Cartilage tissue engineering: Directed differentiation of embryonic stem cells in three-dimensional hydrogel culture.

The clinical goal of tissue engineering is to restore, repair, or replace damaged tissues in the body. Significant advances have been made in recent years using stem cells as a cell source for cartilage tissue engineering and reconstructive surgery applications. Embryonic stem cells have demonstrated the potential to self-renew and differentiate into a wide range of tissues including the chondrogenic lineage, depending on culture conditions. Three-dimensional scaffolds play an important role in tissue regeneration by providing attachment sites as well as bioactive signals for cells to grow and differentiate into specific lineages. The precise microenvironments required for optimal expansion or differentiation of stem cells are only beginning to emerge now, and the controlled differentiation of embryonic stem cells in tissue engineering remains a relatively unexplored field. Hydrogels are a class of polymer-based biomaterials that have been extensively utilized in tissue engineering as scaffolds. We have demonstrated that embryonic stem cells encapsulated within poly(ethylene glycol)-based (PEGDA) photopolymerizing hydrogels and cultured in an appropriate growth factor and medium conditions undergo chondrogenic differentiation with extracellular matrix deposition characteristic of neocartilage (Hwang et al., Stem Cells 24, 284-291). Another hydrogel that has been widely used for encapsulating chondrocytes in cartilage tissue engineering is alginate. This hydrogel also has potential for tissue engineering applications using stem cells. Here, we describe the three-dimensional culture of embryonic stem cell-derived embryoid bodies in hydrogels and their differentiation toward chondrogenic lineage in chemically defined chondrogenic medium in the presence of TGF-beta1 (chondrogenic inducing conditions). We also discuss various tools and assays used for characterizing the tissue-engineered cartilage.

[1]  W. Sufan,et al.  Alginate hydrogel linked with synthetic oligopeptide derived from BMP-2 allows ectopic osteoinduction in vivo. , 2000, Journal of biomedical materials research.

[2]  S. Ichinose,et al.  In vitro cartilage formation of composites of synovium-derived mesenchymal stem cells with collagen gel , 2005, Cell and Tissue Research.

[3]  G. Vunjak‐Novakovic,et al.  Cultivation of cell‐polymer cartilage implants in bioreactors , 1993, Journal of cellular biochemistry.

[4]  Harrie Weinans,et al.  Biochemical and functional modulation of the cartilage collagen network by IGF1, TGFβ2 and FGF2 , 2006 .

[5]  K. Athanasiou,et al.  Rapid phenotypic changes in passaged articular chondrocyte subpopulations , 2005, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[6]  K.,et al.  Perichondral grafting for cartilage lesions of the knee. , 1990, The Journal of bone and joint surgery. British volume.

[7]  J. Mao,et al.  Tissue-engineered Neogenesis of Human-shaped Mandibular Condyle from Rat Mesenchymal Stem Cells , 2003, Journal of dental research.

[8]  T. Quinn,et al.  Surgical Removal of Articular Cartilage Leads to Loss of Chondrocytes from Cartilage Bordering the Wound Edge , 2003, The Journal of bone and joint surgery. American volume.

[9]  A. Miniaci,et al.  Arthroscopic mosaic arthroplasty in the equine third carpal bone. , 2001, Veterinary surgery : VS.

[10]  J. Mason,et al.  Cartilage tissue engineering: current limitations and solutions. , 1999, Clinical orthopaedics and related research.

[11]  J. Williams,et al.  Effect of transforming growth factor beta1 on chondrogenic differentiation of cultured equine mesenchymal stem cells. , 2000, American journal of veterinary research.

[12]  K. Hruska,et al.  In Vitro Generation of Scaffold Independent Neocartilage , 2001, Clinical orthopaedics and related research.

[13]  Lori A. Setton,et al.  Photocrosslinkable Hyaluronan as a Scaffold for Articular Cartilage Repair , 2004, Annals of Biomedical Engineering.

[14]  Christoph Gaissmaier,et al.  Bone morphogenetic protein (BMP)-2 enhances the expression of type II collagen and aggrecan in chondrocytes embedded in alginate beads. , 2004, Osteoarthritis and cartilage.

[15]  Brian J Cole,et al.  Minced cartilage without cell culture serves as an effective intraoperative cell source for cartilage repair , 2006, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[16]  J. Axelman,et al.  Human embryonic germ cell derivatives express a broad range of developmentally distinct markers and proliferate extensively in vitro. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[17]  P. Behrens,et al.  In vivo matrix-guided human mesenchymal stem cells , 2006, Cellular and Molecular Life Sciences.

[18]  Chad A. Cowan,et al.  Derivation of embryonic stem-cell lines from human blastocysts. , 2004, The New England journal of medicine.

[19]  P. Gillet,et al.  The effect of alginate, hyaluronate and hyaluronate derivatives biomaterials on synthesis of non-articular chondrocyte extracellular matrix , 2005, Journal of materials science. Materials in medicine.

[20]  Robert L Sah,et al.  Effect of Growth Factors on Cell Proliferation, Matrix Deposition, and Morphology of Human Nasal Septal Chondrocytes Cultured in Monolayer , 2005, The Laryngoscope.

[21]  R. Eavey,et al.  Expansion of the number of human auricular chondrocytes: recycling of culture media containing floating cells. , 2004, Tissue engineering.

[22]  I. Sekiya,et al.  Comparison of effect of BMP-2, -4, and -6 on in vitro cartilage formation of human adult stem cells from bone marrow stroma , 2005, Cell and Tissue Research.

