 MPa), maximum stress (42 ± 6.1 MPa) and electrical conductivity (1.67 × 10(-7) S/m) were observed in non-aligned MWCNTs compared with the pristine PCL. Cell viability at 14 days was similar in all samples according to the live/dead assay, but the 21 day cell proliferation, measured by MTT was higher in MWCNTs aligned with BGP. Von Kossa and Alizarin red showed larger amounts of mineral deposits on MWCNTs aligned with BGP, indicating that at 21 days, this scaffold promotes osteogenic differentiation of HDPSCs.

  17. Living Bacterial Sacrificial Porogens to Engineer Decellularized Porous Scaffolds

    PubMed Central

    Xu, Feng; Sridharan, BanuPriya; Durmus, Naside Gozde; Wang, ShuQi; Yavuz, Ahmet Sinan; Gurkan, Umut Atakan; Demirci, Utkan

    2011-01-01

    Decellularization and cellularization of organs have emerged as disruptive methods in tissue engineering and regenerative medicine. Porous hydrogel scaffolds have widespread applications in tissue engineering, regenerative medicine and drug discovery as viable tissue mimics. However, the existing hydrogel fabrication techniques suffer from limited control over pore interconnectivity, density and size, which leads to inefficient nutrient and oxygen transport to cells embedded in the scaffolds. Here, we demonstrated an innovative approach to develop a new platform for tissue engineered constructs using live bacteria as sacrificial porogens. E.coli were patterned and cultured in an interconnected three-dimensional (3D) hydrogel network. The growing bacteria created interconnected micropores and microchannels. Then, the scafold was decellularized, and bacteria were eliminated from the scaffold through lysing and washing steps. This 3D porous network method combined with bioprinting has the potential to be broadly applicable and compatible with tissue specific applications allowing seeding of stem cells and other cell types. PMID:21552485

  18. Living bacterial sacrificial porogens to engineer decellularized porous scaffolds.

    PubMed

    Xu, Feng; Sridharan, BanuPriya; Durmus, Naside Gozde; Wang, ShuQi; Yavuz, Ahmet Sinan; Gurkan, Umut Atakan; Demirci, Utkan

    2011-04-28

    Decellularization and cellularization of organs have emerged as disruptive methods in tissue engineering and regenerative medicine. Porous hydrogel scaffolds have widespread applications in tissue engineering, regenerative medicine and drug discovery as viable tissue mimics. However, the existing hydrogel fabrication techniques suffer from limited control over pore interconnectivity, density and size, which leads to inefficient nutrient and oxygen transport to cells embedded in the scaffolds. Here, we demonstrated an innovative approach to develop a new platform for tissue engineered constructs using live bacteria as sacrificial porogens. E.coli were patterned and cultured in an interconnected three-dimensional (3D) hydrogel network. The growing bacteria created interconnected micropores and microchannels. Then, the scafold was decellularized, and bacteria were eliminated from the scaffold through lysing and washing steps. This 3D porous network method combined with bioprinting has the potential to be broadly applicable and compatible with tissue specific applications allowing seeding of stem cells and other cell types.

  19. Long-bone critical-size defects treated with tissue-engineered polycaprolactone-co-lactide scaffolds: a pilot study on rats.

    PubMed

    Rentsch, Claudia; Rentsch, Barbe; Breier, Annette; Spekl, Kathrin; Jung, Roland; Manthey, Suzanne; Scharnweber, Dieter; Zwipp, Hans; Biewener, Achim

    2010-12-01

    The aim of this study was to evaluate the osteogenic potential of embroidered, tissue-engineered polycaprolactone-co-lactide (trade name: PCL) scaffolds for the reconstruction of large bone defects. Ten piled-up PCL scaffolds were implanted in femura with a critical size defect of immunodeficient nude rats for 12 weeks [n = 4, group 1: noncoated, group 2: collagen I (coll I), group 3: collagen I/chondroitin sulfate (coll I/CS), and group 4: collagen I/chondroitin sulfate/human mesenchymal stem cells (coll I/CS/hMSC)]. X-ray examination, computer tomography, and histological analyses of the explanted scaffold pads were performed. The quantification of the bone volume ratio showed a significantly higher rate of new bone formation at coll I/CS-coated scaffolds compared with the other groups. Histological investigations revealed that the defect reconstruction started from the peripheral bone ends and incorporated into the scaffold material. Additionally seeded hMSC on coll I/CS-coated scaffolds showed a higher matrix deposition inside the implant but no higher bone formation was observed. These data imply that the coll I/CS-coated PCL scaffolds have the highest potential for treating critical size defects. The scaffolds, being variable in size and structure, can be adapted to any bone defect.

  20. 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.

  1. Tissue engineering scaffold material of porous nanohydroxyapatite/polyamide 66.

    PubMed

    Xu, Qian; Lu, Hongyan; Zhang, Jingchao; Lu, Guoyu; Deng, Zhennan; Mo, Anchun

    2010-05-13

    The aim of the study was to investigate a porous nanohydroxyapatite/polyamide 66 (n-HA/PA66) scaffold material that was implanted into muscle and tibiae of 16 New Zealand white rabbits to evaluate the biocompatibility and osteogenesis and osteoinductivity of the materials in vivo. The samples were harvested at 2, 4, 12 and 26 weeks respectively, and subjected to histological analysis. At 2 weeks, the experiment showed that osteogenesis was detected in porous n-HA/PA66 composite and the density of new bone formation was similar to the surrounding host bone at 12 weeks. The study indicated that three-dimensional pore structures could facilitate cell adhesion, differentiation and proliferation, and help with fibrovascular and nerve colonization. In conclusion, porous n-HA/PA66 scaffold material could be a good candidate as a bone substitute material used in clinics due to its excellent histocompatibility, osteoconductivity and osteoinductivity.

  2. Integration of 3D Printed and Micropatterned Polycaprolactone Scaffolds for Guidance of Oriented Collagenous Tissue Formation In Vivo.

    PubMed

    Pilipchuk, Sophia P; Monje, Alberto; Jiao, Yizu; Hao, Jie; Kruger, Laura; Flanagan, Colleen L; Hollister, Scott J; Giannobile, William V

    2016-03-01

    Scaffold design incorporating multiscale cues for clinically relevant, aligned tissue regeneration has potential to improve structural and functional integrity of multitissue interfaces. The objective of this preclinical study is to develop poly(ε-caprolactone) (PCL) scaffolds with mesoscale and microscale architectural cues specific to human ligament progenitor cells and assess their ability to form aligned bone-ligament-cementum complexes in vivo. PCL scaffolds are designed to integrate a 3D printed bone region with a micropatterned PCL thin film consisting of grooved pillars. The patterned film region is seeded with human ligament cells, fibroblasts transduced with bone morphogenetic protein-7 genes seeded within the bone region, and a tooth dentin segment positioned on the ligament region prior to subcutaneous implantation into a murine model. Results indicate increased tissue alignment in vivo using micropatterned PCL films, compared to random-porous PCL. At week 6, 30 μm groove depth significantly enhances oriented collagen fiber thickness, overall cell alignment, and nuclear elongation relative to 10 μm groove depth. This study demonstrates for the first time that scaffolds with combined hierarchical mesoscale and microscale features can align cells in vivo for oral tissue repair with potential for improving the regenerative response of other bone-ligament complexes.

  3. Integration of 3D Printed and Micropatterned Polycaprolactone Scaffolds for Guidance of Oriented Collagenous Tissue Formation In vivo

    PubMed Central

    Pilipchuk, Sophia P; Monje, Alberto; Jiao, Yizu; Hao, Jie; Kruger, Laura; Flanagan, Colleen L; Hollister, Scott J

    2016-01-01

    Scaffold design incorporating multi-scale cues for clinically-relevant, aligned tissue regeneration has potential to improve structural and functional integrity of multi-tissue interfaces. The objective of this pre-clinical study was to develop poly(ε-caprolactone) (PCL) scaffolds with mesoscale and microscale architectural cues specific to human ligament progenitor cells and assess their ability to form aligned bone-ligament-cementum complexes in vivo. PCL scaffolds were designed to integrate a 3D printed bone region with a micropatterned PCL thin film consisting of grooved pillars. The patterned film region was seeded with human ligament cells, fibroblasts transduced with BMP-7 genes seeded within the bone region, and a tooth dentin segment positioned on the ligament region prior to subcutaneous implantation into a murine model. Results indicated increased tissue alignment in vivo using micropatterned PCL films, compared to random-porous PCL. At 6 weeks, 30um groove depth significantly enhanced oriented collagen fiber thickness, overall cell alignment, and nuclear elongation relative to 10um groove depth. This study demonstrates for the first time that scaffolds with combined hierarchical mesoscale and microscale features can align cells in vivo for oral tissue repair with potential for improving the regenerative response of other bone-ligament complexes. PMID:26820240

  4. Electrospun Scaffolds for Osteoblast Cells: Peptide-Induced Concentration-Dependent Improvements of Polycaprolactone.

    PubMed

    Dettin, Monica; Zamuner, Annj; Roso, Martina; Gloria, Antonio; Iucci, Giovanna; Messina, Grazia M L; D'Amora, Ugo; Marletta, Giovanni; Modesti, Michele; Castagliuolo, Ignazio; Brun, Paola

    2015-01-01

    The design of hybrid poly-ε-caprolactone (PCL)-self-assembling peptides (SAPs) matrices represents a simple method for the surface functionalization of synthetic scaffolds, which is essential for cell compatibility. This study investigates the influence of increasing concentrations (2.5%, 5%, 10% and 15% w/w SAP compared to PCL) of three different SAPs on the physico-chemical/mechanical and biological properties of PCL fibers. We demonstrated that physico-chemical surface characteristics were slightly improved at increasing SAP concentrations: the fiber diameter increased; surface wettability increased with the first SAP addition (2.5%) and slightly less for the following ones; SAP-surface density increased but no change in the conformation was registered. These results could allow engineering matrices with structural characteristics and desired wettability according to the needs and the cell system used. The biological and mechanical characteristics of these scaffolds showed a particular trend at increasing SAP concentrations suggesting a prevailing correlation between cell behavior and mechanical features of the matrices. As compared with bare PCL, SAP enrichment increased the number of metabolic active h-osteoblast cells, fostered the expression of specific osteoblast-related mRNA transcripts, and guided calcium deposition, revealing the potential application of PCL-SAP scaffolds for the maintenance of osteoblast phenotype.

  5. Electrospun Scaffolds for Osteoblast Cells: Peptide-Induced Concentration-Dependent Improvements of Polycaprolactone

    PubMed Central

    Dettin, Monica; Zamuner, Annj; Roso, Martina; Gloria, Antonio; Iucci, Giovanna; Messina, Grazia M. L.; D'Amora, Ugo; Marletta, Giovanni; Modesti, Michele; Castagliuolo, Ignazio; Brun, Paola

    2015-01-01

    The design of hybrid poly-ε-caprolactone (PCL)-self-assembling peptides (SAPs) matrices represents a simple method for the surface functionalization of synthetic scaffolds, which is essential for cell compatibility. This study investigates the influence of increasing concentrations (2.5%, 5%, 10% and 15% w/w SAP compared to PCL) of three different SAPs on the physico-chemical/mechanical and biological properties of PCL fibers. We demonstrated that physico-chemical surface characteristics were slightly improved at increasing SAP concentrations: the fiber diameter increased; surface wettability increased with the first SAP addition (2.5%) and slightly less for the following ones; SAP-surface density increased but no change in the conformation was registered. These results could allow engineering matrices with structural characteristics and desired wettability according to the needs and the cell system used. The biological and mechanical characteristics of these scaffolds showed a particular trend at increasing SAP concentrations suggesting a prevailing correlation between cell behavior and mechanical features of the matrices. As compared with bare PCL, SAP enrichment increased the number of metabolic active h-osteoblast cells, fostered the expression of specific osteoblast-related mRNA transcripts, and guided calcium deposition, revealing the potential application of PCL-SAP scaffolds for the maintenance of osteoblast phenotype. PMID:26361004

  6. Significant degradability enhancement in multilayer coating of polycaprolactone-bioactive glass/gelatin-bioactive glass on magnesium scaffold for tissue engineering applications

    NASA Astrophysics Data System (ADS)

    Yazdimamaghani, Mostafa; Razavi, Mehdi; Vashaee, Daryoosh; Pothineni, Venkata Raveendra; Rajadas, Jayakumar; Tayebi, Lobat

    2015-05-01

    Magnesium (Mg) is a promising candidate to be used in medical products especially as bone tissue engineering scaffolds. The main challenge for using Mg in biomedical applications is its high degradation rate in the body. For this reason, in this study, a multilayer polymeric layer composed of polycaprolactone (PCL) and gelatin (Gel) reinforced with bioactive glass (BaG) particles has been applied on the surface of Mg scaffolds. The materials characteristics of uncoated Mg scaffold, Mg scaffold coated only with PCL-BaG and Mg scaffold coated with PCL-BaG and Gel-BaG have been analyzed and compared in detail. Scanning electron microscope (SEM) equipped with energy dispersive spectroscopy (EDS), and Fourier transform infrared spectroscopy (FTIR) were utilized for microstructural studies. In vitro bioactivity and biodegradation evaluations were carried out by submerging the scaffolds in simulated body fluid (SBF) at pre-determined time points. The results demonstrated that Mg scaffold coated with PCL-BaG and Gel-BaG exhibited significant improvement in biodegradability.

  7. Evaluation of a thermoresponsive polycaprolactone scaffold for in vitro three-dimensional stem cell differentiation.

    PubMed

    Hruschka, Veronika; Saeed, Aram; Slezak, Paul; Cheikh Al Ghanami, Racha; Feichtinger, Georg Alexander; Alexander, Cameron; Redl, Heinz; Shakesheff, Kevin; Wolbank, Susanne

    2015-01-01

    Tissue engineering (TE) strategies aim at imitating the natural process of regeneration by using bioresorbable scaffolds that support cellular attachment, migration, proliferation, and differentiation. Based on the idea of combining a fully degradable polymer [poly(ɛ-caprolactone)] with a thermoresponsive polymer (polyethylene glycol methacrylate), a scaffold was developed, which liquefies below 20°C and solidifies at 37°C. In this study, this scaffold was evaluated for its ability to support C2C12 cells and human adipose-derived stem cells (ASCs) to generate an expandable three-dimensional (3D) construct for soft or bone TE. As a first step, biomaterial seeding was optimized and cellular attachment, survival, distribution, and persistence within the 3D material were characterized. C2C12 cells were differentiated toward the osteogenic as well as myogenic lineage, while ASCs were cultured in control, adipogenic, or osteogenic differentiation media. Differentiation was examined using quantitative real-time PCR for the expression of osteogenic, myogenic, and adipogenic markers and by enzyme activity and immunoassays. Both cell types attached and were found evenly distributed within the material. C2C12 cells and ASCs demonstrated the potential to differentiate in all tested lineages under 2D conditions. Under 3D osteogenic conditions for C2C12 cells, only osteocalcin expression (fold induction: 16.3±0.2) and alkaline phosphatase (ALP) activity (p<0.001) were increased compared with the control C2C12 cells. Three-dimensional osteogenic differentiation of ASC was limited and donor dependent. Only one donor showed an increase in the osteogenic markers osteocalcin (p=0.027) and osteopontin (p=0.038). In contrast, differentiation toward the myogenic or adipogenic lineage showed expression of specific markers in 3D, at least at the level of the 2D culture. In 3D culture, strong induction of myogenin (p<0.001) as well as myoD (p<0.001) was found in C2C12 cells. The

  8. Hybrid vitronectin-mimicking polycaprolactone scaffolds for human retinal progenitor cell differentiation and transplantation.

    PubMed

    Lawley, Elodie; Baranov, Petr; Young, Michael

    2015-01-01

    Many advances have been made in an attempt to treat retinal degenerative diseases, such as age-related macular degeneration and retinitis pigmentosa. The irreversible loss of photoreceptors is common to both, and currently no restorative clinical treatment exists. It has been shown that retinal progenitor and photoreceptor precursor cell transplantation can rescue the retinal structure and function. Importantly, retinal progenitor cells can be collected from the developing neural retina with further expansion and additional modification in vitro, and the delivery into the degenerative host can be performed as a single-cell suspension injection or as a complex graft transplantation. Previously, we have described several polymer scaffolds for culture and transplantation of retinal progenitor cells of both mouse and human origin. This tissue engineering strategy increases donor cell survival and integration. We have also shown that biodegradable poly(ɛ-caprolactone) induces mature photoreceptor differentiation from human retinal progenitor cells. However, poor adhesive properties limit its use, and therefore it requires additional surface modification. The aim of this work was to study vitronectin-mimicking oligopeptides (Synthemax II-SC) poly(ɛ-caprolactone) films and their effects on human retinal progenitor cell adhesion, proliferation, and differentiation. Here, we show that the incorporation of vitronectin-mimicking oligopeptide into poly(ɛ-caprolactone) leads to dose-dependent increases in cell adhesion; the optimum dose identified as 30 µg/ml. Inhibition of human retinal progenitor cells proliferation was seen on poly(ɛ-caprolactone) and was maintained with the hybrid scaffold. This has been shown to be beneficial for driving cell differentiation. Additionally, we observed equal expression of Nrl, rhodopsin, recoverin, and rod outer membrane 1 after differentiation on the hybrid scaffold as compared to the standard fibronectin coating of poly

  9. Injectable and porous PLGA microspheres that form highly porous scaffolds at body temperature.

    PubMed

    Qutachi, Omar; Vetsch, Jolanda R; Gill, Daniel; Cox, Helen; Scurr, David J; Hofmann, Sandra; Müller, Ralph; Quirk, Robin A; Shakesheff, Kevin M; Rahman, Cheryl V

    2014-12-01

    Injectable scaffolds are of interest in the field of regenerative medicine because of their minimally invasive mode of delivery. For tissue repair applications, it is essential that such scaffolds have the mechanical properties, porosity and pore diameter to support the formation of new tissue. In the current study, porous poly(dl-lactic acid-co-glycolic acid) (PLGA) microspheres were fabricated with an average size of 84±24μm for use as injectable cell carriers. Treatment with ethanolic sodium hydroxide for 2min was observed to increase surface porosity without causing the microsphere structure to disintegrate. This surface treatment also enabled the microspheres to fuse together at 37°C to form scaffold structures. The average compressive strength of the scaffolds after 24h at 37°C was 0.9±0.1MPa, and the average Young's modulus was 9.4±1.2MPa. Scaffold porosity levels were 81.6% on average, with a mean pore diameter of 54±38μm. This study demonstrates a method for fabricating porous PLGA microspheres that form solid porous scaffolds at body temperature, creating an injectable system capable of supporting NIH-3T3 cell attachment and proliferation in vitro.

  10. Injectable and porous PLGA microspheres that form highly porous scaffolds at body temperature

    PubMed Central

    Qutachi, Omar; Vetsch, Jolanda R.; Gill, Daniel; Cox, Helen; Scurr, David J.; Hofmann, Sandra; Müller, Ralph; Quirk, Robin A.; Shakesheff, Kevin M.; Rahman, Cheryl V.

    2014-01-01

    Injectable scaffolds are of interest in the field of regenerative medicine because of their minimally invasive mode of delivery. For tissue repair applications, it is essential that such scaffolds have the mechanical properties, porosity and pore diameter to support the formation of new tissue. In the current study, porous poly(dl-lactic acid-co-glycolic acid) (PLGA) microspheres were fabricated with an average size of 84 ± 24 μm for use as injectable cell carriers. Treatment with ethanolic sodium hydroxide for 2 min was observed to increase surface porosity without causing the microsphere structure to disintegrate. This surface treatment also enabled the microspheres to fuse together at 37 °C to form scaffold structures. The average compressive strength of the scaffolds after 24 h at 37 °C was 0.9 ± 0.1 MPa, and the average Young’s modulus was 9.4 ± 1.2 MPa. Scaffold porosity levels were 81.6% on average, with a mean pore diameter of 54 ± 38 μm. This study demonstrates a method for fabricating porous PLGA microspheres that form solid porous scaffolds at body temperature, creating an injectable system capable of supporting NIH-3T3 cell attachment and proliferation in vitro. PMID:25152354

  11. Fibro-porous poliglecaprone/polycaprolactone conduits: synergistic effect of composition and in vitro degradation on mechanical properties.

    PubMed

    Patel, Harsh N; Garcia, Roman; Schindler, Carrie; Dean, Derrick; Pogwizd, Steven M; Singh, Raj; Vohra, Yogesh K; Thomas, Vinoy

    2015-04-01

    Blends of poliglecaprone (PGC) and polycaprolactone (PCL) of varying compositions were electrospun into tubular conduits and their mechanical, morphological, thermal and in vitro degradation properties were evaluated under simulated physiological conditions. Generally, mechanical strength, modulus and hydrophilic nature were enhanced by the addition of PGC to PCL. An in vitro degradation study in phosphate-buffered saline (pH 7.3) was carried out for up to 1 month to understand the hydrolytic degradation effect on the mechanical properties in both the longitudinal and circumferential directions. Pure PCL and 4:1 PCL/PGC blend scaffolds exhibited considerable elastic stiffening after a 1 month in vitro degradation. Fourier transform infrared spectroscopic and DSC techniques were used to understand the degradation behavior and the changes in structure and crystallinity of the polymeric blends. A 3:1 PCL/PGC blend was concluded to be a judicious blend composition for tubular grafts based on overall results on the mechanical properties and performance after a 1 month in vitro degradation study.

  12. 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.

  13. Protein adsorption and cell adhesion on three-dimensional polycaprolactone scaffolds with respect to plasma modification by etching and deposition techniques

    NASA Astrophysics Data System (ADS)

    Myung, Sung Woon; Ko, Yeong Mu; Kim, Byung Hoon

    2014-11-01

    In this work, protein adsorption and cell adhesion on three-dimensional (3D) polycaprolactone (PCL) scaffolds treated by plasma etching and deposition were performed. The 3D PCL scaffold used as a substrate of a bone tissue was fabricated by recent rapid prototype techniques. To increase surface properties, such as hydrophilicity, roughness, and surface chemistry, through good protein adhesion on scaffolds, oxygen (O2) plasma etching and acrylic acid or allyamine plasma deposition were performed on the 3D PCL scaffolds. The O2 plasma etching induced the formation of random nanoporous structures on the roughened surfaces of the 3D PCL scaffolds. The plasma deposition with acrylic acid and allyamine induced the chemical modification for introducing a functional group. The protein adsorption increased on the O2 plasma-etched surface compared with an untreated 3D PCL scaffold. MC3T3-E1 cells adhered bioactively on the etched and deposited surface compared with the untreated surface. The present plasma modification might be sought as an effective technique for enhancing protein adsorption and cell adhesion.

  14. 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.

  15. Highly porous 3D nanofiber scaffold using an electrospinning technique.

    PubMed

    Kim, Geunhyung; Kim, WanDoo

    2007-04-01

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

  16. Ultra-porous titanium oxide scaffold with high compressive strength

    PubMed Central

    Tiainen, Hanna; Lyngstadaas, S. Petter; Ellingsen, Jan Eirik

    2010-01-01

    Highly porous and well interconnected titanium dioxide (TiO2) scaffolds with compressive strength above 2.5 MPa were fabricated without compromising the desired pore architectural characteristics, such as high porosity, appropriate pore size, surface-to-volume ratio, and interconnectivity. Processing parameters and pore architectural characteristics were investigated in order to identify the key processing steps and morphological properties that contributed to the enhanced strength of the scaffolds. Cleaning of the TiO2 raw powder removed phosphates but introduced sodium into the powder, which was suggested to decrease the slurry stability. Strong correlation was found between compressive strength and both replication times and solid content in the ceramic slurry. Increase in the solid content resulted in more favourable sponge loading, which was achieved due to the more suitable rheological properties of the ceramic slurry. Repeated replication process induced only negligible changes in the pore architectural parameters indicating a reduced flaw size in the scaffold struts. The fabricated TiO2 scaffolds show great promise as load-bearing bone scaffolds for applications where moderate mechanical support is required. PMID:20711636

  17. Exploiting novel sterilization techniques for porous polyurethane scaffolds.

    PubMed

    Bertoldi, Serena; Farè, Silvia; Haugen, Håvard Jostein; Tanzi, Maria Cristina

    2015-05-01

    Porous polyurethane (PU) structures raise increasing interest as scaffolds in tissue engineering applications. Understanding the effects of sterilization on their properties is mandatory to assess their potential use in the clinical practice. The aim of this work is the evaluation of the effects of two innovative sterilization techniques (i.e. plasma, Sterrad(®) system, and ozone) on the morphological, chemico-physical and mechanical properties of a PU foam synthesized by gas foaming, using water as expanding agent. In addition, possible toxic effects of the sterilization were evaluated by in vitro cytotoxicity tests. Plasma sterilization did not affect the morphological and mechanical properties of the PU foam, but caused at some extent degradative phenomena, as detected by infrared spectroscopy. Ozone sterilization had a major effect on foam morphology, causing the formation of new small pores, and stronger degradation and oxidation on the structure of the material. These modifications affected the mechanical properties of the sterilized PU foam too. Even though, no cytotoxic effects were observed after both plasma and ozone sterilization, as confirmed by the good values of cell viability assessed by Alamar Blue assay. The results here obtained can help in understanding the effects of sterilization procedures on porous polymeric scaffolds, and how the scaffold morphology, in particular porosity, can influence the effects of sterilization, and viceversa.

  18. A novel pressed porous silicon-polycaprolactone composite as a dual-purpose implant for the delivery of cells and drugs to the eye.

    PubMed

    Irani, Yazad D; Tian, Yuan; Wang, Mengjia; Klebe, Sonja; McInnes, Steven J; Voelcker, Nicolas H; Coffer, Jeffery L; Williams, Keryn A

    2015-10-01

    Dysfunction of corneal epithelial stem cells can result in painful and blinding disease of the ocular surface. In such cases, treatment may involve transfer of growth factor and normal adult stem cells to the ocular surface. Our purpose was to develop an implantable scaffold for the delivery of drugs and cells to the ocular surface. We examined the potential of novel composite biomaterials fabricated from electrospun polycaprolactone (PCL) fibres into which nanostructured porous silicon (pSi) microparticles of varying sizes (150-250 μm or <40 μm) had been pressed. The PCL fabric provided a flexible support for mammalian cells, whereas the embedded pSi provided a substantial surface area for efficient delivery of adsorbed drugs and growth factors. Measurements of tensile strength of these composites revealed that the pSi did not strongly influence the mechanical properties of the polymer microfiber component for the Si loadings evaluated. Human lens epithelial cells (SRA01/04) attached to the composite materials, and exhibited enhanced attachment and growth when the materials were coated with foetal bovine serum. To examine the ability of the materials to deliver a small-drug payload, pSi microparticles were loaded with fluorescein diacetate prior to cell attachment. After 6 hours (h), cells exhibited intracellular fluorescence, indicative of transfer of the fluorescein diacetate into viable cells and its subsequent enzymatic conversion to fluorescein. To investigate loading of large-molecule biologics, murine BALB/c 3T3 cells, responsive to epidermal growth factor, insulin and transferrin, were seeded on composite materials. The cells showed significantly more proliferation at 48 h when seeded on composites loaded with these biologics, than on unloaded composites. No cell proliferation was observed on PCL alone, indicating the biologics had loaded into the pSi microparticles. Drug release, measured by ELISA for insulin, indicated a burst followed by a slower

  19. Diffusion model to describe osteogenesis within a porous titanium scaffold.

    PubMed

    Schmitt, M; Allena, R; Schouman, T; Frasca, S; Collombet, J M; Holy, X; Rouch, P

    2016-01-01

    In this study, we develop a two-dimensional finite element model, which is derived from an animal experiment and allows simulating osteogenesis within a porous titanium scaffold implanted in ewe's hemi-mandible during 12 weeks. The cell activity is described through diffusion equations and regulated by the stress state of the structure. We compare our model to (i) histological observations and (ii) experimental data obtained from a mechanical test done on sacrificed animal. We show that our mechano-biological approach provides consistent numerical results and constitutes a useful tool to predict osteogenesis pattern.

  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. 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.

  2. Effect of self-assembled nanofibrous silk/polycaprolactone layer on the osteoconductivity and mechanical properties of biphasic calcium phosphate scaffolds.

    PubMed

    Roohani-Esfahani, S I; Lu, Z F; Li, J J; Ellis-Behnke, R; Kaplan, D L; Zreiqat, H

    2012-01-01

    We here present the first successful report on combining nanostructured silk and poly(ε-caprolactone) (PCL) with a ceramic scaffold to produce a composite scaffold that is highly porous (porosity ∼85%, pore size ∼500 μm, ∼100% interconnectivity), strong and non-brittle with a surface that resembles extracellular matrix (ECM). The ECM-like surface was developed by self-assembly of nanofibrous structured silk (20-80 nm diameter, similar to native collagen found in ECM) over a thin PCL layer which is coated on biphasic calcium phosphate (BCP) scaffolds. The effects of different concentrations of silk solution on the mechanical and physical properties of the scaffolds were also comprehensively examined. Our results showed that using silk only (irrespective of concentration) for the modification of ceramic scaffolds could drastically reduce the compressive strength of the modified scaffolds in aqueous media, and the modification made a limited contribution to improving scaffold toughness. Using PCL/nanostructured silk the compressive strength and modulus of the modified scaffolds reached 0.42 MPa (compared with 0.07 MPa for BCP) and ∼25 MPa (compared with 5 MPa for BCP), respectively. The failure strain of the modified scaffold increased more than 6% compared with a BCP scaffold (failure strain of less than 1%), indicating a transformation from brittle to elastic behavior. The cytocompatibility of ECM-like composite scaffolds was investigated by studying the attachment, morphology, proliferation and bone-related gene expression of primary human bone-derived cells. Cells cultured on the developed scaffolds for 7 days had significant up-regulation of cell proliferation (∼1.6-fold higher, P<0.001) and osteogenic gene expression levels (collagen type I, osteocalcin and bone sialoprotein) compared with the other groups tested.

  3. Porous silicon-based scaffolds for tissue engineering and other biomedical applications

    NASA Astrophysics Data System (ADS)

    Coffer, Jeffery L.; Whitehead, Melanie A.; Nagesha, Dattatri K.; Mukherjee, Priyabrata; Akkaraju, Giridhar; Totolici, Mihaela; Saffie, Roghieh S.; Canham, Leigh T.

    2005-06-01

    This work describes the formation of porous composite materials based on a combination of bioactive mesoporous silicon and bioerodible polymers such as poly-caprolactone (PCL). The fabrication of a range of composites prepared by both salt leaching and microemulsion techniques are discussed. Particular attention to the influence of Si content in the composite on in vitro calcification assays are assessed. For each system, cytotoxicity and cellular proliferation are explicitly evaluated through fibroblast cell culture assays.

  4. Inhibitor of PI3K/Akt Signaling Pathway Small Molecule Promotes Motor Neuron Differentiation of Human Endometrial Stem Cells Cultured on Electrospun Biocomposite Polycaprolactone/Collagen Scaffolds.

    PubMed

    Ebrahimi-Barough, Somayeh; Hoveizi, Elham; Yazdankhah, Meysam; Ai, Jafar; Khakbiz, Mehrdad; Faghihi, Faezeh; Tajerian, Roksana; Bayat, Neda

    2017-05-01

    Small molecules as useful chemical tools can affect cell differentiation and even change cell fate. It is demonstrated that LY294002, a small molecule inhibitor of phosphatidylinositol 3-kinase (PI3K)/Akt signal pathway, can inhibit proliferation and promote neuronal differentiation of mesenchymal stem cells (MSCs). The purpose of this study was to investigate the differentiation effect of Ly294002 small molecule on the human endometrial stem cells (hEnSCs) into motor neuron-like cells on polycaprolactone (PCL)/collagen scaffolds. hEnSCs were cultured in a neurogenic inductive medium containing 1 μM LY294002 on the surface of PCL/collagen electrospun fibrous scaffolds. Cell attachment and viability of cells on scaffolds were characterized by scanning electron microscope (SEM) and 3-(4,5-dimethylthiazoyl-2-yl)2,5-diphenyltetrazolium bromide (MTT) assay. The expression of neuron-specific markers was assayed by real-time PCR and immunocytochemistry analysis after 15 days post induction. Results showed that attachment and differentiation of hEnSCs into motor neuron-like cells on the scaffolds with Ly294002 small molecule were higher than that of the cells on tissue culture plates as control group. In conclusion, PCL/collagen electrospun scaffolds with Ly294002 have potential for being used in neural tissue engineering because of its bioactive and three-dimensional structure which enhances viability and differentiation of hEnSCs into neurons through inhibition of the PI3K/Akt pathway. Thus, manipulation of this pathway by small molecules can enhance neural differentiation.

  5. Integrating sol-gel with cold plasmas modified porous polycaprolactone membranes for the drug-release of silver-sulfadiazine and ketoprofen

    NASA Astrophysics Data System (ADS)

    Mangindaan, Dave; Chen, Chao-Ting; Wang, Meng-Jiy

    2012-12-01

    A controlled release system composed of surface modified porous polycaprolactone (PCL) membranes combined with a layer of tetraorthosilicate (TEOS)-chitosan sol-gel was reported in this study. PCL is a hydrophobic, semi-crystalline, and biodegradable polymer with a relatively slow degradation rate. The drugs chosen for release experiments were silver-sulfadiazine (AgSD) and ketoprofen which were impregnated in the TEOS-chitosan sol-gel. The surface modification was achieved by O2 plasma and the surfaces were characterized by water contact angle (WCA) measurements, atomic force microscope (AFM), scanning electron microscope and electron spectroscopy for chemical analysis (ESCA). The results showed that the release of AgSD on O2 plasma treated porous PCL membranes was prolonged when compared with the pristine sample. On the contrary, the release rate of ketoprofen revealed no significant difference on pristine and plasma treated PCL membranes. The prepared PCL membranes showed good biocompatibility for the wound dressing biomaterial applications.

  6. Fabrication of gelatin-hyaluronic acid hybrid scaffolds with tunable porous structures for soft tissue engineering.

    PubMed

    Zhang, Fan; He, Chuanglong; Cao, Lijun; Feng, Wei; Wang, Hongsheng; Mo, Xiumei; Wang, Jinwu

    2011-04-01

    The development of three-dimensional (3-D) scaffolds with highly open porous structure is one of the most important issues in tissue engineering. In this study, 3-D macroporous gelatin/hyaluronic acid (GE/HA) hybrid scaffolds with varying porous morphology were prepared by freeze-drying their blending solutions and subsequent chemical crosslinking by using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). The resulting scaffolds were characterized by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Their swelling, in vitro degradation properties and compressive strength were also investigated. To evaluate in vitro cytocompatibility of scaffolds, mouse L929 fibroblasts were seeded onto the scaffolds for cell morphology and cell viability studies. It was found that the porous structure of scaffolds can be tailored by varying the ratios of gelatin to HA, both the swelling ratios and degradation rate increased with the increase of HA content in hybrid scaffolds, and crosslinking the scaffolds with EDC improved the degradation resistance of the scaffold in culture media and increased the mechanical strength of scaffolds. The in vitro results revealed that the prepared scaffolds do not induce cytotoxic effects and suitable for cell growth, especially in the case of scaffolds with higher gelatin content. The combined results of the physicochemical and biological studies suggested that the developed GE/HA hybrid scaffolds exhibit good potential and biocompatibility for soft tissue engineering applications. Copyright © 2011 Elsevier B.V. All rights reserved.

  7. Preparation and mechanical property of a novel 3D porous magnesium scaffold for bone tissue engineering.

    PubMed

    Zhang, Xue; Li, Xiao-Wu; Li, Ji-Guang; Sun, Xu-Dong

    2014-09-01

    Porous magnesium has been recently recognized as a biodegradable metal for bone substitute applications. A novel porous Mg scaffold with three-dimensional (3D) interconnected pores and with a porosity of 33-54% was produced by the fiber deposition hot pressing (FDHP) technology. The microstructure and morphologies of the porous Mg scaffold were characterized by scanning electron microscopy (SEM), and the effects of porosities on the microstructure and mechanical properties of the porous Mg were investigated. Experimental results indicate that the measured Young's modulus and compressive strength of the Mg scaffold are ranged in 0.10-0.37 GPa, and 11.1-30.3 MPa, respectively, which are fairly comparable to those of cancellous bone. Such a porous Mg scaffold having a 3D interconnected network structure has the potential to be used in bone tissue engineering. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. High porous titanium scaffolds showed higher compatibility than lower porous beta-tricalcium phosphate scaffolds for regulating human osteoblast and osteoclast differentiation.

    PubMed

    Hirota, Makoto; Hayakawa, Tohru; Shima, Takaki; Ametani, Akihiro; Tohnai, Iwai

    2015-04-01

    We compared osteoblast and osteoclast differentiation when using beta-tricalcium phosphate (βTCP) and titanium scaffolds by investigating human mesenchymal stem cells (hMSCs) and osteoclast progenitor cell activities. hMSCs were cultured for 7, 14, and 21days on titanium scaffolds with 60%, 73%, and 87% porosity and on βTCP scaffolds with 60% and 75% porosity. Human osteoclast progenitor cells were cultured with osteoblast for 14 and 21days on 87% titanium and 75% βTCP scaffolds. Viable cell numbers with 60% and 73% titanium were higher than with 87% titanium and βTCP scaffolds (P<0.05). An 87% titanium scaffold resulted in the highest osteocalcin production with calcification on day 14 (P<0.01) in titanium scaffolds. All titanium scaffolds resulted in higher osteocalcin production on days 7 and 14 compared to βTCP scaffolds (P<0.01). Osteoblasts cultured on 87% titanium scaffolds suppressed osteoclast differentiation on day 7 but enhanced osteoclast differentiation on day 14 compared to 75% βTCP scaffolds (P<0.01). These findings concluded that high porosity titanium scaffolds could enhance progression of hMSC/osteoblast differentiation and regulated osteoclast differentiation cooperating with osteoblast differentiation for calcification as compared with lower porous βTCP.

  9. Systematic characterization of porosity and mass transport and mechanical properties of porous polyurethane scaffolds.

    PubMed

    Wang, Yu-Fu; Barrera, Carlos M; Dauer, Edward A; Gu, Weiyong; Andreopoulos, Fotios; Huang, C-Y Charles

    2017-01-01

    One of the key challenges in porous scaffold design is to create a porous structure with desired mechanical function and mass transport properties which support delivery of biofactors and development of function tissue substitute. In recent years, polyurethane (PU) has become one of the most popular biomaterials in various tissue engineering fields. However, there are no studies fully investigating the relations between porosity and both mass transport and mechanical properties of PU porous scaffolds. In this paper, we fabricated PU scaffolds by combining phase inversion and salt (sodium chloride) leaching methods. The tensile and compressive moduli were examined on PU scaffolds fabricated with different PU concentrations (25%, 20% and 15% w/v) and salt/PU weight ratios (9/1, 6/1, 3/1 and 0/1). The mass transport properties of PU scaffolds including hydraulic permeability and glucose diffusivity were also measured. Furthermore, the relationships between the porosity and mass transport and mechanical properties of porous PU scaffold were systemically investigated. The results demonstrated that porosity is a key parameter which governs both mass transport and mechanical properties of porous PU scaffolds. With similar pore sizes, the mass transport and mechanical properties of porous PU scaffold can be described as single functions of porosity regardless of initial PU concentration. The relationships between scaffold porosity and properties can be utilized to facilitate porous PU scaffold fabrication with specific mass transport and mechanical properties. The systematic approach established in this study can be applied to characterization of other biomaterials for scaffold design and fabrication.

  10. The use of a polycaprolactone-tricalcium phosphate scaffold for bone regeneration of tooth socket facial wall defects and simultaneous immediate dental implant placement in Macaca fascicularis.

    PubMed

    Goh, Bee Tin; Chanchareonsook, Nattharee; Tideman, Henk; Teoh, Swee Hin; Chow, James Kwok Fai; Jansen, John A

    2014-05-01

    Bone regeneration and aesthetic outcomes may be compromised when immediate implants are placed at extraction sites with dehiscence defects. The aim of this study was to compare, in a monkey model, peri-implant bone regeneration and implant stability after immediate implant placement into tooth sockets with facial wall defects in two treatment groups. In eight control monkeys, the bony defect was reconstructed with autogenous particulate bone, whereas in 10 test monkeys a polycaprolactone-tricalcium phosphate (PCL-TCP) scaffold was used. The monkeys were sacrificed after 6 months and the specimens were analyzed by histology and histomorphometry. Better maintenance of facial bone contour was noted in the test group; however, bone regeneration was seen only at areas adjacent to a bony wall of the defect. The mean bone-to-implant contact was 27.6 ± 19.1% (control group) versus 6.8 ± 7.9% (test group). The mean bone area percentage was 11.8 ± 10.1% (control group) versus 6.8 ± 6.9% (test group). Implant survival was 100% at 6 months for both the groups. It was concluded that although the use of a PCL-TCP scaffold showed better maintenance of the alveolar contour as compared to autogenous particulate bone at 6 months, there was minimal bone regeneration within the defect. Copyright © 2013 Wiley Periodicals, Inc.

  11. Influence of electrospun scaffolds prepared from distinct polymers on proliferation and viability of endothelial cells

    NASA Astrophysics Data System (ADS)

    Matveeva, V. G.; Antonova, L. V.; Velikanova, E. A.; Sergeeva, E. A.; Krivkina, E. O.; Glushkova, T. V.; Kudryavtseva, Yu. A.; Barbarash, O. L.; Barbarash, L. S.

    2015-10-01

    We compared electrospun nonwoven scaffolds from polylactic acid (PLA), polycaprolactone (PCL), and polyhydroxybutyrate/valerate (PHBV)/polycaprolactone (PHBV/PCL). The surface of PHBV/PCL and PCL scaffolds was highly porous and consisted of randomly distributed fibers, whilst the surface of PLA scaffolds consisted of thin straight fibers, which located more sparsely, forming large pores. Culture of EA.hy 926 endothelial cells on these scaffolds during 7 days and further fluorescent microscopy demonstrated that the surface of PHBV/PCL scaffolds was most favorable for efficient adhesion, proliferation, and viability of endothelial cells. The lowest proliferation rate and cell viability were detected on PLA scaffolds. Therefore, PHBV/PCL electrospun nonwoven scaffolds demonstrated the best results regarding endothelial cell proliferation and viability as compared to PCL and PLA scaffolds.

  12. Influence of electrospun scaffolds prepared from distinct polymers on proliferation and viability of endothelial cells

    SciTech Connect

    Matveeva, V. G. Antonova, L. V. Velikanova, E. A.; Sergeeva, E. A.; Krivkina, E. O.; Glushkova, T. V.; Kudryavtseva, Yu. A.; Barbarash, O. L.; Barbarash, L. S.

    2015-10-27

    We compared electrospun nonwoven scaffolds from polylactic acid (PLA), polycaprolactone (PCL), and polyhydroxybutyrate/valerate (PHBV)/polycaprolactone (PHBV/PCL). The surface of PHBV/PCL and PCL scaffolds was highly porous and consisted of randomly distributed fibers, whilst the surface of PLA scaffolds consisted of thin straight fibers, which located more sparsely, forming large pores. Culture of EA.hy 926 endothelial cells on these scaffolds during 7 days and further fluorescent microscopy demonstrated that the surface of PHBV/PCL scaffolds was most favorable for efficient adhesion, proliferation, and viability of endothelial cells. The lowest proliferation rate and cell viability were detected on PLA scaffolds. Therefore, PHBV/PCL electrospun nonwoven scaffolds demonstrated the best results regarding endothelial cell proliferation and viability as compared to PCL and PLA scaffolds.

  13. A novel method to obtain protein release from porous polymer scaffolds: emulsion coating.

    PubMed

    Sohier, J; Haan, R E; de Groot, K; Bezemer, J M

    2003-02-21

    To obtain the controlled release of proteins from macro-porous polymeric scaffolds, a novel emulsion-coating method has been developed. In this process, a water-in-oil emulsion, from an aqueous protein solution and a polymer solution, is forced through a prefabricated scaffold by applying a vacuum. After solvent evaporation, a polymer film, containing the protein, is then deposited on the porous scaffold surface. This paper reports the effect of processing parameters on the emulsion coating characteristics, scaffold structure, and protein release and stability. Poly(ether-ester) multiblock copolymers were chosen as the polymer matrix for both scaffolds and coating. Macro-porous scaffolds, with a porosity of 77 vol% and pores of approximately 500 microm were prepared by compression moulding/salt leaching. A micro-porous, homogeneous protein-loaded coating could be obtained on the scaffold surface. Due to the coating, the scaffold porosity was decreased, whereas the pore interconnection was increased. A model protein (lysozyme) could effectively be released in a controlled fashion from the scaffolds. Complete lysozyme release could be achieved within 3 days up to more than 2 months by adjusting the coated emulsion parameters. In addition, the coating process did not reduce the enzymatic activity. This new method appears to be promising for tissue engineering applications. Copyright 2002 Elsevier Science B.V.

  14. Nano-TiO2/collagen-chitosan porous scaffold for wound repairing.

    PubMed

    Fan, Xialian; Chen, Keke; He, Xichan; Li, Na; Huang, Jinbao; Tang, Keyong; Li, Yijin; Wang, Fang

    2016-10-01

    Collagen-Chitosan (COL-CS) porous scaffolds have been widely used as a dermal equivalent to induce fibroblasts infiltration and dermal regeneration. To improve the anti-bacterial properties, nano-TiO2 hydrosol was introduced into COL-CS scaffolds. TiO2/COL-CS porous scaffolds were fabricated through a freeze-drying process, and scanning electron microscopy (SEM) was employed to study the micro-structure of the scaffolds. Fourier transform infrared spectroscopy (FT-IR) was used to study the intermolecular interactions in the scaffolds. The swelling property, porosity, degradation, antibacterial behavior, red blood cell aggregation, and cytotoxicity of the composite were investigated. The results showed that the scaffold is good in permeability and it may provide a humid environment for wound repairing. The degradation in lysozyme solution for 4 weeks showed that porous scaffolds are good in stability, which may satisfy the wound coverage protection in the repairing period. An obvious inhibitory effect on Staphylococcus aureus of the porous scaffolds was found, and the red blood cells were easy to form clusters aggregation to stop bleeding. It was suggested that the TiO2/COL-CS composite scaffolds could be a promising candidate for wound repairing dressing. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Mineralized polycaprolactone nanofibrous matrix for odontogenesis of human dental pulp cells.

