Matrix Development in Self-Assembly of Articular Cartilage

Background Articular cartilage is a highly functional tissue which covers the ends of long bones and serves to ensure proper joint movement. A tissue engineering approach that recapitulates the developmental characteristics of articular cartilage can be used to examine the maturation and degeneration of cartilage and produce fully functional neotissue replacements for diseased tissue. Methodology/Principal Findings This study examined the development of articular cartilage neotissue within a self-assembling process in two phases. In the first phase, articular cartilage constructs were examined at 1, 4, 7, 10, 14, 28, 42, and 56 days immunohistochemically, histologically, and through biochemical analysis for total collagen and glycosaminoglycan (GAG) content. Based on statistical changes in GAG and collagen levels, four time points from the first phase (7, 14, 28, and 56 days) were chosen to carry into the second phase, where the constructs were studied in terms of their mechanical characteristics, relative amounts of collagen types II and VI, and specific GAG types (chondroitin 4-sulfate, chondroitin 6-sulfate, dermatan sulfate, and hyaluronan). Collagen type VI was present in initial abundance and then localized to a pericellular distribution at 4 wks. N-cadherin activity also spiked at early stages of neotissue development, suggesting that self-assembly is mediated through a minimization of free energy. The percentage of collagen type II to total collagen significantly increased over time, while the proportion of collagen type VI to total collagen decreased between 1 and 2 wks. The chondroitin 6- to 4- sulfate ratio decreased steadily during construct maturation. In addition, the compressive properties reached a plateau and tensile characteristics peaked at 4 wks. Conclusions/Significance The indices of cartilage formation examined in this study suggest that tissue maturation in self-assembled articular cartilage mirrors known developmental processes for native tissue. In terms of tissue engineering, it is suggested that exogenous stimulation may be necessary after 4 wks to further augment the functionality of developing constructs.

[1]  D. Mooney,et al.  Combining chondrocytes and smooth muscle cells to engineer hybrid soft tissue constructs. , 2000, Tissue engineering.

[2]  Y. Kawakami,et al.  Involvement of Wnt‐5a in chondrogenic pattern formation in the chick limb bud , 1999, Development, growth & differentiation.

[3]  R. Cancedda,et al.  Type VI collagen expression is upregulated in the early events of chondrocyte differentiation. , 1993, Development.

[4]  G E Kempson,et al.  Relationship between the tensile properties of articular cartilage from the human knee and age. , 1982, Annals of the rheumatic diseases.

[5]  R. Mason,et al.  Chondroitin sulphation patterns in synovial fluid in osteoarthritis subsets , 1999, Annals of the rheumatic diseases.

[6]  D. E. Ashhurst,et al.  Development and ageing of the articular cartilage of the rabbit knee joint: distribution of the fibrillar collagens , 1996, Anatomy and Embryology.

[7]  K. J. Grande-Allen,et al.  Glycosaminoglycans and proteoglycans in normal mitral valve leaflets and chordae: association with regions of tensile and compressive loading. , 2004, Glycobiology.

[8]  G. Bentley,et al.  The spatial and temporal pattern of collagens I and II and keratan sulphate in the developing chick metatarsophalangeal joint. , 1987, Development.

[9]  Wilson C. Hayes,et al.  Basic Orthopaedic Biomechanics , 1995 .

[10]  Dan L. Bader,et al.  Mechanical Conditioning Influences the Metabolic Response of Cell-Seeded Constructs , 2003, Cells Tissues Organs.

[11]  C. A. Poole.  Review. Articular cartilage chondrons: form, function and failure , 1997 .

[12]  A. Hollander,et al.  Localization of type VI collagen in tissue-engineered cartilage on polymer scaffolds. , 2006, Tissue engineering.

[13]  S. Ayad,et al.  The Distribution of Type VI Collagen in the Developing Tissues of the Bovine Femoral Head , 1999, The Histochemical Journal.

[14]  Van C. Mow,et al.  Structure and function of articular cartilage and meniscus , 2005 .

[15]  R. Tuan,et al.  Expression and functional involvement of N-cadherin in embryonic limb chondrogenesis. , 1994, Development.

[16]  F. Berenbaum.  The quest for the Holy Grail: a disease-modifying osteoarthritis drug , 2007, Arthritis research & therapy.