[23]  P. Butler,et al.  Injectable Cartilage Using Polyethylene Oxide Polymer Substrates , 1996, Plastic and reconstructive surgery.

[24]  Peter Müller,et al.  Relationship between cell shape and type of collagen synthesised as chondrocytes lose their cartilage phenotype in culture , 1977, Nature.

[25]  J. Elisseeff,et al.  Transdermal photopolymerization of poly(ethylene oxide)-based injectable hydrogels for tissue-engineered cartilage. , 1999, Plastic and reconstructive surgery.

[26]  F. M. Schultz,et al.  Dynamic pressure transmission through agarose gels. , 2000, Tissue engineering.

[27]  S. Bruder,et al.  Osteogenic differentiation of purified, culture‐expanded human mesenchymal stem cells in vitro , 1997, Journal of cellular biochemistry.

[28]  Mina J Bissell,et al.  Modeling tissue-specific signaling and organ function in three dimensions , 2003, Journal of Cell Science.

[29]  Antonios G Mikos,et al.  Transforming growth factor-beta 1 release from oligo(poly(ethylene glycol) fumarate) hydrogels in conditions that model the cartilage wound healing environment. , 2004, Journal of controlled release : official journal of the Controlled Release Society.

[30]  M. McKee,et al.  Chitosan-glycerol phosphate/blood implants elicit hyaline cartilage repair integrated with porous subchondral bone in microdrilled rabbit defects. , 2007, Osteoarthritis and cartilage.

[31]  Jianyi(Jay) Zhang,et al.  Autologous stem cell transplantation for myocardial repair. , 2004, American journal of physiology. Heart and circulatory physiology.

[32]  L. Bonassar,et al.  Injectable Tissue-Engineered Cartilage with Different Chondrocyte Sources , 2004, Plastic and reconstructive surgery.

[33]  J. Glowacki,et al.  Three-dimensional composite of demineralized bone powder and collagen for in vitro analysis of chondroinduction of human dermal fibroblasts. , 1996, Biomaterials.

[34]  J. Vacanti,et al.  De Novo Cartilage Generation Using Calcium Alginate‐Chondrocyte Constructs , 1996, Plastic and reconstructive surgery.

[35]  C. Vacanti,et al.  Tissue-engineered morphogenesis of cartilage and bone by means of cell transplantation using synthetic biodegradable polymer matrices. , 1994, Clinics in plastic surgery.

[36]  I. Martin,et al.  Development and validation of a bioreactor for physical stimulation of engineered cartilage. , 2003, Biorheology.

[37]  Richard Tuli,et al.  Adult mesenchymal stem cells and cell-based tissue engineering , 2002, Arthritis research & therapy.

[38]  D. Lafreniere,et al.  Polyglycolic Acid/Poly‐L‐Lactic Acid Copolymer Use In Laryngotracheal Reconstruction: A Rabbit Model , 2005, The Laryngoscope.

[39]  L. Lohmander,et al.  Tissue engineering of cartilage: do we need it, can we do it, is it good and can we prove it? , 2003, Novartis Foundation symposium.

[40]  V Vécsei,et al.  Dedifferentiation-associated changes in morphology and gene expression in primary human articular chondrocytes in cell culture. , 2002, Osteoarthritis and cartilage.

[41]  W McIntosh,et al.  Transdermal photopolymerization for minimally invasive implantation. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[42]  C. Archer,et al.  Cartilage integration: evaluation of the reasons for failure of integration during cartilage repair. A review. , 2008, European cells & materials.

[43]  Leon Y Lum,et al.  Effect of Gravity on Localization of Chondrocytes Implanted in Cartilage Defects , 2002, Clinical orthopaedics and related research.

[44]  G A Ateshian,et al.  Functional tissue engineering of articular cartilage through dynamic loading of chondrocyte-seeded agarose gels. , 2000, Journal of biomechanical engineering.

[45]  E. Burger,et al.  The role of trabecular demineralized bone in combination with perichondrium in the generation of cartilage grafts. , 1999, Biomaterials.

[46]  D. Heinegård,et al.  Biochemistry and Metabolism of Normal and Osteoarthritic Cartilage , 2003 .

[47]  R. Eavey,et al.  Tissue Engineering of a Human Sized And Shaped Auricle Using a Mold , 2004, The Laryngoscope.

[48]  V. Goldberg,et al.  Hyaluronic acid‐based polymers as cell carriers for tissue‐engineered repair of bone and cartilage , 1999, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[49]  R Langer,et al.  Chondrogenesis in a cell-polymer-bioreactor system. , 1998, Experimental cell research.

[50]  A. Lynn,et al.  Repair of defects in articular joints. Prospects for material-based solutions in tissue engineering. , 2004, The Journal of bone and joint surgery. British volume.

[51]  Takashi Ushida,et al.  Hydrostatic fluid pressure enhances matrix synthesis and accumulation by bovine chondrocytes in three‐dimensional culture , 2002, Journal of cellular physiology.

[52]  Antonios G Mikos,et al.  In vitro cytotoxicity of redox radical initiators for cross-linking of oligo(poly(ethylene glycol) fumarate) macromers. , 2003, Biomacromolecules.

[53]  Maurilio Marcacci,et al.  Articular Cartilage Engineering with Hyalograft® C: 3-Year Clinical Results , 2005, Clinical orthopaedics and related research.