    PubMed

    Kim, Jong-Jin; Bae, Won-Jung; Kim, Joung-Mok; Kim, Jung-Ju; Lee, Eun-Jung; Kim, Hae-Won; Kim, Eun-Cheol

    2014-03-01

    The aim of the present study was to fabricate mineralized polycaprolactone nanofibrous scaffold and investigate its ability to elicit odontogenic differentiation of human dental pulp cells, compared to the pure polycaprolactone scaffold. Polycaprolactone nanofibrous scaffold was produced by electrospinning, and the surface was mineralized with apatite. Cellular behaviors on the mineralized polycaprolactone scaffold were assessed in terms of cell adhesion, growth, and odontoblastic differentiation. To evaluate the signal transduction of human dental pulp cells, mRNA expression was analyzed and Western blotting was performed. Mineralized polycaprolactone showed improved cell proliferation, mineralized nodule formation, and expression of odontoblastic marker genes including alkaline phosphatase, osteopontin, osteocalcin, dentin sialophosphoprotein (DSPP), and dentin matrix protein-1, as compared with pure polycaprolactone. Although the cell adhesion on the mineralized polycaprolactone was similar to that of the polycaprolactone, the expression level of proteins including collagen type I and the key adhesion receptor (integrin components α1, α2, and β1) was upregulated in mineralized polycaprolactone compared to polycaprolactone. Especially, cells seeded onto mineralized polycaprolactone scaffolds showed significantly increased levels of phosphorylated focal adhesion kinase, a marker of integrin activation, and downstream pathways, such as phosphor (p)-Akt, p-extracellular signal regulated kinase, p-c Jun N-terminal kinase, nuclear factor-kappa B, c-fos, and c-jun, compared with pure polycaprolactone. The mineralized polycaprolactone scaffold is attractive for dentin tissue engineering by promoting growth and odontogenic differentiation of human dental pulp cells through the integrin-mediated signaling pathway.

  16. Collagen-grafted porous HDPE/PEAA scaffolds for bone reconstruction.

    PubMed

    Kim, Chang-Shik; Jung, Kyung-Hye; Kim, Hun; Kim, Chan-Bong; Kang, Inn-Kyu

    2016-01-01

    After tumor resection, bone reconstruction such as skull base reconstruction using interconnected porous structure is absolutely necessary. In this study, porous scaffolds for bone reconstruction were prepared using heat-pressing and salt-leaching methods. High-density polyethylene (HDPE) and poly(ethylene-co-acrylic acid) (PEAA) were chosen as the polymer composites for producing a porous scaffold of high mechanical strength and having high reactivity with biomaterials such as collagen, respectively. The porous structure was observed through surface images, and its intrusion volume and porosity were measured. Owing to the carboxylic acids on PEAA, collagen was successfully grafted onto the porous HDPE/PEAA scaffold, which was confirmed by FT-IR spectroscopy and electron spectroscopy for chemical analysis. Osteoblasts were cultured on the collagen-grafted porous scaffold, and their adhesion, proliferation, and differentiation were investigated. The high viability and growth of the osteoblasts suggest that the collagen-grafted porous HDPE/PEAA is a promising scaffold material for bone generation.

  17. 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.

  18. Surface modification on polycaprolactone electrospun mesh and human decalcified bone scaffold with synovium-derived mesenchymal stem cells-affinity peptide for tissue engineering.

    PubMed

    Shao, Zhenxing; Zhang, Xin; Pi, Yanbin; Yin, Ling; Li, La; Chen, Haifeng; Zhou, Chunyan; Ao, Yingfang

    2015-01-01

    Synovium-derived mesenchymal stem cells (SMSC) have been studied for over a decade since first being successfully isolated in 2001. These cells demonstrate the most promising therapeutic efficacy for musculoskeletal regeneration of the MSC family, particularly for cartilage regeneration. However, the mobilization and transfer of MSCs to defective or damaged tissues and organs in vivo with high accuracy and efficiency has been a major problem in tissue engineering (TE). In the present study, we identified a seven amino acid peptide sequence [SMSCs-affinity peptide (LTHPRWP; L7)] through phage display technology that has a high specific affinity to SMSCs. Our analysis suggested that L7 efficiently and specifically interacted with SMSCs without any species specificity. Thereafter, L7 was covalently conjugated onto both polycaprolactone (PCL) electrospun meshes and human decalcified bone scaffolds (hDBSc) to investigate its TE applications. After 24 h coculture with human SMSCs (hSMSCs), L7-conjugated PCL electrospun meshes had significantly more adherent hSMSCs than the control group, and the cells expanded well. Similar results were obtained using hDBSs. These results suggest that the novel L7 peptide sequence has a high specific affinity to SMSCs. Covalently conjugating this peptide to either artificial polymer material (PCL mesh) or natural material (hDBS) significantly enhances the adhesion of SMSCs. This method is applicable to a wide range of potential SMSC-based TE applications, particularly to cartilage regeneration, via surface modification on various type of materials. © 2014 Wiley Periodicals, Inc.

  19. Thermoforming techniques for manufacturing porous scaffolds for application in 3D cell cultivation.

    PubMed

    Borowiec, Justyna; Hampl, Jörg; Gebinoga, Michael; Elsarnagawy, Tarek; Elnakady, Yasser A; Fouad, Hassan; Almajhadi, Fahd; Fernekorn, Uta; Weise, Frank; Singh, Sukhdeep; Elsarnagawy, Dief; Schober, Andreas

    2015-04-01

    Within the scientific community, there is an increasing demand to apply advanced cell cultivation substrates with increased physiological functionalities for studying spatially defined cellular interactions. Porous polymeric scaffolds are utilized for mimicking an organ-like structure or engineering complex tissues and have become a key element for three-dimensional (3D) cell cultivation in the meantime. As a consequence, efficient 3D scaffold fabrication methods play an important role in modern biotechnology. Here, we present a novel thermoforming procedure for manufacturing porous 3D scaffolds from permeable materials. We address the issue of precise thermoforming of porous polymer foils by using multilayer polymer thermoforming technology. This technology offers a new method for structuring porous polymer foils that are otherwise available for non-porous polymers only. We successfully manufactured 3D scaffolds from solvent casted and phase separated polylactic acid (PLA) foils and investigated their biocompatibility and basic cellular performance. The HepG2 cell culture in PLA scaffold has shown enhanced albumin secretion rate in comparison to a previously reported polycarbonate based scaffold with similar geometry.

  20. Porous chitosan scaffold cross-linked by chemical and natural procedure applied to investigate cell regeneration

    NASA Astrophysics Data System (ADS)

    Yao, Chih-Kai; Liao, Jiunn-Der; Chung, Chia-Wei; Sung, Wei-I.; Chang, Nai-Jen

    2012-12-01

    Porous chitosan scaffold is used for tissue engineering and drug delivery, but is limited as a scaffold material due to its mechanical weakness, which restrains cell adhesion on the surface. In this study, a chemical reagent (citrate) and a natural reagent (genipin) are used as cross-linkers for the formation of chitosan-based films. Nanoindentation technique with a continuous stiffness measurement system is particularly applied on the porous scaffold surface to examine the characteristic modulus and nanohardness of a porous scaffold surface. The characteristic modulus of a genipin-cross-linked chitosan surface is ≈2.325 GPa, which is significantly higher than that of an uncross-linked one (≈1.292 GPa). The cell-scaffold surface interaction is assessed. The cell morphology and results of an MTS assay of 3T3-fibroblast cells of a genipin-cross-linked chitosan surface indicate that the enhancement of mechanical properties induced cell adhesion and proliferation on the modified porous scaffold surface. The pore size and mechanical properties of porous chitosan film can be tuned for specific applications such as tissue regeneration.

  1. Gelatin porous scaffolds fabricated using a modified gas foaming technique: characterisation and cytotoxicity assessment.

    PubMed

    Poursamar, S Ali; Hatami, Javad; Lehner, Alexander N; da Silva, Cláudia L; Ferreira, Frederico Castelo; Antunes, A P M

    2015-03-01

    The current study presents an effective and simple strategy to obtain stable porous scaffolds from gelatin via a gas foaming method. The technique exploits the intrinsic foaming ability of gelatin in the presence of CO2 to obtain a porous structure stabilised with glutaraldehyde. The produced scaffolds were characterised using physical and mechanical characterisation methods. The results showed that gas foaming may allow the tailoring of the 3-dimensional structure of the scaffolds with an interconnected porous structure. To assess the effectiveness of the preparation method in mitigating the potential cytotoxicity risk of using glutaraldehyde as a crosslinker, direct and in-direct cytotoxicity assays were performed at different concentrations of glutaraldehyde. The results indicate the potential of the gas foaming method, in the preparation of viable tissue engineering scaffolds.

  2. Construction of biocompatible porous tissue scaffold from the decellularized umbilical artery.

    PubMed

    Xin, Yi; Wu, Guanghui; Wu, Man; Zhang, Xiaoxia; Velot, Emilie; Decot, Véronique; Cui, Wei; Huang, Yimin; Stoltz, Jean-Francois; Du, Jie; Li, Na

    2015-01-01

    The scaffolds prepared from the tissue decellularization conserve the porous 3-D structure and provide an optimal matrix for the tissue regeneration. Since decade, the enzymatic digestion, chemical reagent treatment and mechanical actions such as eversion and abrasion have been used to remove the cells from the intact matrix. In this study, we optimized an enzymatic method to decellularize the umbilical artery to construct a 3-D porous scaffold which is suitable for the culture of mesenchymal stem cells (MSCs). The scaffold maintained the interconnected porous structure. It remained the similar high water content 95.3 ± 1% compared to 94.9 ± 0.6% in the intact umbilical artery (p>0.05). The decellularization process decreased the stress from 0.24 ± 0.05 mPa to 0.15 ± 0.06 mPa (p<0.05). However the decellularization did not change the strain of the artery (45 ± 15% vs. 53 ± 10%, p>0.05). When the scaffold was transplanted to the subcutaneous tissue in the wild type mice, there were less T cells appeared in the surrounding tissue which meant the decreased the immunogenicity by decellularization. This scaffold also supported the adhesion and proliferation of the MSCs. In this study, we constructed a biological compatible porous scaffold from the decellularized umbilical artery which may provide a suitable scaffold for cell-matrix interaction studies and for tissue engineering.

  3. Development of porous Ti6Al4V/chitosan sponge composite scaffold for orthopedic applications.

    PubMed

    Guo, Miao; Li, Xiang

    2016-01-01

    A novel composite scaffold consisting of porous Ti6Al4V part filled with chitosan sponge was fabricated using a combination of electron beam melting and freeze-drying. The mechanical properties of porous Ti6Al4V part were examined via compressive test. The ultimate compressive strength was 85.35 ± 8.68 MPa and the compressive modulus was 2.26 ± 0.42 GPa. The microstructure of composite scaffold was characterized using scanning electron microscopy. The chitosan sponge filled in Ti6Al4V part exhibited highly porous and well-interconnected micro-pore architecture. The osteoblastic cells were seeded on scaffolds to test their seeding efficiency and biocompatibility. Significantly higher cell seeding efficiency was found on composite scaffold. The biological response of osteoblasts on composite scaffolds was superior in terms of improved cell attachment, higher proliferation, and well-spread morphology in relation to porous Ti6Al4V part. These results suggest that the Ti6Al4V/chitosan composite scaffold is potentially useful as a biomedical scaffold for orthopedic applications. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Adipogenic differentiation of stem cells in three-dimensional porous bacterial nanocellulose scaffolds.

    PubMed

    Krontiras, Panagiotis; Gatenholm, Paul; Hägg, Daniel A

    2015-01-01

    There is an increased interest in developing adipose tissue for in vitro and in vivo applications. Current two-dimensional (2D) cell-culture systems of adipocytes are limited, and new methods to culture adipocytes in three-dimensional (3D) are warranted as a more life-like model to study metabolic diseases such as obesity and diabetes. In this study, we have evaluated different porous bacterial nanocellulose scaffolds for 3D adipose tissue. In an initial pilot study, we compared adipogenic differentiation of mice mesenchymal stem cells from a cell line on 2D and 3D scaffolds of bacterial nanocellulose. The 3D scaffolds were engineered by crosslinking homogenized cellulose fibrils using alginate and freeze drying the mixture to obtain a porous structure. Quenching the scaffolds in liquid nitrogen resulted in smaller pores compared to slower freezing using isopropanol. We found that on 2D surfaces, the cells were scarcely distributed and showed limited formation of lipid droplets, whereas cells grown in macroporous 3D scaffolds contained more cells growing in clusters, containing large lipid droplets. All four types of scaffolds contained a lot of adipocytes, but scaffolds with smaller pores contained larger cell clusters than scaffolds with bigger pores, with viable adipocytes present even 4 weeks after differentiation. Scaffolds with lower alginate fractions retained their pore integrity better. We conclude that 3D culturing of adipocytes in bacterial nanocellulose macroporous scaffolds is a promising method for fabrication of adipose tissue as an in vitro model for adipose biology and metabolic disease.

  5. Preparation and Evaluation of Gelatin-Chitosan-Nanobioglass 3D Porous Scaffold for Bone Tissue Engineering

    PubMed Central

    Maji, Kanchan; Dasgupta, Sudip; Pramanik, Krishna; Bissoyi, Akalabya

    2016-01-01

    The aim of the present study was to prepare and characterize bioglass-natural biopolymer based composite scaffold and evaluate its bone regeneration ability. Bioactive glass nanoparticles (58S) in the size range of 20–30 nm were synthesized using sol-gel method. Porous scaffolds with varying bioglass composition from 10 to 30 wt% in chitosan, gelatin matrix were fabricated using the method of freeze drying of its slurry at 40 wt% solids loading. Samples were cross-linked with glutaraldehyde to obtain interconnected porous 3D microstructure with improved mechanical strength. The prepared scaffolds exhibited >80% porosity with a mean pore size range between 100 and 300 microns. Scaffold containing 30 wt% bioglass (GCB 30) showed a maximum compressive strength of 2.2 ± 0.1 MPa. Swelling and degradation studies showed that the scaffold had excellent properties of hydrophilicity and biodegradability. GCB 30 scaffold was shown to be noncytotoxic and supported mesenchymal stem cell attachment, proliferation, and differentiation as indicated by MTT assay and RUNX-2 expression. Higher cellular activity was observed in GCB 30 scaffold as compared to GCB 0 scaffold suggesting the fact that 58S bioglass nanoparticles addition into the scaffold promoted better cell adhesion, proliferation, and differentiation. Thus, the study showed that the developed composite scaffolds are potential candidates for regenerating damaged bone tissue. PMID:26884764

  6. Relationship between osseointegration and superelastic biomechanics in porous NiTi scaffolds.

    PubMed

    Liu, Xiangmei; Wu, Shuilin; Yeung, Kelvin W K; Chan, Y L; Hu, Tao; Xu, Zushun; Liu, Xuanyong; Chung, Jonathan C Y; Cheung, Kenneth M C; Chu, Paul K

    2011-01-01

    The superelastic nature of bones requires matching biomechanical properties from the ideal artificial biomedical implants in order to provide smooth load transfer and foster the growth of new bone tissues. In this work, we determine the biomechanical characteristics of porous NiTi implants and investigate bone ingrowth under actual load-bearing conditions in vivo. In this systematic and comparative study, porous NiTi, porous Ti, dense NiTi, and dense Ti are implanted into 5 mm diameter holes in the distal part of the femur/tibia of rabbits for 15 weeks. The bone ingrowth and interfacial bonding strength are evaluated by histological analysis and push-out test. The porous NiTi materials bond very well to newly formed bone tissues and the highest average strength of 357 N and best ductility are achieved from the porous NiTi materials. The bonding curve obtained from the NiTi scaffold shows similar superelasticity as natural bones with a deflection of 0.30-0.85 mm thus shielding new bone tissues from large load stress. This is believed to be the reason why new bone tissues can penetrate deeply into the porous NiTi scaffold compared to the one made of porous Ti. Histological analysis reveals that new bone tissues adhere and grow well on the external surfaces as well as exposed areas on the inner pores of the NiTi scaffold. The in vitro study indicates that the surface chemical composition and topography of the porous structure leads to good cytocompatibility. Consequently, osteoblasts proliferate smoothly on the entire implant including the flat surface, embossed region, exposed area of the pores, and interconnected channels. In conjunction with the good cytocompatibility, the superelastic biomechanical properties of the porous NiTi scaffold bodes well for fast formation and ingrowth of new bones, and porous NiTi scaffolds are thus suitable for clinical applications under load-bearing conditions.

  7. Fabrication of porous chitosan-polyvinyl pyrrolidone scaffolds from a quaternary system via phase separation.

    PubMed

    Lim, Jin Ik; Im, Heejung; Lee, Woo-Kul

    2015-01-01

    Three-dimensional porous chitosan-polyvinyl pyrrolidone (PVP) scaffolds were fabricated for tissue engineering applications via liquid-liquid or liquid-solid phase separation. A mixture of an acidic aqueous solution with butanol as a non-solvent and a chitosan-PVP quaternary system were freeze-dried. We then studied the homogenous open pore structure and the minute pore distribution in order to improve the mass transfer and cell seeding efficiency while also obtaining the optimal ratio of PVP to provide high interconnectivity and to improve the open-pore structure. The properties of the porous chitosan-PVP scaffolds - including the microstructure, chemical release, water absorption properties, and cell proliferation tests were studied - and the results were compared against those obtained from conventional scaffolds. chitosan-PVP scaffolds with a porosity of over 70% were obtained, and the pore morphology on the surface and within the porous scaffolds showed the presence of homogenous open pores with excellent interconnectivity. As the PVP content increased, main pores (50-100 μm) and minute pores (4-10 μm) could be clearly observed. Also, the porous scaffold showed an improved efficiency for cell adhesion after the cells were cultured for 4 h. After 72 h, the cultured cells presented an increase in the cell proliferation and on the porous scaffolds. These results strongly suggest that the porous chitosan-PVP scaffolds can be widely used in tissue engineering, including for biopatches and artificial skin applications.

  8. 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.

  9. A novel bioactive porous CaSiO3 scaffold for bone tissue engineering.

    PubMed

    Ni, Siyu; Chang, Jiang; Chou, Lee

    2006-01-01

    The aim of this study was to fabricate bioactive porous CaSiO3 scaffolds and examine their effects on proliferation and differentiation of osteoblast-like cells. In this study, porous CaSiO3 scaffolds were obtained by sintering a ceramic slip-coated polymer foam at 1350 degrees C. X-ray diffraction (XRD) of the scaffolds indicated that the products were essentially pure alpha-CaSiO3. The obtained scaffolds had a well-interconnected porous structure with pore sizes ranging from several micrometers to more than 100 microm and porosities of 88.5 +/- 2.8%. The in vitro bioactivity of the scaffolds was investigated by soaking them in simulated body fluid (SBF) for 7 days and then characterizing them by scanning electron microscopy (SEM) and energy-dispersive spectroscopy (EDS) analysis. The results indicated that hydroxyapatite (HAp) was formed on the surface of the scaffolds. In addition, the scaffolds were incubated in Ringer's solution at 37 degrees C to study the in vitro degradation by measurement of weight loss after incubation, which showed that the CaSiO3 scaffolds were degradable. The cellular responses to the scaffolds were assessed in terms of cell proliferation and differentiation. Osteoblast-like cells were seeded into the CaSiO3 scaffolds. SEM observations showed that there was significant cell adhesion, as the cells spread and grew in the scaffolds. In addition, the proliferation rate and alkaline phosphatase (ALP) activity of the cells in the scaffolds were improved as compared to the controls. These studies demonstrate initial in vitro cell compatibility and their potential application to bone tissue engineering. (c) 2005 Wiley Periodicals, Inc

  10. Mechanical, thermal and morphological characterisation of 3D porous Pennisetum purpureum/PLA biocomposites scaffold.

    PubMed

    Revati, R; Abdul Majid, M S; Ridzuan, M J M; Normahira, M; Mohd Nasir, N F; Rahman Y, M N; Gibson, A G

    2017-06-01

    The mechanical, thermal, and morphological properties of a 3D porous Pennisetum purpureum (PP)/polylactic acid (PLA) based scaffold were investigated. In this study, a scaffold containing P. purpureum and PLA was produced using the solvent casting and particulate leaching method. P. purpureum fibre, also locally known as Napier grass, is composed of 46% cellulose, 34% hemicellulose, and 20% lignin. PLA composites with various P. purpureum contents (10%, 20%, and 30%) were prepared and subsequently characterised. The morphologies, structures and thermal behaviours of the prepared composite scaffolds were characterised using field-emission scanning electron microscopy (FESEM), Fourier transform infrared spectroscopy (FTIR), and thermogravimetric analysis (TGA). The morphology was studied using FESEM; the scaffold possessed 70-200μm-sized pores with a high level of interconnectivity. The moisture content and mechanical properties of the developed porous scaffolds were further characterised. The P. purpureum/PLA scaffold had a greater porosity factor (99%) and compression modulus (5.25MPa) than those of the pure PLA scaffold (1.73MPa). From the results, it can be concluded that the properties of the highly porous P. purpureum/PLA scaffold developed in this study can be controlled and optimised. This can be used to facilitate the construction of implantable tissue-engineered cartilage. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Development of a porous PLGA-based scaffold for mastoid air cell regeneration

    PubMed Central

    Gould, Toby W. A.; Birchall, John P.; Mallick, Ali S.; Alliston, Tamara; Lustig, Lawrence R.; Shakesheff, Kevin M.

    2015-01-01

    Objective To develop a porous, biodegradable scaffold for mastoid air cell regeneration. Study Design In vitro development of a temperature-sensitive poly(DL-lactic acid-co-glycolic acid)/poly(ethylene glycol) (PLGA/PEG) scaffold tailored for this application. Methods Human mastoid bone microstructure and porosity was investigated using micro-computed tomography. PLGA/PEG-alginate scaffolds were developed and scaffold porosity was assessed. Human bone marrow mesenchymal stem cells (hBM-MSCs) were cultured on the scaffolds in vitro. Scaffolds were loaded with ciprofloxacin and release of ciprofloxacin over time in vitro was assessed. Results Porosity of human mastoid bone was measured at 83% with an average pore size of 1.3mm. PLGA/PEG-alginate scaffold porosity ranged from 43–78% depending on the alginate bead content. hBM-MSCs proliferate on the scaffolds in vitro, and release of ciprofloxacin from the scaffolds was demonstrated over 7–10 weeks. Conclusion The PLGA/PEG-alginate scaffolds developed in this study demonstrate similar structural features to human mastoid bone, support cell growth and display sustained antibiotic release. These scaffolds may be of potential clinical use in mastoid air cell regeneration. Further in vivo studies to assess the suitability of PLGA/PEG-alginate scaffolds for this application are required. PMID:23670365

  12. Enhanced peripheral nerve regeneration by the combination of a polycaprolactone tubular prosthesis and a scaffold of collagen with supramolecular organization

    PubMed Central

    Maturana, Luiz G; Pierucci, Amauri; Simões, Gustavo F; Vidigal, Mateus; Duek, Eliana A R; Vidal, Benedicto C; Oliveira, Alexandre L R

    2013-01-01

    The purpose of this study was to investigate the influence of implanting collagen with a supramolecular organization on peripheral nerve regeneration, using the sciatic nerve tubulization technique. For this purpose, adult female Sprague Dawley rats were divided into five groups: (1) TP – sciatic nerve repaired with empty polyethylene tubular prothesis (n = 10), (2) TPCL – nerve repair with empty polycaprolactone (PCL) tubing (n = 8), (3) TPCLF – repair with PCL tubing filled with an implant of collagen with a supramolecular organization (n = 10), (4) AG – animals that received a peripheral nerve autograft (n = 8), and (5) Normal nerves (n = 8). The results were assessed by quantification of the regenerated fibers, nerve morphometry, and transmission electron microscopy, 60 days after surgery. Immunohistochemistry and polarization microscopy were also used to analyze the regenerated nerve structure and cellular elements. The results showed that the AG group presented a larger number of regenerated axons. However, the TPCL and TPCLF groups presented more compact regenerated fibers with a morphometric profile closer to normal, both at the tube midpoint and 2 mm distal to the prosthesis. These findings were reinforced by polarization microscopy, which indicated a better collagen/axons suprastructural organization in the TPCLF derived samples. In addition, the immunohistochemical results obtained using the antibody anti-p75NTR as a Schwann cell reactivity marker demonstrated that the Schwann cells were more reactive during the regenerative process in the TPCLF group as compared to the TPCL group and the normal sciatic nerve. Altogether, the results of this study indicated that the implant of collagen with a supramolecular organization positively influenced and stimulated the regeneration process through the nerve gap, resulting in the formation of a better morphologically arranged tissue. PMID:24381812

  13. Porous nanofibrous poly(L-lactic acid) scaffolds supporting cardiovascular progenitor cells for cardiac tissue engineering.

    PubMed

    Liu, Qihai; Tian, Shuo; Zhao, Chao; Chen, Xin; Lei, Ienglam; Wang, Zhong; Ma, Peter X

    2015-10-01

    Myocardial infarction (MI) is the irreversible necrosis of heart with approximately 1.5 million cases every year in the United States. Tissue engineering offers a promising strategy for cardiac repair after MI. However, the optimal cell source for heart tissue regeneration and the ideal scaffolds to support cell survival, differentiation, and integration, remain to be developed. To address these issues, we developed the technology to induce cardiovascular progenitor cells (CPCs) derived from mouse embryonic stem cells (ESCs) towards desired cardiomyocytes as well as smooth muscle cells and endothelial cells. We fabricated extracellular matrix (ECM)-mimicking nanofibrous poly(l-lactic acid) (PLLA) scaffolds with porous structure of high interconnection for cardiac tissue formation. The CPCs were seeded into the scaffolds to engineer cardiac constructs in vitro. Fluorescence staining and RT-PCR assay showed that the scaffolds facilitated cell attachment, extension, and differentiation. Subcutaneous implantation of the cell/scaffold constructs in a nude mouse model showed that the scaffolds favorably supported survival of the grafted cells and their commitment to the three desired lineages in vivo. Thus, our study suggested that the porous nanofibrous PLLA scaffolds support cardiac tissue formation from CPCs. The integration of CPCs with the nanofibrous PLLA scaffolds represents a promising tissue engineering strategy for cardiac repair. Myocardial infarction is the irreversible necrosis of heart with approximately 1.5 million cases every year in the United States. Tissue engineering offers a promising strategy for cardiac repair after MI. However, the optimal cell source for heart tissue regeneration and the ideal scaffolds to support cell survival, differentiation, and integration, remain to be developed. To address these issues, we developed porous nanofibrous PLLA scaffolds that mimic natural extracellular matrix to support cardiac tissue formation from CPCs. The

  14. An improved polymeric sponge replication method for biomedical porous titanium scaffolds.

    PubMed

    Wang, Chunli; Chen, Hongjie; Zhu, Xiangdong; Xiao, Zhanwen; Zhang, Kai; Zhang, Xingdong

    2017-01-01

    Biomedical porous titanium (Ti) scaffolds were fabricated by an improved polymeric sponge replication method. The unique formulations and distinct processing techniques, i.e. a mixture of water and ethanol as solvent, multiple coatings with different viscosities of the Ti slurries and centrifugation for removing the extra slurries were used in the present study. The optimized porous Ti scaffolds had uniform porous structure and completely interconnected macropores (~365.1μm). In addition, two different sizes of micropores (~45.4 and ~6.2μm) were also formed in the skeleton of the scaffold. The addition of ethanol to the Ti slurry increased the compressive strength of the scaffold by improving the compactness of the skeleton. A compressive strength of 83.6±4.0MPa was achieved for a porous Ti scaffold with a porosity of 66.4±1.8%. Our cellular study also revealed that the scaffolds could support the growth and proliferation of mesenchymal stem cells (MSCs).

  15. A novel open-porous magnesium scaffold with controllable microstructures and properties for bone regeneration

    PubMed Central

    Cheng, Meng-qi; Wahafu, Tuerhongjiang; Jiang, Guo-feng; Liu, Wei; Qiao, Yu-qin; Peng, Xiao-chun; Cheng, Tao; Zhang, Xian-long; He, Guo; Liu, Xuan-yong

    2016-01-01

    The traditional production methods of porous magnesium scaffolds are difficult to accurately control the pore morphologies and simultaneously obtain appropriate mechanical properties. In this work, two open-porous magnesium scaffolds with different pore size but in the nearly same porosity are successfully fabricated with high-purity Mg ingots through the titanium wire space holder (TWSH) method. The porosity and pore size can be easily, precisely and individually controlled, as well as the mechanical properties also can be regulated to be within the range of human cancellous bone by changing the orientation of pores without sacrifice the requisite porous structures. In vitro cell tests indicate that the scaffolds have good cytocompatibility and osteoblastic differentiation properties. In vivo findings demonstrate that both scaffolds exhibit acceptable inflammatory responses and can be almost fully degraded and replaced by newly formed bone. More importantly, under the same porosity, the scaffolds with larger pore size can promote early vascularization and up-regulate collagen type 1 and OPN expression, leading to higher bone mass and more mature bone formation. In conclusion, a new method is introduced to develop an open-porous magnesium scaffold with controllable microstructures and mechanical properties, which has great potential clinical application for bone reconstruction in the future. PMID:27071777

  16. A novel open-porous magnesium scaffold with controllable microstructures and properties for bone regeneration

    NASA Astrophysics Data System (ADS)

    Cheng, Meng-Qi; Wahafu, Tuerhongjiang; Jiang, Guo-Feng; Liu, Wei; Qiao, Yu-Qin; Peng, Xiao-Chun; Cheng, Tao; Zhang, Xian-Long; He, Guo; Liu, Xuan-Yong

    2016-04-01

    The traditional production methods of porous magnesium scaffolds are difficult to accurately control the pore morphologies and simultaneously obtain appropriate mechanical properties. In this work, two open-porous magnesium scaffolds with different pore size but in the nearly same porosity are successfully fabricated with high-purity Mg ingots through the titanium wire space holder (TWSH) method. The porosity and pore size can be easily, precisely and individually controlled, as well as the mechanical properties also can be regulated to be within the range of human cancellous bone by changing the orientation of pores without sacrifice the requisite porous structures. In vitro cell tests indicate that the scaffolds have good cytocompatibility and osteoblastic differentiation properties. In vivo findings demonstrate that both scaffolds exhibit acceptable inflammatory responses and can be almost fully degraded and replaced by newly formed bone. More importantly, under the same porosity, the scaffolds with larger pore size can promote early vascularization and up-regulate collagen type 1 and OPN expression, leading to higher bone mass and more mature bone formation. In conclusion, a new method is introduced to develop an open-porous magnesium scaffold with controllable microstructures and mechanical properties, which has great potential clinical application for bone reconstruction in the future.

  17. 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

  18. Collagen/chitosan porous scaffolds with improved biostability for skin tissue engineering.

    PubMed

    Ma, Lie; Gao, Changyou; Mao, Zhengwei; Zhou, Jie; Shen, Jiacong; Hu, Xueqing; Han, Chunmao

    2003-11-01

    Porous scaffolds for skin tissue engineering were fabricated by freeze-drying the mixture of collagen and chitosan solutions. Glutaraldehyde (GA) was used to treat the scaffolds to improve their biostability. Confocal laser scanning microscopy observation confirmed the even distribution of these two constituent materials in the scaffold. The GA concentrations have a slight effect on the cross-section morphology and the swelling ratios of the cross-linked scaffolds. The collagenase digestion test proved that the presence of chitosan can obviously improve the biostability of the collagen/chitosan scaffold under the GA treatment, where chitosan might function as a cross-linking bridge. A detail investigation found that a steady increase of the biostability of the collagen/chitosan scaffold was achieved when GA concentration was lower than 0.1%, then was less influenced at a still higher GA concentration up to 0.25%. In vitro culture of human dermal fibroblasts proved that the GA-treated scaffold could retain the original good cytocompatibility of collagen to effectively accelerate cell infiltration and proliferation. In vivo animal tests further revealed that the scaffold could sufficiently support and accelerate the fibroblasts infiltration from the surrounding tissue. Immunohistochemistry analysis of the scaffold embedded for 28 days indicated that the biodegradation of the 0.25% GA-treated scaffold is a long-term process. All these results suggest that collagen/chitosan scaffold cross-linked by GA is a potential candidate for dermal equivalent with enhanced biostability and good biocompatibility.

  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. 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.

  1. Poly(lactide-co-glycolide) porous scaffolds for tissue engineering and regenerative medicine

    PubMed Central

    Pan, Zhen; Ding, Jiandong

    2012-01-01

    Porous scaffolds fabricated from biocompatible and biodegradable polymers play vital roles in tissue engineering and regenerative medicine. Among various scaffold matrix materials, poly(lactide-co-glycolide) (PLGA) is a very popular and an important biodegradable polyester owing to its tunable degradation rates, good mechanical properties and processibility, etc. This review highlights the progress on PLGA scaffolds. In the latest decade, some facile fabrication approaches at room temperature were put forward; more appropriate pore structures were designed and achieved; the mechanical properties were investigated both for dry and wet scaffolds; a long time biodegradation of the PLGA scaffold was observed and a three-stage model was established; even the effects of pore size and porosity on in vitro biodegradation were revealed; the PLGA scaffolds have also been implanted into animals, and some tissues have been regenerated in vivo after loading cells including stem cells. PMID:23741612

  2. [RESEARCH PROGRESS OF THREE-DIMENSIONAL PRINTING POROUS SCAFFOLDS FOR BONE TISSUE ENGINEERING].

    PubMed

    Wu, Tianqi; Yang, Chunxi

    2016-04-01

    To summarize the research progress of several three-dimensional (3-D)-printing scaffold materials in bone tissue engineering. The recent domestic and international articles about 3-D printing scaffold materials were reviewed and summarized. Compared with conventional manufacturing methods, 3-D printing has distinctive advantages, such as enhancing the controllability of the structure and increasing the productivity. In addition to the traditional metal and ceramic scaffolds, 3-D printing scaffolds carrying seeding cells and tissue factors as well as scaffolds filling particular drugs for special need have been paid more and more attention. The development of 3-D printing porous scaffolds have revealed new perspectives in bone repairing. But it is still at the initial stage, more basic and clinical researches are still needed.

  3. [Research Progress of Collagen-based Three-dimensional Porous Scaffolds Used in Skin Tissue Engineering].

    PubMed

    Zhang, Jing; Tang, Qiwei; Zhou, Aimei; Yang, Shulin

    2015-08-01

    Collagen is a kind of natural biomedical material and collagen based three-dimensional porous scaffolds have been widely used in skin tissue engineering. However, these scaffolds do not meet the requirements for artificial skin substitutes in terms of their poor mechanical properties, short supply, and rejection in the bodies. All of these factors limit their further application in skin tissue engineering. A variety of methods have been chosen to meliorate the situation, such as cross linking and blending other substance for improving mechanical properties. The highly biomimetic scaffolds either in structure or in function can be prepared through culturing cells and loading growth factors. To avoid the drawbacks of unsafety attributing to animals, investigators have fixed their eyes on the recombinant collagen. This paper reviews the the progress of research and application of collagen-based 3-dimensional porous scaffolds in skin tissue engineering.

  4. Development and characterization of novel porous 3D alginate-cockle shell powder nanobiocomposite bone scaffold.

    PubMed

    Bharatham, B Hemabarathy; Abu Bakar, Md Zuki; Perimal, Enoch Kumar; Yusof, Loqman Mohamed; Hamid, Muhajir

    2014-01-01

    A novel porous three-dimensional bone scaffold was developed using a natural polymer (alginate/Alg) in combination with a naturally obtained biomineral (nano cockle shell powder/nCP) through lyophilization techniques. The scaffold was developed in varying composition mixture of Alg-nCP and characterized using various evaluation techniques as well as preliminary in vitro studies on MG63 human osteoblast cells. Morphological observations using SEM revealed variations in structures with the use of different Alg-nCP composition ratios. All the developed scaffolds showed a porous structure with pore sizes ideal for facilitating new bone growth; however, not all combination mixtures showed subsequent favorable characteristics to be used for biological applications. Scaffolds produced using the combination mixture of 40% Alg and 60% nCP produced significantly promising results in terms of mechanical strength, degradation rate, and increased cell proliferation rates making it potentially the optimum composition mixture of Alg-nCP with future application prospects.

  5. Biomimetic formation of apatite on the surface of porous gelatin/bioactive glass nanocomposite scaffolds

    NASA Astrophysics Data System (ADS)

    Mozafari, Masoud; Rabiee, Mohammad; Azami, Mahmoud; Maleknia, Saied

    2010-12-01

    There have been several attempts to combine bioactive glasses (BaGs) with biodegradable polymers to create a scaffold material with excellent biocompatibility, bioactivity, biodegradability and toughness. In the present study, the nanocomposite scaffolds with compositions based on gelatin (Gel) and BaG nanoparticles in the ternary SiO 2-CaO-P 2O 5 system were prepared. In vitro evaluations of the nanocomposite scaffolds were performed, and for investigating their bioactive capacity these scaffolds were soaked in a simulated body fluid (SBF) at different time intervals. The scaffolds showed significant enhancement in bioactivity within few days of immersion in SBF solution. The apatite formation at the surface of the nanocomposite samples confirmed by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX) and X-ray powder diffraction (XRD) analyses. In vitro experiments with osteoblast cells indicated an appropriate penetration of the cells into the scaffold's pores, and also the continuous increase in cell aggregation on the bioactive scaffolds with increase in the incubation time demonstrated the ability of the scaffolds to support cell growth. The SEM observations revealed that the prepared scaffolds were porous with three dimensional (3D) and interconnected microstructure, pore size was 200-500 μm and the porosity was 72-86%. The nanocomposite scaffold made from Gel and BaG nanoparticles could be considered as a highly bioactive and potential bone tissue engineering implant.

  6. Label-free magnetic resonance imaging to locate live cells in three-dimensional porous scaffolds.

    PubMed

    Abarrategi, A; Fernandez-Valle, M E; Desmet, T; Castejón, D; Civantos, A; Moreno-Vicente, C; Ramos, V; Sanz-Casado, J V; Martínez-Vázquez, F J; Dubruel, P; Miranda, P; López-Lacomba, J L

    2012-09-07

    Porous scaffolds are widely tested materials used for various purposes in tissue engineering. A critical feature of a porous scaffold is its ability to allow cell migration and growth on its inner surface. Up to now, there has not been a method to locate live cells deep inside a material, or in an entire structure, using real-time imaging and a non-destructive technique. Herein, we seek to demonstrate the feasibility of the magnetic resonance imaging (MRI) technique as a method to detect and locate in vitro non-labelled live cells in an entire porous material. Our results show that the use of optimized MRI parameters (4.7 T; repetition time = 3000 ms; echo time = 20 ms; resolution 39 × 39 µm) makes it possible to obtain images of the scaffold structure and to locate live non-labelled cells in the entire material, with a signal intensity higher than that obtained in the culture medium. In the current study, cells are visualized and located in different kinds of porous scaffolds. Moreover, further development of this MRI method might be useful in several three-dimensional biomaterial tests such as cell distribution studies, routine qualitative testing methods and in situ monitoring of cells inside scaffolds.

  7. Label-free magnetic resonance imaging to locate live cells in three-dimensional porous scaffolds

    PubMed Central

    Abarrategi, A.; Fernandez-Valle, M. E.; Desmet, T.; Castejón, D.; Civantos, A.; Moreno-Vicente, C.; Ramos, V.; Sanz-Casado, J. V.; Martínez-Vázquez, F. J.; Dubruel, P.; Miranda, P.; López-Lacomba, J. L.

    2012-01-01

    Porous scaffolds are widely tested materials used for various purposes in tissue engineering. A critical feature of a porous scaffold is its ability to allow cell migration and growth on its inner surface. Up to now, there has not been a method to locate live cells deep inside a material, or in an entire structure, using real-time imaging and a non-destructive technique. Herein, we seek to demonstrate the feasibility of the magnetic resonance imaging (MRI) technique as a method to detect and locate in vitro non-labelled live cells in an entire porous material. Our results show that the use of optimized MRI parameters (4.7 T; repetition time = 3000 ms; echo time = 20 ms; resolution 39 × 39 µm) makes it possible to obtain images of the scaffold structure and to locate live non-labelled cells in the entire material, with a signal intensity higher than that obtained in the culture medium. In the current study, cells are visualized and located in different kinds of porous scaffolds. Moreover, further development of this MRI method might be useful in several three-dimensional biomaterial tests such as cell distribution studies, routine qualitative testing methods and in situ monitoring of cells inside scaffolds. PMID:22442095

  8. [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.

  9. 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.

  10. Mechanical properties and shape memory effect of 3D-printed PLA-based porous scaffolds.

    PubMed

    Senatov, F S; Niaza, K V; Zadorozhnyy, M Yu; Maksimkin, A V; Kaloshkin, S D; Estrin, Y Z

    2016-04-01

    In the present work polylactide (PLA)/15wt% hydroxyapatite (HA) porous scaffolds with pre-modeled structure were obtained by 3D-printing by fused filament fabrication. Composite filament was obtained by extrusion. Mechanical properties, structural characteristics and shape memory effect (SME) were studied. Direct heating was used for activation of SME. The average pore size and porosity of the scaffolds were 700μm and 30vol%, respectively. Dispersed particles of HA acted as nucleation centers during the ordering of PLA molecular chains and formed an additional rigid fixed phase that reduced molecular mobility, which led to a shift of the onset of recovery stress growth from 53 to 57°C. A more rapid development of stresses was observed for PLA/HA composites with the maximum recovery stress of 3.0MPa at 70°C. Ceramic particles inhibited the growth of cracks during compression-heating-compression cycles when porous PLA/HA 3D-scaffolds recovered their initial shape. Shape recovery at the last cycle was about 96%. SME during heating may have resulted in "self-healing" of scaffold by narrowing the cracks. PLA/HA 3D-scaffolds were found to withstand up to three compression-heating-compression cycles without delamination. It was shown that PLA/15%HA porous scaffolds obtained by 3D-printing with shape recovery of 98% may be used as self-fitting implant for small bone defect replacement owing to SME.

  11. In vitro analysis and mechanical properties of twin screw extruded single-layered and coextruded multilayered poly(caprolactone) scaffolds seeded with human fetal osteoblasts for bone tissue engineering.

    PubMed

    Ergun, Asli; Yu, Xiaojun; Valdevit, Antonio; Ritter, Arthur; Kalyon, Dilhan M

    2011-12-01

    In vitro culturing and mechanical properties of three types of three-dimensional poly(caprolactone) scaffolds with interconnecting open-foam networks are reported. The scaffolds targeted bone tissue regeneration and were fabricated using twin screw extrusion and coextrusion techniques, for continuous mixing/shaping and formation of single or multilayers with distinct and tailorable porosities and pore sizes. Human fetal preosteoblastic cells, hFOB, were cultured on the extruded and coextruded scaffolds under osteogenic supplements and the samples of the resulting tissue constructs were removed and characterized for cell viability and proliferation using the MTS assay, differentiation, and mineralized matrix synthesis via the alkaline phosphatase, ALP, activity and Alizarin Red staining and cell migration using confocal microscopy and scanning electron microscopy. The hFOB cells formed a confluent lining on scaffold surfaces, migrated to the interior and generated abundant extracellular matrix after 2 weeks of culturing, indicative of the promise of such scaffolds for utilization in tissue engineering. The scaffolds and tissue constructs exhibited compressive fatigue behavior that was similar to that of cancellous bone, suggesting the suitability of their use as bone graft substitutes especially for repair of critical-sized defects or nonunion fractures.

  12. Bioresorbable Ca-phosphate-polymer/metal and Fe-Ag nanocomposites for macro-porous scaffolds with tunable degradation and drug release

    NASA Astrophysics Data System (ADS)

    Gotman, I.; Swain, S. K.; Sharipova, A.; Gutmanas, E. Y.

    2016-11-01

    Bioresorbable implants are increasingly gaining popularity as an attractive alternative to traditional permanent bone healing devices. The advantage of bioresorbable implantable devices is that they slowly degrade over time and disappear once their "mission" is accomplished. Thus, no foreign material is left behind that can cause adverse effects on the host, such as long term local or systemic immune response and stress-shielding related bone atrophy. Resorbable materials considered for surgical implant applications include degradable polymers, Ca phosphate ceramics (CaP) and corrodible metals. Degradable polymers, such as polycaprolactone and lactic acid are weak, lack osteoconductivity and degrade to acidic products that can cause late inflammation. Resorbable CaP ceramics (e.g., β-TCP) are attractive materials for bone regeneration bear close resemblance to the bone mineral, however they are intrinsically brittle and thus unsuitable for use in load-bearing sites. Moreover, introducing high porosity required to encourage better cellular ingrowth into bone regeneration scaffolds is detrimental to the mechanical strength of the material. In present work we review and discuss our results on development of strong bioresorbable Ca-phosphate-polymer/metal nanonocomposites and highly porous scaffolds from them. By introduction of nanoscale ductile polymer or metal phase into CaP ceramic an attempt was made to mimic structure of natural bone, where nanocrystallites of CaP ceramic are bonded by thin collagen layers. Recent results on development of high strength scaffolds from Fe-Ag nanocomposites are also reported. High energy milling of powders followed by cold sintering—high pressure consolidation at ambient temperature in combination with modified porogen leaching method was employed for processing. The developed nanocomposites and scaffolds exhibited high mechanical strength coupled with measurable ductility, gradual lost weight and strength during immersion in

  13. Tailoring properties of porous Poly (vinylidene fluoride) scaffold through nano-sized 58s bioactive glass.

    PubMed

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

    2016-01-01

    The biological properties of porous poly (vinylidene fluoride) (PVDF) scaffolds fabricated by selective laser sintering were tailored through nano-sized 58s bioactive glass. The results showed that 58s bioactive glass distributed evenly in the PVDF matrix. There were some exposed particles on the surface which provided attachment sites for biological response. It was confirmed that the scaffolds had highly bioactivity by the formation of bone-like apatite in simulated body fluid. And the bone-like apatite became dense with the increase in 58s bioactive glass and culture time. Moreover, the scaffolds were suitable for cell adhesion and proliferation compared with the PVDF scaffolds without 58s bioactive glass. The research showed that the PVDF/58s bioactive glass scaffolds had latent application in bone tissue engineering.

