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David L Kaplan - One of the best experts on this subject based on the ideXlab platform.

  • potential of 3 d Tissue constructs engineered from bovine chondrocytes silk fibroin chitosan for in vitro Cartilage Tissue Engineering
    2011
    Co-Authors: Nandana Bhardwaj, David L Kaplan, Quynhhoa T Nguyen, Albert C Chen, Subhas C Kundu
    Abstract:

    The use of cell-scaffold constructs is a promising Tissue Engineering approach to repair Cartilage defects and to study cartilaginous Tissue formation. In this study, silk fibroin/chitosan blended scaffolds were fabricated and studied for Cartilage Tissue Engineering. Silk fibroin served as a substrate for cell adhesion and proliferation while chitosan has a structure similar to that of glycosaminoglycans, and shows promise for Cartilage repair. We compared the formation of cartilaginous Tissue in silk fibroin/chitosan blended scaffolds seeded with bovine chondrocytes and cultured in vitro for 2 weeks. The constructs were analyzed for cell viability, histology, extracellular matrix components glycosaminoglycan and collagen types I and II, and biomechanical properties. Silk fibroin/chitosan scaffolds supported cell attachment and growth, and chondrogenic phenotype as indicated by Alcian Blue histochemistry and relative expression of type II versus type I collagen. Glycosaminoglycan and collagen accumulated in all the scaffolds and was highest in the silk fibroin/chitosan (1:1) blended scaffolds. Static and dynamic stiffness at high frequencies was higher in cell-seeded constructs than non-seeded controls. The results suggest that silk/chitosan scaffolds may be a useful alternative to synthetic cell scaffolds for Cartilage Tissue Engineering.

  • Silk hydrogel for Cartilage Tissue Engineering
    2010
    Co-Authors: Pen G. Chao, Supansa Yodmuang, Xiaoqin Wang, David L Kaplan, Lin Sun, Gordana Vunjak-novakovic
    Abstract:

    Cartilage Tissue Engineering based on cultivation of immature chondrocytes in agarose hydrogel can yield Tissue constructs with biomechanical properties comparable to native Cartilage. However, agarose is immunogenic and nondegradable, and our capability to modify the structure, composition, and mechanical properties of this material is rather limited. In contrast, silk hydrogel is biocompatible and biodegradable, and it can be produced using a water-based method without organic solvents that enables precise control of structural and mechanical properties in a range of interest for Cartilage Tissue Engineering. We observed that one particular preparation of silk hydrogel yielded cartilaginous constructs with biochemical content and mechanical properties matching constructs based on agarose. This finding and the possibility to vary the properties of silk hydrogel motivated this study of the factors underlying the suitability of hydrogels for Cartilage Tissue Engineering. We present data resulting from a systematic variation of silk hydrogel properties, silk extraction method, gel concentration, and gel structure. Data suggest that silk hydrogel can be used as a tool for studies of the hydrogel-related factors and mechanisms involved in Cartilage formation, as well as a tailorable and fully degradable scaffold for Cartilage Tissue Engineering.

  • Cartilage Tissue Engineering with silk scaffolds and human articular chondrocytes
    2006
    Co-Authors: Yongzhong Wang, D J Blasioli, David L Kaplan
    Abstract:

    Adult Cartilage Tissue has poor capability of self-repair, especially in case of severe Cartilage damage due to trauma or age-related degeneration. Autologous cell-based Tissue Engineering using three-dimensional (3-D) porous scaffolds has provided an option for the repair of full thickness defects in adult Cartilage Tissue. Mesenchymal stem cells (MSCs) and chondrocytes are the two major cell sources for Cartilage Tissue Engineering. Silk fibroin as a naturally occurring degradable fibrous protein with unique mechanical properties, excellent biocompatibility and processability has demonstrated strong potential for skeletal Tissue Engineering [Wong Po Foo C, Kaplan DL. Genetic Engineering of fibrous proteins: spider dragline silk and collagen. Adv Drug Deliv Rev 2002; 54: 1131–43; Altman GH, Diaz F, Jakuba C, Calabro T, Horan RL, Chen J, et al. Silk-based biomaterials. Biomaterials 2003; 24: 401–16; Altman GH, Horan RL, Lu HH, Moreau J, Martin I, Richmond JC, et al. Silk matrix for Tissue engineered anterior cruciate ligaments. Biomaterials 2002; 23: 4131–41; Jin HJ, Kaplan DL. Mechanism of silk processing in insects and spiders. Nature 2003; 424: 1057–61; Jin HJ, Fridrikh SV, Rutledge GC, Kaplan DL. Electrospinning Bombyx mori silk with poly(ethylene oxide). Biomacromolecules 2002; 3: 1233–9]. The present study combined adult human chondrocytes (hCHs) with aqueous-derived porous silk fibroin scaffolds for in vitro Cartilage Tissue Engineering. The results were compared with a previous study using the same scaffolds but using MSCs to generate the Cartilage Tissue outcomes. Culture-expanded hCHs attached to, proliferated and redifferentiated in the scaffolds in a serum-free, chemically defined medium containing TGF-β1, based on cell morphology, levels of Cartilage-related gene transcripts, and the presence of a Cartilage-specific ECM. Cell density was critical for the redifferentiation of culture-expanded hCHs in the 3-D aqueous-derived silk fibroin scaffolds. The level of Cartilage-related transcripts (AGC, Col-II, Sox 9 and Col-II/Col-I ratio) and the deposition of Cartilage-specific ECM were significantly upregulated in constructs initiated with higher seeding density. The hCH-based constructs were significantly different than those formed from MSC-based constructs with respect to cell morphology, zonal structure and initial seeding density needed to successfully generate engineered Cartilage-like Tissue. These results suggest fundamental differences between stem cell-based (MSC) and primary cell-based (hCH) Tissue Engineering, as well as the importance of suitable scaffold features, in the optimization of Cartilage-related outcomes in vitro. The present work diversifies cell sources in combination with silk fibroin-based Tissue Engineering applications. Together with our previous studies, the present results show great promise for engineered 3-D silk fibroin scaffolds in autologous cell-based skeletal Tissue Engineering.