[54]  D L Bader,et al.  Compressive strains at physiological frequencies influence the metabolism of chondrocytes seeded in agarose , 1997, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[55]  K. Athanasiou,et al.  Retaining zonal chondrocyte phenotype by means of novel growth environments. , 2005, Tissue engineering.

[56]  Paul N Manson,et al.  Variable cytocompatibility of six cell lines with photoinitiators used for polymerizing hydrogels and cell encapsulation. , 2005, Biomaterials.

[57]  Andre Steinert,et al.  Gene-induced chondrogenesis of primary mesenchymal stem cells in vitro. , 2005, Molecular therapy : the journal of the American Society of Gene Therapy.

[58]  L. Kozak,et al.  Ambulatory and inpatient procedures in the United States, 1996. , 1998, Vital and health statistics. Series 13, Data from the National Health Survey.

[59]  R. J. Koch,et al.  Tissue Engineering with Chondrocytes , 2002, Facial plastic surgery : FPS.

[60]  A. Mikos,et al.  Growing new organs. , 1999, Scientific American.

[61]  G. Vunjak‐Novakovic,et al.  Growth factors for sequential cellular de- and re-differentiation in tissue engineering. , 2002, Biochemical and biophysical research communications.

[62]  Kenneth M. Yamada,et al.  Taking Cell-Matrix Adhesions to the Third Dimension , 2001, Science.

[63]  Eben Alsberg,et al.  Engineering growing tissues , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[64]  Chun Han,et al.  Macroscopic cartilage formation with embryonic stem-cell-derived mesodermal progenitor cells , 2003, Journal of Cell Science.

[65]  J A Thomson,et al.  Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture. , 2000, Developmental biology.

[66]  S. Hung,et al.  Neocartilage from human mesenchymal stem cells in alginate: implied timing of transplantation. , 2005, Journal of biomedical materials research. Part A.

[67]  Moonsoo Jin,et al.  Effects of dynamic compressive loading on chondrocyte biosynthesis in self-assembling peptide scaffolds. , 2004, Journal of biomechanics.

[68]  Gerard A Ateshian,et al.  Synergistic action of growth factors and dynamic loading for articular cartilage tissue engineering. , 2003, Tissue engineering.

[69]  M. Marcacci,et al.  Arthroscopic autologous condrocyte transplantation: technical note , 2002, Knee Surgery, Sports Traumatology, Arthroscopy.

[70]  D J Mooney,et al.  Alginate hydrogels as synthetic extracellular matrix materials. , 1999, Biomaterials.

[71]  M. Brittberg.  Autologous chondrocyte implantation--technique and long-term follow-up. , 2008, Injury.

[72]  A. Jayakrishnan,et al.  Self-cross-linking biopolymers as injectable in situ forming biodegradable scaffolds. , 2005, Biomaterials.

[73]  J. Bonaventure,et al.  Reexpression of cartilage-specific genes by dedifferentiated human articular chondrocytes cultured in alginate beads. , 1994, Experimental cell research.

[74]  Kyriacos A. Athanasiou,et al.  Principles of Cell Mechanics for Cartilage Tissue Engineering , 2004, Annals of Biomedical Engineering.

[75]  K. Wood.  Analytical biotechnology: Imaging: beyond cataloging nucleic acids and proteins , 2005 .

[76]  J. West,et al.  Modification of surfaces with cell adhesion peptides alters extracellular matrix deposition. , 1999, Biomaterials.

[77]  F. Silver,et al.  Cartilage wound healing. An overview. , 1995, Otolaryngologic clinics of North America.

[78]  K. Arden,et al.  Expression of the human chondrocyte phenotype in vitro , 1989, In Vitro Cellular & Developmental Biology.

[79]  W. Lawson,et al.  Validating the Subcutaneous Model of Injectable Autologous Cartilage Using a Fibrin Glue Scaffold , 2004, The Laryngoscope.

[80]  K. Chua,et al.  Insulin-transferrin-selenium prevent human chondrocyte dedifferentiation and promote the formation of high quality tissue engineered human hyaline cartilage. , 2005, European cells & materials.

[81]  J. Leroux,et al.  Novel injectable neutral solutions of chitosan form biodegradable gels in situ. , 2000, Biomaterials.

[82]  B. Obradovic,et al.  Integration of engineered cartilage , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[83]  B. Obradovic,et al.  Bioreactor cultivation conditions modulate the composition and mechanical properties of tissue‐engineered cartilage , 1999, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[84]  J. Kramer,et al.  In vitro differentiation of mouse ES cells: bone and cartilage. , 2003, Methods in enzymology.

[85]  G. Balian,et al.  Marrow stromal cells embedded in alginate for repair of osteochondral defects. , 2000, Arthroscopy : the journal of arthroscopic & related surgery : official publication of the Arthroscopy Association of North America and the International Arthroscopy Association.

[86]  Albert C. Chen,et al.  Depth‐dependent confined compression modulus of full‐thickness bovine articular cartilage , 1997, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[87]  A. Tanigami,et al.  Use of bone morphogenetic protein 2 and diffusion chambers to engineer cartilage tissue for the repair of defects in articular cartilage. , 2005, Arthritis and rheumatism.

[88]  Darwin J. Prockop,et al.  In vitro cartilage formation by human adult stem cells from bone marrow stroma defines the sequence of cellular and molecular events during chondrogenesis , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[89]  C. Ohlsson,et al.  Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. , 1994, The New England journal of medicine.