  14. Effect of silanization on chitosan porous scaffolds for peripheral nerve regeneration.

    PubMed

    Li, Guicai; Zhang, Luzhong; Wang, Caiping; Zhao, Xueying; Zhu, Changlai; Zheng, Yanhong; Wang, Yaling; Zhao, Yahong; Yang, Yumin

    2014-01-30

    The aim of this study was to evaluate the feasibility of using 3-aminopropyltriethoxysilane (APTE) silanization treatment for modification and biocompatibility of lyophilized chitosan porous scaffolds. The process is beneficial for biomaterial development due to its low toxicity and simplicity. The silanization treatment with low APTE concentration showed no significant influence on the morphology of chitosan scaffolds, while a skin-like surface was observed for the silanized scaffolds treated with high APTE concentration. The porosity and surface amino densities were increased after silanization whereas the swelling ratio was reduced, and the degradation ratio in PBS and anti-acid degradation properties of the silanized chitosan scaffolds were significantly improved. The in vitro Schwann cells culture demonstrated that the silanized scaffolds with 8% APTE could obviously facilitate the attachment and proliferation of Schwann cells, indicating great potential for the application in peripheral nerve regeneration.

  15. Silver nanoparticle studded porous polyethylene scaffolds: bacteria struggle to grow on them while mammalian cells thrive.

    PubMed

    D'Britto, Virginia; Kapse, Harsha; Babrekar, Harshada; Prabhune, A A; Bhoraskar, S V; Premnath, V; Prasad, B L V

    2011-07-01

    Silver nanoparticle studded scaffolds were prepared by exploiting the Ag(+) ion reducing activity of sophorolipids--a class of 'glycolipids' that cap the ensuing nanoparticles as well. To achieve this, the porous polyethylene scaffolds are subjected to N(2) + H(2) plasma treatment, in the first step. Subsequently the sophorolipids are covalently attached to the amine groups on the polymer surface through simple amide chemistry to yield sophorolipid grafted polymer scaffolds. These are then exposed to Ag(+) ions under appropriate conditions leading to the formation of silver nanoparticles immobilized on the polymer scaffolds. It has been found that while bacteria do not survive on these silver studded scaffolds, CHO-K1 cells thrive on them making them good candidates for tissue engineering and bio-implant applications.

  16. 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.

  17. Hierarchic micro-patterned porous scaffolds via electrochemical replica-deposition enhance neo-vascularization.

    PubMed

    Varoni, Elena Maria; Altomare, Lina; Cochis, Andrea; GhalayaniEsfahani, Arash; Cigada, Alberto; Rimondini, Lia; De Nardo, Luigi

    2016-04-21

    Neo-vascularization is a key factor in tissue regeneration within porous scaffolds. Here, we tested the hypothesis that micro-patterned scaffolds, with precisely-designed, open micro-channels, might help endothelial cells to produce intra-scaffold vascular networks. Three series of micro-patterned scaffolds were produced via electrochemical replica-deposition of chitosan and cross-linking. All had regularly-oriented micro-channels (ϕ 500 μm), which differed for the inter-channel spacing, at 600, 700, or 900 μm, respectively. Random-pore scaffolds, using the same technique, were taken as controls. Physical-mechanical characterization revealed high water uptake and favorable elastic mechanical behavior for all scaffolds, slightly reduced in the presence of cross-linking and enhanced with the 700 μm-spaced micro-pattern. At MTT assay, mouse endothelial cell viability was >90% at day 1, 3 and 7, confirmed by visual examination with scanning electron microscopy (SEM). Intra-scaffold cell density, at fluorescence analysis, was higher for the 600 μm-spaced and the 700 μm-spaced micro-patterns over the others. The 700 μm-spaced scaffold was selected for the in vivo testing, to be compared to the random-pore one. Neither type produced an inflammatory reaction; both showed excellent tissue ingrowth. Micro-patterned scaffolds enhanced neo-vascularization, demonstrated by immunofluorescent, semi-quantitative analyses. These findings support the use of micro-patterned porous scaffolds, with adequately spaced micro-channels, to promote neo-vascularization.

  18. Design and characterization of a conductive nanostructured polypyrrole-polycaprolactone coated magnesium/PLGA composite for tissue engineering scaffolds.

    PubMed

    Liu, Haixia; Wang, Ran; Chu, Henry K; Sun, Dong

    2015-09-01

    A novel biodegradable and conductive composite consisting of magnesium (Mg), polypyrrole-block-ploycaprolactone (PPy-PCL), and poly(lactic-co-glycolic acid) (PLGA) is synthesized in a core-shell-skeleton manner for tissue engineering applications. Mg particles in the composite are first coated with a conductive nanostructured PPy-PCL layer for corrosion resistance via the UV-induced photopolymerization method. PLGA matrix is then added to tailor the biodegradability of the resultant composite. Composites with different composition ratios are examined through experiments, and their material properties are characterized. The in vitro experiments on culture of 293FT-GFP cells show that the composites are suitable for cell growth and culture. Biodegradability of the composite is also evaluated. By adding PLGA matrix to the composite, the degrading time of the composite can last for more than eight weeks, hence providing a longer period for tissue formation as compared to Mg composites or alloys. The findings of this research will offer a new opportunity to utilize a conductive, nanostructured-coated Mg/PLGA composite as the scaffold material for implants and tissue regeneration. © 2015 Wiley Periodicals, Inc.

  19. Design of porous polymeric scaffolds by gas foaming of heterogeneous blends.

    PubMed

    Salerno, A; Oliviero, M; Di Maio, E; Iannace, S; Netti, P A

    2009-10-01

    One of the challenges in tissue engineering scaffold design is the realization of structures with a pre-defined multi-scaled porous network. Along this line, this study aimed at the design of porous scaffolds with controlled porosity and pore size distribution from blends of poly(epsilon-caprolactone) (PCL) and thermoplastic gelatin (TG), a thermoplastic natural material obtained by de novo thermoplasticization of gelatin. PCL/TG blends with composition in the range from 40/60 to 60/40 (w/w) were prepared by melt mixing process. The multi-phase microstructures of these blends were analyzed by scanning electron microscopy and dynamic mechanical analysis. Furthermore, in order to prepare open porous scaffolds for cell culture and tissue replacement, the TG and PCL were selectively extracted from the blends by the appropriate combination of solvent and extraction parameters. Finally, with the proposed combination of gas foaming and selective polymer extraction technologies, PCL and TG porous materials with multi-scaled and highly interconnected porosities were designed as novel scaffolds for new-tissue regeneration.

  20. Temperature-driven processing techniques for manufacturing fully interconnected porous scaffolds in bone tissue engineering.

    PubMed

    Guarino, V; Ambrosio, L

    2010-12-01

    The development of structures with a predefined multiscale pore network is a major challenge in designing tissue engineering (TE) scaffolds. To address this, several strategies have been investigated to provide biocompatible, biodegradable porous materials that would be suitable for use as scaffolds, and able to guide and facilitate the cell activity involved in the generation of new tissue regeneration. This study seeks to provide an overview of different temperature-driven process technologies for developing scaffolds with tailored porosity, in which pore size distribution is strictly defined and pores are fully interconnected. Here, three-dimensional (3D) porous composite scaffolds based on poly(epsilon-caprolactone) (PCL) were fabricated by thermally induced phase separation (TIPS) and by melt co-continuous polymer blending (MCPB). The combination of these processes with a salt leaching technique enables the establishment of bimodal porosity within the polymer network. This feature may be exploited in the development of substrates with fully interconnected pores, which can be used effectively for tissue regeneration. Various combinations of the proposed techniques provide a range of procedures for the preparation of porous scaffolds with an appropriate combination of morphological and mechanical properties to reproduce the requisite features of the extracellular matrix (ECM) of hard tissues such as bone.

  1. Fabrication of alumina porous scaffolds with aligned oriented pores for bone tissue engineering applications

    NASA Astrophysics Data System (ADS)

    Sarhadi, Fatemeh; Shafiee Afarani, Mahdi; Mohebbi-Kalhori, Davod; Shayesteh, Masoud

    2016-04-01

    In the present study, porous alumina scaffolds with specific orientation and anisotropic properties are fabricated for application in bone tissue repair. The scaffolds with double shape pores, tubular oriented and isotropic rounded pores, were prepared using alumina and silica as starting materials by the slip casting route. Milled polyurethane foam and silk fibers were applied as replica materials as well. The effect of fiber types and diameter and number of fibers on the microstructure and pore size was studied. Moreover, different characteristics such as porosity, density, orientation, flexural strength and compressive strength of the samples were investigated. Results showed that various fibers with different diameters and numbers led to forming the pores with different pore sizes, microstructure and consequently changes in the physical and mechanical properties. In addition, the simultaneous presence of fibers and particles led to more porous scaffolds. The oriented tiny micro-tube and rounded pores were observed in all porous ceramic scaffolds. Mechanical testing showed an anisotropy in the mechanical behaviors such that higher strengths were observed in the oriented pore direction than that of transverse. With increasing the number and diameter of silk fibers, the scaffolds with a high porosity up to 68 vol% and proper flexural strength were obtained.

  2. 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.

  3. Healing of critical-size segmental defects in rat femora using strong porous bioactive glass scaffolds.

    PubMed

    Bi, Lianxiang; Zobell, Brett; Liu, Xin; Rahaman, Mohamed N; Bonewald, Lynda F

    2014-09-01

    The repair of structural bone defects such as segmental defects in the long bones of the limbs is a challenging clinical problem. In this study, the capacity of silicate (13-93) and borate (13-93B3) bioactive glass scaffolds (porosity=47-50%) to heal critical-size segmental defects in rat femurs was evaluated and compared with autografts. Defects were implanted with 13-93 and 13-93B3 scaffolds with a grid-like microstructure (compressive strength=86 MPa and 40 MPa, respectively), 13-93B3 scaffolds with an oriented microstructure (compressive strength=32 MPa) and autografts using intramedullary fixation. Twelve weeks post-implantation, the defects were harvested and evaluated using histomorphometric analysis. The percentage of new bone in the defects implanted with the three groups of glass scaffolds (25-28%) and the total von Kossa-positive area (32-38%) were not significantly different from the autografts (new bone=38%; von Kossa-positive area=40%) (p>0.05). New blood vessel area in the defects implanted with the glass scaffolds (4-8%) and the autografts (5%) showed no significant difference among the four groups. New cartilage formed in the 13-93 grid-like scaffolds (18%) was significantly higher than in 13-93B3 grid-like scaffolds (8%) and in the autografts (8%) (p=0.02). The results indicate that these strong porous bioactive glass scaffolds are promising synthetic implants for structural bone repair.

  4. In vitro degradation of a 3D porous Pennisetum purpureum/PLA biocomposite scaffold.

    PubMed

    Revati, R; Majid, M S Abdul; Ridzuan, M J M; Basaruddin, K S; Rahman Y, M N; Cheng, E M; Gibson, A G

    2017-10-01

    The in vitro degradation and mechanical properties of a 3D porous Pennisetum purpureum (PP)/polylactic acid (PLA)-based scaffold were investigated. In this study, composite scaffolds with PP to PLA ratios of 0%, 10%, 20%, and 30% were immersed in a PBS solution at 37°C for 40 days. Compression tests were conducted to evaluate the compressive strength and modulus of the scaffolds, according to ASTM F451-95. The compression strength of the scaffolds was found to increase from 1.94 to 9.32MPa, while the compressive modulus increased from 1.73 to 5.25MPa as the fillers' content increased from 0wt% to 30wt%. Moreover, field emission scanning electron microscopy (FESEM) and X-ray diffraction were employed to observe and analyse the microstructure and fibre-matrix interface. Interestingly, the degradation rate was reduced for the PLA/PP20 scaffold, though insignificantly, this could be attributed to the improved mechanical properties and stronger fibre-matrix interface. Microstructure changes after degradation were observed using FESEM. The FESEM results indicated that a strong fibre-matrix interface was formed in the PLA/PP20 scaffold, which reflected the addition of P. purpureum into PLA decreasing the degradation rate compared to in pure PLA scaffolds. The results suggest that the P. purpureum/PLA scaffold degradation rate can be altered and controlled to meet requirements imposed by a given tissue engineering application. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Porous ovalbumin scaffolds with tunable properties: a resource-efficient biodegradable material for tissue engineering applications.

    PubMed

    Luo, Baiwen; Choong, Cleo

    2015-01-01

    Natural materials are promising alternatives to synthetic materials used in tissue engineering applications as they have superior biocompatibility and promote better cell attachment and proliferation. Ovalbumin, a natural polymer found in avian egg white, is an example of a nature-derived material. Despite the availability and reported biocompatibility of ovalbumin, limited research has been carried out to investigate the efficacy of ovalbumin-based scaffolds for adipose tissue engineering applications. Hence, the current study was carried out to investigate the effect of different crosslinkers on ovalbumin scaffold properties as first step towards the development of ovalbumin-based scaffolds for adipose tissue engineering applications. In this study, highly porous three-dimensional scaffolds were fabricated by using three different crosslinkers: glutaraldehyde, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and 1,4-butanediol diglycidyl ether. Results showed that the overall scaffold properties such as morphology, pore size and mechanical properties could be modulated based on the type and concentration of crosslinkers used during the fabrication process. Subsequently, the efficacy of the different scaffolds for supporting cell proliferation was investigated. In vitro degradation was also carried on for the best scaffold based on the mechanical and cellular results. Overall, this study is a demonstration of the viability of ovalbumin-based scaffolds as cell carriers for soft tissue engineering applications. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  6. Porous composite scaffolds based on gelatin and partially hydrolyzed alpha-tricalcium phosphate.

    PubMed

    Panzavolta, S; Fini, M; Nicoletti, A; Bracci, B; Rubini, K; Giardino, R; Bigi, A

    2009-02-01

    Porous composite scaffolds of varying compositions were prepared by freeze-drying gelatin foams containing increasing amounts of alpha-tricalcium phosphate (alpha-TCP), up to about 40 wt.%. Due to the presence of gelatin, a partial hydrolysis of alpha-TCP into octacalcium phosphate (OCP) occurs during foaming. As a consequence, the scaffolds contain both alpha-TCP and OCP, in relative amounts of about 74% and 26%, respectively, independent of the initial composition. In physiological conditions the inorganic component of the scaffolds undergoes a further hydrolysis as shown by the finding that after immersion in phosphate-buffered saline at 37 degrees C for 1 week the scaffolds contain poorly crystalline hydroxyapatite together with OCP. The scaffolds display a porous interconnected microstructure. The mean dimensions of the pores decrease from about 350 to about 170 microm as the inorganic phase content increases. Simultaneously, the mean values of the compression strength and Young's modulus increase. Stabilization of the scaffolds was obtained by using a natural, non-toxic, crosslinking agent, genipin, which significantly improves their mechanical properties.

  7. The optimization of porous polymeric scaffolds for chondrocyte/atelocollagen based tissue-engineered cartilage.

    PubMed

    Tanaka, Yoko; Yamaoka, Hisayo; Nishizawa, Satoru; Nagata, Satoru; Ogasawara, Toru; Asawa, Yukiyo; Fujihara, Yuko; Takato, Tsuyoshi; Hoshi, Kazuto

    2010-06-01

    To broaden the clinical application of cartilage regenerative medicine, we should develop an implant-type tissue-engineered cartilage with firmness and 3-D structure. For that, we attempted to use a porous biodegradable polymer scaffold in the combination with atelocollagen hydrogel, and optimized the structure and composition of porous scaffold. We administered chondrocytes/atelocollagen mixture into the scaffolds with various kinds of porosities (80-95%) and pore sizes (0.3-2.0 mm), consisting of PLLA or related polymers (PDLA, PLA/CL and PLGA), and transplanted the constructs in the subcutaneous areas of nude mice. The constructs using scaffolds of excessively large pore sizes (>1 mm) broke out on the skin and impaired the host tissue. The scaffold with the porosity of 95% and pore size of 0.3 mm could effectively retain the cells/gel mixture and indicated a fair cartilage regeneration. Regarding the composition, the tissue-engineered cartilage was superior in PLGA and PLLA to that in PLA/CA and PDLA. The latter two showed the dense accumulation of macrophages, which may deteriorate the cartilage regeneration. Although PLGA or PLLA has been currently recommended for the scaffold of cartilage, the polymer for which biodegradation was exactly synchronized to the cartilage regeneration would improve the quality of the tissue-engineered cartilage.

  8. Processing and characterization of porous structures from chitosan and starch for tissue engineering scaffolds.

    PubMed

    Nakamatsu, Javier; Torres, Fernando G; Troncoso, Omar P; Min-Lin, Yuan; Boccaccini, Aldo R

    2006-12-01

    Natural biodegradable polymers were processed by different techniques for the production of porous structures for tissue engineering scaffolds. Potato, corn, and sweet potato starches and chitosan, as well as blends of these, were characterized and used in the experiments. The techniques used to produce the porous structures included a novel solvent-exchange phase separation technique and the well-established thermally induced phase separation method. Characterization of the open pore structures was performed by measuring pore size distribution, density, and porosity of the samples. A wide range of pore structures ranging from 1 to 400 microm were obtained. The mechanisms of pore formation are discussed for starch and chitosan scaffolds. Pore morphology in starch scaffolds seemed to be determined by the initial freezing temperature/freezing rate, whereas in chitosan scaffolds the shape and size of pores may have been determined by the processing route used. The mechanical properties of the scaffolds were assessed by indentation tests, showing that the indentation collapse strength depends on the pore geometry and the material type. Bioactivity and degradation of the potential scaffolds were assessed by immersion in simulated body fluid.

  9. Bilayer porous scaffold based on poly-(ɛ-caprolactone) nanofibrous membrane and gelatin sponge for favoring cell proliferation

    NASA Astrophysics Data System (ADS)

    Zhou, Zhihua; Zhou, Yang; Chen, Yiwang; Nie, Huarong; Wang, Yang; Li, Fan; Zheng, Yan

    2011-12-01

    Electrospun poly-(ɛ-caprolactone) (PCL) nanofibers has been widely used in the medical prosthesis. However, poor hydrophilicity and the lack of natural recognition sites for covalent cell-recognition signal molecules to promote cell attachment have limited its utility as tissue scaffolds. In this study, Bilayer porous scaffolds based on PCL electrospun membranes and gelatin (GE) sponges were fabricated through soft hydrolysis of PCL electrospun followed by grafting gelatin onto the fiber surface, through crosslinking and freeze drying treatment of additional gelatin coat and grafted gelatin surface. GE sponges were stably anchored on PCL membrane surface with the aid of grafted GE molecules. The morphologies of bilayer porous scaffolds were observed through SEM. The contact angle of the scaffolds was 0°, the mechanical properties of scaffolds were measured by tensile test, Young's moduli of PCL scaffolds before and after hydrolysis are 66-77.3 MPa and 62.3-75.4 MPa, respectively. Thus, the bilayer porous scaffolds showed excellent hydrophilic surface and desirable mechanical strength due to the soft hydrolysis and GE coat. The cell culture results showed that the adipose derived mesenchymal stem cells did more favor to adhere and grow on the bilayer porous scaffolds than on PCL electrospun membranes. The better cell affinity of the final bilayer scaffolds not only attributed to the surface chemistry but also the introduction of bilayer porous structure.

  10. 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

  11. Nanoscale modification of porous gelatin scaffolds with chondroitin sulfate for corneal stromal tissue engineering

    PubMed Central

    Lai, Jui-Yang; Li, Ya-Ting; Cho, Ching-Hsien; Yu, Ting-Chun

    2012-01-01

    Recent studies reflect the importance of using naturally occurring biopolymers as three-dimensional corneal keratocyte scaffolds and suggest that the porous structure of gelatin materials may play an important role in controlling nutrient uptake. In the current study, the authors further consider the application of carbodiimide cross-linked porous gelatin as an alternative to collagen for corneal stromal tissue engineering. The authors developed corneal keratocyte scaffolds by nanoscale modification of porous gelatin materials with chondroitin sulfate (CS) using carbodiimide chemistry. Scanning electron microscopy/energy dispersive X-ray spectroscopy and Fourier transform infrared spectroscopy showed that the amount of covalently incorporated polysaccharide was significantly increased when the CS concentration was increased from 0% to 1.25% (w/v). In addition, as demonstrated by dimethylmethylene blue assays, the CS content in these samples was in the range of 0.078–0.149 nmol per 10 mg scaffold. When compared with their counterparts without CS treatment, various CS-modified porous gelatin membranes exhibited higher levels of water content, light transmittance, and amount of permeated nutrients but possessed lower Young’s modulus and resistance against protease digestion. The hydrophilic and mechanical properties of scaffolds modified with 0.25% CS were comparable with those of native corneas. The samples from this group were biocompatible with the rabbit corneal keratocytes and showed enhanced proliferative and biosynthetic capacity of cultured cells. In summary, the authors found that the nanoscale-level modification has influence on the characteristics and cell-material interactions of CS-containing gelatin hydrogels. Porous membranes with a CS content of 0.112 ± 0.003 nmol per 10 mg scaffold may hold potential for use in corneal stromal tissue engineering. PMID:22403490

  12. Heterogeneous minimal surface porous scaffold design using the distance field and radial basis functions.

    PubMed

    Yoo, Dongjin

    2012-06-01

    This paper presented an effective method for the 3D heterogeneous porous scaffold design of human tissue using triply periodic minimal surface (TPMS) internal pore architectures. First, an implicit solid representing the smooth 3D scalar field for the porosity distribution was reconstructed by interpolating the geometric positions of control points and porosity values defined at those points using an implicit interpolation algorithm based on the thin-plate radial basis function. After generating the implicit solid representing the smooth 3D scalar field for the porosity distribution, a functionally graded tissue scaffold with accurately controlled porosity distribution was designed using the TPMS-based unit cell libraries. Numerical results showed that the proposed scaffold design method has the potential benefits for accurately controlling the spatial porosity distribution within an arbitrarily shaped scaffold while keeping the advantage of the TPMS-based unit cell libraries.

  13. Chronic Label-free Volumetric Photoacoustic Microscopy of Melanoma Cells in Three-Dimensional Porous Scaffolds

    PubMed Central

    Zhang, Yu; Cai, Xin; Choi, Sung-Wook; Kim, Chulhong; Wang, Lihong V.; Xia, Younan

    2010-01-01

    Visualizing cells in three-dimensional (3D) scaffolds has been one of the major challenges in tissue engineering. Most current imaging modalities either suffer from poor penetration depth or require exogenous contrast agents. Here, we demonstrate photoacoustic microscopy (PAM) of the spatial distribution and temporal proliferation of cells inside three-dimensional porous scaffolds with thicknesses over 1 mm. Specifically, we evaluated the effects of seeding and culture methods on the spatial distribution of melanoma cells. Spatial distribution of the cells in the scaffold was well-resolved in PAM images. Moreover, the number of cells in the scaffold was quantitatively measured from the as-obtained volumetric information. The cell proliferation profile obtained from PAM correlated well with what was obtained using the traditional 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. PMID:20727581

  14. Enhanced repair of segmental bone defects in rabbit radius by porous tantalum scaffolds modified with the RGD peptide.

    PubMed

    Wang, Hui; Li, Qijia; Wang, Qian; Zhang, Hui; Shi, Wei; Gan, Hongquan; Song, Huiping; Wang, Zhiqiang

    2017-03-01

    Fast and stable repair of segmental bone defects remains a challenge for clinical orthopedic surgery. In recent years, porous tantalum has been widely applied in clinical orthopedics for low modulus of elasticity, with three-dimensional microstructures similar to cancellous bone and excellent biocompatibility. To further improve bone the repairing ability of porous tantalum, the cyclo(-RGDfK-) peptide was coated on the surface of porous tantalum scaffolds. A model of 15 mm segmental defect was made at the midshaft of right radius in New Zealand White rabbits. In the experimental group, defects were implanted (press-fit) using porous tantalum scaffolds modified with cyclo(-RGDfK-) peptide. Control animals were implanted with non-modified porous tantalum scaffolds or xenogeneic cancellous bone scaffolds, respectively. No implant was provided for the blank group. Bone repair was assessed by X-ray and histological observations at 4, 8, and 16 weeks post-operation, with biomechanical tests and micro-computed tomography performed at 16 weeks post-surgery. The results showed that bone formation was increased at the interface and inside the inner pores of modified porous tantalum scaffolds than those of non-modified porous tantalum scaffolds; biomechanical properties in the modified porous tantalum group were superior to those of the non-modified porous tantalum and xenogeneic cancellous bone groups, while new bone volume fractions using micro-computed tomography analysis were similar between the modified porous tantalum and xenogeneic cancellous bone groups. Our findings suggested that modified porous tantalum scaffolds had enhanced repairing ability in segmental bone defect in rabbit radius, and may serve as a potential material for repairing large bone defects.

  15. 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

  16. Correlation between properties and microstructure of laser sintered porous β-tricalcium phosphate bone scaffolds.

    PubMed

    Shuai, Cijun; Feng, Pei; Zhang, Liyang; Gao, Chengde; Hu, Huanlong; Peng, Shuping; Min, Anjie

    2013-10-01

    A porous β-tricalcium phosphate (β-TCP) bioceramic scaffold was successfully prepared with our homemade selective laser sintering system. Microstructure observation by a scanning electron microscope showed that the grains grew from 0.21 to 1.32 μm with the decrease of laser scanning speed from 250 to 50 mm min(-1). The mechanical properties increased mainly due to the improved apparent density when the laser scanning speed decreased to 150 mm min(-1). When the scanning speed was further decreased, the grain size became larger and the mechanical properties severely decreased. The highest Vickers hardness and fracture toughness of the scaffold were 3.59 GPa and 1.16 MPa m(1/2), respectively, when laser power was 11 W, spot size was 1 mm in diameter, layer thickness was 0.1-0.2 mm and laser scanning speed was 150 mm min(-1). The biocompatibility of these scaffolds was assessed in vitro with MG63 osteoblast-like cells and human bone marrow mesenchymal stem cells. The results showed that all the prepared scaffolds are suitable for cell attachment and differentiation. Moreover, the smaller the grain size, the better the cell biocompatibility. The porous scaffold with a grain size of 0.71 μm was immersed in a simulated body fluid for different days to assess the bioactivity. The surface of the scaffold was covered by a bone-like apatite layer, which indicated that the β-TCP scaffold possesses good bioactivity. These discoveries demonstrated the evolution rule between grain microstructure and the properties that give a useful reference for the fabrication of β-TCP bone scaffolds.

  17. Porous magnesium/PLGA composite scaffolds for enhanced bone regeneration following tooth extraction.

    PubMed

    Brown, Andrew; Zaky, Samer; Ray, Herbert; Sfeir, Charles

    2015-01-01

    Sixty percent of implant-supported dental prostheses require bone grafting to enhance bone quantity and quality prior to implant placement. We have developed a metallic magnesium particle/PLGA composite scaffold to overcome the limitations of currently used dental bone grafting materials. This is the first report of porous metallic magnesium/PLGA scaffolds synthesized using a solvent casting, salt leaching method. We found that incorporation of varying amounts of magnesium into the PLGA scaffolds increased the compressive strength and modulus, as well as provided a porous structure suitable for cell infiltration, as measured by mercury intrusion porosimetry. Additionally, combining basic-degrading magnesium with acidic-degrading PLGA led to an overall pH buffering effect and long-term release of magnesium over the course of a 10-week degradation assay, as measured with inductively coupled plasma-atomic emission spectroscopy. Using an indirect proliferation assay adapted from ISO 10993:5, it was found that extracts of medium from degrading magnesium/PLGA scaffolds increased bone marrow stromal cell proliferation in vitro, a phenomenon observed by other groups investigating magnesium's impact on cells. Finally, magnesium/PLGA scaffold biocompatibility was assessed in a canine socket preservation model. Micro-computed tomography and histological analysis showed the magnesium/PLGA scaffolds to be safer and more effective at preserving bone height than empty controls. Three-dimensional magnesium/PLGA composite scaffolds show promise for dental socket preservation and also, potentially, orthopedic bone regeneration. These scaffolds could decrease inflammation observed with clinically used PLGA devices, as well as enhance osteogenesis, as observed with previously studied magnesium devices.

  18. Correlation between properties and microstructure of laser sintered porous β-tricalcium phosphate bone scaffolds

    NASA Astrophysics Data System (ADS)

    Shuai, Cijun; Feng, Pei; Zhang, Liyang; Gao, Chengde; Hu, Huanlong; Peng, Shuping; Min, Anjie

    2013-10-01

    A porous β-tricalcium phosphate (β-TCP) bioceramic scaffold was successfully prepared with our homemade selective laser sintering system. Microstructure observation by a scanning electron microscope showed that the grains grew from 0.21 to 1.32 μm with the decrease of laser scanning speed from 250 to 50 mm min-1. The mechanical properties increased mainly due to the improved apparent density when the laser scanning speed decreased to 150 mm min-1. When the scanning speed was further decreased, the grain size became larger and the mechanical properties severely decreased. The highest Vickers hardness and fracture toughness of the scaffold were 3.59 GPa and 1.16 MPa m1/2, respectively, when laser power was 11 W, spot size was 1 mm in diameter, layer thickness was 0.1-0.2 mm and laser scanning speed was 150 mm min-1. The biocompatibility of these scaffolds was assessed in vitro with MG63 osteoblast-like cells and human bone marrow mesenchymal stem cells. The results showed that all the prepared scaffolds are suitable for cell attachment and differentiation. Moreover, the smaller the grain size, the better the cell biocompatibility. The porous scaffold with a grain size of 0.71 μm was immersed in a simulated body fluid for different days to assess the bioactivity. The surface of the scaffold was covered by a bone-like apatite layer, which indicated that the β-TCP scaffold possesses good bioactivity. These discoveries demonstrated the evolution rule between grain microstructure and the properties that give a useful reference for the fabrication of β-TCP bone scaffolds.

  19. Porous scaffold design using the distance field and triply periodic minimal surface models.

    PubMed

    Yoo, Dong J

    2011-11-01

    An effective method for the 3D porous scaffold design of human tissue is presented based on a hybrid method of distance field and triply periodic minimal surface (TPMS). By the creative application of traditional distance field algorithm into the Boolean operations of the anatomical model and TPMS-based unit cell library, an almost defects free porous scaffolds having the complicated micro-structure and high quality external surface faithful to a specific anatomic model can be easily obtained without the difficult and time-consuming trimming and re-meshing processes. After generating the distance fields for the given tissue model and required internal micro-structure, a series of simple modifications in distance fields enable us to obtain a complex porous scaffold. Experimental results show that the proposed scaffold design method has the potential to combine the perfectly interconnected pore networks based on the TPMS unit cell libraries and the given external geometry in a consistent framework irrespective of the complexity of the models.

  20. Mechanical characterization of injection-molded macro porous bioceramic bone scaffolds.

    PubMed

    Vivanco, Juan; Aiyangar, Ameet; Araneda, Aldo; Ploeg, Heidi-Lynn

    2012-05-01

    Bioactive ceramic materials like tricalcium phosphate (TCP) have been emerging as viable material alternatives to the current therapies of bone scaffolding to target fracture healing and osteoporosis. Both material and architectural characteristics play a critical role in the osteoconductive capacity and strength of bone scaffolds. Thus, the objective of this research was to investigate the sintering temperature effect of a cost-effective manufacturing process on the architecture and mechanical properties of a controlled macro porous bioceramic bone scaffold. In this study the physical and mechanical properties of β-TCP bioceramic scaffolds were investigated as a function of the sintering temperature in the range of 950-1150 °C. Physical properties investigated included bulk dimensions, pore size, and strut thickness; and, compressive mechanical properties were evaluated in air at room temperature and in saline solution at body temperature. Statistically significant increases in apparent elastic modulus were measured for scaffolds sintered at higher temperatures. Structural stiffness for all the specimens was significantly reduced when tested at body temperature in saline solution. These findings support the development of clinically successful bioceramic scaffolds that may stimulate bone regeneration and scaffold integration while providing structural integrity.

  1. 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.

  2. Biodegradable CSMA/PECA/Graphene Porous Hybrid Scaffold for Cartilage Tissue Engineering.

    PubMed

    Liao, JinFeng; Qu, Ying; Chu, BingYang; Zhang, XiaoNing; Qian, ZhiYong

    2015-05-11

    Owing to the limited repair capacity of articular cartilage, it is essential to develop tissue-engineered cartilage for patients suffering from joint disease and trauma. Herein, we prepared a novel hybrid scaffold composed of methacrylated chondroitin sulfate (CSMA), poly(ethylene glycol) methyl ether-ε-caprolactone-acryloyl chloride (MPEG-PCL-AC, PECA was used as abbreviation for MPEG-PCL-AC) and graphene oxide (GO) and evaluated its potential application in cartilage tissue engineering. To mimic the natural extracellular matrix (ECM) of cartilage, the scaffold had an adequate pore size, porosity, swelling ability, compression modulus and conductivity. Cartilage cells contacted with the scaffold remained viable and showed growth potential. Furthermore, CSMA/PECA/GO scaffold was biocompatible and had a favorable degradation rate. In the cartilage tissue repair of rabbit, Micro-CT and histology observation showed the group of CSMA/PECA/GO scaffold with cellular supplementation had better chondrocyte morphology, integration, continuous subchondral bone, and much thicker newly formed cartilage compared with scaffold group and control group. Our results show that the CSMA/PECA/GO hybrid porous scaffold can be applied in articular cartilage tissue engineering and may have great potential to in other types of tissue engineering applications.

  3. 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

  4. 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.

  5. Highly porous, low elastic modulus 316L stainless steel scaffold prepared by selective laser melting.

    PubMed

    Čapek, Jaroslav; Machová, Markéta; Fousová, Michaela; Kubásek, Jiří; Vojtěch, Dalibor; Fojt, Jaroslav; Jablonská, Eva; Lipov, Jan; Ruml, Tomáš

    2016-12-01

    Recently, porous metallic materials have been extensively studied as candidates for use in the fabrication of scaffolds and augmentations to repair trabecular bone defects, e.g. in surroundings of joint replacements. Fabricating these complex structures by using common approaches (e.g., casting and machining) is very challenging. Therefore, rapid prototyping techniques, such as selective laser melting (SLM), have been investigated for these applications. In this study, we characterized a highly porous (87 vol.%) 316L stainless steel scaffold prepared by SLM. 316L steel was chosen because it presents a biomaterial still widely used for fabrication of joint replacements and, from the practical point of view, use of the same material for fabrication of an augmentation and a joint replacement is beneficial for corrosion prevention. The results are compared to the reported properties of two representative nonporous 316L stainless steels prepared either by SLM or casting and subsequent hot forging. The microstructural and mechanical properties and the surface chemical composition and interaction with the cells were investigated. The studied material exhibited mechanical properties that were similar to those of trabecular bone (compressive modulus of elasticity ~0.15GPa, compressive yield strength ~3MPa) and cytocompatibility after one day that was similar to that of wrought 316L stainless steel, which is a commonly used biomaterial. Based on the obtained results, SLM is a suitable method for the fabrication of porous 316L stainless steel scaffolds with highly porous structures. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. In vivo imaging study of angiogenesis in a channelized porous scaffold.

    PubMed

    Tamplenizza, Margherita; Tocchio, Alessandro; Gerges, Irini; Martello, Federico; Martelli, Cristina; Ottobrini, Luisa; Lucignani, Giovanni; Milani, Paolo; Lenardi, Cristina

    2015-01-01

    The main scientific issue hindering the development of tissue engineering technologies is the lack of proper vascularization. Among the various approaches developed for boosting vascularization, scaffold design has attracted increasing interest over the last few years. The aim of this article is to illustrate a scaffold design strategy for enhancing vascularization based on sacrificial microfabrication of embedded microchannels. This approach was combined with an innovative poly(ether urethane urea) (PEUtU) porous scaffold to provide an alternative graft substitute material for the treatment of tissue defects. Fluorescent and chemiluminescent imaging combined with computed tomography were used to study the behavior of the scaffold composition within living subjects by analyzing angiogenesis and inflammation processes and observing the variation in x-ray absorption, respectively. For this purpose, an IntegriSense 680 probe was used in vivo for the localization and quantification of integrin αvβ3, due to its critical involvement in angiogenesis, and a XenoLight RediJect Inflammation Probe for the study of the decline in inflammation progression during healing. Overall, the collected data suggest the advantages of embedding a synthetic vascular network into a PEUtU porous matrix to enhance in vivo tissue integration, maturation, and regeneration. Moreover, our imaging approach proved to be an efficient and versatile tool for scaffold in vivo testing.

  7. Antimicrobial 3D Porous Scaffolds Prepared by Additive Manufacturing and Breath Figures.

    PubMed

    Vargas Alfredo, Nelson; Dorronsoro, Ane; Cortajarena, Aitziber L; Rodriguez-Hernandez, Juan

    2017-09-21

    We describe herein a novel strategy for the fabrication of efficient 3D printed antibacterial scaffolds. For this purpose, both the surface topography as well as the chemical composition of 3D scaffolds fabricated by additive manufacturing were modified. The scaffolds were fabricated by fused deposition modeling (FDM) using high impact polystyrene (HIPS) filaments. The surface of the objects was then topographically modified providing materials with porous surfaces by means of the Breath Figures approach. The strategy involves the immersion of the scaffold in a polymer solution during a precise period of time. This approach permitted the modification of the pore size varying the immersion time as well as the solution concentration. Moreover, by using polymer blend solutions of polystyrene and polystyrene-b-poly(acrylic acid) (PS23-b-PAA18) and a quaternized polystyrene-b-poly(dimethylaminoethyl methacrylate) (PS42-b-PDMAEMAQ17), the scaffolds were simultaneously chemically modified. The surfaces were characterized by scanning electron microscopy and FT-IR. Finally, the biological response towards bacteria was explored. Porous surfaces prepared using quaternized PDMAEMA as well as those prepared using PAA confer antimicrobial activity to the films, i.e. were able to kill on contact S. aureus employed as model bacteria.

  8. Porous decellularized tissue engineered hypertrophic cartilage as a scaffold for large bone defect healing.

    PubMed

    Cunniffe, Gráinne M; Vinardell, Tatiana; Murphy, J Mary; Thompson, Emmet M; Matsiko, Amos; O'Brien, Fergal J; Kelly, Daniel J

    2015-09-01

    Clinical translation of tissue engineered therapeutics is hampered by the significant logistical and regulatory challenges associated with such products, prompting increased interest in the use of decellularized extracellular matrix (ECM) to enhance endogenous regeneration. Most bones develop and heal by endochondral ossification, the replacement of a hypertrophic cartilaginous intermediary with bone. The hypothesis of this study is that a porous scaffold derived from decellularized tissue engineered hypertrophic cartilage will retain the necessary signals to instruct host cells to accelerate endogenous bone regeneration. Cartilage tissue (CT) and hypertrophic cartilage tissue (HT) were engineered using human bone marrow derived mesenchymal stem cells, decellularized and the remaining ECM was freeze-dried to generate porous scaffolds. When implanted subcutaneously in nude mice, only the decellularized HT-derived scaffolds were found to induce vascularization and de novo mineral accumulation. Furthermore, when implanted into critically-sized femoral defects, full bridging was observed in half of the defects treated with HT scaffolds, while no evidence of such bridging was found in empty controls. Host cells which had migrated throughout the scaffold were capable of producing new bone tissue, in contrast to fibrous tissue formation within empty controls. These results demonstrate the capacity of decellularized engineered tissues as 'off-the-shelf' implants to promote tissue regeneration.

  9. EVA-enhanced embedding medium for histological analysis of 3D porous scaffold material.

    PubMed

    Lim, Jin Ik; Lee, Yong-Keun

    2009-10-01

    When sectioning a 3D porous scaffold made of a soft elastomeric material embedded in paraffin medium, it is not easy to obtain a section because of the different mechanical properties of the paraffin and tissue/scaffold. We describe a new embedding material for histological analysis of various biomaterials that is composed of paraffin and ethylene vinyl acetate (EVA) resin (0, 3, 7, and 13 wt.%). 3D porous poly(L-lactide-epsilon-caprolactone) (PLCL) and chitosan scaffolds were fabricated to test the sectioning efficiency of the paraffin/EVA embedding material. The new embedding material was characterized by rheological analysis and solvent solubility testing in xylene and n-hexane. The hydrophilicity of the new material was assessed by contact angle measurement and its surface roughness was measured using AFM analysis. The staining efficiency of sections embedded in a paraffin/EVA mixture was determined by eosin staining of the chitosan scaffold and chitosan/collagen hybrid scaffold using a fluorescently labeled collagen. Section roughness decreased with increasing EVA content. The softening temperature of the paraffin/EVA mixture was similar to that of paraffin (50-60 degrees C by rheometer). The paraffin/EVA mixture dissolved completely in xylene after 30min at 50 degrees C, and after 30min in n-hexane at 60 degrees C. Therefore, the new embedding medium can be used for histological analysis of various biomaterials and natural tissues.

  10. PLLA-collagen and PLLA-gelatin hybrid scaffolds with funnel-like porous structure for skin tissue engineering

    NASA Astrophysics Data System (ADS)

    Lu, Hongxu; Oh, Hwan Hee; Kawazoe, Naoki; Yamagishi, Kozo; Chen, Guoping

    2012-12-01

    In skin tissue engineering, a three-dimensional porous scaffold is necessary to support cell adhesion and proliferation and to guide cells moving into the repair area in the wound healing process. Structurally, the porous scaffold should have an open and interconnected porous architecture to facilitate homogenous cell distribution. Moreover, the scaffolds should be mechanically strong to protect deformation during the formation of new skin. In this study, the hybrid scaffolds were prepared by forming funnel-like collagen or gelatin sponge on a woven poly(l-lactic acid) (PLLA) mesh. The hybrid scaffolds combined the advantages of both collagen or gelatin (good cell-interactions) and PLLA mesh (high mechanical strength). The hybrid scaffolds were used to culture dermal fibroblasts for dermal tissue engineering. The funnel-like porous structure promoted homogeneous cell distribution and extracellular matrix production. The PLLA mesh reinforced the scaffold to avoid deformation. Subcutaneous implantation showed that the PLLA-collagen and PLLA-gelatin scaffolds promoted the regeneration of dermal tissue and epidermis and reduced contraction during the formation of new tissue. These results indicate that funnel-like hybrid scaffolds can be used for skin tissue regeneration.

  11. PLLA–collagen and PLLA–gelatin hybrid scaffolds with funnel-like porous structure for skin tissue engineering

    PubMed Central

    Lu, Hongxu; Oh, Hwan Hee; Kawazoe, Naoki; Yamagishi, Kozo; Chen, Guoping

    2012-01-01

    In skin tissue engineering, a three-dimensional porous scaffold is necessary to support cell adhesion and proliferation and to guide cells moving into the repair area in the wound healing process. Structurally, the porous scaffold should have an open and interconnected porous architecture to facilitate homogenous cell distribution. Moreover, the scaffolds should be mechanically strong to protect deformation during the formation of new skin. In this study, the hybrid scaffolds were prepared by forming funnel-like collagen or gelatin sponge on a woven poly(l-lactic acid) (PLLA) mesh. The hybrid scaffolds combined the advantages of both collagen or gelatin (good cell-interactions) and PLLA mesh (high mechanical strength). The hybrid scaffolds were used to culture dermal fibroblasts for dermal tissue engineering. The funnel-like porous structure promoted homogeneous cell distribution and extracellular matrix production. The PLLA mesh reinforced the scaffold to avoid deformation. Subcutaneous implantation showed that the PLLA–collagen and PLLA–gelatin scaffolds promoted the regeneration of dermal tissue and epidermis and reduced contraction during the formation of new tissue. These results indicate that funnel-like hybrid scaffolds can be used for skin tissue regeneration. PMID:27877537

  12. 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.

  13. Improving Internal Cell Colonization of Porous Scaffolds with Chemical Gradients Produced by Plasma Assisted Approaches.

    PubMed

    Sardella, Eloisa; Salama, Rania A; Waly, Gihan H; Habib, A Nour; Favia, Pietro; Gristina, Roberto

    2017-02-08

    Cell colonization of the surrounding environment is a very significant process in both physiological and pathological events. In order to understand the tissue regeneration process and thereby provide guidance principles for designing new biomaterials, it is of paramount importance to study the cell colonization in the presence of physical, chemical, and biological cues. Flat "gradient" materials are generally used with this purpose. Three dimensional gradient scaffolds mimicking more precisely the situation in vivo are somewhat more complex to fabricate and characterize. Scaffolds for Tissue Engineering (TE) made of hydrophobic synthetic polymers do not allow good cell colonization: far from their periphery, in fact, internal cell colonization is usually low. In this research poly-ε caprolactone (PCL) scaffolds have been "decorated" with chemical gradients both on top and along their thickness by means of cold plasma processes, in order to improve cell colonization of their core. Plasma treatments with a mixture of argon and oxygen (Ar/O2), as well as plasma deposition of differently cross-linked poly(ethylene oxide) (PEO)-like coatings, have been performed. This study establishes that cross-linked PEO-like domains interspaced with native PCL ones deposited only on top of the scaffold (i.e., coating that penetrates less than 300 μm inside the scaffold) are more effective in promoting cell colonization across the scaffolds than the other tested materials including superhydrophilic samples and that ones produced by tested double step approaches. Last but not least, one result of this research is that, in the case of plasma coatings with low deposition rates and porous materials with a low pore interconnectivity, it is possible to improve penetration of low pressure plasma active species inside the scaffold's core thorough a pretreatment of the porous materials (i.e., penetration up to 4500 mm far from topside).

