An in vitro model for the pathological degradation of articular cartilage in osteoarthritis.

The objective of this study was to develop an in vitro cartilage degradation model that emulates the damage seen in early-stage osteoarthritis. To this end, cartilage explants were collagenase-treated to induce enzymatic degradation of collagen fibers and proteoglycans at the articular surface. To assess changes in mechanical properties, intact and degraded cartilage explants were subjected to a series of confined compression creep tests. Changes in extracellular matrix structure and composition were determined using biochemical and histological approaches. Our results show that collagenase-induced degradation increased the amount of deformation experienced by the cartilage explants under compression. An increase in apparent permeability as well as a decrease in instantaneous and aggregate moduli was measured following collagenase treatment. Histological analysis of degraded explants revealed the presence of surface fibrillation, proteoglycan depletion in the superficial and intermediate zones and loss of the lamina splendens. Collagen cleavage was confirmed by the Col II-3/4Cshort antibody. Degraded specimens experienced a significant decrease in proteoglycan content but maintained total collagen content. Repetitive testing of degraded samples resulted in the gradual collapse of the articular surface and the compaction of the superficial zone. Taken together, our data demonstrates that enzymatic degradation with collagenase can be used to emulate changes seen in early-stage osteoarthritis. Further, our in vitro model provides information on cartilage mechanics and insights on how matrix changes can affect cartilage's functional properties. More importantly, our model can be applied to develop and test treatment options for tissue repair.

[1]  H. Muir,et al.  An experimental model of osteoarthritis; early morphological and biochemical changes. , 1977, The Journal of bone and joint surgery. British volume.

[2]  P. Torzilli,et al.  Increased stromelysin-1 (MMP-3), proteoglycan degradation (3B3- and 7D4) and collagen damage in cyclically load-injured articular cartilage. , 2004, Osteoarthritis and cartilage.

[3]  J. Mort,et al.  Articular cartilage and changes in arthritis: Matrix degradation , 2001, Arthritis research.

[4]  C. Weiss,et al.  An ultrastructural study of osteoarthritis changes in the articular cartilage of human knees. , 1972, The Journal of bone and joint surgery. American volume.

[5]  P. Torzilli,et al.  Fourier transform infrared imaging spectroscopy analysis of collagenase-induced cartilage degradation. , 2005, Journal of biomedical optics.

[6]  J. Mizrahi,et al.  The increased swelling and instantaneous deformation of osteoarthritic cartilage is highly correlated with collagen degradation. , 2000, Arthritis and rheumatism.

[7]  E. Roos,et al.  High prevalence of osteoarthritis 14 years after an anterior cruciate ligament tear in male soccer players: a study of radiographic and patient relevant outcomes , 2004, Annals of the rheumatic diseases.

[8]  V C Mow,et al.  Tensile properties of human knee joint cartilage: I. Influence of ionic conditions, weight bearing, and fibrillation on the tensile modulus , 1986, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[9]  E. Ritman,et al.  Solute transport in cyclically deformed porous tissue scaffolds with controlled pore cross-sectional geometries. , 2009, Tissue engineering. Part A.

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

[11]  D L Bader,et al.  Quantification of sulfated glycosaminoglycans in chondrocyte/alginate cultures, by use of 1,9-dimethylmethylene blue. , 1996, Analytical biochemistry.

[12]  M A Freeman,et al.  The composition of normal and osteoarthritic articular cartilage from human knee joints. With special reference to unicompartmental replacement and osteotomy of the knee. , 1984, The Journal of bone and joint surgery. American volume.

[13]  J. Verhaar,et al.  An osteochondral culture model to study mechanisms involved in articular cartilage repair. , 2012, Tissue engineering. Part C, Methods.

[14]  V C Mow,et al.  Altered mechanics of cartilage with osteoarthritis: human osteoarthritis and an experimental model of joint degeneration. , 1999, Osteoarthritis and cartilage.

[15]  V C Mow,et al.  Variations in the intrinsic mechanical properties of human articular cartilage with age, degeneration, and water content. , 1982, The Journal of bone and joint surgery. American volume.

[16]  V C Mow,et al.  The biphasic poroviscoelastic behavior of articular cartilage: role of the surface zone in governing the compressive behavior. , 1993, Journal of biomechanics.

[17]  Alpo Pelttari,et al.  Articular cartilage superficial zone collagen birefringence reduced and cartilage thickness increased before surface fibrillation in experimental osteoarthritis , 1998, Annals of the rheumatic diseases.

[18]  V C Mow,et al.  Mechanical Properties of Canine Articular Cartilage Are Significantly Altered Following Transection of the Anterior Cruciate Ligament , 1994, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[19]  E. Hedbom,et al.  Molecular aspects of pathogenesis in osteoarthritis: the role of inflammation , 2002, Cellular and Molecular Life Sciences CMLS.

[20]  L. Klareskog,et al.  Characterization of a spontaneously occurring arthritis in male DBA/1 mice. , 1992, Arthritis and rheumatism.

[21]  Smith Rl.  Degradative enzymes in osteoarthritis. , 1999 .