[90]  F. Gustavson,et al.  Bioreactors for tissue engineering of cartilage. , 2009, Advances in biochemical engineering/biotechnology.

[91]  R. Salter,et al.  Biological resurfacing of full-thickness defects in patellar articular cartilage of the rabbit. Investigation of autogenous periosteal grafts subjected to continuous passive motion. , 1992, The Journal of bone and joint surgery. British volume.

[92]  H J Mankin,et al.  Articular cartilage: degeneration and osteoarthritis, repair, regeneration, and transplantation. , 1998, Instructional course lectures.

[93]  M. Moss,et al.  1 – Vertebrate Cartilages* , 1983 .

[94]  Martin Styner,et al.  Computer-Assisted Ankle Joint Arthroplasty Using Bio-engineered Autografts , 2005, MICCAI.

[95]  E B Hunziker,et al.  Mechanical compression modulates matrix biosynthesis in chondrocyte/agarose culture. , 1995, Journal of cell science.

[96]  E. Hunziker.  Articular cartilage repair: problems and perspectives. , 2000, Biorheology.

[97]  U A Stock,et al.  Tissue engineering: current state and prospects. , 2001, Annual review of medicine.

[98]  R. Hirsch,et al.  Prevalence of knee osteoarthritis in the United States: arthritis data from the Third National Health and Nutrition Examination Survey 1991-94. , 2006, The Journal of rheumatology.

[99]  Yilin Cao,et al.  Comparative study of the use of poly(glycolic acid), calcium alginate and pluronics in the engineering of autologous porcine cartilage. , 1998, Journal of biomaterials science. Polymer edition.

[100]  J. Mason,et al.  Stem Cells as Platforms for Delivery of Genes to Enhance Cartilage Repair , 2003, The Journal of bone and joint surgery. American volume.

[101]  L. Peterson,et al.  The repair of experimentally produced defects in rabbit articular cartilage by autologous chondrocyte transplantation , 1989, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[102]  J. Hubbell,et al.  Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering , 2005, Nature Biotechnology.

[103]  Robert Langer,et al.  Controlled‐release of IGF‐I and TGF‐β1 in a photopolymerizing hydrogel for cartilage tissue engineering , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[104]  G. Lister,et al.  Perichondrial resurfacing arthroplasty in the hand. , 1984, The Journal of hand surgery.

[105]  A. Spradling,et al.  Stem cells find their niche , 2001, Nature.

[106]  H. Kleinman,et al.  Complex Extracellular Matrices Promote Tissue‐Specific Stem Cell Differentiation , 2005, Stem cells.

[107]  J. Elisseeff,et al.  Biological Response of Chondrocytes to Hydrogels , 2002, Annals of the New York Academy of Sciences.

[108]  David J Mooney,et al.  Alginate type and RGD density control myoblast phenotype. , 2002, Journal of biomedical materials research.

[109]  Nathaniel S. Hwang,et al.  Musculoskeletal Differentiation of Cells Derived from Human Embryonic Germ Cells , 2005, Stem cells.

[110]  K. Takaoka,et al.  Biodegradable Poly-d,l-Lactic Acid-Polyethylene Glycol Block Copolymers as a BMP Delivery System for Inducing Bone , 2001, The Journal of bone and joint surgery. American volume.

[111]  S. Cannon,et al.  The use of chondrogide membrane in autologous chondrocyte implantation. , 2004, The Knee.

[112]  M D McKee,et al.  Tissue engineering of cartilage using an injectable and adhesive chitosan-based cell-delivery vehicle. , 2005, Osteoarthritis and cartilage.

[113]  M. Ochi,et al.  Effect of glycosaminoglycan production on hardness of cultured cartilage fabricated by the collagen-gel embedding method. , 2002, Tissue engineering.

[114]  Jeremy J Mao,et al.  Tissue-engineered osteochondral constructs in the shape of an articular condyle. , 2005, The Journal of bone and joint surgery. American volume.

[115]  S. Nishimura,et al.  Feasibility of polysaccharide hybrid materials for scaffolds in cartilage tissue engineering: evaluation of chondrocyte adhesion to polyion complex fibers prepared from alginate and chitosan. , 2004, Biomacromolecules.

[116]  Steven A Olson,et al.  The Development of Posttraumatic Arthritis After Articular Fracture , 2006, Journal of orthopaedic trauma.

[117]  J. Yoon,et al.  Tissue‐Engineered Cartilage on Biodegradable Macroporous Scaffolds: Cell Shape and Phenotypic Expression , 2002, The Laryngoscope.

[118]  Borjana Mikic,et al.  Mechanical Modulation of Cartilage Structure and Function During Embryogenesis in the Chick , 2004, Annals of Biomedical Engineering.

[119]  T. Minas.  Autologous Chondrocyte Implantation for Focal Chondral Defects of the Knee , 2001, Clinical orthopaedics and related research.

[120]  K Masuda,et al.  Tissue engineering of stratified articular cartilage from chondrocyte subpopulations. , 2003, Osteoarthritis and cartilage.

[121]  A. Caplan.  Embryonic development and the principles of tissue engineering. , 2003, Novartis Foundation symposium.

[122]  A. Mikos,et al.  Review: tissue engineering for regeneration of articular cartilage. , 2000, Biomaterials.

[123]  N. Lee,et al.  Divergent responses of chondrocytes and endothelial cells to shear stress: cross-talk among COX-2, the phase 2 response, and apoptosis. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[124]  D. Eyre,et al.  Articular cartilage collagen: an irreplaceable framework? , 2006, European cells & materials.