  14. Bone formation in transforming growth factor beta-1-coated porous poly(propylene fumarate) scaffolds.

    PubMed

    Vehof, Johan W M; Fisher, John P; Dean, David; van der Waerden, Jan-Paul C M; Spauwen, Paul H M; Mikos, Antonios G; Jansen, John A

    2002-05-01

    This study determined the bone growth into pretreated poly(propylene fumarate) (PPF) scaffolds implanted into a subcritical size, rabbit cranial defect. PPF scaffolds were constructed by using a photocrosslinking-porogen leaching technique. These scaffolds were then either prewetted (PPF-Pw), treated with RF glow-discharge (PPF-Gd), coated with fibronectin (PPF-Fn), or coated with rhTGF-beta1 (PPF-TGF-beta1). One of each scaffold type was then placed into the cranium of nine rabbits. The rabbits were sacrificed after 8 weeks, and the scaffolds were retrieved for histological analysis. The most bone formation was present in the PPF-TGF-beta1 implants; the newly formed bone had a trabecular appearance together with bone marrow-like tissue. Little or no bone formation was observed in implants without rhTGF-beta1. These histological findings were confirmed by image analysis. Bone surface area, bone area percentage, pore fill percentage, and pore area percentage were significantly higher in the rhTGF-beta1-coated implants than in the noncoated implants. No statistical difference was seen between the PPF-Fn, PPF-Pw, or PPF-Gd scaffolds for these parameters. Quadruple fluorochrome labeling showed that in PPF-TGF-beta1 implants bone formation mainly started in the interior of a pore and proceeded toward the scaffold. We conclude that (a) PPF-TGF-beta1 scaffolds can indeed adequately induce bone formation in porous PPF, and (b) PPF scaffolds prepared by the photocrosslinking-porogen leaching technique are good candidates for the creation of bone graft substitutes.

  15. An animal experimental study of porous magnesium scaffold degradation and osteogenesis

    PubMed Central

    Liu, Y.J.; Yang, Z.Y.; Tan, L.L.; Li, H.; Zhang, Y.Z.

    2014-01-01

    Our objective was to observe the biodegradable and osteogenic properties of magnesium scaffolding under in vivo conditions. Twelve 6-month-old male New Zealand white rabbits were randomly divided into two groups. The chosen operation site was the femoral condyle on the right side. The experimental group was implanted with porous magnesium scaffolds, while the control group was implanted with hydroxyapatite scaffolds. X-ray and blood tests, which included serum magnesium, alanine aminotransferase (ALT), creatinine (CREA), and blood urea nitrogen (BUN) were performed serially at 1, 2, and 3 weeks, and 1, 2, and 3 months. All rabbits were killed 3 months postoperatively, and the heart, kidney, spleen, and liver were analyzed with hematoxylin and eosin (HE) staining. The bone samples were subjected to microcomputed tomography scanning (micro-CT) and hard tissue biopsy. SPSS 13.0 (USA) was used for data analysis, and values of P<0.05 were considered to be significant. Bubbles appeared in the X-ray of the experimental group after 2 weeks, whereas there was no gas in the control group. There were no statistical differences for the serum magnesium concentrations, ALT, BUN, and CREA between the two groups (P>0.05). All HE-stained slices were normal, which suggested good biocompatibility of the scaffold. Micro-CT showed that magnesium scaffolds degraded mainly from the outside to inside, and new bone was ingrown following the degradation of magnesium scaffolds. The hydroxyapatite scaffold was not degraded and had fewer osteoblasts scattered on its surface. There was a significant difference in the new bone formation and scaffold bioabsorption between the two groups (9.29±1.27 vs 1.40±0.49 and 7.80±0.50 vs 0.00±0.00 mm3, respectively; P<0.05). The magnesium scaffold performed well in degradation and osteogenesis, and is a promising material for orthopedics. PMID:25098717

  16. Surface modification of 3D-printed porous scaffolds via mussel-inspired polydopamine and effective immobilization of rhBMP-2 to promote osteogenic differentiation for bone tissue engineering.

    PubMed

    Lee, Sang Jin; Lee, Donghyun; Yoon, Taek Rim; Kim, Hyung Keun; Jo, Ha Hyeon; Park, Ji Sun; Lee, Jun Hee; Kim, Wan Doo; Kwon, Il Keun; Park, Su A

    2016-08-01

    For tissue engineering, a bio-porous scaffold which is applied to bone-tissue regeneration should provide the hydrophilicity for cell attachment as well as provide for the capability to bind a bioactive molecule such as a growth factor in order to improve cell differentiation. In this work, we prepared a three-dimensional (3D) printed polycaprolactone scaffold (PCLS) grafted with recombinant human bone morphogenic protein-2 (rhBMP2) attached via polydopamine (DOPA) chemistry. The DOPA coated PCL scaffold was characterized by contact angle, water uptake, and X-ray photoelectron spectroscopy (XPS) in order to certify that the surface was successfully coated with DOPA. In order to test the loading and release of rhBMP2, we examined the release rate for 28days. For the In vitro cell study, pre-osteoblast MC3T3-E1 cells were seeded onto PCL scaffolds (PCLSs), DOPA coated PCL scaffold (PCLSD), and scaffolds with varying concentrations of rhBMP2 grafted onto the PCLSD 100 and PCLSD 500 (100 and 500ng/ml loaded), respectively. These scaffolds were evaluated by cell proliferation, alkaline phosphatase activity, and real time polymerase chain reaction with immunochemistry in order to verify their osteogenic activity. Through these studies, we demonstrated that our fabricated scaffolds were well coated with DOPA as well as grafted with rhBMP2 at a quantity of 22.7±5ng when treatment with 100ng/ml rhBMP2 and 153.3±2.4ng when treated with 500ng/ml rhBMP2. This grafting enables rhBMP2 to be released in a sustained pattern. In the in vitro results, the cell proliferation and an osteoconductivity of PCLSD 500 groups was greater than any other group. All of these results suggest that our manufactured 3D printed porous scaffold would be a useful construct for application to the bone tissue engineering field. Tissue-engineered scaffolds are not only extremely complex and cumbersome, but also use organic solvents which can negatively influence cellular function. Thus, a rapid

  17. Direct deposited porous scaffolds of calcium phosphate cement with alginate for drug delivery and bone tissue engineering.

    PubMed

    Lee, Gil-Su; Park, Jeong-Hui; Shin, Ueon Sang; Kim, Hae-Won

    2011-08-01

    This study reports the preparation of novel porous scaffolds of calcium phosphate cement (CPC) combined with alginate, and their potential usefulness as a three-dimensional (3-D) matrix for drug delivery and tissue engineering of bone. An α-tricalcium phosphate-based powder was mixed with sodium alginate solution and then directly injected into a fibrous structure in a Ca-containing bath. A rapid hardening reaction of the alginate with Ca(2+) helps to shape the composite into a fibrous form with diameters of hundreds of micrometers, and subsequent pressing in a mold allows the formation of 3-D porous scaffolds with different porosity levels. After transformation of the CPC into a calcium-deficient hydroxyapatite phase in simulated biological fluid the scaffold was shown to retain its mechanical stability. During the process biological proteins, such as bovine serum albumin and lysozyme, used as model proteins, were observed to be effectively loaded onto and released from the scaffolds for up to more than a month, proving the efficacy of the scaffolds as a drug delivering matrix. Mesenchymal stem cells (MSCs) were isolated from rat bone marrow and then cultured on the CPC-alginate porous scaffolds to investigate the ability to support proliferation of cells and their subsequent differentiation along the osteogenic lineage. It was shown that MSCs increasingly actively populated and also permeated into the porous network with time of culture. In particular, cells cultured within a scaffold with a relatively high porosity level showed favorable proliferation and osteogenic differentiation. An in vivo pilot study of the CPC-alginate porous scaffolds after implantation into the rat calvarium for 6 weeks revealed the formation of new bone tissue within the scaffold, closing the defect almost completely. Based on these results, the newly developed CPC-alginate porous scaffolds could be potentially useful as a 3-D matrix for drug delivery and tissue engineering of bone.

  18. Effects of processing parameters in thermally induced phase separation technique on porous architecture of scaffolds for bone tissue engineering.

    PubMed

    Akbarzadeh, Rosa; Yousefi, Azizeh-Mitra

    2014-08-01

    Tissue engineering makes use of 3D scaffolds to sustain three-dimensional growth of cells and guide new tissue formation. To meet the multiple requirements for regeneration of biological tissues and organs, a wide range of scaffold fabrication techniques have been developed, aiming to produce porous constructs with the desired pore size range and pore morphology. Among different scaffold fabrication techniques, thermally induced phase separation (TIPS) method has been widely used in recent years because of its potential to produce highly porous scaffolds with interconnected pore morphology. The scaffold architecture can be closely controlled by adjusting the process parameters, including polymer type and concentration, solvent composition, quenching temperature and time, coarsening process, and incorporation of inorganic particles. The objective of this review is to provide information pertaining to the effect of these parameters on the architecture and properties of the scaffolds fabricated by the TIPS technique. © 2014 Wiley Periodicals, Inc.

  19. Metallizing porous scaffolds as an alternative fabrication method for solid oxide fuel cell anodes

    NASA Astrophysics Data System (ADS)

    Ruiz-Trejo, Enrique; Atkinson, Alan; Brandon, Nigel P.

    2015-04-01

    A combination of electroless and electrolytic techniques is used to incorporate nickel into a porous Ce0.9Gd0.1O1.90 scaffold. First a porous backbone was screen printed into a YSZ electrolyte using an ink that contains sacrificial pore formers. Once sintered, the scaffold was coated with silver using Tollens' reaction followed by electrodeposition of nickel in a Watts bath. At high temperatures the silver forms droplets enabling direct contact between the gadolinia-doped ceria and nickel. Using impedance spectroscopy analysis in a symmetrical cell a total area specific resistance of 1 Ωcm2 at 700 °C in 97% H2 with 3% H2O was found, indicating the potential of this fabrication method for scaling up.

  20. Bioactivity and bone healing properties of biomimetic porous composite scaffold: in vitro and in vivo studies.

    PubMed

    Veronesi, Francesca; Giavaresi, Gianluca; Guarino, Vincenzo; Raucci, Maria Grazia; Sandri, Monica; Tampieri, Anna; Ambrosio, Luigi; Fini, Milena

    2015-09-01

    Tissue engineering (TE) represents a valid alternative to traditional surgical therapies for the management of bone defects that do not regenerate spontaneously. Scaffolds, one of the most important component of TE strategy, should be biocompatible, bioactive, osteoconductive, and osteoinductive. The aim of this study was to evaluate the biological properties and bone regeneration ability of a porous poly(ɛ-caprolactone) (PCL) scaffold, incorporating MgCO3 -doped hydroxyapatite particles, uncoated (PCL_MgCHA) or coated by apatite-like crystals via biomimetic treatment (PCL_MgCHAB). It was observed that both scaffolds are not cytotoxic and, even if cell viability was similar on both scaffolds, PCL_MgCHAB showed higher alkaline phosphatase and collagen I (COLL I) production at day 7. PCL_MgCHA induced more tumor necrosis factor-α release than PCL_MgCHAB, while osteocalcin was produced less by both scaffolds up to 7 days and no significant differences were observed for transforming growth factor-β synthesis. The percentage of new bone trabeculae growth in wide defects carried out in rabbit femoral distal epiphyses was significantly higher in PCL_MgCHAB in comparison with PCL_MgCHA at 4 weeks and even more at 12 weeks after implantation. This study highlighted the role of a biomimetic composite scaffold in bone regeneration and lays the foundations for its future employment in the clinical practice. © 2015 Wiley Periodicals, Inc.

  1. 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.

  2. 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%.

  3. Anode Design Based on Microscale Porous Scaffolds for Advanced Lithium Ion Batteries

    NASA Astrophysics Data System (ADS)

    Park, Hyeji; Choi, Hyelim; Nam, Kyungju; Lee, Sukyung; Um, Ji Hyun; Kim, Kyungbae; Kim, Jae-Hun; Yoon, Won-Sub; Choe, Heeman

    2017-01-01

    Considering the increasing demands for advanced power sources, present-day lithium-ion batteries (LIBs) must provide a higher energy and power density and better cycling stability than conventional LIBs. This study suggests a promising electrode design solution to this problem using Cu, Co, and Ti scaffolds with a microscale porous structure synthesized via freeze-casting. Co3O4 and TiO2 layers are uniformly formed on the Co and Ti scaffolds, respectively, through a simple thermal heat-treatment process, and a SnO2 layer is formed on the Cu scaffold through electroless plating and thermal oxidation. This paper characterizes and evaluates the physical and electrochemical properties of the proposed electrodes using scanning electron microscopy, four-point probe and coin-cell tests to confirm the feasibility of their potential use in LIBs.

  4. Mechanically Enhanced Hierarchically Porous Scaffold Composed of Mesoporous Silica for Host Immune Cell Recruitment.

    PubMed

    Choi, Youngjin; Jeong, Ji Hoon; Kim, Jaeyun

    2017-04-01

    Hierarchically porous materials have been of interest in many diverse fields, including catalysis, separations, and tissue engineering, because the hierarchical porosity of the materials contributes to improvements in mechanical properties, transport properties, and molecule selectivity. In this study, we, for the first time, introduce a new approach to fabricate hierarchical macroporous and mesoporous silica scaffolds based on a salt-leaching process using as-prepared mesoporous silica as a building block. The mechanical strength of the resulting inorganic 3D scaffold was significantly improved by controlling the interfaces of mesoporous silica particles, which allowed for high structural stability during in vivo implantation. Implantation of the scaffold loaded with pro-inflammatory cytokine in mesopores into mice successfully recruited a high number of host immune cells, including dendritic cells, into the macropores, which shows their potential use for immunomodulation. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Anode Design Based on Microscale Porous Scaffolds for Advanced Lithium Ion Batteries

    NASA Astrophysics Data System (ADS)

    Park, Hyeji; Choi, Hyelim; Nam, Kyungju; Lee, Sukyung; Um, Ji Hyun; Kim, Kyungbae; Kim, Jae-Hun; Yoon, Won-Sub; Choe, Heeman

    2017-06-01

    Considering the increasing demands for advanced power sources, present-day lithium-ion batteries (LIBs) must provide a higher energy and power density and better cycling stability than conventional LIBs. This study suggests a promising electrode design solution to this problem using Cu, Co, and Ti scaffolds with a microscale porous structure synthesized via freeze-casting. Co3O4 and TiO2 layers are uniformly formed on the Co and Ti scaffolds, respectively, through a simple thermal heat-treatment process, and a SnO2 layer is formed on the Cu scaffold through electroless plating and thermal oxidation. This paper characterizes and evaluates the physical and electrochemical properties of the proposed electrodes using scanning electron microscopy, four-point probe and coin-cell tests to confirm the feasibility of their potential use in LIBs.

  6. A facile method to determine pore size distribution in porous scaffold by using image processing.

    PubMed

    Lo Re, G; Lopresti, F; Petrucci, G; Scaffaro, R

    2015-09-01

    Image processing permits scientists to investigate morphological properties of three-dimensional structures starting from their bi-dimensional gray-scale representation. In many cases porous structure with complex architecture has to be designed in order to attempt specific properties such in the case of scaffold for tissue engineering. Traditional morphological characterization, like scanning electron microscopy, should be coupled with quantitative information such as pore size distribution (PSD) in order to get a deeper understanding of the influence of the porous structure on tissue regeneration processes and on other related applications, it is remarkable to study a quantitative analysis of porosity and of pores dimension. In this work it was developed as a software able to accomplish the segmentation of images containing pores of any geometry in a semi-automatic way with the aim to measure the PSD. Case study constituted by PLA porous scaffolds with different pore size was adopted. Results indicate that image processing methods well fit the pore size features of PLA scaffolds, overcoming the limits of the more invasive porosimetry techniques.

  7. Direct ink writing of highly porous and strong glass scaffolds for load-bearing bone defects repair and regeneration.

    PubMed

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

    2011-10-01

    The quest for synthetic materials to repair load-bearing bone lost because of trauma, cancer, or congenital bone defects requires the development of porous, high-performance scaffolds with exceptional mechanical strength. However, the low mechanical strength of porous bioactive ceramic and glass scaffolds, compared with that of human cortical bone, has limited their use for these applications. In the present work bioactive 6P53B glass scaffolds with superior mechanical strength were fabricated using a direct ink writing technique. The rheological properties of Pluronic® F-127 (referred to hereafter simply as F-127) hydrogel-based inks were optimized for the printing of features as fine as 30 μm and of three-dimensional scaffolds. The mechanical strength and in vitro degradation of the scaffolds were assessed in a simulated body fluid (SBF). The sintered glass scaffolds showed a compressive strength (136 ± 22 MPa) comparable with that of human cortical bone (100-150 MPa), while the porosity (60%) was in the range of that of trabecular bone (50-90%). The strength is ~100-times that of polymer scaffolds and 4-5-times that of ceramic and glass scaffolds with comparable porosities. Despite the strength decrease resulting from weight loss during immersion in SBF, the value (77 MPa) is still far above that of trabecular bone after 3 weeks. The ability to create both porous and strong structures opens a new avenue for fabricating scaffolds for load-bearing bone defect repair and regeneration.

  8. [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.

  9. Selective laser melting-produced porous titanium scaffolds regenerate bone in critical size cortical bone defects.

    PubMed

    Van der Stok, Johan; Van der Jagt, Olav P; Amin Yavari, Saber; De Haas, Mirthe F P; Waarsing, Jan H; Jahr, Holger; Van Lieshout, Esther M M; Patka, Peter; Verhaar, Jan A N; Zadpoor, Amir A; Weinans, Harrie

    2013-05-01

    Porous titanium scaffolds have good mechanical properties that make them an interesting bone substitute material for large bone defects. These scaffolds can be produced with selective laser melting, which has the advantage of tailoring the structure's architecture. Reducing the strut size reduces the stiffness of the structure and may have a positive effect on bone formation. Two scaffolds with struts of 120-µm (titanium-120) or 230-µm (titanium-230) were studied in a load-bearing critical femoral bone defect in rats. The defect was stabilized with an internal plate and treated with titanium-120, titanium-230, or left empty. In vivo micro-CT scans at 4, 8, and 12 weeks showed more bone in the defects treated with scaffolds. Finally, 18.4 ± 7.1 mm(3) (titanium-120, p = 0.015) and 18.7 ± 8.0 mm(3) (titanium-230, p = 0.012) of bone was formed in those defects, significantly more than in the empty defects (5.8 ± 5.1 mm(3) ). Bending tests on the excised femurs after 12 weeks showed that the fusion strength reached 62% (titanium-120) and 45% (titanium-230) of the intact contralateral femurs, but there was no significant difference between the two scaffolds. This study showed that in addition to adequate mechanical support, porous titanium scaffolds facilitate bone formation, which results in high mechanical integrity of the treated large bone defects.

  10. 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.

  11. Poly(ɛ-caprolactone)/gelatin composite electrospun scaffolds with porous crater-like structures for tissue engineering.

    PubMed

    Hwang, Patrick T J; Murdock, Kyle; Alexander, Grant C; Salaam, Amanee D; Ng, Joshua I; Lim, Dong-Jin; Dean, Derrick; Jun, Ho-Wook

    2016-04-01

    Electrospinning has been widely used to fabricate scaffolds imitating the structure of natural extracellular matrix (ECM). However, conventional electrospinning produces tightly compacted nanofiber layers with only small superficial pores and a lack of bioactivity, which limit the usefulness of electrospinning in biomedical applications. Thus, a porous poly(ε-caprolactone) (PCL)/gelatin composite electrospun scaffold with crater-like structures was developed. Porous crater-like structures were created on the scaffold by a gas foaming/salt leaching process; this unique fiber structure had more large pore areas and higher porosity than the conventional electrospun fiber network. Various ratios of PCL/gelatin (concentration ratios: 100/0, 75/25, and 50/50) composite electrospun scaffolds with and without crater-like structures were characterized by their microstructures, surface chemistry, degradation, mechanical properties, and ability to facilitate cell growth and infiltration. The combination of PCL and gelatin endowed the scaffold with both structural stability of PCL and bioactivity of gelatin. All ratios of scaffolds with crater-like structures showed fairly similar surface chemistry, degradation rates, and mechanical properties to equivalent scaffolds without crater-like structures; however, craterized scaffolds displayed higher human mesenchymal stem cell (hMSC) proliferation and infiltration throughout the scaffolds after 7-day culture. Therefore, these results demonstrated that PCL/gelatin composite electrospun scaffolds with crater-like structures can provide a structurally and biochemically improved three-dimensional ECM-mimicking microenvironment.

  12. Poly(ε-caprolactone)/gelatin composite electrospun scaffolds with porous crater-like structures for tissue engineering

    PubMed Central

    Hwang, Patrick T.J.; Murdock, Kyle; Alexander, Grant C.; Salaam, Amanee D.; Ng, Joshua I.; Lim, Dong-Jin; Dean, Derrick; Jun, Ho-Wook

    2016-01-01

    Electrospinning has been widely used to fabricate scaffolds imitating the structure of natural extracellular matrix (ECM). However, conventional electrospinning produces tightly compacted nanofiber layers with only small superficial pores and a lack of bioactivity, which limit the usefulness of electrospinning in biomedical applications. Thus, a porous poly(ε-caprolactone) (PCL)/gelatin composite electrospun scaffold with crater-like structures was developed. Porous crater-like structures were created on the scaffold by a gas foaming/salt leaching process; this unique fiber structure had more large pore areas and higher porosity than the conventional electrospun fiber network. Various ratios of PCL/gelatin (concentration ratios: 100/0, 75/25, and 50/50) composite electrospun scaffolds with and without crater-like structures were characterized by their microstructures, surface chemistry, degradation, mechanical properties, and ability to facilitate cell growth and infiltration. The combination of PCL and gelatin endowed the scaffold with both structural stability of PCL and bioactivity of gelatin. All ratios of scaffolds with crater-like structures showed fairly similar surface chemistry, degradation rates, and mechanical properties to equivalent scaffolds without crater-like structures; however, craterized scaffolds displayed higher human mesenchymal stem cell (hMSC) proliferation and infiltration throughout the scaffolds after 7-day culture. Therefore, these results demonstrated that PCL/gelatin composite electrospun scaffolds with crater-like structures can provide a structurally and biochemically improved three-dimensional ECM-mimicking microenvironment. PMID:26567028

  13. In vitro cell proliferation evaluation of porous nano-zirconia scaffolds with different porosity for bone tissue engineering.

    PubMed

    Zhu, Yinglan; Zhu, Ruiqiao; Ma, Juan; Weng, Zhiqiang; Wang, Yang; Shi, Xiaolei; Li, Yicai; Yan, Xiaodong; Dong, Zhen; Xu, Jinke; Tang, Chengzhong; Jin, Lei

    2015-09-21

    The selection of scaffold materials and the optimization of scaffold morphological and mechanical properties are critical for successful bone tissue engineering. We fabricated porous scaffolds of nano-sized zirconia using a replication technique. The study aimed to explore the relationship between porosity, pore size, mechanical strength, cell adhesion, and cell proliferation in the zirconia scaffolds. Macro- and micro-structures and compressive strength were comparatively tested. Beagle bone marrow stromal cells were seeded onto the scaffolds to evaluate cell seeding efficiency and cell proliferation profile over 14 d of incubation. The zirconia scaffolds presented a complex porous structure with good interconnectivity of pores. By increasing the sinter cycles, the porosity and pore size of the scaffolds decreased, with mean values ranging from 92.7-68.0% and 830-577 μm, respectively, accompanied by increased compressive strengths of 0.6-4.4 MPa. Cell seeding efficiency and cell proliferation over the first 7 d of incubation increased when the porosity decreased, with cell viability highest in the scaffold with a porosity of 75.2%. After 7 d of incubation, the cell proliferation increased when the porosity increased, highest in the scaffolds with a porosity of 92.7%. These results showed that the zirconia scaffold with a porosity of 75.2% possesses favorable mechanical and biological properties for future applications in bone tissue engineering.

  14. Evaluation of chondrocyte growth in the highly porous scaffolds made by fused deposition manufacturing (FDM) filled with type II collagen.

    PubMed

    Yen, Hung-Jen; Tseng, Ching-Shiow; Hsu, Shan-Hui; Tsai, Ching-Lin

    2009-06-01

    Highly porous poly(D,L-lactide-co-glycolide) (PLGA) scaffolds for cartilage tissue engineering were fabricated in this study using the fused deposition manufacturing (FDM) process and were further modified by type II collagen. The average molecular weight of PLGA decreased to about 60% of the original value after the melt-extrusion process. Type II collagen exhibited sponge-like structure and filled the macroporous FDM scaffolds. An increase of the fiber spacing resulted in an increase of the porosity. The storage modulus of FDM scaffolds with a large fiber spacing was comparable to that of the native porcine articular cartilage. Although the FDM hybrid scaffolds were swollen in various extents after 28 days of in vitro culture, the seeded chondrocytes were well distributed in the interior of the scaffolds with a large fiber spacing and neocartilage was formed around the scaffolds. The study also suggested that a low processing temperature may be required to produce PLGA precision scaffolds using FDM.

  15. Conductive porous scaffolds as potential neural interface materials.

    SciTech Connect

    Hedberg-Dirk, Elizabeth L.; Cicotte, Kirsten N.; Buerger, Stephen P.; Reece, Gregory; Dirk, Shawn M.; Lin, Patrick P.

    2011-11-01

    Our overall intent is to develop improved prosthetic devices with the use of nerve interfaces through which transected nerves may grow, such that small groups of nerve fibers come into close contact with electrode sites, each of which is connected to electronics external to the interface. These interfaces must be physically structured to allow nerve fibers to grow through them, either by being porous or by including specific channels for the axons. They must be mechanically compatible with nerves such that they promote growth and do not harm the nervous system, and biocompatible to promote nerve fiber growth and to allow close integration with biological tissue. They must exhibit selective and structured conductivity to allow the connection of electrode sites with external circuitry, and electrical properties must be tuned to enable the transmission of neural signals. Finally, the interfaces must be capable of being physically connected to external circuitry, e.g. through attached wires. We have utilized electrospinning as a tool to create conductive, porous networks of non-woven biocompatible fibers in order to meet the materials requirements for the neural interface. The biocompatible fibers were based on the known biocompatible material poly(dimethyl siloxane) (PDMS) as well as a newer biomaterial developed in our laboratories, poly(butylene fumarate) (PBF). Both of the polymers cannot be electrospun using conventional electrospinning techniques due to their low glass transition temperatures, so in situ crosslinking methodologies were developed to facilitate micro- and nano-fiber formation during electrospinning. The conductivity of the electrospun fiber mats was controlled by controlling the loading with multi-walled carbon nanotubes (MWNTs). Fabrication, electrical and materials characterization will be discussed along with initial in vivo experimental results.

  16. Performance of PRP Associated with Porous Chitosan as a Composite Scaffold for Regenerative Medicine

    PubMed Central

    Shimojo, Andréa Arruda Martins; Perez, Amanda Gomes Marcelino; Galdames, Sofia Elisa Moraga; Brissac, Isabela Cambraia de Souza; Santana, Maria Helena Andrade

    2015-01-01

    This study aimed to evaluate the in vitro performance of activated platelet-rich plasma associated with porous sponges of chitosan as a composite scaffold for proliferation and osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells. The sponges were prepared by controlled freezing (−20, −80, or −196°C) and lyophilization of chitosan solutions (1, 2, or 3% w/v). The platelet-rich plasma was obtained from controlled centrifugation of whole blood and activated with calcium and autologous serum. The composite scaffolds were prepared by embedding the sponges with the activated platelet-rich plasma. The results showed the performance of the scaffolds was superior to that of activated platelet-rich plasma alone, in terms of delaying the release of growth factors and increased proliferation of the stem cells. The best preparation conditions of chitosan composite scaffolds that coordinated the physicochemical and mechanical properties and cell proliferation were 3% (w/v) chitosan and a −20°C freezing temperature, while −196°C favored osteogenic differentiation. Although the composite scaffolds are promising for regenerative medicine, the structures require stabilization to prevent the collapse observed after five days. PMID:25821851

  17. A Novel Cross-Linked Hyaluronic Acid Porous Scaffold for Cartilage Repair

    PubMed Central

    Bauer, Christoph; Berger, Manuela; Baumgartner, Renate R.; Höller, Sonja; Zwickl, Hannes; Niculescu-Morzsa, Eugenia; Halbwirth, Florian; Nehrer, Stefan

    2015-01-01

    Purpose An important feature of biomaterials used in cartilage regeneration is their influence on the establishment and stabilization of a chondrocytic phenotype of embedded cells. The purpose of this study was to examine the effects of a porous 3-dimensional scaffold made of cross-linked hyaluronic acid on the expression and synthesis performance of human articular chondrocytes. Materials and Methods Osteoarthritic chondrocytes from 5 patients with a mean age of 74 years were passaged twice and cultured within the cross-linked hyaluronic acid scaffolds for 2 weeks. Analyses were performed at 3 different time points. For estimation of cell content within the scaffold, DNA-content (CyQuant cell proliferation assay) was determined. The expression of chondrocyte-specific genes by embedded cells as well as the total amount of sulfated glycosaminoglycans produced during the culture period was analyzed in order to characterize the synthesis performance and differentiation status of the cells. Results Cells showed a homogenous distribution within the scaffold. DNA quantification revealed a reduction of the cell number. This might be attributed to loss of cells from the scaffold during media exchange connected with a stop in cell proliferation. Indeed, the expression of cartilage-specific genes and the production of sulfated glycosaminoglycans were increased and the differentiation index was clearly improved. Conclusions These results suggest that the attachment of osteoarthritic P2 chondrocytes to the investigated material enhanced the chondrogenic phenotype as well as promoted the retention. PMID:27375842

  18. 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.

  19. 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

  20. 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.

  1. Three-dimensional printing of porous ceramic scaffolds for bone tissue engineering.

    PubMed

    Seitz, Hermann; Rieder, Wolfgang; Irsen, Stephan; Leukers, Barbara; Tille, Carsten

    2005-08-01

    This article reports a new process chain for custom-made three-dimensional (3D) porous ceramic scaffolds for bone replacement with fully interconnected channel network for the repair of osseous defects from trauma or disease. Rapid prototyping and especially 3D printing is well suited to generate complex-shaped porous ceramic matrices directly from powder materials. Anatomical information obtained from a patient can be used to design the implant for a target defect. In the 3D printing technique, a box filled with ceramic powder is printed with a polymer-based binder solution layer by layer. Powder is bonded in wetted regions. Unglued powder can be removed and a ceramic green body remains. We use a modified hydroxyapatite (HA) powder for the fabrication of 3D printed scaffolds due to the safety of HA as biocompatible implantable material and efficacy for bone regeneration. The printed ceramic green bodies are consolidated at a temperature of 1250 degrees C in a high temperature furnace in ambient air. The polymeric binder is pyrolysed during sintering. The resulting scaffolds can be used in tissue engineering of bone implants using patient-derived cells that are seeded onto the scaffolds. This article describes the process chain, beginning from data preparation to 3D printing tests and finally sintering of the scaffold. Prototypes were successfully manufactured and characterized. It was demonstrated that it is possible to manufacture parts with inner channels with a dimension down to 450 microm and wall structures with a thickness down to 330 microm. The mechanical strength of dense test parts is up to 22 MPa. Copyright 2005 Wiley Periodicals, Inc.

  2. A biodegradable porous composite scaffold of PGA/beta-TCP for bone tissue engineering.

    PubMed

    Cao, Hong; Kuboyama, Noboru

    2010-02-01

    Polyglycolic acid (PGA) and beta-tricalcium phosphate (beta-TCP) each have many applications as tissue repair materials. In this study, three-dimensional (3D) porous composite scaffolds of PGA/beta-TCP (in 1:1 and 1:3 weight ratios) were fabricated using the solvent casting and particulate leaching method. PGA/beta-TCP scaffolds with high porosity, interconnected 3D pores and rough surfaces were obtained and were observed using scanning electron microscopy (SEM) and micro-computed tomography (micro-CT). The PGA/beta-TCP scaffolds were investigated during the repair of critical bone defects (3 mm diameter, 2 mm depth) in rat femoral medial-epicondyles, compared with hydroxylapatite (HAP) and no implant as controls. Quantitative imageology analysis (volume and density of new bone) and qualitative histological evaluations (hematoxylin and eosin staining; tartrate-resistant acid phosphatase-hematoxylin counterstaining) were characterized using in vivo micro-CT images and histological sections at 0, 14, 30 and 90 days after surgery. Significant differences of all variables were tested by multivariate analysis (p<0.05). The results showed that the bone reformation by using the PGA/beta-TCP scaffolds began within 14 days of surgery, and were healing well at 30 days after surgery. By 90 days after surgery, the bone replacement was almost completed and presented a healthy bone appearance. The new bone mineral densities (mg/cm(3)) with HAP, PGA/beta-TCP (1:1) and PGA/beta-TCP (1:3) at 90 days after surgery were: 390.4+/-18.1, 563.8+/-26.9 and 606.3+/-26.9, respectively. The new bone mineral density with the PGA/beta-TCP scaffold was higher than with HAP (p<0.001), and with the PGA/beta-TCP (1:3) scaffold was higher than with the PGA/beta-TCP (1:1) scaffold at each time examined (p<0.05). The biodegradation percents (%) of HAP, PGA/beta-TCP (1:1) and PGA/beta-TCP (1:3) at 90 days after surgery were: 35.1+/-5.5, 99.0+/-1.0 and 96.2+/-3.3, respectively. The biodegradation

  3. Preparation of a porous conductive scaffold from aniline pentamer-modified polyurethane/PCL blend for cardiac tissue engineering.

    PubMed

    Baheiraei, Nafiseh; Yeganeh, Hamid; Ai, Jafar; Gharibi, Reza; Ebrahimi-Barough, Somayeh; Azami, Mahmoud; Vahdat, Sadaf; Baharvand, Hossein

    2015-10-01

    A novel biodegradable electroactive polyurethane containing aniline pentamer (AP) was blended with polycaprolactone (PCL). The prepared blend (PB) and PCL were further fabricated in to scaffolds using a mixture of poly(ethylene glycol) and salt particles in a double porogen particulate leaching and compression molding methodology. Scaffolds held open and interconnected pores having pore size ranging from several μm to 150 µm. PB scaffolds had compression modulus and strength of 4.1 and 1.3 MPa, respectively. The conductivity of the scaffold was measured as 10(-5) ± 0.09 S .cm(-1) and preserved for at least 100 h post fabrication. Scaffolds supported neonatal cardiomyocytes adhesion and growth with PB showing more extensive effect on the expression of the cardiac genes involved in muscle contraction and relaxation (troponin-T) and cytoskeleton alignment (actinin-4). Our results highlight the potential of incorporation of AP as an electroactive moiety for induction of cardiomyocyte proliferation and repair of damaged heart tissue. © 2015 Wiley Periodicals, Inc.

  4. Preparation and features of polycaprolactone vascular grafts with the incorporated vascular endothelial growth factor

    SciTech Connect

    Sevostyanova, V. V. Khodyrevskaya, Y. I.; Glushkova, T. V.; Antonova, L. V.; Kudryavtseva, Y. A.; Barbarash, O. L.; Barbarash, L. S.

    2015-10-27

    The development of tissue-engineered small-diameter vascular grafts is an urgent issue in cardiovascular surgery. In this study, we assessed how the incorporation of the vascular endothelial growth factor (VEGF) affects morphological and mechanical properties of polycaprolactone (PCL) vascular grafts along with its release kinetics. Vascular grafts were prepared using two-phase electrospinning. In pursuing our aims, we performed scanning electron microscopy, mechanical testing, and enzyme-linked immunosorbent assay. Our results demonstrated the preservation of a highly porous structure and improvement of PCL/VEGF scaffold mechanical properties as compared to PCL grafts. A prolonged VEGF release testifies the use of this construct as a scaffold for tissue-engineered vascular grafts.

  5. Preparation and features of polycaprolactone vascular grafts with the incorporated vascular endothelial growth factor

    NASA Astrophysics Data System (ADS)

    Sevostyanova, V. V.; Khodyrevskaya, Y. I.; Glushkova, T. V.; Antonova, L. V.; Kudryavtseva, Y. A.; Barbarash, O. L.; Barbarash, L. S.

    2015-10-01

    The development of tissue-engineered small-diameter vascular grafts is an urgent issue in cardiovascular surgery. In this study, we assessed how the incorporation of the vascular endothelial growth factor (VEGF) affects morphological and mechanical properties of polycaprolactone (PCL) vascular grafts along with its release kinetics. Vascular grafts were prepared using two-phase electrospinning. In pursuing our aims, we performed scanning electron microscopy, mechanical testing, and enzyme-linked immunosorbent assay. Our results demonstrated the preservation of a highly porous structure and improvement of PCL/VEGF scaffold mechanical properties as compared to PCL grafts. A prolonged VEGF release testifies the use of this construct as a scaffold for tissue-engineered vascular grafts.

  6. Wear mechanism and tribological characteristics of porous NiTi shape memory alloy for bone scaffold.

    PubMed

    Wu, Shuilin; Liu, Xiangmei; Wu, Guosong; Yeung, Kelvin W K; Zheng, Dong; Chung, C Y; Xu, Z S; Chu, Paul K

    2013-09-01

    The abraded debris might cause osteocytic osteolysis on the interface between implants and bone tissues, thus inducing the subsequent mobilization of implants gradually and finally resulting in the failure of bone implants, which imposes restrictions on the applications of porous NiTi shape memory alloys (SMAs) scaffolds for bone tissue engineering. In this work, the effects of the annealing temperature, applied load, and porosity on the tribological behavior and wear resistance of three-dimensional porous NiTi SMA are investigated systematically. The porous structure and phase transformation during the exothermic process affect the tribological properties and wear mechanism significantly. In general, a larger porosity leads to better tribological resistance but sometimes, SMAs with small porosity possess better wear resistance than ones with higher porosity during the initial sliding stage. It can be ascribed to the better superelasticity of the former at the test temperature. The porous NiTi phase during the exothermic reaction also plays an important role in the wear resistance. Generally, porous NiTi has smaller friction coefficients under high loads due to stress-induced superelasticity. The wear mechanism is discussed based on plastic deformation and microcrack propagation.

  7. Topological design and additive manufacturing of porous metals for bone scaffolds and orthopaedic implants: A review.

    PubMed

    Wang, Xiaojian; Xu, Shanqing; Zhou, Shiwei; Xu, Wei; Leary, Martin; Choong, Peter; Qian, M; Brandt, Milan; Xie, Yi Min

    2016-03-01

    One of the critical issues in orthopaedic regenerative medicine is the design of bone scaffolds and implants that replicate the biomechanical properties of the host bones. Porous metals have found themselves to be suitable candidates for repairing or replacing the damaged bones since their stiffness and porosity can be adjusted on demands. Another advantage of porous metals lies in their open space for the in-growth of bone tissue, hence accelerating the osseointegration process. The fabrication of porous metals has been extensively explored over decades, however only limited controls over the internal architecture can be achieved by the conventional processes. Recent advances in additive manufacturing have provided unprecedented opportunities for producing complex structures to meet the increasing demands for implants with customized mechanical performance. At the same time, topology optimization techniques have been developed to enable the internal architecture of porous metals to be designed to achieve specified mechanical properties at will. Thus implants designed via the topology optimization approach and produced by additive manufacturing are of great interest. This paper reviews the state-of-the-art of topological design and manufacturing processes of various types of porous metals, in particular for titanium alloys, biodegradable metals and shape memory alloys. This review also identifies the limitations of current techniques and addresses the directions for future investigations. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Osteogenic differentiation of dura mater stem cells cultured in vitro on three-dimensional porous scaffolds of poly(epsilon-caprolactone) fabricated via co-extrusion and gas foaming.

    PubMed

    Petrie Aronin, C E; Cooper, J A; Sefcik, L S; Tholpady, S S; Ogle, R C; Botchwey, E A

    2008-09-01

    A novel scaffold fabrication method utilizing both polymer blend extrusion and gas foaming techniques to control pore size distribution is presented. Seventy-five per cent of all pores produced using polymer blend extrusion alone were less than 50microm. Introducing a gas technique provided better control of pore size distribution, expanding the range from 0-50 to 0-350microm. Varying sintering time, annealing temperature and foaming pressure also helped to reduce the percentage of pore sizes below 50microm. Scaffolds chosen for in vitro cellular studies had a pore size distribution of 0-300microm, average pore size 66+/-17microm, 0.54+/-0.02% porosity and 98% interconnectivity, measured by micro-computed tomography (microCT) analysis. The ability of the scaffolds to support osteogenic differentiation for subsequent cranial defect repair was evaluated by static and dynamic (0.035+/-0.006ms(-1) terminal velocity) cultivation with dura mater stem cells (DSCs). In vitro studies showed minimal increases in proliferation over 28 days in culture in osteogenic media. Alkaline phosphatase expression remained constant throughout the study. Moderate increases in matrix deposition, as assessed by histochemical staining and microCT analysis, occurred at later time points, days 21 and 28. Although constructs cultured dynamically showed greater mineralization than static conditions, these trends were not significant. It remains unclear whether bioreactor culture of DSCs is advantageous for bone tissue engineering applications. However, these studies show that polycaprolactone (PCL) scaffolds alone, without the addition of other co-polymers or ceramics, support long-term attachment and mineralization of DSCs throughout the entire porous scaffold.

  9. 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.

  10. In vitro and in vivo study of additive manufactured porous Ti6Al4V scaffolds for repairing bone defects

    PubMed Central

    Li, Guoyuan; Wang, Lei; Pan, Wei; Yang, Fei; Jiang, Wenbo; Wu, Xianbo; Kong, Xiangdong; Dai, Kerong; Hao, Yongqiang

    2016-01-01

    Metallic implants with a low effective modulus can provide early load-bearing and reduce stress shielding, which is favorable for increasing in vivo life-span. In this research, porous Ti6Al4V scaffolds with three pore sizes (300~400, 400~500, and 500~700 μm) were manufactured by Electron Beam Melting, with an elastic modulus range of 3.7 to 1.7 GPa. Cytocompatibility in vitro and osseointegration ability in vivo of scaffolds were assessed. hBMSCs numbers increased on all porous scaffolds over time. The group with intended pore sizes of 300 to 400 μm was significantly higher than that of the other two porous scaffolds at days 5 and 7. This group also had higher ALP activity at day 7 in osteogenic differentiation experiment. The scaffold with pore size of 300 to 400 μm was implanted into a 30-mm segmental defect of goat metatarsus. In vivo evaluations indicated that the depth of bone ingrowth increased over time and no implant dislocation occurred during the experiment. Based on its better cytocompatibility and favorable bone ingrowth, the present data showed the capability of the additive manufactured porous Ti6Al4V scaffold with an intended pore size of 300 to 400 μm for large segmental bone defects. PMID:27667204

  11. In vitro and in vivo study of additive manufactured porous Ti6Al4V scaffolds for repairing bone defects.

    PubMed

    Li, Guoyuan; Wang, Lei; Pan, Wei; Yang, Fei; Jiang, Wenbo; Wu, Xianbo; Kong, Xiangdong; Dai, Kerong; Hao, Yongqiang

    2016-09-26

    Metallic implants with a low effective modulus can provide early load-bearing and reduce stress shielding, which is favorable for increasing in vivo life-span. In this research, porous Ti6Al4V scaffolds with three pore sizes (300~400, 400~500, and 500~700 μm) were manufactured by Electron Beam Melting, with an elastic modulus range of 3.7 to 1.7 GPa. Cytocompatibility in vitro and osseointegration ability in vivo of scaffolds were assessed. hBMSCs numbers increased on all porous scaffolds over time. The group with intended pore sizes of 300 to 400 μm was significantly higher than that of the other two porous scaffolds at days 5 and 7. This group also had higher ALP activity at day 7 in osteogenic differentiation experiment. The scaffold with pore size of 300 to 400 μm was implanted into a 30-mm segmental defect of goat metatarsus. In vivo evaluations indicated that the depth of bone ingrowth increased over time and no implant dislocation occurred during the experiment. Based on its better cytocompatibility and favorable bone ingrowth, the present data showed the capability of the additive manufactured porous Ti6Al4V scaffold with an intended pore size of 300 to 400 μm for large segmental bone defects.

  12. In vitro and in vivo study of additive manufactured porous Ti6Al4V scaffolds for repairing bone defects

    NASA Astrophysics Data System (ADS)

    Li, Guoyuan; Wang, Lei; Pan, Wei; Yang, Fei; Jiang, Wenbo; Wu, Xianbo; Kong, Xiangdong; Dai, Kerong; Hao, Yongqiang

    2016-09-01

    Metallic implants with a low effective modulus can provide early load-bearing and reduce stress shielding, which is favorable for increasing in vivo life-span. In this research, porous Ti6Al4V scaffolds with three pore sizes (300~400, 400~500, and 500~700 μm) were manufactured by Electron Beam Melting, with an elastic modulus range of 3.7 to 1.7 GPa. Cytocompatibility in vitro and osseointegration ability in vivo of scaffolds were assessed. hBMSCs numbers increased on all porous scaffolds over time. The group with intended pore sizes of 300 to 400 μm was significantly higher than that of the other two porous scaffolds at days 5 and 7. This group also had higher ALP activity at day 7 in osteogenic differentiation experiment. The scaffold with pore size of 300 to 400 μm was implanted into a 30-mm segmental defect of goat metatarsus. In vivo evaluations indicated that the depth of bone ingrowth increased over time and no implant dislocation occurred during the experiment. Based on its better cytocompatibility and favorable bone ingrowth, the present data showed the capability of the additive manufactured porous Ti6Al4V scaffold with an intended pore size of 300 to 400 μm for large segmental bone defects.

  13. Outer electrospun polycaprolactone shell induces massive foreign body reaction and impairs axonal regeneration through 3D multichannel chitosan nerve guides.