[17]  M. Ferguson,et al.  The development of articular cartilage: II. The spatial and temporal patterns of glycosaminoglycans and small leucine-rich proteoglycans. , 1996, Journal of anatomy.

[18]  Farshid Guilak,et al.  The biomechanical role of the chondrocyte pericellular matrix in articular cartilage. , 2005, Acta biomaterialia.

[19]  Jerry C. Hu,et al.  Structure and Function of Articular Cartilage , 2003 .

[20]  P. Pirttiniemi,et al.  Comparison of Amounts and Properties of Collagen and Proteoglycans in Condylar, Costal and Nasal Cartilages , 1999, Cells Tissues Organs.

[21]  Farshid Guilak,et al.  Alterations in the mechanical properties of the human chondrocyte pericellular matrix with osteoarthritis. , 2003, Journal of biomechanical engineering.

[22]  R. Tuan,et al.  Spatiotemporal profile of N-cadherin expression in the developing limb mesenchyme. , 1994, Cell adhesion and communication.

[23]  M. Stewart,et al.  The role of bone morphogenetic proteins in articular cartilage development, homeostasis and repair , 2007, Veterinary and Comparative Orthopaedics and Traumatology.

[24]  Jerry C. Hu,et al.  The effects of intermittent hydrostatic pressure on self-assembled articular cartilage constructs. , 2006, Tissue engineering.

[25]  F. Guilak,et al.  Zonal variations in the three-dimensional morphology of the chondron measured in situ using confocal microscopy. , 2006, Osteoarthritis and cartilage.

[26]  R. Midura,et al.  Adaptation of FACE methodology for microanalysis of total hyaluronan and chondroitin sulfate composition from cartilage. , 2000, Glycobiology.

[27]  C. A. Poole.  Articular cartilage chondrons: form, function and failure. , 1997, Journal of anatomy.

[28]  J. F. Woessner,et al.  The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. , 1961, Archives of biochemistry and biophysics.

[29]  T. Tokuoka,et al.  Ascorbic acid regulates multiple metabolic activities of cartilage cells. , 1991, The American journal of clinical nutrition.

[30]  S. Ayad,et al.  Chondrons from articular cartilage: I. Immunolocalization of type VI collagen in the pericellular capsule of isolated canine tibial chondrons. , 1988, Journal of cell science.

[31]  Y. Ikada,et al.  Development of artificial articular cartilage , 2000, Proceedings of the Institution of Mechanical Engineers. Part H, Journal of engineering in medicine.

[32]  Yuehuei H. An,et al.  Handbook of Histology Methods for Bone and Cartilage , 2003, Humana Press.

[33]  W A Hing,et al.  The influence of the pericellular microenvironment on the chondrocyte response to osmotic challenge. , 2002, Osteoarthritis and cartilage.

[34]  Kyriacos A Athanasiou,et al.  Effects of confinement on the mechanical properties of self‐assembled articular cartilage constructs in the direction orthogonal to the confinement surface , 2008, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[35]  M. Bayliss,et al.  Age-related changes in the composition and structure of human articular-cartilage proteoglycans. , 1978, The Biochemical journal.

[36]  Erkki Ruoslahti,et al.  Proteoglycans as modulators of growth factor activities , 1991, Cell.

[37]  B. Griffin,et al.  Glycosaminoglycan profiles of myxomatous mitral leaflets and chordae parallel the severity of mechanical alterations. , 2003, Journal of the American College of Cardiology.

[38]  L. Rosenberg.  Chemical basis for the histological use of safranin O in the study of articular cartilage. , 1971, The Journal of bone and joint surgery. American volume.

[39]  F. Guilak,et al.  A Mechano-chemical Model for the Passive Swelling Response of an Isolated Chondron under Osmotic Loading , 2006, Biomechanics and modeling in mechanobiology.

[40]  Koichi Masuda,et al.  Tensile mechanical properties of bovine articular cartilage: Variations with growth and relationships to collagen network components , 2003, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[41]  B. Caterson,et al.  Detection of age-related changes in the distributions of keratan sulfates and chondroitin sulfates in developing chick limbs: an immunocytochemical study. , 1989, Development.

[42]  G E Kempson,et al.  The effects of proteolytic enzymes on the mechanical properties of adult human articular cartilage. , 1976, Biochimica et biophysica acta.

[43]  M. S. Steinberg,et al.  The differential adhesion hypothesis: a direct evaluation. , 2005, Developmental biology.