  • Cartilage Tissue Engineering with silk scaffolds and human articular chondrocytes
    2006
    Co-Authors: Yu-min Wang, D J Blasioli, Hae Jong Kim, H S Kim, David L Kaplan
    Abstract:

    Adult Cartilage Tissue has poor capability of self-repair, especially in case of severe Cartilage damage due to trauma or age-related degeneration. Autologous cell-based Tissue Engineering using three-dimensional (3-D) porous scaffolds has provided an option for the repair of full thickness defects in adult Cartilage Tissue. Mesenchymal stem cells (MSCs) and chondrocytes are the two major cell sources for Cartilage Tissue Engineering. Silk fibroin as a naturally occurring degradable fibrous protein with unique mechanical properties, excellent biocompatibility and process-ability has demonstrated strong potential for skeletal Tissue Engineering. The present study combined adult human chondrocytes (hCHs) with aqueous-derived porous silk fibroin scaffolds for in vitro Cartilage Tissue Engineering. The results were compared with a previous study using the same scaffolds but using MSCs to generate the Cartilage Tissue outcomes. Culture-expanded hCHs attached to, proliferated and re-differentiated in the scaffolds in a serum-free, chemically defined medium containing TGF-beta1, based on cell morphology, levels of Cartilage-related gene transcripts, and the presence of a Cartilage-specific ECM. Cell density was critical for the redifferentiation of culture-expanded hCHs in the 3-D aqueous-derived silk fibroin scaffolds. The level of Cartilage-related transcripts (AGC, Col-II, Sox 9 and Col-II/Col-I ratio) and the deposition of Cartilage-specific ECM were significantly upregulated in constructs initiated with higher seeding density. The hCH-based constructs were significantly different than those formed from MSC-based constructs with respect to cell morphology, zonal structure and initial seeding density needed to successfully generate engineered Cartilage-like Tissue. These results suggest fundamental differences between stem cell-based (MSC) and primary cell-based (hCH) Tissue Engineering, as well as the importance of suitable scaffold features, in the optimization of Cartilage-related outcomes in vitro. The present work diversifies cell sources in combination with silk fibroin-based Tissue Engineering applications. Together with our previous studies, the present results show great promise for engineered 3-D silk fibroin scaffolds in autologous cell-based skeletal Tissue Engineering.

  • in vitro Cartilage Tissue Engineering with 3d porous aqueous derived silk scaffolds and mesenchymal stem cells
    2005
    Co-Authors: Yongzhong Wang, D J Blasioli, Ungjin Kim, Hyeonjoo Kim, David L Kaplan
    Abstract:

    Adult Cartilage Tissue has limited self-repair capacity, especially in the case of severe damages caused by developmental abnormalities, trauma, or aging-related degeneration like osteoarthritis. Adult mesenchymal stem cells (MSCs) have the potential to differentiate into cells of different lineages including bone, Cartilage, and fat. In vitro Cartilage Tissue Engineering using autologous MSCs and three-dimensional (3-D) porous scaffolds has the potential for the successful repair of severe Cartilage damage. Ideally, scaffolds designed for Cartilage Tissue Engineering should have optimal structural and mechanical properties, excellent biocompatibility, controlled degradation rate, and good handling characteristics. In the present work, a novel, highly porous silk scaffold was developed by an aqueous process according to these criteria and subsequently combined with MSCs for in vitro Cartilage Tissue Engineering. Chondrogenesis of MSCs in the silk scaffold was evident by real-time RT-PCR analysis for Cartilage-specific ECM gene markers, histological and immunohistochemical evaluations of Cartilage-specific ECM components. Dexamethasone and TGF-beta3 were essential for the survival, proliferation and chondrogenesis of MSCs in the silk scaffolds. The attachment, proliferation, and differentiation of MSCs in the silk scaffold showed unique characteristics. After 3 weeks of cultivation, the spatial cell arrangement and the collagen type-II distribution in the MSCs-silk scaffold constructs resembles those in native articular Cartilage Tissue, suggesting promise for these novel 3-D degradable silk-based scaffolds in MSC-based Cartilage repair. Further in vivo evaluation is necessary to fully recognize the clinical relevance of these observations.

Stephanie J Bryant - One of the best experts on this subject based on the ideXlab platform.

  • understanding the spatiotemporal degradation behavior of aggrecanase sensitive poly ethylene glycol hydrogels for use in Cartilage Tissue Engineering
    2017
    Co-Authors: Shankar Lalitha Sridhar, Stacey C Skaalure, Umut Akalp, Franck J Vernerey, Stephanie J Bryant
    Abstract:

    Enzyme-sensitive hydrogels are promising cell delivery vehicles for Cartilage Tissue Engineering. However, a better understanding of their spatiotemporal degradation behavior and its impact on Tissue growth is needed. The goal of this study was to combine experimental and computational approaches to provide new insights into spatiotemporal changes in hydrogel crosslink density and extracellular matrix (ECM) growth and how these changes influence the evolving macroscopic properties as a function of time. Hydrogels were designed from aggrecanase-sensitive peptide crosslinks using a simple and robust thiol–norbornene photoclick reaction. To study the influence of variations in cellular activity of different donors, chondrocytes were isolated from either juvenile or adult bovine donors. Initial studies were performed to validate and calibrate the model against experiments. Through this process, two key features were identified. These included spatial variations in the hydrogel crosslink density in the immedia...

  • an enzyme sensitive peg hydrogel based on aggrecan catabolism for Cartilage Tissue Engineering
    2015
    Co-Authors: Stacey C Skaalure, Stephanie J Bryant
    Abstract:

    A new Cartilage-specific degradable hydrogel based on photoclickable thiol-ene PEG hydrogels is presented. The hydrogel crosslinks are composed of the peptide, CRDTEGE-ARGSVIDRC, derived from the aggrecanase-cleavable site in aggrecan. This new hydrogel is evaluated for use in Cartilage Tissue Engineering by encapsulating bovine chondrocytes from different cell sources (skeletally immature (juvenile) and mature (adult) donors and adult cells stimulated with pro-inflammatory lipopolysaccharide (LPS)) and culturing for 12 weeks. Regardless of cell source, a two-fold decrease in compressive modulus is observed by 12 weeks, but without significant hydrogel swelling indicating limited bulk degradation. For juvenile cells, a connected matrix rich in aggrecan and collagen II, but minimal collagens I and X is observed. For adult cells, less matrix, but similar quality, is deposited. Aggrecanase activity is elevated, although without accelerating bulk hydrogel degradation. LPS further decreased matrix production, but did not affect aggrecanase activity. In contrast, matrix deposition in the non-degradable hydrogels consisted of aggrecan and collagens I, II and X, indicative of hypertrophic Cartilage. Lastly, no inflammatory response in chondrocytes is observed by the aggrecanase-sensitive hydrogels. Overall, we demonstrate that this new aggrecanase-sensitive hydrogel, which is degradable by chondrocytes and promotes a hyaline-like engineered Cartilage, is promising for Cartilage regeneration.

  • an enzyme sensitive peg hydrogel based on aggrecan catabolism for Cartilage Tissue Engineering
    2015
    Co-Authors: Stacey C Skaalure, Stanley Chu, Stephanie J Bryant
    Abstract:

    A new Cartilage-specific degradable hydrogel based on photoclickable thiol-ene poly(ethylene glycol) (PEG) hydrogels is presented. The hydrogel crosslinks are composed of the peptide, CRDTEGE-ARGSVIDRC, derived from the aggrecanase-cleavable site in aggrecan. This new hydrogel is evaluated for use in Cartilage Tissue Engineering by encapsulating bovine chondrocytes from different cell sources (skeletally immature (juvenile) and mature (adult) donors and adult cells stimulated with proinflammatory lipopolysaccharide (LPS)) and culturing for 12 weeks. Regardless of cell source, a twofold decrease in compressive modulus is observed by 12 weeks, but without significant hydrogel swelling indicating limited bulk degradation. For juvenile cells, a connected matrix rich in aggrecan and collagen II, but minimal collagens I and X is observed. For adult cells, less matrix, but similar quality, is deposited. Aggrecanase activity is elevated, although without accelerating bulk hydrogel degradation. LPS further decreases matrix production, but does not affect aggrecanase activity. In contrast, matrix deposition in the nondegradable hydrogels consists of aggrecan and collagens I, II, and X, indicative of hypertrophic Cartilage. Lastly, no inflammatory response in chondrocytes is observed by the aggrecanase-sensitive hydrogels. Overall, it is demonstrated that this new aggrecanase-sensitive hydrogel, which is degradable by chondrocytes and promotes a hyaline-like engineered Cartilage, is promising for Cartilage regeneration.

  • tailoring the degradation of hydrogels formed from multivinyl poly ethylene glycol and poly vinyl alcohol macromers for Cartilage Tissue Engineering
    2003
    Co-Authors: Penny J Martens, Stephanie J Bryant, Kristi S Anseth
    Abstract:

    Tuning the degradation profiles of polymer cell carriers to match cell and Tissue growth is an important design parameter for (Cartilage) Tissue Engineering. In this study, degradable hydrogels wer...

Yongzhong Wang - One of the best experts on this subject based on the ideXlab platform.

  • Cartilage Tissue Engineering with silk scaffolds and human articular chondrocytes
    2006
    Co-Authors: Yongzhong Wang, D J Blasioli, David L Kaplan
    Abstract:

    Adult Cartilage Tissue has poor capability of self-repair, especially in case of severe Cartilage damage due to trauma or age-related degeneration. Autologous cell-based Tissue Engineering using three-dimensional (3-D) porous scaffolds has provided an option for the repair of full thickness defects in adult Cartilage Tissue. Mesenchymal stem cells (MSCs) and chondrocytes are the two major cell sources for Cartilage Tissue Engineering. Silk fibroin as a naturally occurring degradable fibrous protein with unique mechanical properties, excellent biocompatibility and processability has demonstrated strong potential for skeletal Tissue Engineering [Wong Po Foo C, Kaplan DL. Genetic Engineering of fibrous proteins: spider dragline silk and collagen. Adv Drug Deliv Rev 2002; 54: 1131–43; Altman GH, Diaz F, Jakuba C, Calabro T, Horan RL, Chen J, et al. Silk-based biomaterials. Biomaterials 2003; 24: 401–16; Altman GH, Horan RL, Lu HH, Moreau J, Martin I, Richmond JC, et al. Silk matrix for Tissue engineered anterior cruciate ligaments. Biomaterials 2002; 23: 4131–41; Jin HJ, Kaplan DL. Mechanism of silk processing in insects and spiders. Nature 2003; 424: 1057–61; Jin HJ, Fridrikh SV, Rutledge GC, Kaplan DL. Electrospinning Bombyx mori silk with poly(ethylene oxide). Biomacromolecules 2002; 3: 1233–9]. The present study combined adult human chondrocytes (hCHs) with aqueous-derived porous silk fibroin scaffolds for in vitro Cartilage Tissue Engineering. The results were compared with a previous study using the same scaffolds but using MSCs to generate the Cartilage Tissue outcomes. Culture-expanded hCHs attached to, proliferated and redifferentiated in the scaffolds in a serum-free, chemically defined medium containing TGF-β1, based on cell morphology, levels of Cartilage-related gene transcripts, and the presence of a Cartilage-specific ECM. Cell density was critical for the redifferentiation of culture-expanded hCHs in the 3-D aqueous-derived silk fibroin scaffolds. The level of Cartilage-related transcripts (AGC, Col-II, Sox 9 and Col-II/Col-I ratio) and the deposition of Cartilage-specific ECM were significantly upregulated in constructs initiated with higher seeding density. The hCH-based constructs were significantly different than those formed from MSC-based constructs with respect to cell morphology, zonal structure and initial seeding density needed to successfully generate engineered Cartilage-like Tissue. These results suggest fundamental differences between stem cell-based (MSC) and primary cell-based (hCH) Tissue Engineering, as well as the importance of suitable scaffold features, in the optimization of Cartilage-related outcomes in vitro. The present work diversifies cell sources in combination with silk fibroin-based Tissue Engineering applications. Together with our previous studies, the present results show great promise for engineered 3-D silk fibroin scaffolds in autologous cell-based skeletal Tissue Engineering.