[125]  R A Good,et al.  Cellular engineering. , 1977, Clinical bulletin.

[126]  R. Eavey,et al.  The effect of fibroblast growth factor and transforming growth factor-beta on porcine chondrocytes and tissue-engineered autologous elastic cartilage. , 2001, Tissue engineering.

[127]  Shyni Varghese,et al.  Multifunctional chondroitin sulphate for cartilage tissue-biomaterial integration. , 2007, Nature materials.

[128]  David J. Mooney,et al.  Cyclic mechanical strain regulates the development of engineered smooth muscle tissue , 1999, Nature Biotechnology.

[129]  F. Barry,et al.  Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation-dependent gene expression of matrix components. , 2001, Experimental cell research.

[130]  A. Lumsden,et al.  Roles of Hemodynamic Forces in Vascular Cell Differentiation , 2005, Annals of Biomedical Engineering.

[131]  A. Poole,et al.  Chondrogenesis in periosteal explants. An organ culture model for in vitro study. , 1994, The Journal of bone and joint surgery. American volume.

[132]  D. Mooney,et al.  Hydrogels for tissue engineering: scaffold design variables and applications. , 2003, Biomaterials.

[133]  D. Chepeha,et al.  Tissue engineering: the current status of this futuristic modality in head neck reconstruction , 2004, Current opinion in otolaryngology & head and neck surgery.

[134]  Virginie Sottile,et al.  In vitro osteogenic differentiation of human ES cells. , 2003, Cloning and stem cells.

[135]  Wan-Ju Li,et al.  Chondrocyte phenotype in engineered fibrous matrix is regulated by fiber size. , 2006, Tissue engineering.

[136]  Miqin Zhang,et al.  Chitosan-alginate as scaffolding material for cartilage tissue engineering. , 2005, Journal of biomedical materials research. Part A.

[137]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[138]  J. Ralphs,et al.  Biology of fibrocartilage cells. , 2004, International review of cytology.

[139]  Steven A. Goldstein,et al.  Chondrocyte Differentiation is Modulated by Frequency and Duration of Cyclic Compressive Loading , 2001, Annals of Biomedical Engineering.

[140]  Naoki Ishiguro,et al.  Chondrogenesis enhanced by overexpression of sox9 gene in mouse bone marrow-derived mesenchymal stem cells. , 2003, Biochemical and biophysical research communications.

[141]  D. Buttle,et al.  Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. , 1986, Biochimica et biophysica acta.

[142]  J. Vacanti,et al.  Tissue Engineering and Its Potential Impact on Surgery , 2001, World journal of surgery.

[143]  C. Streuli,et al.  Extracellular matrix remodelling and cellular differentiation. , 1999, Current opinion in cell biology.

[144]  Jennifer H Elisseeff,et al.  An extracellular matrix extract for tissue-engineered cartilage. , 2003, IEEE engineering in medicine and biology magazine : the quarterly magazine of the Engineering in Medicine & Biology Society.

[145]  J. Hubbell,et al.  Incorporation of adhesion peptides into nonadhesive hydrogels useful for tissue resurfacing. , 1998, Journal of biomedical materials research.

[146]  P. Person.  2 – Invertebrate Cartilages , 1983 .

[147]  M. Bhargava,et al.  Effect of serum and platelet-derived growth factor on chondrocytes grown in collagen gels. , 1999, Tissue engineering.

[148]  M. Saito,et al.  Human autologous culture expanded bone marrow mesenchymal cell transplantation for repair of cartilage defects in osteoarthritic knees. , 2002, Osteoarthritis and cartilage.

[149]  D. Seliktar,et al.  Extracellular Stimulation in Tissue Engineering , 2005, Annals of the New York Academy of Sciences.

[150]  S. Hsu,et al.  Evaluation of chitosan-alginate-hyaluronate complexes modified by an RGD-containing protein as tissue-engineering scaffolds for cartilage regeneration. , 2004, Artificial organs.

[151]  Lars Engebretsen,et al.  Clinical application of scaffolds for cartilage tissue engineering , 2008, Knee Surgery, Sports Traumatology, Arthroscopy.

[152]  Linda G Griffith,et al.  Engineering principles of clinical cell-based tissue engineering. , 2004, The Journal of bone and joint surgery. American volume.

[153]  T. Holmes,et al.  Novel peptide-based biomaterial scaffolds for tissue engineering. , 2002, Trends in biotechnology.

[154]  G. Verbruggen,et al.  Culture of chondrocytes in alginate surrounded by fibrin gel: characteristics of the cells over a period of eight weeks , 2001, Annals of the rheumatic diseases.

[155]  S J Bryant,et al.  Cytocompatibility of UV and visible light photoinitiating systems on cultured NIH/3T3 fibroblasts in vitro , 2000, Journal of biomaterials science. Polymer edition.

[156]  Kyriacos A. Athanasiou,et al.  Biomechanical Strategies for Articular Cartilage Regeneration , 2003, Annals of Biomedical Engineering.

[157]  L. Hangody,et al.  Mosaicplasty for the treatment of articular cartilage defects: application in clinical practice. , 1998, Orthopedics.

[158]  Derrick E Rancourt,et al.  Induction of chondro-, osteo- and adipogenesis in embryonic stem cells by bone morphogenetic protein-2: Effect of cofactors on differentiating lineages , 2005, BMC Developmental Biology.