[22]  F. Guilak,et al.  Simultaneous changes in the mechanical properties, quantitative collagen organization, and proteoglycan concentration of articular cartilage following canine meniscectomy , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[23]  T. Fields,et al.  Chick cartilage chondroitin sulfate proteoglycan core protein. I. Generation and characterization of peptides and specificity for glycosaminoglycan attachment. , 1990, The Journal of biological chemistry.

[24]  C. Enwemeka,et al.  A simplified method for the analysis of hydroxyproline in biological tissues. , 1996, Clinical biochemistry.

[25]  Lotke Pa,et al.  Changes in the permeability of human articular cartilage in early degenerative osteoarthritis. , 1971 .

[26]  Y. Zheng,et al.  Real-time ultrasonic assessment of progressive proteoglycan depletion in articular cartilage. , 2008, Ultrasound in medicine & biology.

[27]  A. Jeng,et al.  Elevated substance P and accelerated cartilage degradation in rabbit knees injected with interleukin-1 and tumor necrosis factor. , 1990, Arthritis and rheumatism.

[28]  C. Rorabeck,et al.  Damage to type II collagen in aging and osteoarthritis starts at the articular surface, originates around chondrocytes, and extends into the cartilage with progressive degeneration. , 1995, The Journal of clinical investigation.

[29]  H. Ohtsu,et al.  Induction of swelling, synovial hyperplasia and cartilage proteoglycan loss upon intra-articular injection of transforming growth factor beta-2 in the rabbit. , 1992, Cytokine.

[30]  Marc Thibault,et al.  Cyclic compression of cartilage/bone explants in vitro leads to physical weakening, mechanical breakdown of collagen and release of matrix fragments , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[31]  N. Schachar,et al.  Collagen fibril structure of normal, aging, and osteoarthritic cartilage , 1992, The Journal of pathology.

[32]  V. Mow,et al.  Tensile properties of human knee joint cartilage. II. Correlations between weight bearing and tissue pathology and the kinetics of swelling , 1987, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[33]  R. Dalziel,et al.  Articular cartilage. , 1971, Lancet.

[34]  C. Rorabeck,et al.  Enhanced cleavage of type II collagen by collagenases in osteoarthritic articular cartilage. , 1997, The Journal of clinical investigation.

[35]  P. Torzilli,et al.  Movement of interstitial water through loaded articular cartilage. , 1983, Journal of biomechanics.

[36]  H. Muir,et al.  Biochemical changes in the cartilage of the knee in experimental and natural osteoarthritis in the dog. , 1976, The Journal of bone and joint surgery. British volume.

[37]  K. Pritzker,et al.  Animal models for osteoarthritis: processes, problems and prospects. , 1994, Annals of the rheumatic diseases.

[38]  V C Mow,et al.  Altered structure-function relationships for articular cartilage in human osteoarthritis and an experimental canine model. , 1993, Agents and actions. Supplements.

[39]  D. Kelly,et al.  The role of the superficial region in determining the dynamic properties of articular cartilage. , 2012, Osteoarthritis and cartilage.

[40]  J. Jurvelin,et al.  Ultrasonic quantitation of superficial degradation of articular cartilage. , 2004, Ultrasound in medicine & biology.

[41]  S. Jimenez,et al.  Osteoarthritis cartilage histopathology: grading and staging. , 2006, Osteoarthritis and cartilage.

[42]  N. Broom,et al.  Articular cartilage collagen and proteoglycans. Their functional interdependency. , 1983, Arthritis and rheumatism.

[43]  Jerry C. Hu,et al.  A proposed model of naturally occurring osteoarthritis in the domestic rabbit , 2011, Lab Animal.

[44]  Martin Englund,et al.  The role of biomechanics in the initiation and progression of OA of the knee. , 2010, Best practice & research. Clinical rheumatology.

[45]  M. Hyttinen,et al.  Transgenic mouse models for studying the role of cartilage macromolecules in osteoarthritis. , 2002, Rheumatology.

[46]  R. G. Johnson,et al.  Degenerative changes in dog articular cartilage induced by a unilateral tibial valgus osteotomy. , 1988, Experimental pathology.

[47]  R. L. Smith.  Degradative enzymes in osteoarthritis. , 1999, Frontiers in bioscience : a journal and virtual library.

[48]  D. Zukor,et al.  Sites of collagenase cleavage and denaturation of type II collagen in aging and osteoarthritic articular cartilage and their relationship to the distribution of matrix metalloproteinase 1 and matrix metalloproteinase 13. , 2002, Arthritis and rheumatism.

[49]  J. Granda,et al.  Changes in the permeability of human articular cartilage in early degenerative osteoarthritis. , 1971, Surgical forum.

[50]  V. He,et al.  Clostridium histolyticum collagenases: a new look at some old enzymes. , 1992 .

[51]  W. B. van den Berg,et al.  Degenerative knee joint lesions in mice after a single intra-articular collagenase injection. A new model of osteoarthritis. , 1990, Journal of experimental pathology.

[52]  I. Reimann.  Experimental osteoarthritis of the knee in rabbits induced by alteration of the load-bearing. , 1973, Acta Orthopaedica Scandinavica.

[53]  A. Werner,et al.  Visualization of collagenase-induced cartilage degradation using NMR microscopy. , 1999, Investigative radiology.

[55]  G. M. Herring.  Methods for the study of the glycoproteins and proteoglycans of bone using bacterial collagenase , 1977, Calcified Tissue Research.

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