    PubMed

    Duda, Sven; Dreyer, Lutz; Behrens, Peter; Wienecke, Soenke; Chakradeo, Tanmay; Glasmacher, Birgit; Haastert-Talini, Kirsten

    2014-01-01

    We report on the performance of composite nerve grafts with an inner 3D multichannel porous chitosan core and an outer electrospun polycaprolactone shell. The inner chitosan core provided multiple guidance channels for regrowing axons. To analyze the in vivo properties of the bare chitosan cores, we separately implanted them into an epineural sheath. The effects of both graft types on structural and functional regeneration across a 10 mm rat sciatic nerve gap were compared to autologous nerve transplantation (ANT). The mechanical biomaterial properties and the immunological impact of the grafts were assessed with histological techniques before and after transplantation in vivo. Furthermore during a 13-week examination period functional tests and electrophysiological recordings were performed and supplemented by nerve morphometry. The sheathing of the chitosan core with a polycaprolactone shell induced massive foreign body reaction and impairment of nerve regeneration. Although the isolated novel chitosan core did allow regeneration of axons in a similar size distribution as the ANT, the ANT was superior in terms of functional regeneration. We conclude that an outer polycaprolactone shell should not be used for the purpose of bioartificial nerve grafting, while 3D multichannel porous chitosan cores could be candidate scaffolds for structured nerve grafts.

  14. 3D Printing Bioceramic Porous Scaffolds with Good Mechanical Property and Cell Affinity.

    PubMed

    Chang, Chih-Hao; Lin, Chih-Yang; Liu, Fwu-Hsing; Chen, Mark Hung-Chih; Lin, Chun-Pin; Ho, Hong-Nerng; Liao, Yunn-Shiuan

    2015-01-01

    Artificial bone grafting is widely used in current orthopedic surgery for bone defect problems. Unfortunately, surgeons remain unsatisfied with the current commercially available products. One of the major complaints is that these products cannot provide sufficient mechanical strength to support the human skeletal structure. In this study, we aimed to develop a bone scaffold with better mechanical property and good cell affinity by 3D printing (3DP) techniques. A self-developed 3D printer with laser-aided gelling (LAG) process was used to fabricate bioceramic scaffolds with inter-porous structures. To improve the mechanical property of the bioceramic parts after heating, CaCO3 was added to the silica ceramic slurry. CaCO3 was blended into a homogenous SiO2-sol dispersion at weight ratios varying from 0/100 to 5/95 to 9/91 (w/w). Bi-component CaCO3/SiO2-sol was prepared as a biocomposite for the 3DP scaffold. The well-mixed biocomposite was used to fabricate the bioceramic green part using the LAG method. The varied scaffolds were sintered at different temperatures ranging from 900 to 1500°C, and the mechanical property was subsequently analyzed. The scaffolds showed good property with the composite ratio of 5:95 CaCO3:SiO2 at a sintering temperature of 1300°C. The compressive strength was 47 MPa, and the porosity was 34%. The topography of the sintered 3DP bioceramic scaffold was examined by SEM, EDS and XRD. The silica bioceramic presented no cytotoxicity and good MG-63 osteoblast-like cell affinity, demonstrating good biocompatibility. Therefore, the new silica biocomposite is viable for fabricating 3DP bone bioceramics with improved mechanical property and good cell affinity.

  15. 3D Printing Bioceramic Porous Scaffolds with Good Mechanical Property and Cell Affinity

    PubMed Central

    Chang, Chih-Hao; Lin, Chih-Yang; Liu, Fwu-Hsing; Chen, Mark Hung-Chih; Lin, Chun-Pin; Ho, Hong-Nerng; Liao, Yunn-Shiuan

    2015-01-01

    Artificial bone grafting is widely used in current orthopedic surgery for bone defect problems. Unfortunately, surgeons remain unsatisfied with the current commercially available products. One of the major complaints is that these products cannot provide sufficient mechanical strength to support the human skeletal structure. In this study, we aimed to develop a bone scaffold with better mechanical property and good cell affinity by 3D printing (3DP) techniques. A self-developed 3D printer with laser-aided gelling (LAG) process was used to fabricate bioceramic scaffolds with inter-porous structures. To improve the mechanical property of the bioceramic parts after heating, CaCO3 was added to the silica ceramic slurry. CaCO3 was blended into a homogenous SiO2-sol dispersion at weight ratios varying from 0/100 to 5/95 to 9/91 (w/w). Bi-component CaCO3/SiO2-sol was prepared as a biocomposite for the 3DP scaffold. The well-mixed biocomposite was used to fabricate the bioceramic green part using the LAG method. The varied scaffolds were sintered at different temperatures ranging from 900 to 1500°C, and the mechanical property was subsequently analyzed. The scaffolds showed good property with the composite ratio of 5:95 CaCO3:SiO2 at a sintering temperature of 1300°C. The compressive strength was 47 MPa, and the porosity was 34%. The topography of the sintered 3DP bioceramic scaffold was examined by SEM, EDS and XRD. The silica bioceramic presented no cytotoxicity and good MG-63 osteoblast-like cell affinity, demonstrating good biocompatibility. Therefore, the new silica biocomposite is viable for fabricating 3DP bone bioceramics with improved mechanical property and good cell affinity. PMID:26618362

  16. Porous CaP/silk composite scaffolds to repair femur defects in an osteoporotic model.

    PubMed

    Cheng, Ning; Dai, Jing; Cheng, Xiangrong; Li, Shu'e; Miron, Richard J; Wu, Tao; Chen, Wenli; Zhang, Yufeng; Shi, Bin

    2013-08-01

    The most common complication for patients with postmenopausal osteoporosis is bone-related defects and fractures. While routine medication has a high probability of undesirable side effects, new approaches have aimed to develop regeneration procedures that stimulate new bone formation while reversing bone loss. Recently, we have synthesized a new hybrid CaP/silk scaffold with a CaP-phase distribution and pore architecture better suited to facilitate cell differentiation and bone formation. The aim of the present study was to compare the involved remodeling process and therapeutic effect of porous CaP/silk composite scaffolds upon local implantation into osteoporotic defects. Wistar rats were used to induce postmenopausal osteoporotic model by bilateral ovariectomy. The pure silk and hybrid CaP/silk scaffolds were implanted into critical sized defects created in distal femoral epiphysis. After 14 and 28 days, the in vivo osteogenetic efficiency was evaluated by μCT analysis, hematoxylin and eosin staining, Safranin O staining, tartrate-resistant acid phosphatase staining, and immunohistochemical assessment. Animals with or without critical-sized defects were used as drill or blank controls, respectively. The osteoporotic defect model was well established with significantly decreased μCT parameters of BV/TV, Tb.N and increased Tb.Sp, porosity, combined with changes in histological observations. During the healing process, the critical-sized drill control defects failed to regenerate appreciable bone tissue, while more significantly increased bone formation and mineralization with dynamic scaffold degradation and decreased osteoclastic bone resorption could be detected within defects with hybrid CaP/silk scaffolds compared to pure silk scaffolds.

  17. Characterization of Silk Fibroin/Chitosan 3D Porous Scaffold and In Vitro Cytology

    PubMed Central

    Zeng, Shuguang; Liu, Lei; Shi, Yong; Qiu, Junqi; Fang, Wei; Rong, Mingdeng; Guo, Zehong; Gao, Wenfeng

    2015-01-01

    Bone tissue engineering is a powerful tool to treat bone defects caused by trauma, infection, tumors and other factors. Both silk fibroin (SF) and chitosan (CS) are non-toxic and have good biocompatibility, but are poor biological scaffolds when used alone. In this study, the microscopic structure and related properties of SF/CS composite scaffolds with different component ratios were examined. The scaffold material most suitable for osteoblast growth was determined, and these results offer an experimental basis for the future reconstruction of bone defects. First, via freeze-drying and chemical crosslinking methods, SF/CS composites with different component ratios were prepared and their structure was characterized. Changes in the internal structure of the SF and CS mixture were observed, confirming that the mutual modification between the two components was complete and stable. The internal structure of the composite material was porous and three-dimensional with a porosity above 90%. We next studied the pore size, swelling ratio, water absorption ratio, degradation and in vitro cell proliferation. For the 40% SF-60% CS group, the pore size of the scaffold was suitable for the growth of osteoblasts, and the rate of degradation was steady. This favors the early adhesion, growth and proliferation of MG-63 cells. In addition to good biocompatibility and satisfactory cell affinity, this material promotes the secretion of extracellular matrix materials by osteoblasts. Thus, 40% SF-60% CS is a good material for bone tissue engineering. PMID:26083846

  18. 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.

  19. Polyurethane biomaterials for fabricating 3D porous scaffolds and supporting vascular cells.

    PubMed

    Grenier, Stéphanie; Sandig, Martin; Mequanint, Kibret

    2007-09-15

    Successful tissue engineering of vascular grafts largely depends on synthetic scaffolds that support the survival, proliferation, and differentiation of seeded cells. To investigate the utility of polyurethanes for vascular tissue engineering, three-dimensional porous polyurethane scaffolds with highly interconnected pore structures were fabricated by a pressure differential/particulate leaching technique. Ammonium chloride and paraffin porogens were prepared to fabricate the scaffolds. Grinding of ammonium chloride resulted in particulates with uniform particle sizes but irregular shapes. Paraffin particulates made by a dispersion method, on the other hand, had spherical shapes and uniform particle sizes. Polyurethane scaffolds fabricated from these particulates had open faced, highly interconnected channels that could allow cellular infiltration and nutrient delivery. Human coronary artery smooth muscle and endothelial cell interactions with polyurethane surfaces revealed these biomaterials to maintain the contractile phenotype of human coronary artery smooth muscle cells and the formation of endothelial monolayers. During longer culture times, surface modification with cell adhesive extracellular matrix (ECM) protein promoted vascular cell proliferation, maintenance of the differentiated phenotype and endothelial monolayer integrity. Our results suggest that these polyurethanes, in conjunction with cell adhesive ECM proteins, could also support vascular cells in three-dimensional bioreactor-based culture conditions. Copyright 2007 Wiley Periodicals, Inc.

  20. Highly Porous Gelatin Reinforced 3D Scaffolds for Articular Cartilage Regeneration.

    PubMed

    Amadori, Sofia; Torricelli, Paola; Panzavolta, Silvia; Parrilli, Annapaola; Fini, Milena; Bigi, Adriana

    2015-07-01

    3D highly porous (93% total porosity) gelatin scaffolds were prepared according to a novel, simple method, which implies gelatin foaming, gelification, soaking into ethanol and successive freeze-drying. Reinforcement of the as-prepared scaffolds (GEL) was performed through immersion in aqueous solutions at different gelatin concentrations. Reinforcement solutions with and without genipin addition allowed to prepare two series of samples:cross-linked and uncross-linked samples, respectively. The amount of gelatin adsorbed onto the reinforced samples increases as a function of gelatin concentration in solution and provokes a drastic improvement of the compressive modulus and collapse strength up to values of about 30 and 4 MPa, respectively. The open and interconnected porosity, although slightly reduced, is still of the order of 80% in the samples reinforced with the highest concentration of gelatin. Water uptake ability evaluated after immersion in PBS for 20 s decreases with gelatin reinforcement. The presence of genipin in cross-linked samples reduces gelatin release and stabilizes the scaffolds in solution. Chondrocytes from human articular cartilage adhere, proliferate, and penetrate into the scaffolds. The evaluation of differentiation markers both on the supernatants of cell culture and by means of quantitative polymerase chain reaction (qPCR) indicates a dose-dependent promotion of cell differentiation.

  1. Biocompatible, biodegradable and porous liquid crystal elastomer scaffolds for spatial cell cultures.

    PubMed

    Sharma, Anshul; Neshat, Abdollah; Mahnen, Cory J; Nielsen, Alek D; Snyder, Jacob; Stankovich, Tory L; Daum, Benjamin G; LaSpina, Emily M; Beltrano, Gabrielle; Gao, Yunxiang; Li, Shuo; Park, Byung-Wook; Clements, Robert J; Freeman, Ernest J; Malcuit, Christopher; McDonough, Jennifer A; Korley, LaShanda T J; Hegmann, Torsten; Hegmann, Elda

    2015-02-01

    Here we report on the modular synthesis and characterization of biodegradable, controlled porous, liquid crystal elastomers (LCE) and their use as three-dimensional cell culture scaffolds. The elastomers were prepared by cross-linking of star block-co-polymers with pendant cholesterol units resulting in the formation of smectic-A LCEs as determined by polarized optical microscopy, DSC, and X-ray diffraction. Scanning electron microscopy revealed the porosity of the as-prepared biocompatible LCEs, making them suitable as 3D cell culture scaffolds. Biodegradability studies in physiological buffers at varying pH show that these scaffolds are intact for about 11 weeks after which degradation sets in at an exponential rate. Initial results from cell culture studies indicate that these smectic LCEs are compatible with growth, survival, and expansion of cultured neuroblastomas and myoblasts when grown on the LCEs for extended time periods (about a month). These preliminary cell studies focused on characterizing the elastomer-based scaffolds' biocompatibility and the successful 3D incorporation as well as growth of cells in 60 to 150-μm thick elastomer sheets.

  2. 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.

  3. Surface functionalization of 3D glass-ceramic porous scaffolds for enhanced mineralization in vitro

    NASA Astrophysics Data System (ADS)

    Ferraris, Sara; Vitale-Brovarone, Chiara; Bretcanu, Oana; Cassinelli, Clara; Vernè, Enrica

    2013-04-01

    Bone reconstruction after tissue loosening due to traumatic, pathological or surgical causes is in increasing demand. 3D scaffolds are a widely studied solution for supporting new bone growth. Bioactive glass-ceramic porous materials can offer a three-dimensional structure that is able to chemically bond to bone. The ability to surface modify these devices by grafting biologically active molecules represents a challenge, with the aim of stimulating physiological bone regeneration with both inorganic and organic signals. In this research work glass ceramic scaffolds with very high mechanical properties and moderate bioactivity have been functionalized with the enzyme alkaline phosphatase (ALP). The material surface was activated in order to expose hydroxyl groups. The activated surface was further grafted with ALP both via silanization and also via direct grafting to the surface active hydroxyl groups. Enzymatic activity of grafted samples were measured by means of UV-vis spectroscopy before and after ultrasonic washing in TRIS-HCl buffer solution. In vitro inorganic bioactivity was investigated by soaking the scaffolds after the different steps of functionalization in a simulated body fluid (SBF). SEM observations allowed the monitoring of the scaffold morphology and surface chemical composition after soaking in SBF. The presence of ALP enhanced the in vitro inorganic bioactivity of the tested material.

  4. Modeling vascularized bone regeneration within a porous biodegradable CaP scaffold loaded with growth factors.

    PubMed

    Sun, Xiaoqiang; Kang, Yunqing; Bao, Jiguang; Zhang, Yuanyuan; Yang, Yunzhi; Zhou, Xiaobo

    2013-07-01

    Osteogenetic microenvironment is a complex constitution in which extracellular matrix (ECM) molecules, stem cells and growth factors each interact to direct the coordinate regulation of bone tissue development. Importantly, angiogenesis improvement and revascularization are critical for osteogenesis during bone tissue regeneration processes. In this study, we developed a three-dimensional (3D) multi-scale system model to study cell response to growth factors released from a 3D biodegradable porous calcium phosphate (CaP) scaffold. Our model reconstructed the 3D bone regeneration system and examined the effects of pore size and porosity on bone formation and angiogenesis. The results suggested that scaffold porosity played a more dominant role in affecting bone formation and angiogenesis compared with pore size, while the pore size could be controlled to tailor the growth factor release rate and release fraction. Furthermore, a combination of gradient VEGF with BMP2 and Wnt released from the multi-layer scaffold promoted angiogenesis and bone formation more readily than single growth factors. These results demonstrated that the developed model can be potentially applied to predict vascularized bone regeneration with specific scaffold and growth factors.

  5. Pd nanoparticles formation inside porous polymeric scaffolds followed by in situ XANES/SAXS

    NASA Astrophysics Data System (ADS)

    Longo, A.; Lamberti, C.; Agostini, G.; Borfecchia, E.; Lazzarini, A.; Liu, W.; Giannici, F.; Portale, G.; Groppo, E.

    2016-05-01

    Simultaneous time-resolved SAXS and XANES techniques were employed to follow in situ the formation of Pd nanoparticles from palladium acetate precursor in two porous polymeric supports: polystyrene (PS) and poly(4-vinyl-pyridine) (P4VP). In this study we have investigated the effect of the use of different reducing agents (H2 and CO) from the gas phase. These results, in conjunction with data obtained by diffuse reflectance IR (DRIFT) spectroscopy and TEM measurements, allowed us to unravel the different roles played by gaseous H2 and CO in the formation of the Pd nanoparticles for both PS and P4VP hosting scaffolds.

  6. 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

  7. 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

  8. A first approach to evaluate the cell dose in highly porous scaffolds by using a nondestructive metabolic method

    PubMed Central

    Divieto, Carla; Sassi, Maria Paola

    2015-01-01

    Background: In cell-based therapies, in vitro studies on biomimetic cell–scaffold constructs can facilitate the determination of the cell dose, a key factor in guaranteeing the effectiveness of the treatment. However, highly porous scaffolds do not allow a nondestructive evaluation of the cell number. Our objective was to develop a nondestructive method for human mesenchymal stem cells dose evaluation in a highly porous scaffold for bone regeneration. Materials & measurement method: Proliferation trend of human mesenchymal stem cells on Biocoral® scaffolds was measured by a resazurin-based assay here optimized for 3D cultures. The method allows to noninvasively follow the cell proliferation on biocorals over 3 weeks with very high reproducibility. Conclusion: This reliable method could be a powerful tool in cell-based therapies for cell dose determination. PMID:28031911

  9. Stress-strain analysis of porous scaffolds made from titanium alloys synthesized via SLS method

    NASA Astrophysics Data System (ADS)

    Shishkovsky, I.

    2009-09-01

    A layer-by-layer selective laser sintering (SLS) technology seems to be greatly promising for solving the plastic surgery problems, particularly those pertaining to the facial reconstruction. Made from titanium-based alloys (titanium or nitinol, i.e. NiTi-intermetallic phase), the porous scaffolds for cranioplasty are an efficient tool for rectifying the face defects and for the dental orthopedic surgery. The progress in the oral surgery and teeth implantation is caused by the problem of an osteointegration on the one hand, and by achievements of the implant synthesis techniques, on the other hand. An important problem thereby is a profound study of the stress-strain behavior of porous implants under the masticatory load or pressure. In the present study the ways for the optimization of the porous implant structural and strength properties as the function of the laser synthesis parameters are described. The finite element approach (ANSYS) was used here for a complex dowel description and numerical simulations. In order to evaluate the processes in the porous implant under the external loading, a CAD 3D model was built for different internal and external configurations of the implant and/or initial shape of powdered particles. The stress-strain dependences were calculated that displayed the irregularity of the stress distribution by the implant volume in the bone tissue. Most of the values are concentrated in places of object contact.

  10. Laser 3D printing with sub-microscale resolution of porous elastomeric scaffolds for supporting human bone stem cells.

    PubMed

    Petrochenko, Peter E; Torgersen, Jan; Gruber, Peter; Hicks, Lucas A; Zheng, Jiwen; Kumar, Girish; Narayan, Roger J; Goering, Peter L; Liska, Robert; Stampfl, Jürgen; Ovsianikov, Aleksandr

    2015-04-02

    A reproducible method is needed to fabricate 3D scaffold constructs that results in periodic and uniform structures with precise control at sub-micrometer and micrometer length scales. In this study, fabrication of scaffolds by two-photon polymerization (2PP) of a biodegradable urethane and acrylate-based photoelastomer is demonstrated. This material supports 2PP processing with sub-micrometer spatial resolution. The high photoreactivity of the biophotoelastomer permits 2PP processing at a scanning speed of 1000 mm s(-1), facilitating rapid fabrication of relatively large structures (>5 mm(3)). These structures are custom printed for in vitro assay screening in 96-well plates and are sufficiently flexible to enable facile handling and transplantation. These results indicate that stable scaffolds with porosities of greater than 60% can be produced using 2PP. Human bone marrow stromal cells grown on 3D scaffolds exhibit increased growth and proliferation compared to smooth 2D scaffold controls. 3D scaffolds adsorb larger amounts of protein than smooth 2D scaffolds due to their larger surface area; the scaffolds also allow cells to attach in multiple planes and to completely infiltrate the porous scaffolds. The flexible photoelastomer material is biocompatible in vitro and is associated with facile handling, making it a viable candidate for further study of complex 3D-printed scaffolds.

  11. Novel calcified gum Arabic porous nano-composite scaffold for bone tissue regeneration.

    PubMed

    Hadavi, M; Hasannia, S; Faghihi, Sh; Mashayekhi, F; Zadeh, H H; Mostofi, S B

    2017-07-08

    The aim of this study was to investigate the biomechanical and biological properties of a nanocomposite scaffold containing both mineral and polysaccharide constituents. Hydroxyapatite nanoparticles (n-HA) was synthesized from dead abra ovata shells using wet chemical methods and was used in different ratios in concert with gum Arabic, a branched plant polysaccharide. N-HA/gum nanocomposite was fabricated with freeze-drying process and characterized by FTIR and SEM for chemical structure and morphology. Porosity was estimated using liquid substitution method. The scaffold mechanical properties were evaluated by compressive test measurement. Osteogenic differentiation was assessed using alkaline phosphatase production and biomineralization was evaluated using Alizarin red assay. Results demonstrated that the hydroxyapatite/gum Arabic nanocomposite had favorable biocompatibility and a similar structure to natural bone matrix. Porous nanocomposite possessed macropore networks with a porosity 87-93% and mean pore size ranging between 164 and 230 μm. The gum/HA with a ratio of 50% w/w HA had the highest compressive modulus of ∼75.3 MPa Pa (MPa) and the ultimate compressive stress of ∼16.6 MPa. C2C12 cells cultured on a scaffold with higher percentage (40 and 50 w/w) of HA demonstrated increased ALP levels and calcium deposition. The data from the present study demonstrated significant changes to the biomechanical properties and osteoconductivity of the nanocomposite scaffold by modulating its mineral content. Nanocomposite scaffolds containing gum and n-HA of 40-50% exhibited highest mechanical properties, as well as supported increased biomineralization. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Strategies for the chemical analysis of highly porous bone scaffolds using secondary ion mass spectrometry.

    PubMed

    Wang, Daming; Poologasundarampillai, Gowsihan; van den Bergh, Wouter; Chater, Richard J; Kasuga, Toshihiro; Jones, Julian R; McPhail, David S

    2014-02-01

    Understanding the distribution of critical elements (e.g. silicon and calcium) within silica-based bone scaffolds synthesized by different methods is central to the optimization of these materials. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) has been used to determine this information due to its very high surface sensitivity and its ability to map all the elements and compounds in the periodic table with high spatial resolution. The SIMS image data can also be combined with depth profiles to construct three-dimensional chemical maps. However, the scaffolds have interconnected pore networks, which are very challenging structures for the SIMS technique. To overcome this problem two experimental methodologies have been developed. The first method involved the use of the focused ion beam technique to obtain clear images of the regions of interest and subsequently mark them by introducing fiducial marks; the samples were then analysed using the ToF-SIMS technique to yield the chemical analyses of the regions of interest. The second method involved impregnating the pores using a suitable reagent so that a flat surface could be achieved, and this was followed by secondary ion mapping and 3D chemical imaging with ToF-SIMS. The samples used in this work were sol-gel 70S30C foam and electrospun fibres and calcium-containing silica/gelatin hybrid scaffolds. The results demonstrate the feasibility of both these experimental methodologies and indicate that these methods can provide an opportunity to compare various artificial bone scaffolds, which will be of help in improving scaffold synthesis and processing routes. The techniques are also transferable to many other types of porous material.

  13. Improved dimensional stability with bioactive glass fibre skeleton in poly(lactide-co-glycolide) porous scaffolds for tissue engineering.

    PubMed

    Haaparanta, Anne-Marie; Uppstu, Peter; Hannula, Markus; Ellä, Ville; Rosling, Ari; Kellomäki, Minna

    2015-11-01

    Bone tissue engineering requires highly porous three-dimensional (3D) scaffolds with preferable osteoconductive properties, controlled degradation, and good dimensional stability. In this study, highly porous 3D poly(d,l-lactide-co-glycolide) (PLGA) - bioactive glass (BG) composites (PLGA/BG) were manufactured by combining highly porous 3D fibrous BG mesh skeleton with porous PLGA in a freeze-drying process. The 3D structure of the scaffolds was investigated as well as in vitro hydrolytic degradation for 10weeks. The effect of BG on the dimensional stability, scaffold composition, pore structure, and degradation behaviour of the scaffolds was evaluated. The composites showed superior pore structure as the BG fibres inhibited shrinkage of the scaffolds. The BG was also shown to buffer the acidic degradation products of PLGA. These results demonstrate the potential of these PLGA/BG composites for bone tissue engineering, but the ability of this kind of PLGA/BG composites to promote bone regeneration will be studied in forthcoming in vivo studies.

  14. Fabrication of three-dimensional porous scaffold based on collagen fiber and bioglass for bone tissue engineering.

    PubMed

    Long, Teng; Yang, Jun; Shi, Shan-Shan; Guo, Ya-Ping; Ke, Qin-Fei; Zhu, Zhen-An

    2015-10-01

    An ideal scaffold for bone tissue engineering should have interconnected porous structure, good biocompatibility, and mechanical properties well-matched with natural bones. Collagen is the key component in the extracellular matrix (ECM) of natural bones, and plays an important role in bone regeneration. The biological activity of collagen has promoted it to be an advantageous biomaterial for bone tissue engineering; however, the mechanical properties of these scaffolds are insufficient and the porous structures are not stable in the wet state. An effective strategy to solve this problem is to fabricate a hybrid scaffold of biologically derived and synthetic material, which have the necessary bioactivity and mechanical stability needed for bone synthesis. In this work, a three-dimensional macroporous bone scaffold based on collagen (CO) fiber and bioglass (BG) is fabricated by a slurry-dipping technique, and its relevant mechanical and biological properties are evaluated. The CO/BG scaffold is interconnected with a porosity of 81 ± 4.6% and pore size of 40-200 μm. Compared with CO scaffold, water absorption value of CO/BG scaffold decreases greatly from 889% to 52%, which significantly alleviates the swelling behavior of collagen and improves the stability of scaffold structure. The CO/BG scaffold has a compression strength of 5.8 ± 1.6 MPa and an elastic modulus of 0.35 ± 0.01 Gpa, which are well-matched with the mechanical properties of trabecular bones. In vitro cell assays demonstrate that the CO/BG scaffold has good biocompatibility to facilitate the spreading and proliferation of human bone marrow stromal cells. Hence, the CO/BG scaffold is promising for bone tissue engineering application.

  15. Preparation and characterization of bimodal porous poly(γ-benzyl-L-glutamate) scaffolds for bone tissue engineering.

    PubMed

    Qian, Junmin; Yong, Xueqing; Xu, Weijun; Jin, Xinxia

    2013-12-01

    An ideal scaffold in bone tissue-engineering strategy should provide biomimetic extracellular matrix-like architecture and biological properties. Poly(γ-benzyl-L-glutamate) (PBLG) has been a popular model polypeptide for various potential biomedical applications due to its good biocompatibility and biodegradability. This study developed novel bimodal porous PBLG polypeptide scaffolds via a combination of biotemplating method and in situ ring-opening polymerization of γ-benzyl-L-gIutamate N-carboxyanhydride (BLG-NCA). The PBLG scaffolds were characterized by proton nuclear magnetic resonance spectroscopy, X-ray diffraction, differential scanning calorimetry, scanning electron microscope (SEM) and mechanical test. The results showed that the semi-crystalline PBLG scaffolds exhibited an anisotropic porous structure composed of honeycomb-like channels (100-200 μm in diameter) and micropores (5-20 μm), with a very high porosity of 97.4±1.6%. The compressive modulus and glass transition temperature were 402.8±20.6 kPa and 20.2°C, respectively. The in vitro biocompatibility evaluation with MC3T3-E1 cells using SEM, fluorescent staining and MTT assay revealed that the PBLG scaffolds had good biocompatibility and favored cell attachment, spread and proliferation. Therefore, the bimodal porous polypeptide scaffolds are promising for bone tissue engineering.

  16. Direct Ink Writing of Highly Porous and Strong Glass Scaffolds for Load-bearing Bone Defects Repair and Regeneration

    PubMed Central

    Fu, Qiang; Saiz, Eduardo; Tomsia, Antoni P.

    2011-01-01

    The quest for synthetic materials to repair load-bearing bone lost because of trauma, cancer, or congenital bone defects requires development of porous and high-performance scaffolds with exceptional mechanical strength. However, the low mechanical strength of porous bioactive ceramic and glass scaffolds, compared with that of human cortical bone, has limited their use for these applications. In the present work, bioactive 6P53B glass scaffolds with superior mechanical strength were fabricated using a direct ink writing technique. The rheological properties of Pluronic® F-127 (referred to hereafter simply as F-127) hydrogel-based inkswere optimized for the printing of features as fine as 30 μm and of the three-dimensional scaffolds. The mechanical strength and in vitro degradation of the scaffolds were assessed in a simulated body fluid (SBF). The sintered glass scaffolds show a compressive strength (136 ± 22 MPa) comparable to that of human cortical bone (100-150 MPa), while the porosity (60%) is in the range of that of trabecular bone (50-90%).The strength is ~100 times that of polymer scaffolds and 4–5 times that of ceramic and glass scaffolds with comparable porosities. Despite the strength decrease resulting from weight loss during immersion in an SBF, the value (77 MPa) is still far above that of trabecular bone after three weeks. The ability to create both porous and strong structures opens a new avenue for fabricating scaffolds for load-bearing bone defect repair and regeneration. PMID:21745606

  17. Design, construction and mechanical testing of digital 3D anatomical data-based PCL-HA bone tissue engineering scaffold.

    PubMed

    Yao, Qingqiang; Wei, Bo; Guo, Yang; Jin, Chengzhe; Du, Xiaotao; Yan, Chao; Yan, Junwei; Hu, Wenhao; Xu, Yan; Zhou, Zhi; Wang, Yijin; Wang, Liming

    2015-01-01

    The study aims to investigate the techniques of design and construction of CT 3D reconstructional data-based polycaprolactone (PCL)-hydroxyapatite (HA) scaffold. Femoral and lumbar spinal specimens of eight male New Zealand white rabbits were performed CT and laser scanning data-based 3D printing scaffold processing using PCL-HA powder. Each group was performed eight scaffolds. The CAD-based 3D printed porous cylindrical stents were 16 piece × 3 groups, including the orthogonal scaffold, the Pozi-hole scaffold and the triangular hole scaffold. The gross forms, fiber scaffold diameters and porosities of the scaffolds were measured, and the mechanical testing was performed towards eight pieces of the three kinds of cylindrical scaffolds, respectively. The loading force, deformation, maximum-affordable pressure and deformation value were recorded. The pore-connection rate of each scaffold was 100 % within each group, there was no significant difference in the gross parameters and micro-structural parameters of each scaffold when compared with the design values (P > 0.05). There was no significant difference in the loading force, deformation and deformation value under the maximum-affordable pressure of the three different cylinder scaffolds when the load was above 320 N. The combination of CT and CAD reverse technology could accomplish the design and manufacturing of complex bone tissue engineering scaffolds, with no significant difference in the impacts of the microstructures towards the physical properties of different porous scaffolds under large load.

  18. New Method for the Deposition of Nickel Oxide in Porous Scaffolds for Electrodes in Solid Oxide Fuel Cells and Electrolyzers.

    PubMed

    Ruiz-Trejo, Enrique; Puolamaa, Milla; Sum, Brian; Tariq, Farid; Yufit, Vladimir; Brandon, Nigel P

    2017-01-10

    A simple chemical bath deposition is used to coat a complex porous ceramic scaffold with a conformal Ni layer. The resulting composite is used as a solid oxide fuel cell electrode, and its electrochemical response is measured in humidified hydrogen. X-ray tomography is used to determine the microstructural characteristics of the uncoated and Ni-coated porous structure, which include the surface area to total volume, the radial pore size, and the size of the necks between the pores.

  19. Response of murine bone marrow-derived mesenchymal stromal cells to dry-etched porous silicon scaffolds.

    PubMed

    Hajj-Hassan, Mohamad; Khayyat-Kholghi, Maedeh; Wang, Huifen; Chodavarapu, Vamsy; Henderson, Janet E

    2011-11-01

    Porous silicon shows great promise as a bio-interface material due to its large surface to volume ratio, its stability in aqueous solutions and to the ability to precisely regulate its pore characteristics. In the current study, porous silicon scaffolds were fabricated from single crystalline silicon wafers by a novel xenon difluoride dry etching technique. This simplified dry etch fabrication process allows selective formation of porous silicon using a standard photoresist as mask material and eliminates the post-formation drying step typically required for the wet etching techniques, thereby reducing the risk of damaging the newly formed porous silicon. The porous silicon scaffolds supported the growth of primary cultures of bone marrow derived mesenchymal stromal cells (MSC) plated at high density for up to 21 days in culture with no significant loss of viability, assessed using Alamar Blue. Scanning electron micrographs confirmed a dense lawn of cells at 9 days of culture and the presence of MSC within the pores of the porous silicon scaffolds. Copyright © 2011 Wiley Periodicals, Inc.

  20. Influence of Parathyroid Hormone-Loaded PLGA Nanoparticles in Porous Scaffolds for Bone Regeneration

    PubMed Central

    Gentile, Piergiorgio; Nandagiri, Vijay Kumar; Pabari, Ritesh; Daly, Jacqueline; Tonda-Turo, Chiara; Ciardelli, Gianluca; Ramtoola, Zebunnissa

    2015-01-01

    Biodegradable poly(lactide-co-glycolide) (PLGA) nanoparticles, containing human parathyroid hormone (PTH (1–34)), prepared by a modified double emulsion-solvent diffusion-evaporation method, were incorporated in porous freeze-dried chitosan-gelatin (CH-G) scaffolds. The PTH-loaded nanoparticles (NPTH) were characterised in terms of morphology, size, protein loading, release kinetics and in vitro assessment of biological activity of released PTH and cytocompatibility studies against clonal human osteoblast (hFOB) cells. Structural integrity of incorporated and released PTH from nanoparticles was found to be intact by using Tris-tricine SDS-PAGE. In vitro PTH release kinetics from PLGA nanoparticles were characterised by a burst release followed by a slow release phase for 3–4 weeks. The released PTH was biologically active as evidenced by the stimulated release of cyclic AMP from hFOB cells as well as increased mineralisation studies. Both in vitro and cell studies demonstrated that the PTH bioactivity was maintained during the fabrication of PLGA nanoparticles and upon release. Finally, a content of 33.3% w/w NPTHs was incorporated in CH-G scaffolds, showing an intermittent release during the first 10 days and, followed by a controlled release over 28 days of observation time. The increased expression of Alkaline Phosphatase levels on hFOB cells further confirmed the activity of intermittently released PTH from scaffolds. PMID:26343649

  1. Liposome combined porous beta-TCP scaffold: preparation, characterization, and anti-biofilm activity.

    PubMed

    Zhu, Chong-Tao; Xu, Yong-Qing; Shi, Jian; Li, Jun; Ding, Jing

    2010-08-01

    The objective of this study was to design a novel artificial bone scaffold for therapy and prevention of refractory bacterial infection. Porous beta-tricalcium phosphate (beta-TCP) scaffold was combined with liposomal gentamicin (GS) to form a novel complex drug carrier. The liposome combined beta-TCP scaffold (LCS) was characterized for its liposome binding rate, drug loading, and micromorphology. The anti-biofilm activity of LCS was evaluated by Staphylococcus aureus biofilm in vitro. The drug release from LCS was recognized as an initial high dose of liposomal GS released from the matrix and a further sustained release of free GS from the liposome, respectively, and it is an ideal release pattern for treatment and prevention of post-operative osteomyelitis. The release kinetics was influenced by variation of particle size of liposome. LCS displayed a potential anti-biofilm activity even in the lowest GS concentration (2.5 microg/mL), and the regrowth time was extended from 5.0 h to 9.5 h. At a higher dosage range, the highest anti-biofilm activity was achieved by LCS with liposomal particle size of 800 nm. In conclusion, the development of LCS showed a new pathway for controlled delivery of liposomal antibiotics for treatment of osteomyelitis caused by persistent bacterial infection.

  2. Polyester type polyHIPE scaffolds with an interconnected porous structure for cartilage regeneration

    PubMed Central

    Naranda, Jakob; Sušec, Maja; Maver, Uroš; Gradišnik, Lidija; Gorenjak, Mario; Vukasović, Andreja; Ivković, Alan; Rupnik, Marjan Slak; Vogrin, Matjaž; Krajnc, Peter

    2016-01-01

    Development of artificial materials for the facilitation of cartilage regeneration remains an important challenge in orthopedic practice. Our study investigates the potential for neocartilage formation within a synthetic polyester scaffold based on the polymerization of high internal phase emulsions. The fabrication of polyHIPE polymer (PHP) was specifically tailored to produce a highly porous (85%) structure with the primary pore size in the range of 50–170 μm for cartilage tissue engineering. The resulting PHP scaffold was proven biocompatible with human articular chondrocytes and viable cells were observed within the materials as evaluated using the Live/Dead assay and histological analysis. Chondrocytes with round nuclei were organized into multicellular layers on the PHP surface and were observed to grow approximately 300 μm into the scaffold interior. The accumulation of collagen type 2 was detected using immunohistochemistry and chondrogenic specific genes were expressed with favorable collagen type 2 to 1 ratio. In addition, PHP samples are biodegradable and their baseline mechanical properties are similar to those of native cartilage, which enhance chondrocyte cell growth and proliferation. PMID:27340110

  3. Polyester type polyHIPE scaffolds with an interconnected porous structure for cartilage regeneration

    NASA Astrophysics Data System (ADS)

    Naranda, Jakob; Sušec, Maja; Maver, Uroš; Gradišnik, Lidija; Gorenjak, Mario; Vukasović, Andreja; Ivković, Alan; Rupnik, Marjan Slak; Vogrin, Matjaž; Krajnc, Peter

    2016-06-01

    Development of artificial materials for the facilitation of cartilage regeneration remains an important challenge in orthopedic practice. Our study investigates the potential for neocartilage formation within a synthetic polyester scaffold based on the polymerization of high internal phase emulsions. The fabrication of polyHIPE polymer (PHP) was specifically tailored to produce a highly porous (85%) structure with the primary pore size in the range of 50–170 μm for cartilage tissue engineering. The resulting PHP scaffold was proven biocompatible with human articular chondrocytes and viable cells were observed within the materials as evaluated using the Live/Dead assay and histological analysis. Chondrocytes with round nuclei were organized into multicellular layers on the PHP surface and were observed to grow approximately 300 μm into the scaffold interior. The accumulation of collagen type 2 was detected using immunohistochemistry and chondrogenic specific genes were expressed with favorable collagen type 2 to 1 ratio. In addition, PHP samples are biodegradable and their baseline mechanical properties are similar to those of native cartilage, which enhance chondrocyte cell growth and proliferation.

  4. Icariin delivery porous PHBV scaffolds for promoting osteoblast expansion in vitro.

    PubMed

    Xia, Leilei; Li, Yongsheng; Zhou, Zheng; Dai, Yao; Liu, Hongbo; Liu, Hairong

    2013-08-01

    How cells could proliferate quickly and maintain their biological activity by using appropriate scaffolds remains a big challenge for tissue engineering. By integrating icariin, a bioactive ingredient of traditional Chinese medicine (TCM) Epimedii herba, with PHBV scaffolds, novel icariin delivery porous PHBV scaffolds (IDPPSs) were fabricated with a combination of the solvent casting and salt leaching techniques. IDPPSs displayed a high porosity, 88.80%, and appropriate mechanical properties which were required for 3-D cell culture. IDPPSs significantly promoted the proliferation of human osteoblast-like MG-63 cells and the pre-osteoblast MC3T3-E1 cells, while IDPPSs containing 0.1% icariin (wt.%) showed the highest effect compared with other samples. Although IDPPSs continuously released icariin to the solution in 28 days, cells attached to IDPPSs received an enhanced growth stimulation compared with which were not physically in contact with IDPPSs. Up-regulated transcription of growth factor genes and extracellular matrix genes, including BMP2, BMP6, BMP7 and BGN, was observed in IDPPS-cultured MG-63 cells, illustrating that enhanced cellular proliferation results from alteration of gene transcription. These results implied the potential commercial use of IDPPSs in tissue engineering.

  5. Biodegradable and bioactive porous scaffold structures prepared using fused deposition modeling.

    PubMed

    Korpela, Jyrki; Kokkari, Anne; Korhonen, Harri; Malin, Minna; Närhi, Timo; Seppälä, Jukka

    2013-05-01

    Three-dimensional printing (3DP) refers to a group of additive manufacturing techniques that can be utilized in tissue engineering applications. Fused deposition modeling (FDM) is a 3DP method capable of using common thermoplastic polymers. However, the scope of materials applicable for FDM has not been fully recognized. The purpose of this study was to examine the creation of biodegradable porous scaffold structures using different materials in FDM and to determine the compressive properties and the fibroblast cell response of the structures. To the best of our knowledge, the printability of a poly(ε-caprolactone)/bioactive glass (PCL/BAG) composite and L-lactide/ε-caprolactone 75/25 mol % copolymer (PLC) was demonstrated for the first time. Scanning electron microscope (SEM) images showed BAG particles at the surface of the printed PCL/BAG scaffolds. Compressive testing showed the possibility of altering the compressive stiffness of a scaffold without changing the compressive modulus. Compressive properties were significantly dependent on porosity level and structural geometry. Fibroblast proliferation was significantly higher in polylactide than in PCL or PCL/BAG composite. Optical microscope images and SEM images showed the viability of the cells, which demonstrated the biocompatibility of the structures. Copyright © 2012 Wiley Periodicals, Inc.

  6. Promoted dermis healing from full-thickness skin defect by porous silk fibroin scaffolds (PSFSs).

    PubMed

    Guan, Guoping; Bai, Lun; Zuo, Baoqi; Li, Mingzhong; Wu, Zhengyu; Li, Yonglin; Wang, Lu

    2010-01-01

    Studies on skin substitutes and dermal scaffolds have been extensively carried out in the past several decades and some commercial products derived from collagen and polymers have been in marketing. Yet little research on silk fibroin based dermal scaffolds and products has been reported so far. In the present study, therefore, porous silk fibroin scaffolds (PSFSs) have been prepared by freeze drying method. The effects of PSFSs on skin recovery from full thickness defect have been examined by histological evaluation with respect to neovascularization, dermal regeneration and infiltration of inflammatory cells. In addition, tissue compatibility between PSFSs and polyvinyl alcohol (PVA) sponges (as control) has been semiquantitatively compared by scoring method. The results showed that at day 18 after implantation, new tissues formed in PSFSs whose structure was almost equal to normal skin structure where proportional distribution of functional blood vessels could be found. Furthermore, infiltration of inflammatory cells in PSFSs disappeared within 7 days. By contrast, a variety of interstices, fibrous encapsulization and moderate infiltration of inflammatory cells could be found in PVA sponges at day 18 after implantation. In summary, PSFSs has significantly promoted the skin recovery from full thickness defect, showing fibroin's outstanding tissue compatibility.

  7. A comparison study of different physical treatments on cartilage matrix derived porous scaffolds for tissue engineering applications

    PubMed Central

    Moradi, Ali; Pramanik, Sumit; Ataollahi, Forough; Abdul Khalil, Alizan; Kamarul, Tunku; Pingguan-Murphy, Belinda

    2014-01-01

    Native cartilage matrix derived (CMD) scaffolds from various animal and human sources have drawn attention in cartilage tissue engineering due to the demonstrable presence of bioactive components. Different chemical and physical treatments have been employed to enhance the micro-architecture of CMD scaffolds. In this study we have assessed the typical effects of physical cross-linking methods, namely ultraviolet (UV) light, dehydrothermal (DHT) treatment, and combinations of them on bovine articular CMD porous scaffolds with three different matrix concentrations (5%, 15% and 30%) to assess the relative strengths of each treatment. Our findings suggest that UV and UV–DHT treatments on 15% CMD scaffolds can yield architecturally optimal scaffolds for cartilage tissue engineering. PMID:27877731

  8. A comparison study of different physical treatments on cartilage matrix derived porous scaffolds for tissue engineering applications

    NASA Astrophysics Data System (ADS)

    Moradi, Ali; Pramanik, Sumit; Ataollahi, Forough; Khalil, Alizan Abdul; Kamarul, Tunku; Pingguan-Murphy, Belinda

    2014-12-01

    Native cartilage matrix derived (CMD) scaffolds from various animal and human sources have drawn attention in cartilage tissue engineering due to the demonstrable presence of bioactive components. Different chemical and physical treatments have been employed to enhance the micro-architecture of CMD scaffolds. In this study we have assessed the typical effects of physical cross-linking methods, namely ultraviolet (UV) light, dehydrothermal (DHT) treatment, and combinations of them on bovine articular CMD porous scaffolds with three different matrix concentrations (5%, 15% and 30%) to assess the relative strengths of each treatment. Our findings suggest that UV and UV-DHT treatments on 15% CMD scaffolds can yield architecturally optimal scaffolds for cartilage tissue engineering.