[44]  E. Thonar,et al.  Regulation of immature cartilage growth by IGF-I, TGF-β1, BMP-7, and PDGF-AB: role of metabolic balance between fixed charge and collagen network , 2008, Biomechanics and modeling in mechanobiology.

[45]  B. Swoboda,et al.  Expression of type VI collagen in normal and osteoarthritic human cartilage. , 1999, Osteoarthritis and cartilage.

[46]  Jerry C. Hu,et al.  A self-assembling process in articular cartilage tissue engineering. , 2006, Tissue engineering.

[47]  M. Ferguson,et al.  The development of articular cartilage: I. The spatial and temporal patterns of collagen types. , 1996, Journal of anatomy.

[48]  Koichi Masuda,et al.  Mechanisms of cartilage growth: modulation of balance between proteoglycan and collagen in vitro using chondroitinase ABC. , 2007, Arthritis and rheumatism.

[49]  A. Grodzinsky,et al.  Structure of pericellular matrix around agarose-embedded chondrocytes. , 2007, Osteoarthritis and cartilage.

[50]  Albert C. Chen,et al.  Compressive properties and function—composition relationships of developing bovine articular cartilage , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[51]  W M Lai,et al.  Biphasic indentation of articular cartilage--II. A numerical algorithm and an experimental study. , 1989, Journal of biomechanics.

[52]  P. Pavasant,et al.  Hyaluronan synthesis by epiphysial chondrocytes is regulated by growth hormone, insulin-like growth factor-1, parathyroid hormone and transforming growth factor-beta 1. , 1996, Matrix biology : journal of the International Society for Matrix Biology.

[53]  J. Buckwalter,et al.  Interspecies comparisons of in situ intrinsic mechanical properties of distal femoral cartilage , 1991, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[54]  S. Ayad,et al.  Chondrons from articular cartilage. V. Immunohistochemical evaluation of type VI collagen organisation in isolated chondrons by light, confocal and electron microscopy. , 1992, Journal of cell science.

[55]  R. Tuan,et al.  Long‐term in vitro analysis of limb cartilage development: Involvement of Wnt signaling , 2004, Journal of cellular biochemistry.

[56]  V. Mow,et al.  The mechanical environment of the chondrocyte: a biphasic finite element model of cell-matrix interactions in articular cartilage. , 2000, Journal of biomechanics.

[57]  B. Caterson,et al.  Macromolecular Organization and In Vitro Growth Characteristics of Scaffold-free Neocartilage Grafts , 2007, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[58]  D. Platt,et al.  Ageing of equine articular cartilage: structure and composition of aggrecan and decorin. , 1998, Equine veterinary journal.

[59]  K. Athanasiou,et al.  Tissue Engineering with Chondrogenically Differentiated Human Embryonic Stem Cells , 2007, Stem cells.

[60]  T. Glant,et al.  Chondrons from articular cartilage. (IV). Immunolocalization of proteoglycan epitopes in isolated canine tibial chondrons. , 1991, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[61]  C. Hamanishi,et al.  Hypoxia-induced hyaluronan synthesis by articular chondrocytes: the role of nitric oxide , 2006, Inflammation Research.

[62]  F. J. Dzida,et al.  Comparative study of the intrinsic mechanical properties of the human acetabular and femoral head cartilage , 1991, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[63]  E. Koyama,et al.  Development of Articular Cartilage: What Do We Know About it and How May It Occur? , 2000, Connective tissue research.

[64]  M. Simpson,et al.  Inducible Hyaluronan Production Reveals Differential Effects on Prostate Tumor Cell Growth and Tumor Angiogenesis* , 2007, Journal of Biological Chemistry.

[65]  V. Mow,et al.  Biphasic creep and stress relaxation of articular cartilage in compression? Theory and experiments. , 1980, Journal of biomechanical engineering.

[66]  D L Bader,et al.  The influence of elaborated pericellular matrix on the deformation of isolated articular chondrocytes cultured in agarose. , 1998, Biochimica et biophysica acta.

[67]  Farshid Guilak,et al.  Osteoarthritic changes in the biphasic mechanical properties of the chondrocyte pericellular matrix in articular cartilage. , 2005, Journal of biomechanics.

[68]  W. R. Jones,et al.  The deformation behavior and mechanical properties of chondrocytes in articular cartilage. , 1999, Osteoarthritis and cartilage.