  • in vitro Cartilage Tissue Engineering with 3d porous aqueous derived silk scaffolds and mesenchymal stem cells
    2005
    Co-Authors: Yongzhong Wang, D J Blasioli, Ungjin Kim, Hyeonjoo Kim, David L Kaplan
    Abstract:

    Adult Cartilage Tissue has limited self-repair capacity, especially in the case of severe damages caused by developmental abnormalities, trauma, or aging-related degeneration like osteoarthritis. Adult mesenchymal stem cells (MSCs) have the potential to differentiate into cells of different lineages including bone, Cartilage, and fat. In vitro Cartilage Tissue Engineering using autologous MSCs and three-dimensional (3-D) porous scaffolds has the potential for the successful repair of severe Cartilage damage. Ideally, scaffolds designed for Cartilage Tissue Engineering should have optimal structural and mechanical properties, excellent biocompatibility, controlled degradation rate, and good handling characteristics. In the present work, a novel, highly porous silk scaffold was developed by an aqueous process according to these criteria and subsequently combined with MSCs for in vitro Cartilage Tissue Engineering. Chondrogenesis of MSCs in the silk scaffold was evident by real-time RT-PCR analysis for Cartilage-specific ECM gene markers, histological and immunohistochemical evaluations of Cartilage-specific ECM components. Dexamethasone and TGF-beta3 were essential for the survival, proliferation and chondrogenesis of MSCs in the silk scaffolds. The attachment, proliferation, and differentiation of MSCs in the silk scaffold showed unique characteristics. After 3 weeks of cultivation, the spatial cell arrangement and the collagen type-II distribution in the MSCs-silk scaffold constructs resembles those in native articular Cartilage Tissue, suggesting promise for these novel 3-D degradable silk-based scaffolds in MSC-based Cartilage repair. Further in vivo evaluation is necessary to fully recognize the clinical relevance of these observations.

  • in vitro Cartilage Tissue Engineering with 3d porous aqueous derived silk scaffolds and mesenchymal stem cells
    2005
    Co-Authors: Yongzhong Wang, D J Blasioli, Ungjin Kim, Hyeonjoo Kim, David L Kaplan
    Abstract:

    Abstract Adult Cartilage Tissue has limited self-repair capacity, especially in the case of severe damages caused by developmental abnormalities, trauma, or aging-related degeneration like osteoarthritis. Adult mesenchymal stem cells (MSCs) have the potential to differentiate into cells of different lineages including bone, Cartilage, and fat. In vitro Cartilage Tissue Engineering using autologous MSCs and three-dimensional (3-D) porous scaffolds has the potential for the successful repair of severe Cartilage damage. Ideally, scaffolds designed for Cartilage Tissue Engineering should have optimal structural and mechanical properties, excellent biocompatibility, controlled degradation rate, and good handling characteristics. In the present work, a novel, highly porous silk scaffold was developed by an aqueous process according to these criteria and subsequently combined with MSCs for in vitro Cartilage Tissue Engineering. Chondrogenesis of MSCs in the silk scaffold was evident by real-time RT-PCR analysis for Cartilage-specific ECM gene markers, histological and immunohistochemical evaluations of Cartilage-specific ECM components. Dexamethasone and TGF-β3 were essential for the survival, proliferation and chondrogenesis of MSCs in the silk scaffolds. The attachment, proliferation, and differentiation of MSCs in the silk scaffold showed unique characteristics. After 3 weeks of cultivation, the spatial cell arrangement and the collagen type-II distribution in the MSCs-silk scaffold constructs resembles those in native articular Cartilage Tissue, suggesting promise for these novel 3-D degradable silk-based scaffolds in MSC-based Cartilage repair. Further in vivo evaluation is necessary to fully recognize the clinical relevance of these observations.

D J Blasioli - One of the best experts on this subject based on the ideXlab platform.

  • Cartilage Tissue Engineering with silk scaffolds and human articular chondrocytes
    2006
    Co-Authors: Yongzhong Wang, D J Blasioli, David L Kaplan
    Abstract:

    Adult Cartilage Tissue has poor capability of self-repair, especially in case of severe Cartilage damage due to trauma or age-related degeneration. Autologous cell-based Tissue Engineering using three-dimensional (3-D) porous scaffolds has provided an option for the repair of full thickness defects in adult Cartilage Tissue. Mesenchymal stem cells (MSCs) and chondrocytes are the two major cell sources for Cartilage Tissue Engineering. Silk fibroin as a naturally occurring degradable fibrous protein with unique mechanical properties, excellent biocompatibility and processability has demonstrated strong potential for skeletal Tissue Engineering [Wong Po Foo C, Kaplan DL. Genetic Engineering of fibrous proteins: spider dragline silk and collagen. Adv Drug Deliv Rev 2002; 54: 1131–43; Altman GH, Diaz F, Jakuba C, Calabro T, Horan RL, Chen J, et al. Silk-based biomaterials. Biomaterials 2003; 24: 401–16; Altman GH, Horan RL, Lu HH, Moreau J, Martin I, Richmond JC, et al. Silk matrix for Tissue engineered anterior cruciate ligaments. Biomaterials 2002; 23: 4131–41; Jin HJ, Kaplan DL. Mechanism of silk processing in insects and spiders. Nature 2003; 424: 1057–61; Jin HJ, Fridrikh SV, Rutledge GC, Kaplan DL. Electrospinning Bombyx mori silk with poly(ethylene oxide). Biomacromolecules 2002; 3: 1233–9]. The present study combined adult human chondrocytes (hCHs) with aqueous-derived porous silk fibroin scaffolds for in vitro Cartilage Tissue Engineering. The results were compared with a previous study using the same scaffolds but using MSCs to generate the Cartilage Tissue outcomes. Culture-expanded hCHs attached to, proliferated and redifferentiated in the scaffolds in a serum-free, chemically defined medium containing TGF-β1, based on cell morphology, levels of Cartilage-related gene transcripts, and the presence of a Cartilage-specific ECM. Cell density was critical for the redifferentiation of culture-expanded hCHs in the 3-D aqueous-derived silk fibroin scaffolds. The level of Cartilage-related transcripts (AGC, Col-II, Sox 9 and Col-II/Col-I ratio) and the deposition of Cartilage-specific ECM were significantly upregulated in constructs initiated with higher seeding density. The hCH-based constructs were significantly different than those formed from MSC-based constructs with respect to cell morphology, zonal structure and initial seeding density needed to successfully generate engineered Cartilage-like Tissue. These results suggest fundamental differences between stem cell-based (MSC) and primary cell-based (hCH) Tissue Engineering, as well as the importance of suitable scaffold features, in the optimization of Cartilage-related outcomes in vitro. The present work diversifies cell sources in combination with silk fibroin-based Tissue Engineering applications. Together with our previous studies, the present results show great promise for engineered 3-D silk fibroin scaffolds in autologous cell-based skeletal Tissue Engineering.

  • Cartilage Tissue Engineering with silk scaffolds and human articular chondrocytes
    2006
    Co-Authors: Yu-min Wang, D J Blasioli, Hae Jong Kim, H S Kim, David L Kaplan
    Abstract:

    Adult Cartilage Tissue has poor capability of self-repair, especially in case of severe Cartilage damage due to trauma or age-related degeneration. Autologous cell-based Tissue Engineering using three-dimensional (3-D) porous scaffolds has provided an option for the repair of full thickness defects in adult Cartilage Tissue. Mesenchymal stem cells (MSCs) and chondrocytes are the two major cell sources for Cartilage Tissue Engineering. Silk fibroin as a naturally occurring degradable fibrous protein with unique mechanical properties, excellent biocompatibility and process-ability has demonstrated strong potential for skeletal Tissue Engineering. The present study combined adult human chondrocytes (hCHs) with aqueous-derived porous silk fibroin scaffolds for in vitro Cartilage Tissue Engineering. The results were compared with a previous study using the same scaffolds but using MSCs to generate the Cartilage Tissue outcomes. Culture-expanded hCHs attached to, proliferated and re-differentiated in the scaffolds in a serum-free, chemically defined medium containing TGF-beta1, based on cell morphology, levels of Cartilage-related gene transcripts, and the presence of a Cartilage-specific ECM. Cell density was critical for the redifferentiation of culture-expanded hCHs in the 3-D aqueous-derived silk fibroin scaffolds. The level of Cartilage-related transcripts (AGC, Col-II, Sox 9 and Col-II/Col-I ratio) and the deposition of Cartilage-specific ECM were significantly upregulated in constructs initiated with higher seeding density. The hCH-based constructs were significantly different than those formed from MSC-based constructs with respect to cell morphology, zonal structure and initial seeding density needed to successfully generate engineered Cartilage-like Tissue. These results suggest fundamental differences between stem cell-based (MSC) and primary cell-based (hCH) Tissue Engineering, as well as the importance of suitable scaffold features, in the optimization of Cartilage-related outcomes in vitro. The present work diversifies cell sources in combination with silk fibroin-based Tissue Engineering applications. Together with our previous studies, the present results show great promise for engineered 3-D silk fibroin scaffolds in autologous cell-based skeletal Tissue Engineering.