  9. Adhesion and proliferation of human mesenchymal stem cells from dental pulp on porous silicon scaffolds.

    PubMed

    Collart-Dutilleul, Pierre-Yves; Secret, Emilie; Panayotov, Ivan; Deville de Périère, Dominique; Martín-Palma, Raúl J; Torres-Costa, Vicente; Martin, Marta; Gergely, Csilla; Durand, Jean-Olivier; Cunin, Frédérique; Cuisinier, Frédéric J

    2014-02-12

    In regenerative medicine, stem-cell-based therapy often requires a scaffold to deliver cells and/or growth factors to the injured site. Porous silicon (pSi) is a promising biomaterial for tissue engineering as it is both nontoxic and bioresorbable. Moreover, surface modification can offer control over the degradation rate of pSi and can also promote cell adhesion. Dental pulp stem cells (DPSC) are pluripotent mesenchymal stem cells found within the teeth and constitute a readily source of stem cells. Thus, coupling the good proliferation and differentiation capacities of DPSC with the textural and chemical properties of the pSi substrates provides an interesting approach for therapeutic use. In this study, the behavior of human DPSC is analyzed on pSi substrates presenting pores of various sizes, 10 ± 2 nm, 36 ± 4 nm, and 1.0 ± 0.1 μm, and undergoing different chemical treatments, thermal oxidation, silanization with aminopropyltriethoxysilane (APTES), and hydrosilylation with undecenoic acid or semicarbazide. DPSC adhesion and proliferation were followed for up to 72 h by fluorescence microscopy, scanning electron microscopy (SEM), enzymatic activity assay, and BrdU assay for mitotic activity. Porous silicon with 36 nm pore size was found to offer the best adhesion and the fastest growth rate for DPSC compared to pSi comporting smaller pore size (10 nm) or larger pore size (1 μm), especially after silanization with APTES. Hydrosilylation with semicarbazide favored cell adhesion and proliferation, especially mitosis after cell adhesion, but such chemical modification has been found to led to a scaffold that is stable for only 24-48 h in culture medium. Thus, semicarbazide-treated pSi appeared to be an appropriate scaffold for stem cell adhesion and immediate in vivo transplantation, whereas APTES-treated pSi was found to be more suitable for long-term in vitro culture, for stem cell proliferation and differentiation.

  10. Biosensors based on porous cellulose nanocrystal-poly(vinyl alcohol) scaffolds.

    PubMed

    Schyrr, Bastien; Pasche, Stéphanie; Voirin, Guy; Weder, Christoph; Simon, Yoan C; Foster, E Johan

    2014-08-13

    Cellulose nanocrystals (CNCs), which offer a high aspect ratio, large specific surface area, and large number of reactive surface groups, are well suited for the facile immobilization of high density biological probes. We here report functional high surface area scaffolds based on cellulose nanocrystals (CNCs) and poly(vinyl alcohol) (PVA) and demonstrate that this platform is useful for fluorescence-based sensing schemes. Porous CNC/PVA nanocomposite films with a thickness of 25-70 nm were deposited on glass substrates by dip-coating with an aqueous mixture of the CNCs and PVA, and the porous nanostructure was fixated by heat treatment. In a subsequent step, a portion of the scaffold's hydroxyl surface groups was reacted with 2-(acryloxy)ethyl (3-isocyanato-4-methylphenyl)carbamate to permit the immobilization of thiolated fluorescein-substituted lysine, which was used as a first sensing motif, via nucleophile-based thiol-ene Michael addition. The resulting sensor films exhibit a nearly instantaneous and pronounced change of their fluorescence emission intensity in response to changes in pH. The approach was further extended to the detection of protease activity by immobilizing a Förster-type resonance energy transfer chromophore pair via a labile peptide sequence to the scaffold. This sensing scheme is based on the degradation of the protein linker in the presence of appropriate enzymes, which separate the chromophores and causes a turn-on of the originally quenched fluorescence. Using a standard benchtop spectrometer to monitor the increase in fluorescence intensity, trypsin was detected at a concentration of 250 μg/mL, i.e., in a concentration that is typical for abnormal proteolytic activity in wound fluids.

  11. In vivo guided vascular regeneration with a non-porous elastin-like polypeptide hydrogel tubular scaffold.

    PubMed

    Mahara, Atsushi; Kiick, Kristi L; Yamaoka, Tetsuji

    2017-01-28

    Herein, we demonstrate a new approach for small-caliber vascular reconstruction using a non-porous elastin-like polypeptide hydrogel tubular scaffold, based on the concept of guided vascular regeneration (GVR). The scaffolds are composed of elastin-like polypeptide, (Val-Pro-Gly-Ile-Gly)n , for compliance matching and antithrombogenicity and an Arg-Gly-Asp (RGD) motif for connective tissue regeneration. When the polypeptide was mixed with an aqueous solution of β-[Tris(hydroxymethyl)phosphino]propionic acid at 37°C, the polypeptide hydrogel was rapidly formed. The elastic modulus of the hydrogel was 4.4kPa. The hydrogel tubular scaffold was formed in a mold and reinforced with poly(lactic acid) nanofibers. When tubular scaffolds with an inner diameter of 1 mm and length of 5 mm were implanted into rat abdominal aortae, connective tissue grew along the scaffold luminal surface from the flanking native tissues, resulting in new blood vessel tissue with a thickness of 200 μm in 1 month. In contrast, rats implanted with control scaffolds without the RGD motif died. These results indicate that the non-porous hydrogel tubular scaffold containing the RGD motif effectively induced rapid tissue regeneration and that GVR is a promising strategy for the regeneration of small-diameter blood vessels. This article is protected by copyright. All rights reserved.

  12. 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.

  13. 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

  14. 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.

  15. Evaluation of the growth of chondrocytes and osteoblasts seeded into precision scaffolds fabricated by fused deposition manufacturing.

    PubMed

    Hsu, Shan-hui; Yen, Hung-Jen; Tseng, Ching-Shiow; Cheng, Chia-Sheng; Tsai, Ching-Lin

    2007-02-01

    In this study, fused deposition manufacturing (FDM) was utilized to fabricate the precision scaffolds for cartilage and bone regeneration. Cell seeding into such scaffolds was evaluated. For poly(D,l-lactide) (PLA) scaffolds used for cartilage regeneration, the structure with larger inner space, four direction stacking (4D) and small interval of fibers were better. Chondrocyte proliferated well with matrix accumulation in precision scaffolds coated with type II collagen at 4 weeks of in vitro culture. The seeding efficiency of osteoblasts in most polycaprolactone (PCL) scaffolds used for bone regeneration could arrive 50% of original cell seeding density, and the amount of cells in scaffolds increased to double fold after 2 weeks of in vitro culture. The histological cross-section also revealed proliferation and mineralization of osteoblasts among the PCL fibers. The results indicated that the highly porous and interconnected structure of precision scaffolds could benefit cell ingrowth.

  16. Fabrication and durable antibacterial properties of 3D porous wet electrospun RCSC/PCL nanofibrous scaffold with silver nanoparticles

    NASA Astrophysics Data System (ADS)

    Zhang, Mei; Lin, Han; Wang, Yilong; Yang, Guang; Zhao, He; Sun, Dahui

    2017-08-01

    Electrospunnanofibers are used as three-dimensional (3D) scaffold materials that can alter cell attachment and cell proliferation, change the antibacterial properties of materials, and can be used as wound dressings. But the fabrication of porous 3D scaffold structure and the antibacterial properties enhancing are challenges remained to improve. With the states here, a Ranachensinensis skin collagen (RCSC)/poly(ɛ-caprolactone) (PCL)AgNP-loaded3D nanofiber scaffold is fabricated as a wound dressing material by using an improved wet electrospinning method (blending). The nanoscale of the AgNPs is proved. The 3D porous morphologies of the materials with different AgNP loadings, are determined with field emission scanning electron microscopy (FESEM) and the presence and uniformity distribution of AgNPs is confirmed by Energy dispersive X-ray (EDX) spectroscopy. The silver-ion release rates, antibacterial properties, and cytotoxicities of dressing materials with different AgNP contents are evaluated using ICP-AES, the zone inhibition method, and MTT testing. These results showed that the improved wet electrospun is an effective way to fabricate AgNP loaded 3D scaffold materials with porous structure and nearly 90% porosity and the presence of AgNPs in dressing materials strengthen the antibacterial properties. The RCSC/PCL 3D scaffold materials containing 2.0%AgNP would be promising for dressing materials application nearly without cytotoxicities.

  17. Glycerol-mediated nanostructure modification leading to improved transparency of porous polymeric scaffolds for high performance 3D cell imaging.

    PubMed

    Zhao, Shan; Shen, Zhiyuan; Wang, Jingyu; Li, Xiaokang; Zeng, Yang; Wang, Bingjie; He, Yonghong; Du, Yanan

    2014-07-14

    Glycerol is among the most commonly used optical clearing agents for tissues clearance largely due to refractive index (RI) matching between glycerol and the submerged tissues. Here we applied glycerol as structure modifier at both macroscopic (as porogen) and nanoscopic (as nanostructure ameliorant) scales to fabricate transparent porous scaffolds made from poly(ethylene glycol) (PEG) as well as other widely used biomaterials (e.g., PLGA, PA, or gelatin), whose nanostructures, in the scale of light wavelength, dominantly improved the optical transmittance of the scaffolds even when immersed in RI mismatched medium (e.g., culture medium or water). We further exploited the clearing mechanisms based on Mie scattering theory, illustrating that conformational changes of polymer chains induced by solvent effects of glycerol enhanced the anisotropy (i.e., directional alignment) of the nanostructures, leading to reduced crystallinity and scattering of the resulted PEG scaffolds. Our findings represent the first and systematic demonstration with both experimental and theoretical evidence in effectively clearing porous polymeric scaffolds by mechanisms other than RI matching, which could tackle the limitations of current optical imaging of cells cultured within three-dimensional (3D) opaque porous scaffolds, such as poor visibility, low spatial resolution, and small penetration depth.

  18. Tailor-made biopolymers porous scaffold fabrication for tissue engineering: application of radiant energy in the form of microwave under vacuum.

    PubMed

    Jaya, S; Durance, T D

    2008-01-01

    Many methods are available for developing three-dimensional porous scaffolds using various polymeric materials for tissue-engineering applications. Each has its own advantages and disadvantages. Some of the available methods and their limitations were discussed briefly. This paper focuses on the scope of novel technology called radiant energy application under vacuum for the fabrication of three-dimensional porous scaffolds for tissue engineering applications. Radiant energy application in the form of microwave under vacuum has been shown to develop and maintain the porous structure in fruits and vegetables after dehydration, which produced the microstructure similar to the freeze dried materials. Same principle of applying radiant energy under vacuum was used on the biopolymeric gels to create tailor-made, porous scaffolds for biomedical purposes. It has many advantages over the other existing methods of scaffold fabrication. This paper also reviews the scaffolds design recently fabricated by the authors using radiant energy under vacuum.

  19. Bone regeneration in strong porous bioactive glass (13-93) scaffolds with an oriented microstructure implanted in rat calvarial defects.

    PubMed

    Liu, Xin; Rahaman, Mohamed N; Fu, Qiang

    2013-01-01

    There is a need for synthetic bone graft substitutes to repair large bone defects resulting from trauma, malignancy and congenital diseases. Bioactive glass has attractive properties as a scaffold material but factors that influence its ability to regenerate bone in vivo are not well understood. In the present work, the ability of strong porous scaffolds of 13-93 bioactive glass with an oriented microstructure to regenerate bone was evaluated in vivo using a rat calvarial defect model. Scaffolds with an oriented microstructure of columnar pores (porosity=50%; pore diameter=50-150 μm) showed mostly osteoconductive bone regeneration, and new bone formation, normalized to the available pore area (volume) of the scaffolds, increased from 37% at 12 weeks to 55% at 24 weeks. Scaffolds of the same glass with a trabecular microstructure (porosity=80%; pore width=100-500 μm), used as the positive control, showed bone regeneration in the pores of 25% and 46% at 12 and 24 weeks, respectively. The brittle mechanical response of the as-fabricated scaffolds changed markedly to an elastoplastic response in vivo at both implantation times. These results indicate that both groups of 13-93 bioactive glass scaffolds could potentially be used to repair large bone defects, but scaffolds with the oriented microstructure could also be considered for the repair of loaded bone.

  20. An endothelial cultured condition medium embedded porous PLGA scaffold for the enhancement of mouse embryonic stem cell differentiation.

    PubMed

    Li, Ching-Wen; Pan, Wei-Ting; Ju, Jyh-Cherng; Wang, Gou-Jen

    2016-04-12

    In this study, we have developed a microporous poly(lactic-co-glycolic acid) (PLGA) scaffold that combines a continuous release property and a three-dimensional (3D) scaffolding technique for the precise and efficient formation of endothelial cell lineage from embryonic stem cells (ESCs). Eight PLGA scaffolds (14.29%, 16.67%, 20% and 25% concentrations of PLGA solutions) mixed with two crystal sizes of sodium chloride (NaCl) were fabricated by leaching. Then, vascular endothelial cell conditioned medium (ECCM) mixed with gelatin was embedded into the scaffold for culturing of mouse embryonic stem cells (mESCs). The 14.29% PLGA scaffolds fabricated using non-ground NaCl particles (NG-PLGA) and the 25% PLGA containing scaffolds fabricated using ground NaCl particles (G-PLGA) possessed minimum and maximum moisture content and bovine serum albumin (BSA) content properties, respectively. These two groups of scaffolds were used for future experiments in this study. Cell culture results demonstrated that the proposed porous scaffolds without growth factors were sufficient to induce mouse ESCs to differentiate into endothelial-like cells in the early culture stages, and combined with embedded ECCM could provide a long-term inducing system for ESC differentiation.

  1. Bone regeneration in strong porous bioactive glass (13–93) scaffolds with an oriented microstructure implanted in rat calvarial defects

    PubMed Central

    Liu, Xin; Rahaman, Mohamed N.; Fu, Qiang

    2012-01-01

    There is a need for synthetic bone graft substitutes to repair large bone defects resulting from trauma, malignancy, and congenital diseases. Bioactive glass has attractive properties as a scaffold material but factors that influence its ability to regenerate bone in vivo are not well understood. In the present work, the ability of strong porous scaffolds of 13–93 bioactive glass with an oriented microstructure to regenerate bone was evaluated in vivo using a rat calvarial defect model. Scaffolds with an oriented microstructure of columnar pores (porosity = 50%; pore diameter = 50–150 µm) showed mostly osteoconductive bone regeneration, and new bone formation, normalized to the available pore area (volume) of the scaffolds, increased from 37% at 12 weeks to 55% at 24 weeks. Scaffolds of the same glass with a trabecular microstructure (porosity = 80%; pore width = 100–500 µm), used as the positive control, showed bone regeneration in the pores of 25% and 46% at 12 and 24 weeks, respectively. The brittle mechanical response of the as-fabricated scaffolds changed markedly to an elasto-plastic response in vivo at both implantation times. These results indicate that both groups of 13–93 bioactive glass scaffolds could potentially be used to repair large bone defects, but scaffolds with the oriented microstructure could also be considered for the repair of loaded bone. PMID:22922251

  2. 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.

  3. The effect of collagen-chitosan porous scaffold thickness on dermal regeneration in a one-stage grafting procedure.

    PubMed

    Haifei, Shi; Xingang, Wang; Shoucheng, Wu; Zhengwei, Mao; Chuangang, You; Chunmao, Han

    2014-01-01

    Dermal substitutes are used as dermal regeneration templates to reduce scar formation and improve wound healing. Unlike autografts, dermal substitutes lack normal vascular networks. The increased distance required for diffusion of oxygen and nutrients to the autograft following interpositioning of the substitute dramatically affects graft survival. To evaluate the effect of collagen-chitosan scaffold thickness on dermal regeneration, single-layer collagen-chitosan porous scaffolds of 0.5-, 1- and 2-mm thicknesses were fabricated and used to treat full-thickness wounds in a one-stage grafting procedure in a rat model. Skin-graft viability, wound contraction, histological changes, and wound tensile strength were evaluated. The results indicated that the distance for the diffusion of oxygen and nutrients to the autograft in the 2-mm-thick scaffold provided less support for graft take, which resulted in graft necrosis, extensive inflammatory reaction, marked foreign-body reaction (FBR), rapid scaffold degradation, and abnormal collagen deposition and remodeling. In contrast, the thinner scaffolds, especially of that 0.5-mm thickness, promoted earlier angiogenesis, ensuring skin-graft viability with a mild FBR, and ordered fibroblast infiltration and better collagen remodeling. It can be concluded that collagen-chitosan porous scaffolds with a thickness of <1mm are more suitable for dermal regeneration and can be used as dermal templates for treatment of dermal defects using a one-stage grafting procedure. © 2013 Elsevier Ltd. All rights reserved.

  4. 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.

  5. Chondrogenic Differentiation of Adipose-Derived Adult Stem Cells by a Porous Scaffold Derived from Native Articular Cartilage Extracellular Matrix

    PubMed Central

    Cheng, Nai-Chen; Estes, Bradley T.; Awad, Hani A.

    2009-01-01

    Adipose-derived adult stem cells (ASCs) have the ability to differentiate into a chondrogenic phenotype in response to specific environmental signals such as growth factors or artificial biomaterial scaffolds. In this study, we examined the hypothesis that a porous scaffold derived exclusively from articular cartilage can induce chondrogenesis of ASCs. Human ASCs were seeded on porous scaffolds derived from adult porcine articular cartilage and cultured in standard medium without exogenous growth factors. Chondrogenesis of ASCs seeded within the scaffold was evident by quantitative RT-PCR analysis for cartilage-specific extracellular matrix (ECM) genes. Histological and immunohistochemical examination showed abundant production of cartilage-specific ECM components—particularly, type II collagen—after 4 or 6 weeks of culture. After 6 weeks of culture, the cellular morphology in the ASC-seeded constructs resembled those in native articular cartilage tissue, with rounded cells residing in the glycosaminoglycan-rich regions of the scaffolds. Biphasic mechanical testing showed that the aggregate modulus of the ASC-seeded constructs increased over time, reaching 150 kPa by day 42, more than threefold higher than that of the unseeded controls. These results suggest that a porous scaffold derived from articular cartilage has the ability to induce chondrogenic differentiation of ASCs without exogenous growth factors, with significant synthesis and accumulation of ECM macromolecules, and the development of mechanical properties approaching those of native cartilage. These findings support the potential for a processed cartilage ECM as a biomaterial scaffold for cartilage tissue engineering. Additional in vivo evaluation is necessary to fully recognize the clinical implication of these observations. PMID:18950290

  6. Construction of mesenchymal stem cell-containing collagen gel with a macrochanneled polycaprolactone scaffold and the flow perfusion culturing for bone tissue engineering.

    PubMed

    Yu, Hye-Sun; Won, Jong-Eun; Jin, Guang-Zhen; Kim, Hae-Won

    2012-06-01

    A novel bone tissue-engineering construct was developed by using poly(ɛ-caprolactone) (PCL)-macrochanneled scaffolds combined with stem cell-seeded collagen hydrogels and then applying flow perfusion culture. Rat mesenchymal stem cells (MSCs) were loaded into collagen hydrogels, which were then combined with macrochanneled PCL scaffolds. Collagen hydrogels were demonstrated to provide favorable growth environments for MSCs and to foster proliferation. Cell number determination identified retention of substantially fewer (50-60%) cells when they were seeded directly onto macrochanneled PCL than of cells engineered within collagen hydrogels. Additionally, the cells actively proliferated within the combined scaffold for up to 7 days. MSC-loaded collagen-PCL scaffolds were subsequently cultured under flow perfusion to promote proliferation and osteogenic differentiation. Cells proliferated to levels significantly higher in flow perfusion culture than that under static conditions during 21 days. A quantitative polymerase chain reaction (QPCR) assay revealed significant alterations in the transcription of bone-related genes such as osteopontin (OPN), osteocalcin (OCN), and bone sialoprotein (BSP), such as 8-, 2.5-, and 3-fold induction, respectively, after 10 days of flow perfusion relative to those in static culture. OPN and OCN protein levels, as determined by Western blot, increased under flow perfusion. Cellular mineralization was significantly enhanced by the flow perfusion during 21 and 28 days. Analyses of mechanosensitive gene expression induced by flow perfusion shear stress revealed significant upregulation of c-fos and cyclooxygenase-2 (COX-2) during the initial culture period (3-5 days), suggesting that osteogenic stimulation was possible as a result of mechanical force-driven transduction. These results provide valuable information for the design of a new bone tissue-engineering system by combining stem cell-loaded collagen hydrogels with macrochanneled

  7. Construction of Mesenchymal Stem Cell–Containing Collagen Gel with a Macrochanneled Polycaprolactone Scaffold and the Flow Perfusion Culturing for Bone Tissue Engineering

    PubMed Central

    Yu, Hye-Sun; Won, Jong-Eun; Jin, Guang-Zhen

    2012-01-01

    Abstract A novel bone tissue-engineering construct was developed by using poly(ɛ-caprolactone) (PCL)-macrochanneled scaffolds combined with stem cell-seeded collagen hydrogels and then applying flow perfusion culture. Rat mesenchymal stem cells (MSCs) were loaded into collagen hydrogels, which were then combined with macrochanneled PCL scaffolds. Collagen hydrogels were demonstrated to provide favorable growth environments for MSCs and to foster proliferation. Cell number determination identified retention of substantially fewer (50–60%) cells when they were seeded directly onto macrochanneled PCL than of cells engineered within collagen hydrogels. Additionally, the cells actively proliferated within the combined scaffold for up to 7 days. MSC-loaded collagen–PCL scaffolds were subsequently cultured under flow perfusion to promote proliferation and osteogenic differentiation. Cells proliferated to levels significantly higher in flow perfusion culture than that under static conditions during 21 days. A quantitative polymerase chain reaction (QPCR) assay revealed significant alterations in the transcription of bone-related genes such as osteopontin (OPN), osteocalcin (OCN), and bone sialoprotein (BSP), such as 8-, 2.5-, and 3-fold induction, respectively, after 10 days of flow perfusion relative to those in static culture. OPN and OCN protein levels, as determined by Western blot, increased under flow perfusion. Cellular mineralization was significantly enhanced by the flow perfusion during 21 and 28 days. Analyses of mechanosensitive gene expression induced by flow perfusion shear stress revealed significant upregulation of c-fos and cyclooxygenase-2 (COX-2) during the initial culture period (3–5 days), suggesting that osteogenic stimulation was possible as a result of mechanical force-driven transduction. These results provide valuable information for the design of a new bone tissue-engineering system by combining stem cell-loaded collagen hydrogels with

  8. Reinforcement of a porous collagen scaffold with surface-activated PLA fibers.

    PubMed

    Liu, Xi; Huang, Changbin; Feng, Yujie; Liang, Jie; Fan, Yujiang; Gu, Zhongwei; Zhang, Xingdong

    2010-01-01

    A hybrid porous collagen scaffold mechanically reinforced with surface-activated poly(lactic acid) (PLA) fiber was prepared. PLA fibers, 20 mum in diameter and 1 mm in length, were aminolyzed with hexanediamine to introduce free amino groups on the surfaces. After the amino groups were transferred to aldehyde groups by treatment with glutaraldehyde, different amounts (1.5, 3, 5 and 8 mg) of surface-activated PLA fibers were homogeneously mixed with 2 ml type-I collagen solution (pH 2.8, 0.6 wt%). This mixture solution was then freeze-dried and cross-linked to obtain collagen sponges with surface-activated PLA fiber. Scanning electron microscopy observation indicated that the collagen sponges had a highly interconnected porous structure with an average pore size of 170 mum, irrespective of PLA fiber incorporation. The dispersion of surface-activated PLA fibers was homogeneous in collagen sponge, in contrast to unactivated PLA fibers. The compression modulus test results showed that, compared with unactivated PLA fibers, the surface-activated PLA fibers enhanced the resistance of collagen sponge to compression more significantly. Cytotoxicity assay by MTT test showed no cytotoxicity of these collagen sponges. L929 mouse fibroblast cell-culture studies in vitro revealed that the number of L929 cells attached to the collagen sponge with surface-activated PLA fibers, both 6 h and 24 h after seeding, was higher than that in pure collagen sponge and sponge with unactivated PLA fibers. In addition, a better distribution of cells infiltrated in collagen sponge with surface-activated PLA fibers was observed by histological staining. These results indicated that the collagen sponge reinforced with surface-activated PLA fibers is a promising biocompatible scaffold for tissue engineering.

  9. 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

  10. Preparation and characterization of cross-linked carboxymethyl chitin porous membrane scaffold for biomedical applications.

    PubMed

    Zhao, Liqing; Wu, Yiguang; Chen, Shu; Xing, Tao

    2015-08-01

    Porous dermal scaffold membrane (PDSM) was successfully prepared by using a so-called sol-gel freeze-drying method. In this method, the carboxymethyl chitin (CMC) hydrosol was first cross-linked by 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), and then lyophilized to form the PDSM. For the first time, this research focused on the cross-linked CMC as the only component for three-dimensional PDSM. The effects of cross-linking conditions on the performance of the PDSM were investigated. And PDSM with optimal performance was obtained through 4-h cross-linking at 4 wt% of CMC concentration in the hydrosol, where the mass ratio of EDC to NHS to CMC was 5:3:10. The porosity of the optimized PDSM was more than 90% and the water swelling rate was above 4000%. The pore size was well distributed and was between 100 μm and 200 μm. And the tensile strength was above 0.09 MPa. The as-made PDSM could be degraded above 80% in 12 days in the presence of a 0.2mg/mL lysozyme solution. Very importantly, the PDSM had no cytotoxicity and good biocompatibility from MTT assays. Our results showed the application possibility of the as-prepared PDSM as dermal scaffold for skin tissue engineering.

  11. Towards a methodology for the effective surface modification of porous polymer scaffolds.

    PubMed

    Safinia, Laleh; Datan, Nathalie; Höhse, Marek; Mantalaris, Athanassios; Bismarck, Alexander

    2005-12-01

    A novel low-pressure radio-frequency plasma treatment protocol was developed to achieve the effective through-thickness surface modification of large porous poly (D,L-lactide) (PDLLA) polymer scaffolds using air or water: ammonia plasma treatments. Polymer films were modified as controls. Scanning electron micrographs and maximum bubble point measurements demonstrated that the PDLLA foams have the high porosity, void fraction and interconnected pores required for use as tissue engineering scaffolds. The polymer surface of the virgin polymer does contain acidic functional groups but is hydrophobic. Following exposure to air or water: ammonia plasma, an increased number of polar functional groups and improved wetting behaviour, i.e. hydrophilicity, of wet surfaces was detected. The number of polar surface functional groups increased (hence the decrease in water contact angles) with increasing exposure time to plasma. The change in surface composition and wettablility of wet polymer constructs was characterised by zeta potential and contact angle measurements. The hydrophobic recovery of the treated PDLLA polymer surfaces was also studied. Storage of the treated polymer constructs in ambient air caused an appreciable hydrophobic recovery, whereas in water only partial hydrophobic recovery occurred. However, in both cases the initial surface characteristics decay as function of time.

  12. Chondrogenesis in perfusion bioreactors using porous silk scaffolds and hESC-derived MSCs.

    PubMed

    Tiğli, R Seda; Cannizaro, Chris; Gümüşderelioğlu, Menemşe; Kaplan, David L

    2011-01-01

    Tissue engineered cartilage can be grown in vitro with the use of cell-scaffold constructs and bioreactors. The present study was designed to investigate the effects of perfusion bioreactors on the chondrogenic potential of engineered constructs prepared from porous silk fibroin scaffolds seeded with human embryonic stem cell (hESC)-derived mesencyhmal stem cells (MSCs). After four weeks of incubation, constructs cultured in perfusion bioreactors showed significantly higher amounts of glycosaminoglycans (GAGs) (p < 0.001), DNA (p < 0.001), total collagen (p < 0.01), and collagen II (p < 0.01) in comparison to static culture. Mechanical stiffness of constructs increased 3.7-fold under dynamic culture conditions and RT-PCR results concluded that cells cultured in perfusion bioreactors highly expressed (p < 0.001) cartilage-related genes when compared with static culture. Distinct differences were noted in tissue morphology, including polygonal extracellular matrix structure of engineered constructs in thin superficial zones and an inner zone under static and dynamic conditions, respectively. The results suggest that the utility of perfusion bioreactors to modulate the growth of tissue-engineered cartilage and enhance tissue growth in vitro.

  13. Development of a porous 3D graphene-PDMS scaffold for improved osseointegration.

    PubMed

    Li, Jianfeng; Liu, Xiao; Crook, Jeremy M; Wallace, Gordon G

    2017-08-01

    Osseointegration in orthopedic surgery plays an important role for bone implantation success. Traditional treatment of implant surface aimed at improved osseointegration has limited capability for its poor performance in supporting cell growth and proliferation. Polydimethylsiloxane (PDMS) is a widely used silicon-based organic polymer material with properties that are useful in cosmetics, domestic applications and mechanical engineering. In addition, the biocompatibility of PDMS, in part due to the high solubility of oxygen, makes it an ideal material for cell-based implants. Notwithstanding its potential, a property that can inhibit PDMS bioactivity is the high hydrophobicity, limiting its use to date in tissue engineering. Here, we describe an efficient approach to produce porous, durable and cytocompatible PDMS-based 3D structures, coated with reduced graphene oxide (RGO). The RGO/PDMS scaffold has good mechanical strength and with pore sizes ranging from 10 to 600μm. Importantly, the scaffold is able to support growth and differentiation of human adipose stem cells (ADSCs) to an osteogenic cell lineage, indicative of its potential as a transition structure of an osseointegrated implant. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. The effect of poly(lactic-co-glycolic acid) (PLGA) coating on the mechanical, biodegradable, bioactive properties and drug release of porous calcium silicate scaffolds.

    PubMed

    Zhao, Lang; Wu, Chengtie; Lin, Kaili; Chang, Jiang

    2012-01-01

    Ideal scaffolds for bone tissue engineering require 3D interconnected porous structures, enough mechanical strength for hand of treatment as well as proper bioactivity and biodegradability. Calcium silicate (CaSiO3, CS) scaffolds have been studied for bone tissue engineering application due to their excellent bioactivity. However, the main disadvantages of CS scaffolds are their low mechanical strength and high alkaline ionic products. In this study, sintered CS scaffolds were prepared and coated with poly(lactic-co-glycolic acid) (PLGA), and the effect of PLGA coating on the mechanical, biodegradable, bioactive properties and drug release of porous CS scaffolds were investigated. The results showed that the PLGA-coated CS scaffolds maintained large pore size and high porosity. The compressive strength of PLGA/CS scaffolds was significantly improved compared to pure CS scaffolds, and increased with the increase of intrinsic viscosity and concentration of PLGA. In addition, the PLGA coating neutralized alkaline level resulted from the ionic products of CS scaffolds and reduced the pH value of biological solution during the degradation of scaffolds. It was found that PLGA/CS scaffolds still maintained excellent apatite-mineralization ability in SBF. Furthermore, the PLGA coating effectively inhibited the burst release and maintained a sustained release of drugs from the CS scaffolds. Our results indicate that the PLGA/CS scaffolds have great potential for bone tissue engineering application by the virtue of improved mechanical strength, and excellent bioactivity, degradation as well as drug-delivery property.

  15. Monotonic and cyclic loading behavior of porous scaffolds made from poly(para-phenylene) for orthopedic applications.

    PubMed

    Hoyt, Anthony J; Yakacki, Christopher M; Fertig, Ray S; Dana Carpenter, R; Frick, Carl P

    2015-01-01

    Porous poly(para-phenylene) (PPP) scaffolds have tremendous potential as an orthopedic biomaterial; however, the underlying mechanisms controlling the monotonic and cyclic behavior are poorly understood. The purpose of this study was to develop a method to integrate micro-computed tomography (μCT), finite-element analysis (FEA), and experimental results to uncover the relationships between the porous structure and mechanical behavior. The μCT images were taken from porous PPP scaffolds with a porosity of 75vol% and pore size distribution between 420 and 500µm. Representative sections of the image were segmented and converted into finite-element meshes. It was shown through FEA that localized stresses within the microstructure were approximately 100 times higher than the applied global stress during the linear loading regime. Experimental analysis revealed the S-N fatigue curves for fully dense and porous PPP samples were parallel on log-log plots, with the endurance limit for porous samples in tension being approximately 100 times lower than their solid PPP counterparts (0.3-35MPa) due to the extreme stress concentrations caused by the porous microarchitecture. The endurance limit for porous samples in compression was much higher than in tension (1.60MPa). Through optical, laser-scanning, and scanning-electron microscopy it was found that porous tensile samples failed under Mode I fracture in both monotonic and cyclic loading. By comparison, porous compressive samples failed via strut buckling/pore collapse monotonically and by shearing fracture during cyclic loading. Monotonic loading showed that deformation behavior was strongly correlated with pore volume fraction, matching foam theory well; however, fatigue behavior was much more sensitive to local stresses believed to cause crack nucleation.

  16. Fabrication of three-dimensional porous scaffolds with controlled filament orientation and large pore size via an improved E-jetting technique.

    PubMed

    Li, Jin Lan; Cai, Yan Li; Guo, Yi Lin; Fuh, Jerry Ying Hsi; Sun, Jie; Hong, Geok Soon; Lam, Ruey Na; Wong, Yoke San; Wang, Wilson; Tay, Bee Yen; Thian, Eng San

    2014-05-01

    Biodegradable polymeric scaffolds have been widely used in tissue engineering as a platform for cell proliferation and subsequent tissue regeneration. Conventional microextrusion methods for three-dimensional (3D) scaffold fabrication were limited by their low resolution. Electrospinning, a form of electrohydrodynamic (EHD) printing, is an attractive method due to its capability of fabricating high-resolution scaffolds at the nanometer/micrometer scale level. However, the scaffold was composed of randomly orientated filaments which could not guide the cells in a specific direction. Furthermore, the pores of the electrospun scaffold were small, thus preventing cell infiltration. In this study, an alternative EHD jet printing (E-jetting) technique has been developed and employed to fabricate 3D polycaprolactone (PCL) scaffolds with desired filament orientation and pore size. The effect of PCL solution concentration was evaluated. Results showed that solidified filaments were achieved at concentration >70% (w/v). Uniform filaments of diameter 20 μm were produced via the E-jetting technique, and X-ray diffraction and attenuated total reflectance Fourier transform infrared spectroscopic analyses revealed that there was no physicochemical changes toward PCL. Scaffold with a pore size of 450 μm and porosity level of 92%, was achieved. A preliminary in vitro study illustrated that live chondrocytes were attaching on the outer and inner surfaces of collagen-coated E-jetted PCL scaffolds. E-jetted scaffolds increased chondrocytes extracellular matrix secretion, and newly formed matrices from chondrocytes contributed significantly to the mechanical strength of the scaffolds. All these results suggested that E-jetting is an alternative scaffold fabrication technique, which has the capability to construct 3D scaffolds with aligned filaments and large pore sizes for tissue engineering applications.

  17. Nanostructured Porous Silicon: The Winding Road from Photonics to Cell Scaffolds – A Review

    PubMed Central

    Hernández-Montelongo, Jacobo; Muñoz-Noval, Alvaro; García-Ruíz, Josefa Predestinación; Torres-Costa, Vicente; Martín-Palma, Raul J.; Manso-Silván, Miguel

    2015-01-01

    For over 20 years, nanostructured porous silicon (nanoPS) has found a vast number of applications in the broad fields of photonics and optoelectronics, triggered by the discovery of its photoluminescent behavior in 1990. Besides, its biocompatibility, biodegradability, and bioresorbability make porous silicon (PSi) an appealing biomaterial. These properties are largely a consequence of its particular susceptibility to oxidation, leading to the formation of silicon oxide, which is readily dissolved by body fluids. This paper reviews the evolution of the applications of PSi and nanoPS from photonics through biophotonics, to their use as cell scaffolds, whether as an implantable substitute biomaterial, mainly for bony and ophthalmological tissues, or as an in vitro cell conditioning support, especially for pluripotent cells. For any of these applications, PSi/nanoPS can be used directly after synthesis from Si wafers, upon appropriate surface modification processes, or as a composite biomaterial. Unedited studies of fluorescently active PSi structures for cell culture are brought to evidence the margin for new developments. PMID:26029688

  18. Nanogel tectonic porous gel loading biologics, nanocarriers, and cells for advanced scaffold.

    PubMed

    Hashimoto, Yoshihide; Mukai, Sada-atsu; Sawada, Shin-ichi; Sasaki, Yoshihiro; Akiyoshi, Kazunari

    2015-01-01

    We developed a new self-assembled amphiphilic nanogel-crosslinked porous (NanoCliP) gel that can trap proteins, liposomes, and cells. The NanoCliP gel was prepared by Michael addition of a self-assembled nanogel of acryloyl group-modified cholesterol-bearing pullulan to pentaerythritol tetra (mercaptoethyl) polyoxyethylene, followed by freezing-induced phase separation. Dynamic rheological analysis revealed that the storage modulus (G') of the NanoCliP gel was approximately 10 times greater than that of a nonporous nanogel-crosslinked gel. Two-photon excitation deep imaging revealed that the NanoCliP gel comprises interconnected pores of several hundred micrometers in diameter. The NanoCliP gel trapped proteins and liposomes via hydrophobic interactions because its amphiphilic nanogels exhibit chaperone-like activity. Mouse embryonic fibroblasts penetrated the interconnected pores and adhered to the porous surface of fibronectin-complexed NanoCliP gel. In vivo, the NanoCliP gel enhanced cell infiltration, tissue ingrowth, and neovascularization without requiring exogenous growth factors, suggesting that the NanoCliP gel is a promising scaffold for tissue engineering. Copyright © 2014 Elsevier Ltd. All rights reserved.

  19. Preparation and Reinforcement of Dual‐Porous Biocompatible Cellulose Scaffolds for Tissue Engineering

    PubMed Central

    Pircher, Nicole; Fischhuber, David; Carbajal, Leticia; Strauß, Christine; Nedelec, Jean‐Marie; Kasper, Cornelia; Rosenau, Thomas

    2015-01-01

    1 Biocompatible cellulose‐based aerogels composed of nanoporous struts, which embed interconnected voids of controlled micron‐size, have been prepared employing temporary templates of fused porogens, reinforcement by interpenetrating PMMA networks and supercritical carbon dioxide drying. Different combinations of cellulose solvent (Ca(SCN)2/H2O/LiCl or [EMIm][OAc]/DMSO) and anti‐solvent (EtOH), porogen type (paraffin wax or PMMA spheres) and porogen size (various fractions in the range of 100–500 μm) as well as intensity of PMMA reinforcement have been investigated to tailor the materials for cell scaffolding applications. All aerogels exhibited an open and dual porosity (micronporosity >100 μm and nanoporosity extending to the low micrometer range). Mechanical properties of the dual‐porous aerogels under compressive stress were considerably improved by introduction of interpenetrating PMMA networks. The effect of the reinforcing polymer on attachment, spreading, and proliferation of NIH 3T3 fibroblast cells, cultivated on selected dual‐porous aerogels to pre‐evaluate their biocompatibility was similarly positive. PMID:26941565

  20. Preparation and Reinforcement of Dual-Porous Biocompatible Cellulose Scaffolds for Tissue Engineering.

    PubMed

    Pircher, Nicole; Fischhuber, David; Carbajal, Leticia; Strauß, Christine; Nedelec, Jean-Marie; Kasper, Cornelia; Rosenau, Thomas; Liebner, Falk

    2015-09-01

    1Biocompatible cellulose-based aerogels composed of nanoporous struts, which embed interconnected voids of controlled micron-size, have been prepared employing temporary templates of fused porogens, reinforcement by interpenetrating PMMA networks and supercritical carbon dioxide drying. Different combinations of cellulose solvent (Ca(SCN)2/H2O/LiCl or [EMIm][OAc]/DMSO) and anti-solvent (EtOH), porogen type (paraffin wax or PMMA spheres) and porogen size (various fractions in the range of 100-500 μm) as well as intensity of PMMA reinforcement have been investigated to tailor the materials for cell scaffolding applications. All aerogels exhibited an open and dual porosity (micronporosity >100 μm and nanoporosity extending to the low micrometer range). Mechanical properties of the dual-porous aerogels under compressive stress were considerably improved by introduction of interpenetrating PMMA networks. The effect of the reinforcing polymer on attachment, spreading, and proliferation of NIH 3T3 fibroblast cells, cultivated on selected dual-porous aerogels to pre-evaluate their biocompatibility was similarly positive.

  1. Differentiation of Induced Pluripotent Stem Cells to Neural Retinal Precursor Cells on Porous Poly-Lactic-co-Glycolic Acid Scaffolds

    PubMed Central

    Worthington, Kristan S.; Wiley, Luke A.; Guymon, C. Allan; Salem, Aliasger K.

    2016-01-01

    Abstract Purpose: Cell replacement therapy for the treatment of retinal degeneration is an increasingly feasible approach, but one that still requires optimization of the transplantation strategy. To this end, various polymer substrates can increase cell survival and integration, although the effect of their pore size on cell behavior, particularly differentiation, has yet to be explored. Methods: Salt crystals of varying known size were used to impart structure to poly(lactic-co-glycolic acid) (PLGA) scaffolds by a salt leaching/solvent evaporation process. Mouse induced pluripotent stem cells (miPSCs) were seeded to the polymer scaffolds and supplemented with retinal differentiation media for up to 2 weeks. Proliferation was measured during the course of 2 weeks, while differentiation was evaluated using cell morphology and expression of early retinal development markers. Results: The salt leaching method of porous PLGA fabrication resulted in amorphous smooth pores. Cells attached to these scaffolds and proliferated, reaching a maximum cell number at 10 days postseeding that was 5 times higher on porous PLGA than on nonporous controls. The morphology of many of these cells, including their formation of neurites, was suggestive of neural phenotypes, while their expression of Sox2, Pax6, and Otx2 indicates early retinal development. Conclusions: The use of porous PLGA scaffolds to differentiate iPSCs to retinal phenotypes is a feasible pretransplantation approach. This adds to an important knowledge base; understanding how developing retinal cells interact with polymer substrates with varying structure is a crucial component of optimizing cell therapy strategies. PMID:26692377

  2. Optimization and evaluation of silk fibroin-chitosan freeze-dried porous scaffolds for cartilage tissue engineering application.

    PubMed

    Vishwanath, Varshini; Pramanik, Krishna; Biswas, Amit

    2016-01-01

    Silk fibroin/chitosan blend has been reported to be an attractive biomaterial that provides a 3D porous structure with controllable pore size and mechanical property suitable for tissue engineering applications. However, there is no systematic study for optimizing the ratio of silk fibroin (SF) and chitosan (CS) which seems to influence the scaffold property to a great extent. The present research, therefore, investigates the effect of blend ratio of SF and CS on scaffold property and establishes the optimum value of blend ratio. Among the various blends, the scaffolds with blend ratio of SF/CS (80:20) were found to be superior. The scaffold possesses pore size in the range 71-210 μm and porosity of 82.2 ± 1.3%. The compressive strength of the scaffold was measured as 190 ± 0.2 kPa. The cell supportive property of the scaffold in terms of cell attachment, cell viability, and proliferation was confirmed by cell culture study using mesenchymal stem cells derived from umbilical cord blood. Furthermore, the assessment of glycosaminoglycan secretion on the scaffolds indicates its potentiality toward cartilage tissue regeneration.

  3. Protein growth factors loaded highly porous chitosan scaffold: a comparison of bone healing properties.

    PubMed

    Nandi, Samit K; Kundu, Biswanath; Basu, Debabrata

    2013-04-01

    Present study aimed to investigate and compare effectiveness of porous chitosan alone and in combination with insulin like growth factor-1 (IGF-1) and bone morphogenetic protein-2 (BMP-2) in bone healing. Highly porous (85±2%) with wide distribution of macroporous (70-900 μm) chitosan scaffolds were fabricated as bone substitutes by employing a simple liquid hardening method using 2% (w/v) chitosan suspension. IGF-1 and BMP-2 were infiltrated using vacuum infiltration with freeze drying method. Adsorption efficiency was found to be 87±2 and 90±2% for BMP-2 and IGF-1 respectively. After thorough material characterization (pore details, FTIR and SEM), samples were used for subsequent in vivo animal trial. Eighteen rabbit models were used to evaluate and compare control (chitosan) (group A), chitosan with IGF-1 (group B) and chitosan with BMP-2 (group C) in the repair of critical size bone defect in tibia. Radiologically, there was evidence of radiodensity in defect area from 60th day (initiated on 30th day) in groups B and C as compared to group A and attaining nearly bony density in most of the part at day 90. Histological results depicted well developed osteoblastic proliferation around haversian canal along with proliferating fibroblast, vascularization and reticular network which was more pronounced in group B followed by groups C and A. Fluorochrome labeling and SEM studies in all groups showed similar outcome. Hence, porous chitosan alone and in combination with growth factors (GFs) can be successfully used for bone defect healing with slight advantage of IGF-1 in chitosan samples.

  4. Modeling the fluid-dynamics and oxygen consumption in a porous scaffold stimulated by cyclic squeeze pressure.

    PubMed

    Ferroni, Marco; Giusti, Serena; Nascimento, Diana; Silva, Ana; Boschetti, Federica; Ahluwalia, Arti

    2016-08-01

    The architecture and dynamic physical environment of tissues can be recreated in-vitro by combining 3D porous scaffolds and bioreactors able to apply controlled mechanical stimuli on cells. In such systems, the entity of the stimuli and the distribution of nutrients within the engineered construct depend on the micro-structure of the scaffolds. In this work, we present a new approach for optimizing computational fluid-dynamics (CFD) models for the investigation of fluid-induced forces generated by cyclic squeeze pressure within a porous construct, coupled with oxygen consumption of cardiomyocytes. A 2D axial symmetric macro-scaled model of a squeeze pressure bioreactor chamber was used as starting point for generating time dependent pressure profiles. Subsequently the fluid movement generated by the pressure fields was coupled with a complete 3D micro-scaled model of a porous protein cryogel. Oxygen transport and consumption inside the scaffold was evaluated considering a homogeneous distribution of cardiomyocytes throughout the structure, as confirmed by preliminary cell culture experiments. The results show that a 3D description of the system, coupling a porous geometry and time dependent pressure driven flow with fluid-structure-interaction provides an accurate and meaningful description of the microenvironment in terms of shear stress and oxygen distribution than simple stationary 2D models. Copyright © 2016 IPEM. Published by Elsevier Ltd. All rights reserved.

  5. Urethral reconstruction with a 3D porous bacterial cellulose scaffold seeded with lingual keratinocytes in a rabbit model.