[159]  Michael Sittinger,et al.  A tissue-engineering model for the manufacture of auricular-shaped cartilage implants , 2002, European Archives of Oto-Rhino-Laryngology.

[160]  R. Tuan,et al.  Concepts in gene therapy for cartilage repair. , 2008, Injury.

[161]  Guoping Chen,et al.  The use of a novel PLGA fiber/collagen composite web as a scaffold for engineering of articular cartilage tissue with adjustable thickness. , 2003, Journal of biomedical materials research. Part A.

[162]  V. Goldberg,et al.  Cartilage regeneration using principles of tissue engineering. , 2001, Clinical orthopaedics and related research.

[163]  A. Nixon,et al.  Gene-based approaches for the repair of articular cartilage , 2004, Gene Therapy.

[164]  J. Thumboo,et al.  Direct and indirect costs of osteoarthritis in Singapore: a comparative study among multiethnic Asian patients with osteoarthritis. , 2007, The Journal of rheumatology.

[165]  Ranieri Cancedda,et al.  Serum-free growth medium sustains commitment of human articular chondrocyte through maintenance of Sox9 expression. , 2004, Tissue engineering.

[166]  Christian Clausen,et al.  Aging is associated with decreased maximal life span and accelerated senescence of bone marrow stromal cells. , 2003, Bone.

[167]  L. Hangody,et al.  Mosaicplasty for the Treatment of Articular Defects of the Knee and Ankle , 2001, Clinical orthopaedics and related research.

[168]  G. B. Fiore,et al.  Mechanobiology of engineered cartilage cultured under a quantified fluid-dynamic environment , 2002, Biomechanics and modeling in mechanobiology.

[169]  P. D. Di Cesare,et al.  Scaffolds for Articular Cartilage Repair , 2004, Annals of Biomedical Engineering.

[170]  L. Bonassar,et al.  A Novel Injectable Approach for Cartilage Formation in Vivo Using PLG Microspheres , 2004, Annals of Biomedical Engineering.

[171]  Gordana Vunjak-Novakovic,et al.  Bioreactors mediate the effectiveness of tissue engineering scaffolds , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[172]  P. Manson,et al.  The effect of incorporating RGD adhesive peptide in polyethylene glycol diacrylate hydrogel on osteogenesis of bone marrow stromal cells. , 2005, Biomaterials.

[173]  Aguilar Ef.  Auricular reconstruction in congenital anomalies of the ear. , 2001 .

[174]  S. Kawai,et al.  Chondrogenic differentiation of murine embryonic stem cells: Effects of culture conditions and dexamethasone , 2004, Journal of cellular biochemistry.

[175]  Jennifer L. West,et al.  Tethered-TGF-β increases extracellular matrix production of vascular smooth muscle cells , 2001 .

[176]  S. Trippel.  Growth factor actions on articular cartilage. , 1995, The Journal of rheumatology. Supplement.

[177]  R. Tuan,et al.  Mesenchymal stem cell-based cartilage tissue engineering: cells, scaffold and biology. , 2004, Cytotherapy.

[178]  G. Lust,et al.  Insulin-like growth factor-I enhances cell-based repair of articular cartilage. , 2002, The Journal of bone and joint surgery. British volume.

[179]  K. Yao,et al.  Biomimetic surface modification of poly(L-lactic acid) with chitosan and its effects on articular chondrocytes in vitro. , 2003, Biomaterials.

[180]  D. Eyre.  Articular cartilage and changes in Arthritis: Collagen of articular cartilage , 2001, Arthritis research.

[181]  Gerard A. Ateshian,et al.  A Paradigm for Functional Tissue Engineering of Articular Cartilage via Applied Physiologic Deformational Loading , 2004, Annals of Biomedical Engineering.

[182]  W. B. van den Berg,et al.  Crosslinked type II collagen matrices: preparation, characterization, and potential for cartilage engineering. , 2002, Biomaterials.

[183]  Junzo Tanaka,et al.  Growth factor combination for chondrogenic induction from human mesenchymal stem cell. , 2004, Biochemical and biophysical research communications.

[184]  Jerry C. Hu,et al.  Zonal and topographical differences in articular cartilage gene expression , 2004, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[185]  E B Hunziker,et al.  Articular cartilage repair: basic science and clinical progress. A review of the current status and prospects. , 2002, Osteoarthritis and cartilage.

[186]  Tae Gwan Park,et al.  Hyaluronic acid modified biodegradable scaffolds for cartilage tissue engineering. , 2005, Biomaterials.

[187]  Jeffrey A Hubbell,et al.  Materials as morphogenetic guides in tissue engineering. , 2003, Current opinion in biotechnology.

[188]  F. Guilak,et al.  Multipotent Stromal Cells Derived From the Infrapatellar Fat Pad of the Knee , 2003, Clinical orthopaedics and related research.

[189]  T. Speaker,et al.  A method to attach lectins to the surface of spermine alginate microcapsules based on the avidin biotin interaction. , 1996, Journal of microencapsulation.

[190]  K W Anderson,et al.  Cell-interactive Alginate Hydrogels for Bone Tissue Engineering , 2001, Journal of dental research.

[191]  Edward C. Jones,et al.  Chondral resurfacing of articular cartilage defects in the knee with the microfracture technique. Surgical technique. , 2006, The Journal of bone and joint surgery. American volume.

[192]  P. Benya,et al.  Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels , 1982, Cell.

[193]  C. Heath,et al.  Influence of intermittent pressure, fluid flow, and mixing on the regenerative properties of articular chondrocytes. , 1999, Biotechnology and bioengineering.