  • in vitro Cartilage Tissue Engineering with 3d porous aqueous derived silk scaffolds and mesenchymal stem cells
    2005
    Co-Authors: Yongzhong Wang, D J Blasioli, Ungjin Kim, Hyeonjoo Kim, David L Kaplan
    Abstract:

    Adult Cartilage Tissue has limited self-repair capacity, especially in the case of severe damages caused by developmental abnormalities, trauma, or aging-related degeneration like osteoarthritis. Adult mesenchymal stem cells (MSCs) have the potential to differentiate into cells of different lineages including bone, Cartilage, and fat. In vitro Cartilage Tissue Engineering using autologous MSCs and three-dimensional (3-D) porous scaffolds has the potential for the successful repair of severe Cartilage damage. Ideally, scaffolds designed for Cartilage Tissue Engineering should have optimal structural and mechanical properties, excellent biocompatibility, controlled degradation rate, and good handling characteristics. In the present work, a novel, highly porous silk scaffold was developed by an aqueous process according to these criteria and subsequently combined with MSCs for in vitro Cartilage Tissue Engineering. Chondrogenesis of MSCs in the silk scaffold was evident by real-time RT-PCR analysis for Cartilage-specific ECM gene markers, histological and immunohistochemical evaluations of Cartilage-specific ECM components. Dexamethasone and TGF-beta3 were essential for the survival, proliferation and chondrogenesis of MSCs in the silk scaffolds. The attachment, proliferation, and differentiation of MSCs in the silk scaffold showed unique characteristics. After 3 weeks of cultivation, the spatial cell arrangement and the collagen type-II distribution in the MSCs-silk scaffold constructs resembles those in native articular Cartilage Tissue, suggesting promise for these novel 3-D degradable silk-based scaffolds in MSC-based Cartilage repair. Further in vivo evaluation is necessary to fully recognize the clinical relevance of these observations.

  • in vitro Cartilage Tissue Engineering with 3d porous aqueous derived silk scaffolds and mesenchymal stem cells
    2005
    Co-Authors: Yongzhong Wang, D J Blasioli, Ungjin Kim, Hyeonjoo Kim, David L Kaplan
    Abstract:

    Abstract Adult Cartilage Tissue has limited self-repair capacity, especially in the case of severe damages caused by developmental abnormalities, trauma, or aging-related degeneration like osteoarthritis. Adult mesenchymal stem cells (MSCs) have the potential to differentiate into cells of different lineages including bone, Cartilage, and fat. In vitro Cartilage Tissue Engineering using autologous MSCs and three-dimensional (3-D) porous scaffolds has the potential for the successful repair of severe Cartilage damage. Ideally, scaffolds designed for Cartilage Tissue Engineering should have optimal structural and mechanical properties, excellent biocompatibility, controlled degradation rate, and good handling characteristics. In the present work, a novel, highly porous silk scaffold was developed by an aqueous process according to these criteria and subsequently combined with MSCs for in vitro Cartilage Tissue Engineering. Chondrogenesis of MSCs in the silk scaffold was evident by real-time RT-PCR analysis for Cartilage-specific ECM gene markers, histological and immunohistochemical evaluations of Cartilage-specific ECM components. Dexamethasone and TGF-β3 were essential for the survival, proliferation and chondrogenesis of MSCs in the silk scaffolds. The attachment, proliferation, and differentiation of MSCs in the silk scaffold showed unique characteristics. After 3 weeks of cultivation, the spatial cell arrangement and the collagen type-II distribution in the MSCs-silk scaffold constructs resembles those in native articular Cartilage Tissue, suggesting promise for these novel 3-D degradable silk-based scaffolds in MSC-based Cartilage repair. Further in vivo evaluation is necessary to fully recognize the clinical relevance of these observations.

Antonios G Mikos - One of the best experts on this subject based on the ideXlab platform.

  • applications of decellularized extracellular matrix in bone and Cartilage Tissue Engineering
    2019
    Co-Authors: Yu Seon Kim, Marjan Majid, Anthony J Melchiorri, Antonios G Mikos
    Abstract:

    Regenerative therapies for bone and Cartilage injuries are currently unable to replicate the complex microenvironment of native Tissue. There are many Tissue Engineering approaches attempting to address this issue through the use of synthetic materials. Although synthetic materials can be modified to simulate the mechanical and biochemical properties of the cell microenvironment, they do not mimic in full the multitude of interactions that take place within Tissue. Decellularized extracellular matrix (dECM) has been established as a biomaterial that preserves a Tissue's native environment, promotes cell proliferation, and provides cues for cell differentiation. The potential of dECM as a therapeutic agent is rising, but there are many limitations of dECM restricting its use. This review discusses the recent progress in the utilization of bone and Cartilage dECM through applications as scaffolds, particles, and supplementary factors in bone and Cartilage Tissue Engineering.