    PubMed

    Huang, Jian-Wen; Lv, Xiang-Guo; Li, Zhe; Song, Lu-Jie; Feng, Chao; Xie, Min-Kai; Li, Chao; Li, Hong-Bin; Wang, Ji-Hong; Zhu, Wei-Dong; Chen, Shi-Yan; Wang, Hua-Ping; Xu, Yue-Min

    2015-09-11

    The goal of this study was to evaluate the effects of urethral reconstruction with a three-dimensional (3D) porous bacterial cellulose (BC) scaffold seeded with lingual keratinocytes in a rabbit model. A novel 3D porous BC scaffold was prepared by gelatin sponge interfering in the BC fermentation process. Rabbit lingual keratinocytes were isolated, expanded, and seeded onto 3D porous BC. BC alone (group 1, N  =  10), 3D porous BC alone (group 2, N  =  10), and 3D porous BC seeded with lingual keratinocytes (group 3, N  =  10) were used to repair rabbit ventral urethral defects (2.0   ×   0.8 cm). Scanning electron microscopy revealed that BC consisted of a compact laminate while 3D porous BC was composed of a porous sheet buttressed by a dense outer layer. The average pore diameter and porosity of the 3D porous BC were 4.23   ±   1.14 μm and 67.00   ±   6.80%, respectively. At 3 months postoperatively, macroscopic examinations and retrograde urethrograms of urethras revealed that all urethras maintained wide calibers in group 3. Strictures were found in all rabbits in groups 1 and 2. Histologically, at 1 month postoperatively, intact epithelium occurred in group 3, and discontinued epithelium was found in groups 1 and 2. However, groups 2 and 3 exhibited similar epithelial regeneration, which was superior to that of group 1 at 3 months (p  <  0.05). Comparisons of smooth muscle content and endothelia density among the three groups revealed a significant increase at each time point (p  <  0.05). Our results demonstrated that 3D porous BC seeded with lingual keratinocytes enhanced urethral tissue regeneration. 3D porous BC could potentially be used as an optimized scaffold for urethral reconstruction.

  6. Osteochondral tissue formation through adipose-derived stromal cell differentiation on biomimetic polycaprolactone nanofibrous scaffolds with graded insulin and Beta-glycerophosphate concentrations.

    PubMed

    Erisken, Cevat; Kalyon, Dilhan M; Wang, Hongjun; Ornek-Ballanco, Ceren; Xu, Jiahua

    2011-05-01

    The ability to fabricate tissue engineering scaffolds containing systematic gradients in the distributions of stimulators provides additional means for the mimicking of the important gradients observed in native tissues. Here the concentration distributions of two bioactive agents were varied concomitantly for the first time (one increasing, whereas the other decreasing monotonically) in between the two sides of a nanofibrous scaffold. This was achieved via the application of a new processing method, that is, the twin-screw extrusion and electrospinning method, to generate gradients of insulin, a stimulator of chondrogenic differentiation, and β-glycerophosphate (β-GP), for mineralization. The graded poly(ɛ-caprolactone) mesh was seeded with human adipose-derived stromal cells and cultured over 8 weeks. The resulting tissue constructs were analyzed for and revealed indications of selective differentiation of human adipose-derived stromal cells toward chondrogenic lineage and mineralization as functions of position as a result of the corresponding concentrations of insulin and β-GP. Chondrogenic differentiation of the stem cells increased at insulin-rich locations and mineralization increased at β-GP-rich locations.

  7. Biological performance of a polycaprolactone-based scaffold used as fusion cage device in a large animal model of spinal reconstructive surgery.

    PubMed

    Abbah, Sunny A; Lam, Christopher X L; Hutmacher, Dietmar W; Goh, James C H; Wong, Hee-Kit

    2009-10-01

    A bioactive and bioresorbable scaffold fabricated from medical grade poly (epsilon-caprolactone) and incorporating 20% beta-tricalcium phosphate (mPCL-TCP) was recently developed for bone regeneration at load bearing sites. In the present study, we aimed to evaluate bone ingrowth into mPCL-TCP in a large animal model of lumbar interbody fusion. Six pigs underwent a 2-level (L3/4; L5/6) anterior lumbar interbody fusion (ALIF) implanted with mPCL-TCP + 0.6 mg rhBMP-2 as treatment group while four other pigs implanted with autogenous bone graft served as control. Computed tomographic scanning and histology revealed complete defect bridging in all (100%) specimen from the treatment group as early as 3 months. Histological evidence of continuing bone remodeling and maturation was observed at 6 months. In the control group, only partial bridging was observed at 3 months and only 50% of segments in this group showed complete defect bridging at 6 months. Furthermore, 25% of segments in the control group showed evidence of graft fracture, resorption and pseudoarthrosis. In contrast, no evidence of graft fractures, pseudoarthrosis or foreign body reaction was observed in the treatment group. These results reveal that mPCL-TCP scaffolds could act as bone graft substitutes by providing a suitable environment for bone regeneration in a dynamic load bearing setting such as in a porcine model of interbody spine fusion.

  8. New Method for the Deposition of Nickel Oxide in Porous Scaffolds for Electrodes in Solid Oxide Fuel Cells and Electrolyzers

    PubMed Central

    Puolamaa, Milla; Sum, Brian; Tariq, Farid; Yufit, Vladimir; Brandon, Nigel P.

    2016-01-01

    Abstract A simple chemical bath deposition is used to coat a complex porous ceramic scaffold with a conformal Ni layer. The resulting composite is used as a solid oxide fuel cell electrode, and its electrochemical response is measured in humidified hydrogen. X‐ray tomography is used to determine the microstructural characteristics of the uncoated and Ni‐coated porous structure, which include the surface area to total volume, the radial pore size, and the size of the necks between the pores. PMID:27739632

  9. Doped Tricalcium Phosphate Scaffolds by Thermal Decomposition of Naphthalene: Mechanical Properties and In vivo Osteogenesis in a Rabbit Femur Model

    PubMed Central

    Ke, Dongxu; Dernell, William; Bandyopadhyay, Amit; Bose, Susmita

    2015-01-01

    Tricalcium phosphate (TCP) is a bioceramic that is widely used in orthopedic and dental applications. TCP structures show excellent biocompatibility as well as biodegradability. In this study, porous β-TCP scaffolds were prepared by thermal decomposition of naphthalene. Scaffolds with 57.64 ± 3.54 % density and a maximum pore size around 100 μm were fabricated via removing 30% naphthalene at 1150°C. The compressive strength for these scaffolds was 32.85 ± 1.41 MPa. Furthermore, by mixing 1 wt % SrO and 0.5 wt % SiO2, pore interconnectivity improved, but the compressive strength decreased to 22.40 ± 2.70 MPa. However, after addition of polycaprolactone (PCL) coating layers, the compressive strength of doped scaffolds increased to 29.57 ± 3.77 MPa. Porous scaffolds were implanted in rabbit femur defects to evaluate their biological property. The addition of dopants triggered osteoinduction by enhancing osteoid formation, osteocalcin expression and bone regeneration, especially at the interface of the scaffold and host bone. This study showed processing flexibility to make interconnected porous scaffolds with different pore size and volume fraction porosity with high compressive mechanical strength and better bioactivity. Results show that SrO/SiO2 doped porous TCP scaffolds have excellent potential to be used in bone tissue engineering applications. PMID:25504889

  10. Doped tricalcium phosphate scaffolds by thermal decomposition of naphthalene: Mechanical properties and in vivo osteogenesis in a rabbit femur model.

    PubMed

    Ke, Dongxu; Dernell, William; Bandyopadhyay, Amit; Bose, Susmita

    2015-11-01

    Tricalcium phosphate (TCP) is a bioceramic that is widely used in orthopedic and dental applications. TCP structures show excellent biocompatibility as well as biodegradability. In this study, porous β-TCP scaffolds were prepared by thermal decomposition of naphthalene. Scaffolds with 57.64% ± 3.54% density and a maximum pore size around 100 μm were fabricated via removing 30% naphthalene at 1150°C. The compressive strength for these scaffolds was 32.85 ± 1.41 MPa. Furthermore, by mixing 1 wt % SrO and 0.5 wt % SiO2 , pore interconnectivity improved, but the compressive strength decreased to 22.40 ± 2.70 MPa. However, after addition of polycaprolactone coating layers, the compressive strength of doped scaffolds increased to 29.57 ± 3.77 MPa. Porous scaffolds were implanted in rabbit femur defects to evaluate their biological property. The addition of dopants triggered osteoinduction by enhancing osteoid formation, osteocalcin expression, and bone regeneration, especially at the interface of the scaffold and host bone. This study showed processing flexibility to make interconnected porous scaffolds with different pore size and volume fraction porosity, while maintaining high compressive mechanical strength and excellent bioactivity. Results show that SrO/SiO2 -doped porous TCP scaffolds have excellent potential to be used in bone tissue engineering applications. © 2014 Wiley Periodicals, Inc.

  11. Anti-biofouling 3D porous systems: the blend effect of oxazoline-based oligomers on chitosan scaffolds.

    PubMed

    Correia, Vanessa G; Coelho, Margarida; Barroso, Telma; Raje, Vivek P; Bonifácio, Vasco D B; Casimiro, Teresa; Pinho, Mariana G; Aguiar-Ricardo, Ana

    2013-01-01

    The production, characterization and anti-biofouling activity of 3D porous scaffolds combining different blends of chitosan and oxazoline-based antimicrobial oligomers is reported. The incorporation of ammonium quaternized oligo(2-oxazoline)s into the composition of the scaffold enhances the stability of the chitosan scaffold under physiological conditions as well as its ability to repel protein adsorption. The blended scaffolds showed mean pore sizes in the range of 18-32 μm, a good pore interconnectivity and high porosity, as well as a large surface area, ultimate key features for anti-biofouling applications. Bovine serum albumin (BSA) adhesion profiles showed that the composition of the scaffolds plays a critical role in the chitosan-oligooxazoline system. Oligobisoxazoline-enriched scaffolds (20% w/w, CB8020) decreased protein adsorption (BSA) by up to 70%. Moreover, 1 mg of CB8020 was able to kill 99.9% of Escherichia coli cells upon contact, demonstrating its potential as promising material for production of tailored non-fouling 3D structures to be used in the construction of novel devices with applications in the biomedical field and water treatment processes.

  12. Comparison of different fabrication techniques used for processing 3-dimensional, porous, biodegradable scaffolds from modified starch for bone tissue engineering.

    PubMed

    Kunjachan, V; Subramanian, A; Hanna, M; Guan, J J

    2004-01-01

    3 dimensional, porous, biodegradable scaffolds were fabricated using modified starch of varying degree of substitution (DS) by extrusion processing. Freeze drying/lyophilization was also employed to fabricate scaffolds from modified starch. The research efforts have been focused on the comparison of the above-mentioned techniques by comparing the properties of the fabricated scaffolds in the paradigm of bone tissue engineering. The physicomechanical properties like porosity, compressive strength and modulus, pore size and microstructure were tested and analyzed by liquid replacement, mechanical testing and scanning electron microscopy respectively. The biodegradability of scaffolds was evaluated by soaking the samples in aqueous medium and Hank's balanced salt solution at 37-degree invitro. The cytotoxicity studies on these scaffolds were also conducted. The scaffolds have a 3D structure consisting of interconnected pores with good porosity, pore size, adequate compressive strength and modulus and exhibit good biodegradability as well as biocompatibility. After further optimization in the processing conditions and parameters they could be made useful for bone tissue engineering.

  13. Controlling protein release from scaffolds using polymer blends and composites.

    PubMed

    Ginty, Patrick J; Barry, John J A; White, Lisa J; Howdle, Steve M; Shakesheff, Kevin M

    2008-01-01

    We report the development of three protein loaded polymer blend and composite materials that modify the release kinetics of the protein from poly(dl-lactic acid) (P(dl)LA) scaffolds. P(dl)LA has been combined with either poly(ethylene glycol) (PEG), poly(caprolactone) (PCL) microparticles or calcium alginate fibres using supercritical CO(2) (scCO(2)) processing to form single and dual protein release scaffolds. P(dl)LA was blended with the hydrophilic polymer PEG using scCO(2) to increase the water uptake of the resultant scaffold and modify the release kinetics of an encapsulated protein. This was demonstrated by the more rapid release of the protein when compared to the release rate from P(dl)LA only scaffolds. For the P(dl)LA/alginate scaffolds, the protein loaded alginate fibres were processed into porous protein loaded P(dl)LA scaffolds using scCO(2) to produce dual release kinetics from the scaffolds. Protein release from the hydrophilic alginate fibres was more rapid in the initial stages, complementing the slower release from the slower degrading P(dl)LA scaffolds. In contrast, when protein loaded PCL particles were loaded into P(dl)LA scaffolds, the rate of protein release was retarded from the slow degrading PCL phase.

  14. Relationship between micro-porosity, water permeability and mechanical behavior in scaffolds for cartilage engineering.

    PubMed

    Vikingsson, L; Claessens, B; Gómez-Tejedor, J A; Gallego Ferrer, G; Gómez Ribelles, J L

    2015-08-01

    In tissue engineering the design and optimization of biodegradable polymeric scaffolds with a 3D-structure is an important field. The porous scaffold provide the cells with an adequate biomechanical environment that allows mechanotransduction signals for cell differentiation and the scaffolds also protect the cells from initial compressive loading. The scaffold have interconnected macro-pores that host the cells and newly formed tissue, while the pore walls should be micro-porous to transport nutrients and waste products. Polycaprolactone (PCL) scaffolds with a double micro- and macro-pore architecture have been proposed for cartilage regeneration. This work explores the influence of the micro-porosity of the pore walls on water permeability and scaffold compliance. A Poly(Vinyl Alcohol) with tailored mechanical properties has been used to simulate the growing cartilage tissue inside the scaffold pores. Unconfined and confined compression tests were performed to characterize both the water permeability and the mechanical response of scaffolds with varying size of micro-porosity while volume fraction of the macro-pores remains constant. The stress relaxation tests show that the stress response of the scaffold/hydrogel construct is a synergic effect determined by the performance of the both components. This is interesting since it suggests that the in vivo outcome of the scaffold is not only dependent upon the material architecture but also the growing tissue inside the scaffold׳s pores. On the other hand, confined compression results show that compliance of the scaffold is mainly controlled by the micro-porosity of the scaffold and less by hydrogel density in the scaffold pores. These conclusions bring together valuable information for customizing the optimal scaffold and to predict the in vivo mechanical behavior. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. 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

  16. A Route to Aliphatic Poly(ester)s with Thiol Pendant Groups: From Monomer Design to Editable Porous Scaffolds.

    PubMed

    Fuoco, Tiziana; Finne-Wistrand, Anna; Pappalardo, Daniela

    2016-04-11

    Biodegradable aliphatic polyesters such as poly(lactide) and poly(ε-caprolactone), largely used in tissue engineering applications, lack suitable functional groups and biological cues to enable interactions with cells. Because of the ubiquity of thiol groups in the biological environment and the pliability of thiol chemistry, we aimed to design and synthesize poly(ester) chains bearing pendant thiol-protected groups. To achieve this, 3-methyl-6-(tritylthiomethyl)-1,4-dioxane-2,5-dione, a lactide-type monomer possessing a pendant thiol-protected group, was synthesized. This molecule, when used as a monomer in controlled ring-opening polymerization in combination with lactide and ε-caprolactone, appeared to be a convenient "building block" for the preparation of functionalized aliphatic copolyesters, which were easily modified further. A polymeric sample bearing pyridyl disulfide groups, able to bind a cysteine-containing peptide, was efficiently obtained from a two-step modification reaction. Porous scaffolds were then prepared by blending this latter copolymer sample with poly(L-lactide-co-ε-caprolactone) followed by salt leaching. A further disulfide exchange reaction performed in aqueous medium formed porous scaffolds with covalently linked arginine-glycine-aspartic acid sequences. The scaffolds were characterized by thermal and mechanical tests, and scanning electron microscopy surface images revealed a highly porous morphology. Moreover, a cytotoxicity test indicated good cell viability.

  17. Effect of porous YSZ scaffold microstructure on the long-term performance of infiltrated Ni-YSZ anodes

    NASA Astrophysics Data System (ADS)

    Buyukaksoy, Aligul; Kammampata, Sanoop P.; Birss, Viola I.

    2015-08-01

    Ni infiltration into porous YSZ scaffolds is a promising route for the construction of high performing and redox-stable Ni-YSZ anodes for application in solid oxide fuel cells (SOFCs). However, the long-term instability of this type of anode is a critical problem. Here, it is shown that an interconnected Ni film, rather than discrete Ni particles, can be formed inside a porous, pre-sintered YSZ scaffold by using a polymeric Ni-based precursor as the infiltration medium. To understand the effect of the YSZ microstructure on the long-term stability and the electrochemical performance of the resulting composites, two types of Ni-YSZ anodes were investigated. Anodes prepared by polymeric Ni infiltration into a YSZ scaffold with large grains (0.5 μm) and pores (0.5 μm and 5 μm) showed extensive agglomeration in the Ni phase, resulting in poor stability and poor activity. In contrast, Ni infiltration into YSZ scaffolds with finer particle and pore sizes (∼200 nm each) produced anodes with a very small polarization resistance of ca. 0.1 Ω cm2 per electrode at 800 °C. An increase of only ∼5% was seen in the resistance after ca. 110 h at this temperature, achieved by preventing Ni agglomeration.

  18. Endothelial pattern formation in hybrid constructs of additive manufactured porous rigid scaffolds and cell-laden hydrogels for orthopedic applications.

    PubMed

    Shanjani, Yaser; Kang, Yunqing; Zarnescu, Livia; Ellerbee Bowden, Audrey K; Koh, Jeong-Tae; Ker, Dai Fei Elmer; Yang, Yunzhi

    2017-01-01

    Vascularization of tissue engineering constructs (TECs) in vitro is of critical importance for ensuring effective and satisfactory clinical outcomes upon implantation of TECs. Biomechanical properties of TECs have remarkable influence on the in vitro vascularization of TECs. This work utilized in vitro experiments and finite element analysis to investigate endothelial patterns in hybrid constructs of soft collagen gels and rigid macroporous poly(ε-caprolactone)-β-tricalcium phosphate (PCL-β-TCP) scaffold seeded/embedded with human umbilical vein endothelial cells (HUVECs) for bone tissue engineering applications. We first fabricated and characterized well-defined porous PCL-β-TCP scaffolds with identical pore size (500µm) but different strut sizes (200 and 400µm) using additive manufacturing (AM) technology, and then assessed the HUVEC׳s proliferation and morphogenesis within collagen, PCL-β-TCP scaffold, and the collagen-scaffold hybrid construct. Results showed that, in the hybrid construct, the cell population in the collagen component dropped by day 7 but then increased by day 14. Also, cells migrated onto the struts of the scaffold component, proliferated over time, and formed networks on the thinner struts (i.e., 200µm). Also, the thinner struts resulted in formation of long linear cellular cords structures within the pores. Finite element simulation demonstrated principal stress patterns similar to the observed cell-network pattern. It is probable that the scaffold component modulated patterns of principal stresses in the collagen component as biomechanical cues for reorganization of cell network patterns. Also, the scaffold component significantly improved the mechanical integrity of hydrogel component in the hybrid construct for weight-bearing applications. These results have collectively indicated that the manipulation of micro-architecture of scaffold could be an effective means to further regulate and guide desired cellular response in hybrid

  19. Comprehensive Genetic Analysis of Early Host Body Reactions to the Bioactive and Bio-Inert Porous Scaffolds

    PubMed Central

    Ehashi, Tomo; Takemura, Taro; Hanagata, Nobutaka; Minowa, Takashi; Kobayashi, Hisatoshi; Ishihara, Kazuhiko; Yamaoka, Tetsuji

    2014-01-01

    To design scaffolds for tissue regeneration, details of the host body reaction to the scaffolds must be studied. Host body reactions have been investigated mainly by immunohistological observations for a long time. Despite of recent dramatic development in genetic analysis technologies, genetically comprehensive changes in host body reactions are hardly studied. There is no information about host body reactions that can predict successful tissue regeneration in the future. In the present study, porous polyethylene scaffolds were coated with bioactive collagen or bio-inert poly(2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate) (PMB) and were implanted subcutaneously and compared the host body reaction to those substrates by normalizing the result using control non-coat polyethylene scaffold. The comprehensive analyses of early host body reactions to the scaffolds were carried out using a DNA microarray assay. Within numerous genes which were expressed differently among these scaffolds, particular genes related to inflammation, wound healing, and angiogenesis were focused upon. Interleukin (IL)-1β and IL-10 are important cytokines in tissue responses to biomaterials because IL-1β promotes both inflammation and wound healing and IL-10 suppresses both of them. IL-1β was up-regulated in the collagen-coated scaffold. Collagen-specifically up-regulated genes contained both M1- and M2-macrophage-related genes. Marked vessel formation in the collagen-coated scaffold was occurred in accordance with the up-regulation of many angiogenesis-inducible factors. The DNA microarray assay provided global information regarding the host body reaction. Interestingly, several up-regulated genes were detected even on the very bio-inert PMB-coated surfaces and those genes include inflammation-suppressive and wound healing-suppressive IL-10, suggesting that not only active tissue response but also the inert response may relates to these genetic regulations. PMID:24454803

  20. Finite-element design and optimization of a three-dimensional tetrahedral porous titanium scaffold for the reconstruction of mandibular defects.

    PubMed

    Luo, Danmei; Rong, Qiguo; Chen, Quan

    2017-09-01

    Reconstruction of segmental defects in the mandible remains a challenge for maxillofacial surgery. The use of porous scaffolds is a potential method for repairing these defects. Now, additive manufacturing techniques provide a solution for the fabrication of porous scaffolds with specific geometrical shapes and complex structures. The goal of this study was to design and optimize a three-dimensional tetrahedral titanium scaffold for the reconstruction of mandibular defects. With a fixed strut diameter of 0.45mm and a mean cell size of 2.2mm, a tetrahedral structural porous scaffold was designed for a simulated anatomical defect derived from computed tomography (CT) data of a human mandible. An optimization method based on the concept of uniform stress was performed on the initial scaffold to realize a minimal-weight design. Geometric and mechanical comparisons between the initial and optimized scaffold show that the optimized scaffold exhibits a larger porosity, 81.90%, as well as a more homogeneous stress distribution. These results demonstrate that tetrahedral structural titanium scaffolds are feasible structures for repairing mandibular defects, and that the proposed optimization scheme has the ability to produce superior scaffolds for mandibular reconstruction with better stability, higher porosity, and less weight. Copyright © 2017 IPEM. Published by Elsevier Ltd. All rights reserved.

  1. Biodegradable HA-PLA 3-D porous scaffolds: effect of nano-sized filler content on scaffold properties.

    PubMed

    Kothapalli, Chandrasekhar R; Shaw, Montgomery T; Wei, Mei

    2005-11-01

    Scaffolds comprising poly(lactic acid) and nano-hydroxyapatite (HA) were prepared using the solvent-casting/salt-leaching technique. NaCl was used as the leaching agent. Nano-sized HA was synthesized by a hydrothermal method at 170 degrees C and autogenous pressure. High-resolution TEM imaging revealed that the HA particles were ellipsoidal-shaped with needle-like morphologies. The particles had an average size of approximately 25 nm in width and 150 nm in length with aspect ratios ranging from 6 to 8. As the HA content increased in the scaffold from 0 to 50 wt%, the compression modulus of the scaffolds increased from 4.72+/-1.2 to 9.87+/-1.8 MPa, while the yield strength from 0.29+/-0.03 to 0.44+/-0.01 MPa. Such polymeric scaffolds should be suitable materials for non-load sharing tissue-engineering applications.

  2. Porous-Nickel-Scaffolded Tin-Antimony Anodes with Enhanced Electrochemical Properties for Li/Na-Ion Batteries.

    PubMed

    Li, Jiachen; Pu, Jun; Liu, Ziqiang; Wang, Jian; Wu, Wenlu; Zhang, Huigang; Ma, Haixia

    2017-08-02

    The energy and power densities of rechargeable batteries urgently need to be increased to meet the ever-increasing demands of consumer electronics and electric vehicles. Alloy anodes are among the most promising candidates for next-generation high-capacity battery materials. However, the high capacities of alloy anodes usually suffer from some serious difficulties related to the volume changes of active materials. Porous supports and nanostructured alloy materials have been explored to address these issues. However, these approaches seemingly increase the active material-based properties and actually decrease the electrode-based capacity because of the oversized pores and heavy mass of mechanical supports. In this study, we developed an ultralight porous nickel to scaffold with high-capacity SnSb alloy anodes. The porous-nickel-supported SnSb alloy demonstrates a high specific capacity and good cyclability for both Li-ion and Na-ion batteries. Its capacity retains 580 mA h g(-1) at 2 A g(-1) after 100 cycles in Li-ion batteries. For a Na-ion battery, the composite electrode can even deliver a capacity of 275 mA h g(-1) at 1 A g(-1) after 1000 cycles. This study demonstrates that combining the scaffolding function of ultralight porous nickel and the high capacity of the SnSb alloy can significantly enhance the electrochemical performances of Li/Na-ion batteries.

  3. Production, characterisation, and cytocompatibility of porous titanium-based particulate scaffolds.

    PubMed

    Luthringer, B J C; Ali, F; Akaichi, H; Feyerabend, F; Ebel, T; Willumeit, R

    2013-10-01

    Despite its non-matching mechanical properties titanium remains the preferred metal implant material in orthopaedics. As a consequence in some cases stress shielding effect occurs, leading to implant loosening, osteopenia, and finally revision surgery. Porous metal scaffolds to allow easier specialised cells ingrowth with mechanical properties closer to the ones of bone can overcome this problem. This should improve healing processes, implant integration, and dynamic strength of implants retaining. Three Ti-6Al-4V materials were metal injection moulded and tailored porosities were effectively achieved. After microstructural and mechanical characterisation, two different primary cells of mesenchymal origin (human umbilical cord perivascular cells and human bone derived cells which revealed to be two pertinent models) as well as one cell line originated from primary osteogenic sarcoma, Saos-2, were bestowed to investigate cell-material interaction on genomic and proteome levels. Biological examinations disclosed that no material has negative impact on early adhesion, proliferation or cell viability. An efficient cell ingrowth into material with an average porosity of 25-50 μm was proved.

  4. Morphological examination of highly porous polylactic acid/Bioglass(®) scaffolds produced via nonsolvent induced phase separation.

    PubMed

    Rezabeigi, Ehsan; Wood-Adams, Paula M; Drew, Robin A L

    2016-09-19

    In this study, we produce highly porous (up to ∼91%) composite scaffolds of polylactic acid (PLA) containing 2 wt % sol-gel-derived 45S5 Bioglass(®) particles via nonsolvent induced phase separation at -23°C with no sacrificial phases involved. Before the incorporation of the bioglass with PLA, the particles are surface modified with a silane coupling agent which effectively diminishes agglomeration between them leading to a better dispersion of bioactive particles throughout the scaffold. Interestingly, the incorporation route (via solvent dichloromethane or nonsolvent hexane) of the surface modified particles in the foaming process has the greatest impact on porosity, crystallinity, and morphology of the scaffolds. The composite scaffolds with a morphology consisting of both mesopores and large macropores, which is potentially beneficial for bone regeneration applications, are examined further. SEM images show that the surface modified bioglass particles take-up a unique configuration within the mesoporous structure of these scaffolds ensuring that the particles are well interlocked but not completely covered by PLA such that they can be in contact with physiological fluids. The results of preliminary in vitro tests confirm that this PLA/bioglass configuration promotes the interaction of the bioactive phase with physiological fluids. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2016.

  5. Study on surface modification of porous apatite-wollastonite bioactive glass ceramic scaffold

    NASA Astrophysics Data System (ADS)

    Cao, Bin; Zhou, Dali; Xue, Ming; Li, Guangda; Yang, Weizhong; Long, Qin; Ji, Li

    2008-11-01

    Chitosan (CS) was used to modify the surface of apatite-wollastonite bioactive glass ceramic (AW GC) scaffold to prepare AW/CS composite scaffold. The in vitro bioactivity of the AW/CS composite scaffold was investigated by simulated body fluid (SBF) soaking experiment. Cell growth on the surface of the material was evaluated by co-culturing osteogenic marrow stromal cells (MSCs) of rabbits with the scaffold. The results showed that the compressive strength of AW GC scaffold was improved dramatically after being modified by CS, whereas the mineralization rate was delayed. MSCs can attach well on the surface of the composite scaffold.

  6. Stabilization of porous chitosan improves the performance of its association with platelet-rich plasma as a composite scaffold.

    PubMed

    Shimojo, A A M; Perez, A G M; Galdames, S E M; Brissac, I C S; Santana, M H A

    2016-03-01

    This study offers innovative perspectives for optimizing of scaffolds based on correlation structure-function aimed the regenerative medicine. Thus, we evaluated in vitro performance of stabilized porous chitosan (SPCHTs) associated with activated platelet-rich plasma (aP-PRP) as a composite scaffold for the proliferation and osteogenic differentiation of human adipose-derived mesenchymal stem cells (h-AdMSCs). The porous structure of chitosan (PCHT) was prepared similarly to solid sponges by controlled freezing (-20 °C) and lyophilization of a 3% (w/v) chitosan solution. Stabilization was performed by treating the PCHT with sodium hydroxide (TNaOH), an ethanol series (TEtOH) or by crosslinking with tripolyphosphate (CTPP). The aP-PRP was obtained from the controlled centrifugation of whole blood and activated with autologous serum and calcium. Imaging of the structures showed fibrin networks inside and on the surface of SPCHTs as a consequence of electrostatic interactions. SPCHTs were non-cytotoxic, and the porosity, pore size and Young's modulus were approximately 96%, 145 μm and 1.5 MPa for TNaOH and TEtOH and 94%, 110 μm and 1.8 MPa for CTPP, respectively. Stabilization maintained the integrity of the SPCHTs for at least 10 days of cultivation. SPCHTs showed controlled release of the growth factors TGF-β1 and PDGF-AB. Although generating different patterns, all of the stabilization treatments improved the proliferation of seeded h-AdMSCs on the composite scaffold compared to aP-PRP alone, and differentiation of the composite scaffold treated with TEtOH was significantly higher than for non-stabilized PCHT. We conclude that the composite scaffolds improved the in vitro performance of PRP and have potential in regenerative medicine.

  7. Image-Based Three-Dimensional Analysis to Characterize the Texture of Porous Scaffolds

    PubMed Central

    Pennella, Francesco; Gallo, Diego; Ciardelli, Gianluca; Bignardi, Cristina; Audenino, Alberto; Morbiducci, Umberto

    2014-01-01

    The aim of the present study is to characterize the microstructure of composite scaffolds for bone tissue regeneration containing different ratios of chitosan/gelatin blend and bioactive glasses. Starting from realistic 3D models of the scaffolds reconstructed from micro-CT images, the level of heterogeneity of scaffold architecture is evaluated performing a lacunarity analysis. The results demonstrate that the presence of the bioactive glass component affects not only macroscopic features such as porosity, but mainly scaffold microarchitecture giving rise to structural heterogeneity, which could have an impact on the local cell-scaffold interaction and scaffold performances. The adopted approach allows to investigate the scale-dependent pore distribution within the scaffold and the related structural heterogeneity features, providing a comprehensive characterization of the scaffold texture. PMID:24995272

  8. Gas anti-solvent precipitation assisted salt leaching for generation of micro- and nano-porous wall in bio-polymeric 3D scaffolds.

    PubMed

    Flaibani, Marina; Elvassore, Nicola

    2012-08-01

    The mass transport through biocompatible and biodegradable polymeric 3D porous scaffolds may be depleted by non-porous impermeable internal walls. As consequence the concentration of metabolites and growth factors within the scaffold may be heterogeneous leading to different cell fate depending on spatial cell location, and in some cases it may compromise cell survival. In this work, we fabricated polymeric scaffolds with micro- and nano-scale porosity by developing a new technique that couples two conventional scaffold production methods: solvent casting-salt leaching and gas antisolvent precipitation. 10-15 w/w solutions of a hyaluronic benzyl esters (HYAFF11) and poly-(lactic acid) (PLA) were used to fill packed beds of 0.177-0.425 mm NaCl crystals. The polymer precipitation in micro and nano-porous structures between the salt crystals was induced by high-pressure gas, then its flushing extracted the residual solvent. The salt was removed by water-wash. Morphological analysis by scanning electron microscopy showed a uniform porosity (~70%) and a high interconnectivity between porous. The polymeric walls were porous themselves counting for 30% of the total porosity. This wall porosity did not lead to a remarkable change in compressive modulus, deformation, and rupture pressure. Scaffold biocompatibility was tested with murine muscle cell line C2C12 for 4 and 7 days. Viability analysis and histology showed that micro- and nano-porous scaffolds are biocompatible and suitable for 3D cell culture promoting cell adhesion on the polymeric wall and allowing their proliferation in layers. Micro- and nano-scale porosities enhance cell migration and growth in the inner part of the scaffold.

  9. Novel biodegradable poly(propylene fumarate)-co-poly(l-lactic acid) porous scaffolds fabricated by phase separation for tissue engineering applications

    PubMed Central

    Liu, Xifeng; Miller, A. Lee; Waletzki, Brian E.; Yaszemski, Michael J.

    2015-01-01

    Scaffolds with intrinsically interconnected porous structures are highly desirable in tissue engineering and regenerative medicine. In this study, three-dimensional polymer scaffolds with highly interconnected porous structures were fabricated by thermally induced phase separation of novel synthesized biodegradable poly(propylene fumarate)-co-poly(l-lactic acid) in a dioxane/water binary system. Defined porous scaffolds were achieved by optimizing conditions to attain interconnected porous structures. The effect of phase separation parameters on scaffold morphology were investigated, including polymer concentration (1, 3, 5, 7, and 9%), quench time (1, 4, and 8 min), dioxane/water ratio (83/17, 85/15, and 87/13 wt/wt), and freeze temperature (−20, −80, and −196 °C). Interesting pore morphologies were created by adjusting these processing parameters, e.g., flower-shaped (5%; 85/15; 1 min; −80 °C), spherulite-like (5%; 85/15; 8 min; −80 °C), and bead-like (5%; 87/13; 1 min; −80 °C) morphology. Modulation of phase separation conditions also resulted in remarkable differences in scaffold porosities (81% to 91%) and thermal properties. Furthermore, scaffolds with varied mechanic strengths, degradation rates, and protein adsorption capabilities could be fabricated using the phase separation method. In summary, this work provides an effective route to generate multi-dimensional porous scaffolds that can be applied to a variety of hydrophobic polymers and copolymers. The generated scaffolds could potentially be useful for various tissue engineering applications including bone tissue engineering. PMID:26989483

  10. Ingrowth of human mesenchymal stem cells into porous silk particle reinforced silk composite scaffolds: An in vitro study.

    PubMed

    Rockwood, Danielle N; Gil, Eun Seok; Park, Sang-Hyug; Kluge, Jonathan A; Grayson, Warren; Bhumiratana, Sarindr; Rajkhowa, Rangam; Wang, Xungai; Kim, Sung Jun; Vunjak-Novakovic, Gordana; Kaplan, David L

    2011-01-01

    Silk fibroin protein is biodegradable and biocompatible, exhibiting excellent mechanical properties for various biomedical applications. However, porous three-dimensional (3-D) silk fibroin scaffolds, or silk sponges, usually fall short in matching the initial mechanical requirements for bone tissue engineering. In the present study, silk sponge matrices were reinforced with silk microparticles to generate protein-protein composite scaffolds with desirable mechanical properties for in vitro osteogenic tissue formation. It was found that increasing the silk microparticle loading led to a substantial increase in the scaffold compressive modulus from 0.3 MPa (non-reinforced) to 1.9 MPa for 1:2 (matrix:particle) reinforcement loading by dry mass. Biochemical, gene expression, and histological assays were employed to study the possible effects of increasing composite scaffold stiffness, due to microparticle reinforcement, on in vitro osteogenic differentiation of human mesenchymal stem cells (hMSCs). Increasing silk microparticle loading increased the osteogenic capability of hMSCs in the presence of bone morphogenic protein-2 (BMP-2) and other osteogenic factors in static culture for up to 6 weeks. The calcium adsorption increased dramatically with increasing loading, as observed from biochemical assays, histological staining, and microcomputer tomography (μCT) analysis. Specifically, calcium content in the scaffolds increased by 0.57, 0.71, and 1.27 mg (per μg of DNA) from 3 to 6 weeks for matrix to particle dry mass loading ratios of 1:0, 1:1, and 1:2, respectively. In addition, μCT imaging revealed that at 6 weeks, bone volume fraction increased from 0.78% for non-reinforced to 7.1% and 6.7% for 1:1 and 1:2 loading, respectively. Our results support the hypothesis that scaffold stiffness may strongly influence the 3-D in vitro differentiation capabilities of hMSCs, providing a means to improve osteogenic outcomes.

  11. Hyaluronic acid doped-poly(3,4-ethylenedioxythiophene)/chitosan/gelatin (PEDOT-HA/Cs/Gel) porous conductive scaffold for nerve regeneration.

    PubMed

    Wang, Shuping; Guan, Shui; Zhu, Zhibo; Li, Wenfang; Liu, Tianqing; Ma, Xuehu

    2017-02-01

    Conducting polymer, as a "smart" biomaterial, has been increasingly used to construct tissue engineered scaffold for nerve tissue regeneration. In this study, a novel porous conductive scaffold was prepared by incorporating conductive hyaluronic acid (HA) doped-poly(3,4-ethylenedioxythiophene) (PEDOT-HA) nanoparticles into a chitosan/gelatin (Cs/Gel) matrix. The physicochemical characteristics of Cs/Gel scaffold with 0-10wt% PEDOT-HA were analyzed and the results indicated that the incorporation of PEDOT-HA into scaffold increased the electrical and mechanical properties while decreasing the porosity and water absorption. Moreover, in vitro biodegradation of scaffold displayed a declining trend with the PEDOT-HA content increased. About the biocompatibility of conductive scaffold, neuron-like rat phaeochromocytoma (PC12) cells were cultured in scaffold to evaluate cell adhesion and growth. 8% PEDOT-HA/Cs/Gel scaffold had a higher cell adhesive efficiency and cell viability than the other conductive scaffolds. Furthermore, cells in the scaffold with 8wt% PEDOT-HA expressed higher synapse growth gene of GAP43 and SYP compared with Cs/Gel control group. These results suggest that 8%PEDOT-HA/Cs/Gel scaffold is an attractive cell culture conductive substrate which could support cell adhesion, survival, proliferation, and synapse growth for the application in nerve tissue regeneration. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. 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.

  13. In vitro and in vivo evaluation of biodegradable, open-porous scaffolds made of sintered magnesium W4 short fibres.

    PubMed

    Bobe, K; Willbold, E; Morgenthal, I; Andersen, O; Studnitzky, T; Nellesen, J; Tillmann, W; Vogt, C; Vano, K; Witte, F

    2013-11-01

    A cytocompatible and biocompatible, degradable, open-porous, mechanically adaptable metal scaffold made of magnesium alloy W4 melt-extracted short fibres was fabricated by liquid phase sintering. Cylindrical samples (3×5 mm) of sintered W4 short fibres were evaluated under in vitro (L929, HOB, eudiometer, weight loss) and in vivo conditions (rabbits: 6 and 12 weeks). The in vitro corrosion environment (e.g., temperature, flow, composition of corrosion solution, exposure time) significantly influenced the corrosion rates of W4 scaffolds compared with corrosion in vivo. Corrosion rates under cell culture conditions for 72 h varied from 1.05 to 3.43 mm y(-1) depending on the media composition. Corrosion rates measured in eudiometric systems for 24 h were ~24-27 times higher (3.88-4.43 mm y(-1)) than corrosion in vivo after 6 weeks (0.16 mm y(-1)). Moreover, it was found that the cell culture media composition significantly influences the ionic composition of the extract by selectively dissolving ions from W4 samples or their corrosion products. A pilot in vivo study for 6 and 12 weeks demonstrated active bone remodelling, no foreign body reaction and no clinical observation of gas formation during W4 scaffold implantation. Long-term in vivo studies need to be conducted to prove complete degradation of the W4 scaffold and total replacement by the host tissue.

  14. Production of Hollow Bacterial Cellulose Microspheres Using Microfluidics to Form an Injectable Porous Scaffold for Wound Healing.

    PubMed

    Yu, Jiaqing; Huang, Tzu-Rung; Lim, Zhen Han; Luo, Rongcong; Pasula, Rupali Reddy; Liao, Lun-De; Lim, Sierin; Chen, Chia-Hung

    2016-12-01

    Bacterial cellulose (BC) is a biocompatible material with high purity and robust mechanical strength used to fabricate desirable scaffolds for 3D cell culture and wound healing. However, the chemical resistance of BC and its insolubility in the majority of solutions make it difficult to manipulate using standard chemical methods. In this study, a microfluidic process is developed to produce hollow BC microspheres with desirable internal structures and morphology. Microfluidics is used to generate a core-shell structured microparticle with an alginate core and agarose shell as a template to encapsulate Gluconacetobacter xylinus for long-term static culture. G. xylinus then secretes BC, which becomes entangled within the shell of the structured hydrogel microparticles and forms BC microspheres. The removal of the hydrogel template via thermal-chemical treatments yields robust BC microspheres exhibiting a hollow morphology. These hollow microspheres spontaneously assemble as functional units to form a novel injectable scaffold. In vitro, a highly porous scaffold is created to enable effective 3D cell culture with a high cell proliferation rate and better depth distribution. In vivo, this injectable scaffold facilitates tissue regeneration, resulting in rapid wound-healing in a Sprague Dawley rat skin model. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. 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.

  16. Preparation of nano/macroporous polycaprolactone microspheres for an injectable cell delivery system using room temperature ionic liquid and camphene.

    PubMed

    Kim, Seong Yeol; Hwang, Ji-Young; Shin, Ueon Sang

    2016-03-01

    The nano/macroporous polycaprolactone (PCL) microspheres with cell active surfaces were developed as an injectable cell delivery system. Room temperature ionic liquid (RTIL) and camphene were used as a liquid mold and a porogen, respectively. Various-sized spheres of 244-601μm with pores of various size and shape of 0.02-100μm, were formed depending on the camphene/RTIL ratio (0.8-2.6). To give cell activity, the surface of porous microspheres were further modified with nerve growth factors (NGF) containing gelatin to give a thin NGF/gelatin layer, to which the neural progenitor cells (PC-12) attached and extended their neurites on to the surface layers of the microspheres. The developed microspheres may be potentially applicable as a neuronal cell delivery scaffold for neuron tissue engineering.

  17. Microfabrication of complex porous tissue engineering scaffolds using 3D projection stereolithography

    PubMed Central

    Gauvin, Robert; Chen, Ying-Chieh; Lee, Jin Woo; Soman, Pranav; Zorlutuna, Pinar; Nichol, Jason W.; Bae, Hojae; Chen, Shaochen; Khademhosseini, Ali

    2013-01-01

    The success of tissue engineering will rely on the ability to generate complex, cell seeded three-dimensional (3D) structures. Therefore, methods that can be used to precisely engineer the architecture and topography of scaffolding materials will represent a critical aspect of functional tissue engineering. Previous approaches for 3D scaffold fabrication based on top-down and process driven methods are often not adequate to produce complex structures due to the lack of control on scaffold architecture, porosity, and cellular interactions. The proposed projection stereolithography (PSL) platform can be used to design intricate 3D tissue scaffolds that can be engineered to mimic the microarchitecture of tissues, based on computer aided design (CAD). The PSL system was developed, programmed and optimized to fabricate 3D scaffolds using gelatin methacrylate (GelMA). Variation of the structure and prepolymer concentration enabled tailoring the mechanical properties of the scaffolds. A dynamic cell seeding method was utilized to improve the coverage of the scaffold throughout its thickness. The results demonstrated that the interconnectivity of pores allowed for uniform human umbilical vein endothelial cells (HUVECs) distribution and proliferation in the scaffolds, leading to high cell density and confluency at the end of the culture period. Moreover, immunohistochemistry results showed that cells seeded on the scaffold maintained their endothelial phenotype, demonstrating the biological functionality of the microfabricated GelMA scaffolds. PMID:22365811

  18. The generation of biomolecular patterns in highly porous collagen-GAG scaffolds using direct photolithography.

    PubMed

    Martin, Teresa A; Caliari, Steven R; Williford, Paul D; Harley, Brendan A; Bailey, Ryan C

    2011-06-01

    The extracellular matrix (ECM) is a complex organization of structural proteins found within tissues and organs. Heterogeneous tissues with spatially and temporally modulated properties play an important role in organism physiology. Here we present a benzophenone (BP) based direct, photolithographic approach to spatially pattern solution phase biomolecules within collagen-GAG (CG) scaffolds and demonstrate creation of a wide range of patterns composed of multiple biomolecular species in a manner independent from scaffold fabrication steps. We demonstrate the ability to immobilize biomolecules at surface densities of up to 1000 ligands per square micron on the scaffold strut surface and to depths limited by the penetration depth of the excitation source into the scaffold structure. Importantly, while BP photopatterning does further crosslink the CG scaffold, evidenced by increased mechanical properties and collagen crystallinity, it does not affect scaffold microstructural or compositional properties or negatively influence cell adhesion, viability, or proliferation. We show that covalently photoimmobilized fibronectin within a CG scaffold significantly increases the speed of MC3T3-E1 cell attachment relative to the bare CG scaffold or non-specifically adsorbed fibronectin, suggesting that this approach can be used to improve scaffold bioactivity. Our findings, on the whole, establish the use of direct, BP photolithography as a methodology for covalently incorporating activity-improving biochemical cues within 3D collagen biomaterial scaffolds with spatial control over biomolecular deposition. Copyright © 2011 Elsevier Ltd. All rights reserved.

  19. 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.

  20. Kinetics and isotherm of fibronectin adsorption to three-dimensional porous chitosan scaffolds explored by 125I-radiolabelling

    PubMed Central

    Amaral, Isabel F.; Sousa, Susana R.; Neiva, Ismael; Marcos-Silva, Lara; Kirkpatrick, Charles J.; Barbosa, Mário A.; Pêgo, Ana P.