[194]  Jennifer H. Elisseeff,et al.  Engineering Structurally Organized Cartilage and Bone Tissues , 2004, Annals of Biomedical Engineering.

[195]  K. Ono,et al.  Chondrocytes embedded in collagen gels maintain cartilage phenotype during long-term cultures. , 1984, Clinical orthopaedics and related research.

[196]  E. Thonar,et al.  Adult human chondrocytes cultured in alginate form a matrix similar to native human articular cartilage. , 1996, The American journal of physiology.

[197]  E. Thonar,et al.  Human Septal Chondrocyte Redifferentiation in Alginate, Polyglycolic Acid Scaffold, and Monolayer Culture , 2003, The Laryngoscope.

[198]  L. Niswander,et al.  Requirement for BMP Signaling in Interdigital Apoptosis and Scale Formation , 1996, Science.

[199]  J. Thomson,et al.  Embryonic stem cell lines derived from human blastocysts. , 1998, Science.

[200]  X. Edward Guo,et al.  Mechano-electrochemical properties of articular cartilage: their inhomogeneities and anisotropies. , 2002, Annual review of biomedical engineering.

[201]  A. Goodship,et al.  The effects of hyaluronic acid on articular chondrocytes. , 2005, Journal of Bone and Joint Surgery-british Volume.

[202]  A. J. Grodzinsky,et al.  Self-assembling peptide hydrogel fosters chondrocyte extracellular matrix production and cell division: Implications for cartilage tissue repair , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[203]  A. U. Daniels,et al.  Effects of scaffold composition and architecture on human nasal chondrocyte redifferentiation and cartilaginous matrix deposition. , 2005, Biomaterials.

[204]  Cato T Laurencin,et al.  Electrospun nanofibrous structure: a novel scaffold for tissue engineering. , 2002, Journal of biomedical materials research.

[205]  R Langer,et al.  Selective differentiation of mammalian bone marrow stromal cells cultured on three-dimensional polymer foams. , 2001, Journal of biomedical materials research.

[206]  Somponnat Sampattavanich,et al.  Effects of Three‐Dimensional Culture and Growth Factors on the Chondrogenic Differentiation of Murine Embryonic Stem Cells , 2006, Stem cells.

[207]  H. Weinans,et al.  Improved cartilage integration and interfacial strength after enzymatic treatment in a cartilage transplantation model , 2004, Arthritis research & therapy.

[208]  G. Naughton,et al.  Evaluation of matrix scaffolds for tissue engineering of articular cartilage grafts. , 1997, Journal of biomedical materials research.

[209]  C. M. Agrawal,et al.  Biodegradable polymeric scaffolds for musculoskeletal tissue engineering. , 2001, Journal of biomedical materials research.

[210]  G. Gibson,et al.  Effects of matrix macromolecules on chondrocyte gene expression: synthesis of a low molecular weight collagen species by cells cultured within collagen gels , 1982, The Journal of cell biology.

[211]  C. L. Murphy,et al.  Effect of oxygen tension and alginate encapsulation on restoration of the differentiated phenotype of passaged chondrocytes. , 2001, Tissue engineering.

[212]  B L Currier,et al.  Biodegradable Polymer Scaffolds for Cartilage Tissue Engineering , 2001, Clinical orthopaedics and related research.

[213]  H. Lorenz,et al.  Multilineage cells from human adipose tissue: implications for cell-based therapies. , 2001, Tissue engineering.

[214]  S. Waldman,et al.  Effect of Biomechanical Conditioning on Cartilaginous Tissue Formation in Vitro , 2003, The Journal of bone and joint surgery. American volume.

[215]  J. Vacanti,et al.  Bone and cartilage reconstruction with tissue engineering approaches. , 1994, Otolaryngologic clinics of North America.

[216]  A. Zaninelli,et al.  Osteoarthritis: an overview of the disease and its treatment strategies. , 2005, Seminars in arthritis and rheumatism.

[217]  R Langer,et al.  Neocartilage formation in vitro and in vivo using cells cultured on synthetic biodegradable polymers. , 1993, Journal of biomedical materials research.

[218]  N. Adachi,et al.  Optimum combination of monolayer and three-dimensional cultures for cartilage-like tissue engineering. , 2003, Tissue engineering.

[219]  Johnny Huard,et al.  Gene therapy and tissue engineering based on muscle-derived stem cells. , 2002, Current opinion in molecular therapeutics.

[220]  S. Nishimura,et al.  Feasibility of chitosan-based hyaluronic acid hybrid biomaterial for a novel scaffold in cartilage tissue engineering. , 2005, Biomaterials.

[221]  N. Socci,et al.  Derivation of Multipotent Mesenchymal Precursors from Human Embryonic Stem Cells , 2005, PLoS medicine.

[222]  M. Heidaran,et al.  Molecular Signaling in Bioengineered Tissue Microenvironments , 2002, Annals of the New York Academy of Sciences.

[223]  S. Bhatia,et al.  Three-Dimensional Photopatterning of Hydrogels Containing Living Cells , 2002 .

[224]  B. A. Byers,et al.  Transient exposure to transforming growth factor beta 3 under serum-free conditions enhances the biomechanical and biochemical maturation of tissue-engineered cartilage. , 2008, Tissue engineering. Part A.

[225]  A Ratcliffe,et al.  Formulation of PEG-based hydrogels affects tissue-engineered cartilage construct characteristics , 2001, Journal of materials science. Materials in medicine.