  • bioactive polymer extracellular matrix scaffolds fabricated with a flow perfusion bioreactor for Cartilage Tissue Engineering
    2010
    Co-Authors: Jiehong Liao, Xua Guo, Jane K Grandealle, Kurtis F Kaspe, Antonios G Mikos
    Abstract:

    In this study, electrospun poly(ɛ-caprolactone) (PCL) microfiber scaffolds, coated with cartilaginous extracellular matrix (ECM), were fabricated by first culturing chondrocytes under dynamic conditions in a flow perfusion bioreactor and then decellularizing the cellular constructs. The decellularization procedure yielded acellular PCL/ECM composite scaffolds containing glycosaminoglycan and collagen. PCL/ECM composite scaffolds were evaluated for their ability to support the chondrogenic differentiation of mesenchymal stem cells (MSCs) in vitro using serum-free medium with or without the addition of transforming growth factor-β1 (TGF-β1). PCL/ECM composite scaffolds supported chondrogenic differentiation induced by TGF-β1 exposure, as evidenced in the up-regulation of aggrecan (11.6 ± 3.8 fold) and collagen type II (668.4 ± 317.7 fold) gene expression. The presence of cartilaginous matrix alone reduced collagen type I gene expression to levels observed with TGF-β1 treatment. Cartilaginous matrix further enhanced the effects of growth factor treatment on MSC chondrogenesis as evidenced in the higher glycosaminoglycan synthetic activity for cells cultured on PCL/ECM composite scaffolds. Therefore, flow perfusion culture of chondrocytes on electrospun microfiber scaffolds is a promising method to fabricate polymer/extracellular matrix composite scaffolds that incorporate both natural and synthetic components to provide biological signals for Cartilage Tissue Engineering applications.

  • bioactive polymer extracellular matrix scaffolds fabricated with a flow perfusion bioreactor for Cartilage Tissue Engineering
    2010
    Co-Authors: Jiehong Liao, Jane K Grandeallen, Kurtis F Kasper, Antonios G Mikos
    Abstract:

    In this study, electrospun poly(e-caprolactone) (PCL) microfiber scaffolds, coated with cartilaginous extracellular matrix (ECM), were fabricated by first culturing chondrocytes under dynamic conditions in a flow perfusion bioreactor and then decellularizing the cellular constructs. The decellularization procedure yielded acellular PCL/ECM composite scaffolds containing glycosaminoglycan and collagen. PCL/ECM composite scaffolds were evaluated for their ability to support the chondrogenic differentiation of mesenchymal stem cells (MSCs) in vitro using serum-free medium with or without the addition of transforming growth factor-β1 (TGF-β1). PCL/ECM composite scaffolds supported chondrogenic differentiation induced by TGF-β1 exposure, as evidenced in the up-regulation of aggrecan (11.6 ± 3.8 fold) and collagen type II (668.4 ± 317.7 fold) gene expression. The presence of cartilaginous matrix alone reduced collagen type I gene expression to levels observed with TGF-β1 treatment. Cartilaginous matrix further enhanced the effects of growth factor treatment on MSC chondrogenesis as evidenced in the higher glycosaminoglycan synthetic activity for cells cultured on PCL/ECM composite scaffolds. Therefore, flow perfusion culture of chondrocytes on electrospun microfiber scaffolds is a promising method to fabricate polymer/extracellular matrix composite scaffolds that incorporate both natural and synthetic components to provide biological signals for Cartilage Tissue Engineering applications.

  • delivery of tgf β1 and chondrocytes via injectable biodegradable hydrogels for Cartilage Tissue Engineering applications
    2005
    Co-Authors: Hansoo Park, Johnna S Temenoff, Theresa A Holland, Yasuhiko Tabata, Antonios G Mikos
    Abstract:

    In this work, novel hydrogel composites, based on the biodegradable polymer, oligo(poly(ethylene glycol) fumarate) (OPF) and gelatin microparticles (MPs) were utilized as injectable cell and growth factor carriers for Cartilage Tissue Engineering applications. Specifically, bovine chondrocytes were embedded in composite hydrogels co-encapsulating gelatin MPs loaded with transforming growth factor-β1 (TGF-β1). Hydrogels with embedded cells co-encapsulating unloaded MPs and those with no MPs served as controls in order to assess the effects of MPs and TGF-β1 on chondrocyte function. Samples were cultured up to 28 days in vitro. By 14 days, cell attachment to embedded gelatin MPs within the constructs was observed via light microscopy. Bioassay results showed that, over the 21 day period, there was a statistically significant increase in cellular proliferation for samples containing gelatin MPs, but no increase was exhibited in samples without MPs over the culture period. The release of TGF-β1 further increased cell construct cellularity. Over the same time period, glycosaminoglycan content per cell remained constant for all formulations, suggesting that the dramatic increase in cell number for samples with TGF-β1-loaded MPs was accompanied by maintenance of the cell phenotype. Overall, these data indicate the potential of OPF hydrogel composites containing embedded chondrocytes and TGF-β1-loaded gelatin MPs as a novel strategy for Cartilage Tissue Engineering.