    2013-01-01

    In this study, 125I-radiolabelling was explored to follow the kinetics and isotherm of fibronectin (FN) adsorption to porous polymeric scaffolds, as well as to assess the elution and exchangeability of pre-adsorbed FN following incubation in serum-containing culture medium. Chitosan (CH) porous scaffolds with two different degrees of acetylation (DA 4% and 15%) were incubated in FN solutions with concentrations ranging from 5 to 50 µg/mL. The kinetic and isotherm of FN adsorption to CH were successfully followed using 125I-FN as a tracer molecule. While on DA 4% the levels of adsorbed FN increased linearly with FN solution concentration, on DA 15% a saturation plateau was attained, and FN adsorbed amounts were significantly lower. These findings were supported by immunofluorescent studies that revealed, for the same FN solution concentration, higher levels of exposed cell-binding domains on DA 4% as compared with DA 15%. Following incubation in serum containing medium, DA 4% also revealed higher ability to exchange pre-adsorbed FN by new FN molecules from serum than DA 15%. In accordance, when assessing the efficacy of passively adsorbed FN to promote endothelial cell (EC) adhesion to CH, ECs were found to adhere at higher levels to DA 4% as compared with DA 15%, 5 µg/mL of FN being already efficient in promoting cell adhesion and cytoskeletal organization on CH with DA 4%. Taken together the results show that protein radiolabelling can be used as an effective tool to study protein adsorption to porous polymeric scaffolds, both from single and complex protein solutions. PMID:23635535

  1. Load-adaptive scaffold architecturing: a bioinspired approach to the design of porous additively manufactured scaffolds with optimized mechanical properties.

    PubMed

    Rainer, Alberto; Giannitelli, Sara M; Accoto, Dino; De Porcellinis, Stefano; Guglielmelli, Eugenio; Trombetta, Marcella

    2012-04-01

    Computer-Aided Tissue Engineering (CATE) is based on a set of additive manufacturing techniques for the fabrication of patient-specific scaffolds, with geometries obtained from medical imaging. One of the main issues regarding the application of CATE concerns the definition of the internal architecture of the fabricated scaffolds, which, in turn, influences their porosity and mechanical strength. The present study envisages an innovative strategy for the fabrication of highly optimized structures, based on the a priori finite element analysis (FEA) of the physiological load set at the implant site. The resulting scaffold micro-architecture does not follow a regular geometrical pattern; on the contrary, it is based on the results of a numerical study. The algorithm was applied to a solid free-form fabrication process, using poly(ε-caprolactone) as the starting material for the processing of additive manufactured structures. A simple and intuitive geometry was chosen as a proof-of-principle application, on which finite element simulations and mechanical testing were performed. Then, to demonstrate the capability in creating mechanically biomimetic structures, the proximal femur subjected to physiological loading conditions was considered and a construct fitting a femur head portion was designed and manufactured.

  2. Novel real function based method to construct heterogeneous porous scaffolds and additive manufacturing for use in medical engineering.

    PubMed

    Yang, Nan; Tian, Yanling; Zhang, Dawei

    2015-11-01

    Heterogeneous porous scaffolds have important applications in biomedical engineering, as they can mimic the structures of natural tissues to achieve the corresponding properties. Here, we introduce a new and easy to implement real function based method for constructing complex, heterogeneous porous structures, including hybrid structures, stochastic structures, functionally gradient structures, and multi-scale structures, or their combinations (e.g., hybrid multi-scale structures). Based on micro-CT data, a femur-mimetic structure with gradient morphology was constructed using our method and fabricated using stereolithography. Results showed that our method could generate gradient porosity or gradient specific surfaces and be sufficiently flexible for use with micro-CT data and additive manufacturing (AM) techniques.

  3. Complex flow characterisation of a porous tissue scaffold measured by Doppler optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Tomlins, Peter H.; Wang, Yiwei; Zhang, Bufa; Tedaldi, Matthew; Tomlins, Paul E.

    2008-02-01

    The flow of culture medium through a mechanically stimulated cell-seeded tissue scaffold is a factor influencing not only the transport of essential nutrients and waste product removal but also impacting on the degradation kinetics of the scaffold. Being able to map spatial and temporal changes in fluid flow behaviour is key to the development of improved bioreactors and tissue scaffold designs, especially for the new generation of multiple tissue reactors. In this paper we demonstrate the excellent metrological benefits of fast Doppler optical coherence tomography for time-lapse characterisation of tissue scaffolds placed in a dynamic flow environment.

  4. Fabrication and characterization of a rapid prototyped tissue engineering scaffold with embedded multicomponent matrix for controlled drug release.

    PubMed

    Chen, Muwan; Le, Dang Q S; Hein, San; Li, Pengcheng; Nygaard, Jens V; Kassem, Moustapha; Kjems, Jørgen; Besenbacher, Flemming; Bünger, Cody

    2012-01-01

    Bone tissue engineering implants with sustained local drug delivery provide an opportunity for better postoperative care for bone tumor patients because these implants offer sustained drug release at the tumor site and reduce systemic side effects. A rapid prototyped macroporous polycaprolactone scaffold was embedded with a porous matrix composed of chitosan, nanoclay, and β-tricalcium phosphate by freeze-drying. This composite scaffold was evaluated on its ability to deliver an anthracycline antibiotic and to promote formation of mineralized matrix in vitro. Scanning electronic microscopy, confocal imaging, and DNA quantification confirmed that immortalized human bone marrow-derived mesenchymal stem cells (hMSC-TERT) cultured in the scaffold showed high cell viability and growth, and good cell infiltration to the pores of the scaffold. Alkaline phosphatase activity and osteocalcin staining showed that the scaffold was osteoinductive. The drug-release kinetics was investigated by loading doxorubicin into the scaffold. The scaffolds comprising nanoclay released up to 45% of the drug for up to 2 months, while the scaffold without nanoclay released 95% of the drug within 4 days. Therefore, this scaffold can fulfill the requirements for both bone tissue engineering and local sustained release of an anticancer drug in vitro. These results suggest that the scaffold can be used clinically in reconstructive surgery after bone tumor resection. Moreover, by changing the composition and amount of individual components, the scaffold can find application in other tissue engineering areas that need local sustained release of drug.

  5. Fabrication and characterization of a rapid prototyped tissue engineering scaffold with embedded multicomponent matrix for controlled drug release

    PubMed Central

    Chen, Muwan; Le, Dang QS; Hein, San; Li, Pengcheng; Nygaard, Jens V; Kassem, Moustapha; Kjems, Jørgen; Besenbacher, Flemming; Bünger, Cody

    2012-01-01

    Bone tissue engineering implants with sustained local drug delivery provide an opportunity for better postoperative care for bone tumor patients because these implants offer sustained drug release at the tumor site and reduce systemic side effects. A rapid prototyped macroporous polycaprolactone scaffold was embedded with a porous matrix composed of chitosan, nanoclay, and β-tricalcium phosphate by freeze-drying. This composite scaffold was evaluated on its ability to deliver an anthracycline antibiotic and to promote formation of mineralized matrix in vitro. Scanning electronic microscopy, confocal imaging, and DNA quantification confirmed that immortalized human bone marrow-derived mesenchymal stem cells (hMSC-TERT) cultured in the scaffold showed high cell viability and growth, and good cell infiltration to the pores of the scaffold. Alkaline phosphatase activity and osteocalcin staining showed that the scaffold was osteoinductive. The drug-release kinetics was investigated by loading doxorubicin into the scaffold. The scaffolds comprising nanoclay released up to 45% of the drug for up to 2 months, while the scaffold without nanoclay released 95% of the drug within 4 days. Therefore, this scaffold can fulfill the requirements for both bone tissue engineering and local sustained release of an anticancer drug in vitro. These results suggest that the scaffold can be used clinically in reconstructive surgery after bone tumor resection. Moreover, by changing the composition and amount of individual components, the scaffold can find application in other tissue engineering areas that need local sustained release of drug. PMID:22904634

  6. 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.

  7. 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.

  8. A study on the effect of degradation media on the physical and mechanical properties of porous PLGA 85/15 scaffolds.

    PubMed

    Perron, Josee K; Naguib, Hani E; Daka, Joseph; Chawla, Attar; Wilkins, Ruth

    2009-11-01

    This study investigates the effect of PLGA 85/15 scaffold on the cell growth and viability of a cell line, and the degradation of the scaffold in different media. The cell line used was human promyelocytic leukemia cells (HL-60). Three different media were considered: distilled water, a phosphate buffered saline (PBS) solution, and HL-60 cell line. Porous PLGA 85/15 scaffolds were prepared with an optimized gas foaming/salt leaching technique using a NaCl/polymer mass ratio of five, a saturation pressure of 5.52 MPa and a saturation time of 12 h. The cell growth and viability were not impaired by the presence of the scaffold. The mass change of the scaffold due to degradation over the period was varied only by 4% across all three media. The average macropore size and molecular weight decreased as the degradation time increased in each medium. The scaffolds maintained mechanical and structural integrity throughout the study in all three media over the degradation period studied, and the change of Young's modulus of the scaffold under wet condition was not significant. Overall, PBS solution most strongly affected physical and mechanical properties, followed by dH(2)O and HL-60 cells. The distinct variations of the scaffold's properties using different media, demonstrated the importance of carefully selecting the medium to perform in vitro studies. The medium must replicate the actual environment where the scaffold would be used, to represent accurately the changes in properties that the scaffold would be undergoing.

  9. A Porous Tissue Engineering Scaffold Selectively Degraded by Cell-Generated Reactive Oxygen Species

    PubMed Central

    Martin, John R.; Gupta, Mukesh K.; Page, Jonathan M.; Yu, Fang; Davidson, Jeffrey M.; Guelcher, Scott A.

    2014-01-01

    Biodegradable tissue engineering scaffolds are commonly fabricated from poly(lactide-co-glycolide) (PLGA) or similar polyesters that degrade by hydrolysis. PLGA hydrolysis generates acidic breakdown products that trigger an accelerated, autocatalytic degradation mechanism that can create mismatched rates of biomaterial breakdown and tissue formation. Reactive oxygen species (ROS) are key mediators of cell function in both health and disease, especially at sites of inflammation and tissue healing, and induction of inflammation and ROS are natural components of the in vivo response to biomaterial implantation. Thus, polymeric biomaterials that are selectively degraded by cell-generated ROS may have potential for creating tissue engineering scaffolds with better matched rates of tissue in-growth and cell-mediated scaffold biodegradation. To explore this approach, a series of poly(thioketal) (PTK) urethane (PTK-UR) biomaterial scaffolds were synthesized that degrade specifically by an ROS-dependent mechanism. PTK-UR scaffolds had significantly higher compressive moduli than analogous poly(ester urethane) (PEUR) scaffolds formed from hydrolytically-degradable ester-based diols (p < 0.05). Unlike PEUR scaffolds, the PTK-UR scaffolds were stable under aqueous conditions out to 25 weeks but were selectively degraded by ROS, indicating that their biodegradation would be exclusively cell-mediated. The in vitro oxidative degradation rates of the PTK-URs followed first-order degradation kinetics, were significantly dependent on PTK composition (p < 0.05), and correlated to ROS concentration. In subcutaneous rat wounds, PTK-UR scaffolds supported cellular infiltration and granulation tissue formation, followed first-order degradation kinetics over 7 weeks, and produced significantly greater stenting of subcutaneous wounds compared to PEUR scaffolds. These combined results indicate that ROS-degradable PTK-UR tissue engineering scaffolds have significant advantages over analogous

  10. Preparation of tissue engineering porous scaffold with poly(lactic acid) and polyethylene glycol solution blend by solvent-casting/particulate-leaching

    NASA Astrophysics Data System (ADS)

    Huang, Ran; Zhu, Xiaomin; Zhao, Tingting; Wan, Ajun

    2014-12-01

    Polyethylene glycol/poly(lactic acid) solution blend is employed as the raw materials to prepare porous scaffold of potential usage in tissue engineering. The solution blend can be naturally introduced in the classical solvent casting/particular leaching technique in porous matrix preparation. The PEG presence is to modify the degradation behavior of scaffolds to fit particular requirements in tissue engineering. The porous matrix of PEG/PLA with various weight ratios are made with pores size ˜ 250 μ m. The SEM characterizations have been done to investigate the porous morphology of products, the results indicate that though with the clear semi-miscibility feature of PEG/PLA blends, the macro-structure is not significantly affected by the PEG content percentage. The degradation results show an enhanced weight loss rate with the presence of PEG as expected.

  11. Development of organic solvent-free micro-/nano-porous polymer scaffolds for musculoskeletal regeneration.

    PubMed

    Lin, S T C; Musson, D S; Amirapu, S; Cornish, J; Bhattacharyya, D

    2017-05-01

    The use of biomaterial scaffolds has been an enormous field of research in tissue engineering, where the aim is to use graft materials for assisting the human body in recovering lost functions. Currently, there are many ways biomaterial scaffolds can be fabricated; however, many of these techniques involve the use of toxic organic solvents during the process. As biocompatibility is one of the mandatory requirements in designing a successful scaffold, there is an interest in fabricating scaffolds that are completely organic solvent-free. This paper describes the development and characterization of novel micro-/nano-fibrillar composites (MFC/NFC) that can produce scaffolds which are completely free from organic solvents. In this research, the cytocompatibility of these materials have been tested in vitro using mouse osteoblast-like cells and primary rat tenocytes, where cell numbers increase over the culture period, demonstrating the material viability. Gene expression analysis of primary rat tenocytes on MFC/NFC scaffolds demonstrate tenocytic behavior, and histology studies show an increase in cell formation on NFC scaffolds. This study establishes the potential of using the MFC/NFC technique to produce completely organic solvent-free scaffolds capable of hosting musculoskeletal cells, in the hope of providing a graft material for non-union skeletal fractures and rotator cuff repairs. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1393-1404, 2017.

  12. Degradability, cytocompatibility, and osteogenesis of porous scaffolds of nanobredigite and PCL–PEG–PCL composite

    PubMed Central

    Hou, Jun; Fan, Donghui; Zhao, Lingming; Yu, Baoqin; Su, Jiacan; Wei, Jie; Shin, Jung-Woog

    2016-01-01

    Biocomposite scaffolds were fabricated by incorporation of nanobredigite (n-BD) into the polymer of poly(ε-caprolactone)–poly(ethyleneglycol)–poly(ε-caprolactone) (PCL–PEG–PCL). The results revealed that the addition of n-BD into PCL–PEG–PCL significantly improved water absorption, compressive strength, and degradability of the scaffolds of n-BD/PCL–PEG–PCL composite (n-BPC) compared with PCL–PEG–PCL scaffolds alone. In addition, the proliferation and alkaline phosphatase activity of MG63 cells cultured on n-BPC scaffolds were obviously higher than that cultured on PCL–PEG–PCL scaffolds. Moreover, the results of the histological evaluation from the animal model revealed that the n-BPC scaffolds significantly improved new bone formation compared with the PCL–PEG–PCL scaffolds, indicating good osteogenesis. The n-BPC scaffolds with good biocompatibility could stimulate cell proliferation, differentiation, and bone tissue regeneration and would be an excellent candidate for bone defect repair. PMID:27555774

  13. Development of highly porous scaffolds based on bioactive silicates for dental tissue engineering

    SciTech Connect

    Goudouri, O.M.; Theodosoglou, E.; Kontonasaki, E.; Will, J.; Chrissafis, K.; Koidis, P.; Paraskevopoulos, K.M.; Boccaccini, A.R.

    2014-01-01

    Graphical abstract: - Highlights: • Synthesis of an Mg-based glass-ceramic via the sol–gel technique. • The heat treatment of the glass-ceramic promoted the crystallization of akermanite. • Akermanite scaffolds coated with gelatin were successfully fabricated. • An HCAp layer was developed on the surface of all scaffolds after 9 days in SBF. - Abstract: Various scaffolding materials, ceramics and especially Mg-based ceramic materials, including akermanite (Ca{sub 2}MgSi{sub 2}O{sub 7}) and diopside (CaMgSi{sub 2}O{sub 6}), have attracted interest for dental tissue regeneration because of their improved mechanical properties and controllable biodegradation. The aim of the present work was the synthesis of an Mg-based glass-ceramic, which would be used for the construction of workable akermanite scaffolds. The characterization of the synthesized material was performed by Fourier Transform Infrared Spectroscopy (FTIR) X-Ray Diffractometry (XRD) and Scanning Electron Microscopy (SEM). Finally, the apatite forming ability of the scaffolds was assessed by immersion in simulated body fluid. The scaffolds were fabricated by the foam replica technique and were subsequently coated with gelatin to provide a functional surface for increased cell attachment. Finally, SEM microphotographs and FTIR spectra of the scaffolds after immersion in SBF solution indicated the inorganic bioactive character of the scaffolds suitable for the intended applications in dental tissue engineering.

  14. In vitro degradation of porous PLLA/pearl powder composite scaffolds.

    PubMed

    Liu, Y S; Huang, Q L; Kienzle, A; Müller, W E G; Feng, Q L

    2014-05-01

    The in vitro degradation behavior of poly-L-lactide (PLLA), PLLA/aragonite pearl powder and PLLA/vaterite pearl powder scaffolds was investigated. The scaffolds were soaked in phosphate buffer solution (PBS) up to 200 days. Scanning electron microscopy (SEM), gel permeation chromatography (GPC), and differential scanning calorimetry (DSC) were used to observe any degradation of the scaffolds. Degradation behaviors such as changes in pH, porosity, bulk density, water absorption, weight loss and mechanical properties were discussed. The results show that a gradual increase of the pH in composite scaffolds can decrease the rate of hydrolysis of PLLA. PLLA/vaterite and PLLA/aragonite scaffolds have a similar degradation behavior but a slower rate of degradation than PLLA.

  15. Electrospun Starch-Polycaprolactone Nanofiber-Based Constructs for Tissue Engineering

    NASA Astrophysics Data System (ADS)

    Jukola, H.; Nikkola, L.; Gomes, M. E.; Reis, R. L.; Ashammakhi, N.

    2008-02-01

    In the field of biomaterials starch-based polymers have been widely studied for several different applications, including scaffolds for tissue engineering. Recently, electrospinning has been gaining interest as a promising method to manufacture highly porous 3D structures. Such structures provide a high surface area for cell attachment and proliferation, being adequate for several uses in tissue engineering. The aim of the current work is to develop nanofiber-based constructs from starch-polycaprolactone (SPCL 30/70 wt%) blends by means of electrospinning and to study the effect of different solvents. Solutions of 5-15 wt% either in acetic acid or chloroform were electrospun to aluminum foil. The voltage used was 30 kV and the counter-electrode distance was 25 cm. The microstructure of the obtained constructs was characterized by using scanning electron microscopy (SEM). It was possible to obtain highly porous 3D scaffolds with a typical nanofiber-mesh structure by using electrospinning from different SPCL-solvent solutions. Electrospinning was most successful when using higher concentrations (15 wt%). With lower concentrations the process was not very feasible and at a concentration of 5 wt% it was not possible to obtain fibers. The diameter of the fibers obtained was 130-180 nm. SEM analysis revealed the presence of particles which are assumed to be starch. The particles were interconnected by the nanofibers. It is possible to produce highly porous nanofiber-based constructs from SPCL by using electrospinning. Such constructs may have applications in tissue engineering of different tissues, such as bone, skin and cartilage.

  16. Computer aided-designed, 3-dimensionally printed porous tissue bioscaffolds for craniofacial soft tissue reconstruction.

    PubMed

    Zopf, David A; Mitsak, Anna G; Flanagan, Colleen L; Wheeler, Matthew; Green, Glenn E; Hollister, Scott J

    2015-01-01

    To determine the potential of an integrated, image-based computer-aided design (CAD) and 3-dimensional (3D) printing approach to engineer scaffolds for head and neck cartilaginous reconstruction for auricular and nasal reconstruction. Proof of concept revealing novel methods for bioscaffold production with in vitro and in vivo animal data. Multidisciplinary effort encompassing 2 academic institutions. Digital Imaging and Communications in Medicine (DICOM) computed tomography scans were segmented and utilized in image-based CAD to create porous, anatomic structures. Bioresorbable polycaprolactone scaffolds with spherical and random porous architecture were produced using a laser-based 3D printing process. Subcutaneous in vivo implantation of auricular and nasal scaffolds was performed in a porcine model. Auricular scaffolds were seeded with chondrogenic growth factors in a hyaluronic acid/collagen hydrogel and cultured in vitro over 2 months' duration. Auricular and nasal constructs with several types of microporous architecture were rapidly manufactured with high fidelity to human patient anatomy. Subcutaneous in vivo implantation of auricular and nasal scaffolds resulted in an excellent appearance and complete soft tissue ingrowth. Histological analysis of in vitro scaffolds demonstrated native-appearing cartilaginous growth that respected the boundaries of the scaffold. Integrated, image-based CAD and 3D printing processes generated patient-specific nasal and auricular scaffolds that supported cartilage regeneration. © American Academy of Otolaryngology—Head and Neck Surgery Foundation 2014.

  17. Honeycomb-shaped surface topography induces differentiation of human mesenchymal stem cells (hMSCs): uniform porous polymer scaffolds prepared by the breath figure technique.

    PubMed

    Kawano, Takahito; Sato, Madoka; Yabu, Hiroshi; Shimomura, Masatsugu

    2013-11-29

    Polystyrene honeycomb scaffolds with different pore sizes were successfully fabricated by casting a polymer solution under humid conditions in order to investigate the effect of porous microtopography on hMSC differentiation. We have used honeycomb scaffolds to achieve the microtopography-induced differentiation of hMSCs. Honeycomb scaffolds led hMSCs to osteospecific and myospecific differentiations depending on the size of pores. This selective differentiation suggested that surface microtopography alone may be effective for using hMSCs in regenerative medicine and tissue engineering.

  18. Anti-inflammatory effects of sodium alginate/gelatine porous scaffolds merged with fucoidan in murine microglial BV2 cells.

    PubMed

    Nguyen, Van-Tinh; Ko, Seok-Chun; Oh, Gun-Woo; Heo, Seong-Yeong; Jeon, You-Jin; Park, Won Sun; Choi, Il-Whan; Choi, Sung-Wook; Jung, Won-Kyo

    2016-12-01

    Microglia are the immune cells of the central nervous system (CNS). Overexpression of inflammatory mediators by microglia can induce several neurological diseases. Thus, the underlying basic requirement for neural tissue engineering is to develop materials that exhibit little or no neuro-inflammatory effects. In this study, we have developed a method to create porous scaffolds by adding fucoidan (Fu) into porous sodium alginate (Sa)/gelatine (G) (SaGFu). For mechanical characterization, in vitro degradation, stress/strain, swelling, and pore size were measured. Furthermore, the biocompatibility was evaluated by assessing the adhesion and proliferation of BV2 microglial cells on the SaGFu porous scaffolds using scanning electron microscopy (SEM) and lactate dehydrogenase (LDH) assay, respectively. Moreover, we studied the neuro-inflammatory effects of SaGFu on BV2 microglial cells. The effect of gelatine and fucoidan content on the various properties of the scaffold was investigated and the results showed that mechanical properties increased porosity and swelling ratio with an increase in the gelatine and fucoidan, while the in vitro biodegradability decreased. The average SaGFu diameter attained by fabrication of SaGFu ranged from 60 to 120μm with high porosity (74.44%-88.30%). Cell culture using gelatine 2.0% (SaG2Fu) and 4.0% (SaG4Fu), showed good cell proliferation; more than 60-80% that with Sa alone. Following stimulation with 0.5μg/mL LPS, microglia cultured in porous SaGFu decreased their expression of nitric oxide (NO), prostaglandin E2 (PGE2), and reactive oxygen species (ROS). SaG2Fu and SaG4Fu also inhibited the activation and translocation of p65 NF-κB protein levels, resulting in reduction of NO, ROS, and PGE2 production. These results provide insights into the diverse biological effects and opens new avenues for the applications of SaGFu in neuroscience. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. A bioactive and bioresorbable porous cubic composite scaffold loaded with bone marrow aspirate: a potential alternative to autogenous bone grafting.

    PubMed

    Tanaka, Kojiro; Takemoto, Mitsuru; Fujibayashi, Shunsuke; Neo, Masashi; Shikinami, Yasuo; Nakamura, Takashi

    2011-03-15

    Experimental animal study. To investigate the osteogenic properties of a particulate uncalcined, unsintered hydroxyapatite/polydllactide (u-HA/PdlLA) composite scaffold loaded with bone marrow aspirate (BMA). Because of the high morbidity associated with bone graft harvesting, current research in spine surgery has largely focused on bone graft alternatives involving a combination of scaffolds and osteogenic substances. BMA is obtained by a simple and relatively noninvasive method and can easily be clinically applied as an osteogenic material. However, few studies have reported successful posterolateral spinal fusion (PLF) with BMA-loaded synthetic materials. Porous u-HA/PdlLA composites loaded with BMA were used as bone graft substitutes. In experiment 1, porous u-HA/PdlLA cylinders containing or lacking BMA were implanted in rabbit muscles. They were retrieved 4, 8, and 12 weeks after implantation, and ectopic bone formation was histologically evaluated. In experiment 2, 48 rabbits underwent PLF with 1 of 4 bone grafts: autogenous bone (group 1); single-strip u-HA/PdlLA alone (group 2); morselized u-HA/PdlLA + BMA (group 3); or single-strip u-HA/PdlLA + BMA (group 4). After 12 weeks, fusion was assessed by manual palpation, microcomputed tomography, mechanical tests, and histologic examination. In experiment 1, ectopic bone formation was observed in BMA-loaded u-HA/PdlLA, and the new bone area increased until 12 weeks after implantation. In experiment 2, the fusion rates in groups 1, 2, 3, and 4 were 58.3%, 16.7%, 66.7%, and 91.7%, respectively, as determined by manual palpation, and 66.7%, 16.7%, 75.0%, and 91.7%, respectively, as determined by microcomputed tomography. The mechanical strength was significantly greater in group 4 than in the other groups (P < 0.05). Conclusion. Our results indicate that BMA-loaded porous μ-HA/PdlLA is an effective alternative to autogenous bone grafts. The structure and composition of porous u-HA/PdlLA render it an effective

  20. Chondrocytes culture in three-dimensional porous alginate scaffolds enhanced cell proliferation, matrix synthesis and gene expression.

    PubMed

    Lin, Yu-Ju; Yen, Chi-Nan; Hu, Yu-Chen; Wu, Yung-Chih; Liao, Chun-Jen; Chu, I-Ming

    2009-01-01

    For the limited availability of autologous chondrocytes, a cultured system for expansion in vitro until sufficient cells are obtained must be developed. These cells must maintain their chondrocyte phenotype in vitro as well as in vivo, following implantation to ensure that differentiated chondrocytes synthesize a normal hyaline cartilage matrix and not a fibro-cartilage matrix. This study uses porous three-dimensional (3-D) alginate scaffolds within a perfusion system to culture low-density (5 x 10(5) cells) primary porcine chondrocytes for 1-4 weeks to study their proliferation and differentiation. The results of RT-PCR reveal that most cells could maintain their differentiation state for up to 4 weeks of culturing. Chondrocytes proliferated to 3 x 10(7) cells after 4 weeks in culture. Alginate scaffolds induced the formation of chondrocyte clusters and stimulated the synthesis of matrix, which effects were evaluated using histology and electron microscopy. These findings demonstrate that culturing chondrocytes in alginate scaffolds may effectively prevent the dedifferentiation and improve autologous chondrocyte transplantation. 2008 Wiley Periodicals, Inc.

  1. A transferrin variant as the targeting ligand for polymeric nanoparticles incorporated in 3-D PLGA porous scaffolds.

    PubMed

    Lopes, André M; Chen, Kevin Y; Kamei, Daniel T

    2017-04-01

    We have developed doxorubicin (DOX)-loaded poly(lactide-co-glycolide) (PLGA) nanoparticles (DP) conjugated with polyethylene glycol (PEG) and transferrin (Tf) to form Tf-PEG-DPs (TPDPs), and incorporated these TPDPs into three-dimensional (3-D) PLGA porous scaffolds to form a controlled delivery system. To our knowledge, this represents the first use of a Tf variant (oxalate Tf) to improve the targeted delivery of drug-encapsulated nanoparticles (NPs) in PLGA scaffolds to PC3 prostate cancer cells. The PLGA scaffolds with TPDPs incorporated have been shown to release drugs for sustained delivery and provided a continuous release of DOX. The MTS assay was also performed to determine the potency of native and oxalate TPDPs, and a 3.0-fold decrease in IC50 values were observed between the native and oxalate TPDPs. The lower IC50 value for the oxalate version signifies greater potency compared to the native version, since a lower concentration of drug was required to achieve the same therapeutic effect. These results suggest that this technology has potential to become a new implantable polymeric device to improve the controlled and targeted drug delivery of Tf-conjugated NPs for cancer therapy.

  2. Compatibility of Porous Chitosan Scaffold with the Attachment and Proliferation of human Adipose-Derived Stem Cells In Vitro

    PubMed Central

    Gomathysankar, Sankaralakshmi; Halim, Ahmad Sukari; Yaacob, Nik Soriani; Noor, Norhayati Mohd; Mohamed, Mohaini

    2016-01-01

    Adipose-derived stem cells (ASCs) have potential applications in the repair and regeneration of various tissues and organs. The use of various scaffold materials as an excellent template for mimicking the extracellular matrix to induce the attachment and proliferation of different cell types has always been of interest in the field of tissue engineering because ideal biomaterials are in great demand. Chitosan, a marine polysaccharide, have wide clinical applications and it acts as a promising scaffold for cell migration and proliferation. ASCs, with their multi-differentiation potential, and chitosan, with its great biocompatibility with ASCs, were investigated in the present study. ASCs were isolated and were characterized by two different methods: immunocytochemistry and flow cytometry, using the mesenchymal stem cell markers CD90, CD105, CD73 and CD29. The ASCs were then induced to differentiate into adipogenic, osteogenic and chondrogenic lineages. These ASCs were incorporated into a porous chitosan scaffold (PCS), and their structural morphology was studied using a scanning electron microscope and hematoxylin and eosin staining. The proliferation rate of the ASCs on the PCS was assessed using a PrestoBlue viability assay. The results indicated that the PCS provides an excellent template for the adhesion and proliferation of ASCs. Thus, this study revealed that PCS is a promising biomaterial for inducing the proliferation of ASCs, which could lead to successful tissue reconstruction in the field of tissue engineering. PMID:28096632

  3. Compatibility of Porous Chitosan Scaffold with the Attachment and Proliferation of human Adipose-Derived Stem Cells In Vitro.

    PubMed

    Gomathysankar, Sankaralakshmi; Halim, Ahmad Sukari; Yaacob, Nik Soriani; Noor, Norhayati Mohd; Mohamed, Mohaini

    2016-01-01

    Adipose-derived stem cells (ASCs) have potential applications in the repair and regeneration of various tissues and organs. The use of various scaffold materials as an excellent template for mimicking the extracellular matrix to induce the attachment and proliferation of different cell types has always been of interest in the field of tissue engineering because ideal biomaterials are in great demand. Chitosan, a marine polysaccharide, have wide clinical applications and it acts as a promising scaffold for cell migration and proliferation. ASCs, with their multi-differentiation potential, and chitosan, with its great biocompatibility with ASCs, were investigated in the present study. ASCs were isolated and were characterized by two different methods: immunocytochemistry and flow cytometry, using the mesenchymal stem cell markers CD90, CD105, CD73 and CD29. The ASCs were then induced to differentiate into adipogenic, osteogenic and chondrogenic lineages. These ASCs were incorporated into a porous chitosan scaffold (PCS), and their structural morphology was studied using a scanning electron microscope and hematoxylin and eosin staining. The proliferation rate of the ASCs on the PCS was assessed using a PrestoBlue viability assay. The results indicated that the PCS provides an excellent template for the adhesion and proliferation of ASCs. Thus, this study revealed that PCS is a promising biomaterial for inducing the proliferation of ASCs, which could lead to successful tissue reconstruction in the field of tissue engineering.

  4. Seeding of mesenchymal stem cells into inner part of interconnected porous biodegradable scaffold by a new method with a filter paper.

    PubMed

    Yamanaka, Katsuyuki; Yamamoto, Katsushi; Sakai, Yuhiro; Suda, Youko; Shigemitsu, Yusuke; Kaneko, Tadashi; Kato, Koichi; Kumagai, Tomohiro; Kato, Yukio

    2015-01-01

    An appropriate physical support provided by scaffolds creates a supportive environment that directs proliferation and differentiation of stem cells. However, it is difficult to homogenously inoculate stem cells into the inner part of scaffolds at high cell densities. In this study, mesenchymal stem cells were seeded into a hydroxyapatite/poly (D, L-lactic-co-glycolic acid) (HAP/PLGA) scaffold that had enough mechanical strength and porous 3-D structure. With an aid of a filter paper placed under the bottom of a HAP/PLGA block, the cells suspended in a culture medium flowed from the top to the bottom through interconnected pores in the scaffold, and distributed almost homogenously, as compared to cell distribution near the surface of the block by the conventional method using centrifugation or reduced pressure. This simple method with a filter paper may be useful in preparation of cell-scaffold complexes for tissue engineering.

  5. 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).

  6. Improvement of mechanical and biological properties of porous CaSiO3 scaffolds by poly(D,L-lactic acid) modification.

    PubMed

    Wu, Chengtie; Ramaswamy, Yogambha; Boughton, Philip; Zreiqat, Hala

    2008-03-01

    Porous calcium silicates (CaSiO3, WT) are regarded as a potential bioactive material for bone tissue engineering. However, their insufficient mechanical strength and high dissolution (degradation) limit their biological applications. The aim of this study is to surface modify WT scaffolds with poly(d,l-lactic acid) (PDLLA) to improve their mechanical and biological properties. The phase composition, microstructure, porosity and interconnectivity of WT and PDLLA-modified WT (WTPL) scaffolds were analyzed by X-ray diffraction, scanning electron microscopy and micro-computerized tomography. The WTPL scaffolds maintained a more uniform and continuous inner network, compared to that of the WT scaffolds, while maintaining the pore size, porosity and interconnectivity of the original materials. The compressive strength, compressive modulus and percentage strain of the WT and WTPL scaffolds were assessed in air and phosphate-buffered saline. PDLLA modification significantly improved the compressive strength and decreased the brittleness of the WT scaffolds. The weight loss and apatite-forming ability of the two scaffolds were evaluated by soaking them in simulated body fluid (SBF) for 1, 3, 7, 14 and 28days. PDLLA modification decreased the dissolution of the WT scaffolds while maintaining their apatite-forming ability in SBF. In addition, PDLLA modification improved the spreading and viability of human bone-derived cells. Our results indicate that PDLLA-modified CaSiO3 scaffolds possess improved mechanical and biological properties, suggesting their potential application for bone tissue regeneration.

  7. Surface biofunctionalization of three-dimensional porous poly(lactic acid) scaffold using chitosan/OGP coating for bone tissue engineering.

    PubMed

    Zeng, Sen; Ye, Jianhua; Cui, Zhixiang; Si, Junhui; Wang, Qianting; Wang, Xiaofeng; Peng, Kaiping; Chen, Wenzhe

    2017-08-01

    As one of the stimulators on bone formation, osteogenic growth peptide (OGP) improves both proliferation and differentiation of the bone cells in vitro and in vivo. The aim of this work was the preparation of three dimensional porous poly(lactic acid) (PLA) scaffold with high porosity from PLA-dioxane-water ternary system with the use of vacuum-assisted solvent casting, phase separation, solvent extraction and particle leaching methods. Then, by surface coating of PLA scaffold with chitosan (CS)/OGP solution, biofunctionalization of PLA scaffold had been completed for application in bone regeneration. The effects of frozen temperature (-20, -50, -80°C) and PLA solution concentration (10, 12, 14wt%) on the microstructure, water absorption, porosity, hydrophilicity, mechanical properties, and biocompatibility of PLA and CS/OGP/PLA scaffold were investigated. Results showed that both PLA and CS/OGP/PLA scaffolds have an interconnected network structure and a porosity of up to 96.1% and 91.5%, respectively. The CS/OGP/PLA scaffold exhibited better hydrophilicity and mechanical properties than that of uncoated PLA scaffold. Moreover, the results of cell culture test showed that CS/OGP coating could stimulate the proliferation and growth of osteoblast cells on CS/OGP/PLA scaffold. These finding suggested that the surface biofunctionalization by CS/OGP coating layer could be an effective method on enhancing cell adhesion to synthetic polymer-based scaffolds in tissue engineering application and the developed porous CS/OGP/PLA scaffold should be considered as alternative biomaterials for bone regeneration. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. 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.

  9. Use of Clotted Human Plasma and Aprotinin in Skin Tissue Engineering: A Novel Approach to Engineering Composite Skin on a Porous Scaffold.

    PubMed

    Paul, Michelle; Kaur, Pritinder; Herson, Marisa; Cheshire, Perdita; Cleland, Heather; Akbarzadeh, Shiva

    2015-10-01

    Tissue-engineered composite skin is a promising therapy for the treatment of chronic and acute wounds, including burns. Providing the wound bed with a dermal scaffold populated by autologous dermal and epidermal cellular components can further entice host cell infiltration and vascularization to achieve permanent wound closure in a single stage. However, the high porosity and the lack of a supportive basement membrane in most commercially available dermal scaffolds hinders organized keratinocyte proliferation and stratification in vitro and may delay re-epithelization in vivo. The objective of this study was to develop a method to enable the in vitro production of a human skin equivalent (HSE) that included a porous scaffold and dermal and epidermal cells expanded ex vivo, with the potential to be used for definitive treatment of skin defects in a single procedure. A collagen-glycosaminoglycan dermal scaffold (Integra(®)) was populated with adult fibroblasts. A near-normal skin architecture was achieved by the addition of coagulated human plasma to the fibroblast-populated scaffold before seeding cultured keratinocytes. This resulted in reducing scaffold pore size and improving contact surfaces. Skin architecture and basement membrane formation was further improved by the addition of aprotinin (a serine protease inhibitor) to the culture media to inhibit premature clot digestion. Histological assessment of the novel HSE revealed expression of keratin 14 and keratin 10 similar to native skin, with a multilayered neoepidermis morphologically comparable to human skin. Furthermore, deposition of collagen IV and laminin-511 were detected by immunofluorescence, indicating the formation of a continuous basement membrane at the dermal-epidermal junction. The proposed method was efficient in producing an in vitro near native HSE using the chosen off-the-shelf porous scaffold (Integra). The same principles and promising outcomes should be applicable to other biodegradable

  10. 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

  11. Four-Dimensional Printing Hierarchy Scaffolds with Highly Biocompatible Smart Polymers for Tissue Engineering Applications.

    PubMed

    Miao, Shida; Zhu, Wei; Castro, Nathan J; Leng, Jinsong; Zhang, Lijie Grace

    2016-10-01

    The objective of this study was to four-dimensional (4D) print novel biomimetic gradient tissue scaffolds with highly biocompatible naturally derived smart polymers. The term "4D printing" refers to the inherent smart shape transformation of fabricated constructs when implanted minimally invasively for seamless and dynamic integration. For this purpose, a series of novel shape memory polymers with excellent biocompatibility and tunable shape changing effects were synthesized and cured in the presence of three-dimensional printed sacrificial molds, which were subsequently dissolved to create controllable and graded porosity within the scaffold. Surface morphology, thermal, mechanical, and biocompatible properties as well as shape memory effects of the synthesized smart polymers and resultant porous scaffolds were characterized. Fourier transform infrared spectroscopy and gel content analysis confirmed the formation of chemical crosslinking by reacting polycaprolactone triol and castor oil with multi-isocyanate groups. Differential scanning calorimetry revealed an adjustable glass transition temperature in a range from -8°C to 35°C. Uniaxial compression testing indicated that the obtained polymers, possessing a highly crosslinked interpenetrating polymeric networks, have similar compressive modulus to polycaprolactone. Shape memory tests revealed that the smart polymers display finely tunable recovery speed and exhibit greater than 92% shape fixing at -18°C or 0°C and full shape recovery at physiological temperature. Scanning electron microscopy analysis of fabricated scaffolds revealed a graded microporous structure, which mimics the nonuniform distribution of porosity found within natural tissues. With polycaprolactone serving as a control, human bone marrow-derived mesenchymal stem cell adhesion, proliferation, and differentiation greatly increased on our novel smart polymers. The current work will significantly advance the future design and development of

  12. Four-Dimensional Printing Hierarchy Scaffolds with Highly Biocompatible Smart Polymers for Tissue Engineering Applications

    PubMed Central

    Miao, Shida; Zhu, Wei; Castro, Nathan J.; Leng, Jinsong

    2016-01-01

    The objective of this study was to four-dimensional (4D) print novel biomimetic gradient tissue scaffolds with highly biocompatible naturally derived smart polymers. The term “4D printing” refers to the inherent smart shape transformation of fabricated constructs when implanted minimally invasively for seamless and dynamic integration. For this purpose, a series of novel shape memory polymers with excellent biocompatibility and tunable shape changing effects were synthesized and cured in the presence of three-dimensional printed sacrificial molds, which were subsequently dissolved to create controllable and graded porosity within the scaffold. Surface morphology, thermal, mechanical, and biocompatible properties as well as shape memory effects of the synthesized smart polymers and resultant porous scaffolds were characterized. Fourier transform infrared spectroscopy and gel content analysis confirmed the formation of chemical crosslinking by reacting polycaprolactone triol and castor oil with multi-isocyanate groups. Differential scanning calorimetry revealed an adjustable glass transition temperature in a range from −8°C to 35°C. Uniaxial compression testing indicated that the obtained polymers, possessing a highly crosslinked interpenetrating polymeric networks, have similar compressive modulus to polycaprolactone. Shape memory tests revealed that the smart polymers display finely tunable recovery speed and exhibit greater than 92% shape fixing at −18°C or 0°C and full shape recovery at physiological temperature. Scanning electron microscopy analysis of fabricated scaffolds revealed a graded microporous structure, which mimics the nonuniform distribution of porosity found within natural tissues. With polycaprolactone serving as a control, human bone marrow-derived mesenchymal stem cell adhesion, proliferation, and differentiation greatly increased on our novel smart polymers. The current work will significantly advance the future design and

  13. Micro-CT of Porous Apatite Fiber Scaffolds Studied by Projection X-ray Microscopy

    NASA Astrophysics Data System (ADS)

    Moriya, J.; Aizawa, M.; Yoshimura, H.

    2011-09-01

    Hydroxyapatite (HAp) has been widely used as a scaffold for repairing fractured bone. For bone regeneration, the crystal structure, crystal orientation, and composition of HAp as well as the morphology of apatite scaffold are considered to be important. The apatite scaffold constructed by single-crystal fibers with pores showed good results for cellular response. Especially, apatite fiber scaffold (AFS) with large pores, 100 to 250 μm, was found to enhance cell activities such as cell proliferation and differentiation. Here, the three-dimensional (3-D) structure of apatite scaffolds was investigated by means of x-ray computed tomography (x-ray CT) using a scanning electron microscope (SEM) modified projection x-ray microscope. The 3-D structures of apatite fiber scaffolds (AFS) were reconstructed from a series of 180 x-ray projection images taken around a single rotation axis using the Feldkamp-based cone-beam reconstruction method. Extracted cross sections from CT data revealed a network-structure of apatite fibers. The distribution of pores inside the AFS in different preparations was compared.

  14. 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.

  15. 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.

  16. Cryopreservation of Cell/Scaffold Tissue-Engineered Constructs

    PubMed Central

    Costa, Pedro F.; Dias, Ana F.; Reis, Rui L.

    2012-01-01

    The aim of this work was to study the effect of cryopreservation over the functionality of tissue-engineered constructs, analyzing the survival and viability of cells seeded, cultured, and cryopreserved onto 3D scaffolds. Further, it also evaluated the effect of cryopreservation over the properties of the scaffold material itself since these are critical for the engineering of most tissues and in particular, tissues such as bone. For this purpose, porous scaffolds, namely fiber meshes based on a starch and poly(caprolactone) blend were seeded with goat bone marrow stem cells (GBMSCs) and cryopreserved for 7 days. Discs of the same material seeded with GBMSCs were also used as controls. After this period, these samples were analyzed and compared to samples collected before the cryopreservation process. The obtained results demonstrate that it is possible to maintain cell viability and scaffolds properties upon cryopreservation of tissue-engineered constructs based on starch scaffolds and goat bone marrow mesenchymal cells using standard cryopreservation methods. In addition, the outcomes of this study suggest that the greater porosity and interconnectivity of scaffolds favor the retention of cellular content and cellular viability during cryopreservation processes, when compared with nonporous discs. These findings indicate that it might be possible to prepare off-the-shelf engineered tissue substitutes and preserve them to be immediately available upon request for patients' needs. PMID:22676448

  17. (Citric acid–co–polycaprolactone triol) polyester

    PubMed Central

    Thomas, Lynda V.; Nair, Prabha D.

    2011-01-01

    Tissue engineering holds enormous challenges for materials science, wherein the ideal scaffold to be used is expected to be biocompatible, biodegradable and possess mechanical and physical properties that are suitable for target application. In this context, we have prepared degradable polyesters in different ratios by a simple polycondensation technique with citric acid and polycaprolactone triol. Differential scanning calorimetry indicated that the materials were amorphous based the absence of a crystalline melting peak and the presence of a glass transition temperature below 37°C. These polyesters were found to be hydrophilic and could be tailor-made into tubes and films. Porosity could also be introduced by addition of porogens. All the materials were non-cytotoxic in an in vitro cytotoxicity assay and may degrade via hydrolysis to non-toxic degradation products. These polyesters have potential implications in the field of soft tissue engineering on account of their similarity of properties. PMID:23507730

  18. 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

  19. Computational modeling of flow-induced shear stresses within 3D salt-leached porous scaffolds imaged via micro-CT.

    PubMed

    Voronov, Roman; Vangordon, Samuel; Sikavitsas, Vassilios I; Papavassiliou, Dimitrios V

    2010-05-07

    Flow-induced shear stresses have been found to be a stimulatory factor