[226]  E. Thonar,et al.  Characterization of human nasal septal chondrocytes cultured in alginate. , 2005, Journal of the American College of Surgeons.

[227]  Christopher G Williams,et al.  In vitro chondrogenesis of bone marrow-derived mesenchymal stem cells in a photopolymerizing hydrogel. , 2003, Tissue engineering.

[228]  Catherine M. Verfaillie,et al.  Pluripotency of mesenchymal stem cells derived from adult marrow , 2002, Nature.

[229]  P. Collas,et al.  Induction of dedifferentiation, genomewide transcriptional programming, and epigenetic reprogramming by extracts of carcinoma and embryonic stem cells. , 2005, Molecular biology of the cell.

[230]  Guoping Chen,et al.  Tissue engineering of cartilage using a hybrid scaffold of synthetic polymer and collagen. , 2004, Tissue engineering.

[231]  Krishnendu Roy,et al.  Biomimetic three-dimensional cultures significantly increase hematopoietic differentiation efficacy of embryonic stem cells. , 2005, Tissue engineering.

[232]  P. Benya,et al.  Independent regulation of collagen types by chondrocytes during the loss of differentiated function in culture , 1978, Cell.

[233]  G. Vunjak‐Novakovic,et al.  Bioreactor studies of native and tissue engineered cartilage. , 2002, Biorheology.

[234]  Lorenzo Moroni,et al.  Biomaterials engineered for integration , 2008 .

[235]  R. A. Mashelkar,et al.  Novel Macroscopic Self-Organization in Polymer Gels , 2001 .

[236]  M. Shive,et al.  Chitosan-glycerol phosphate/blood implants improve hyaline cartilage repair in ovine microfracture defects. , 2005, The Journal of bone and joint surgery. American volume.

[237]  Adel Alhadlaq,et al.  Adult Stem Cell Driven Genesis of Human-Shaped Articular Condyle , 2004, Annals of Biomedical Engineering.

[238]  J. Itskovitz‐Eldor,et al.  Differentiation of human embryonic stem cells on three-dimensional polymer scaffolds , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[239]  M. Neumeister,et al.  A tissue-engineering technique for vascularized laryngotracheal reconstruction. , 2003, Archives of otolaryngology--head & neck surgery.

[240]  S. Bruder,et al.  Mesenchymal stem cells: building blocks for molecular medicine in the 21st century. , 2001, Trends in molecular medicine.

[241]  E. Kastenbauer,et al.  Clinical aspects and strategy for biomaterial engineering of an auricle based on three-dimensional stereolithography , 2003, European Archives of Oto-Rhino-Laryngology.

[242]  A. Seifalian,et al.  Advancing cartilage tissue engineering: the application of stem cell technology. , 2005, Current opinion in biotechnology.

[243]  M. Ochi,et al.  Current concepts in tissue engineering technique for repair of cartilage defect. , 2001, Artificial organs.

[244]  Ung-Jin Kim,et al.  In vitro cartilage tissue engineering with 3D porous aqueous-derived silk scaffolds and mesenchymal stem cells. , 2005, Biomaterials.

[245]  S. Hsu,et al.  Evaluation of biodegradable polyesters modified by type II collagen and Arg-Gly-Asp as tissue engineering scaffolding materials for cartilage regeneration. , 2006, Artificial organs.

[246]  H. Mankin,et al.  The response of articular cartilage to mechanical injury. , 1982, The Journal of bone and joint surgery. American volume.

[247]  E. Caterson,et al.  Three-dimensional cartilage formation by bone marrow-derived cells seeded in polylactide/alginate amalgam. , 2001, Journal of biomedical materials research.

[248]  D. Krause,et al.  Plasticity of marrow-derived stem cells. , 2003, Blood.

[249]  Ick Chan Kwon,et al.  Effects of a chitosan scaffold containing TGF-beta1 encapsulated chitosan microspheres on in vitro chondrocyte culture. , 2004, Artificial organs.

[250]  Hans Hauner,et al.  Cartilage-like gene expression in differentiated human stem cell spheroids: a comparison of bone marrow-derived and adipose tissue-derived stromal cells. , 2003, Arthritis and rheumatism.

[251]  H. Stegemann,et al.  Determination of hydroxyproline. , 1967, Clinica chimica acta; international journal of clinical chemistry.

[252]  R. Sellers,et al.  Repair of Articular Cartilage Defects One Year After Treatment with Recombinant Human Bone Morphogenetic Protein-2 (rhBMP-2)* , 2000, The Journal of bone and joint surgery. American volume.

[253]  M. Brittberg,et al.  Autologous chondrocytes used for articular cartilage repair: an update. , 2001, Clinical orthopaedics and related research.

[254]  A. Jayakrishnan,et al.  Evaluation of an in situ forming hydrogel wound dressing based on oxidized alginate and gelatin. , 2005, Biomaterials.

[255]  M. McKee,et al.  Aged bovine chondrocytes display a diminished capacity to produce a collagen‐rich, mechanically functional cartilage extracellular matrix , 2005, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[256]  S. Bryant,et al.  Crosslinking density influences the morphology of chondrocytes photoencapsulated in PEG hydrogels during the application of compressive strain , 2004, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[257]  Peter W Zandstra,et al.  Efficiency of embryoid body formation and hematopoietic development from embryonic stem cells in different culture systems. , 2002, Biotechnology and bioengineering.