Mechanical injury potentiates proteoglycan catabolism induced by interleukin-6 with soluble interleukin-6 receptor and tumor necrosis factor alpha in immature bovine and adult human articular cartilage.

OBJECTIVE Traumatic joint injury can damage cartilage and release inflammatory cytokines from adjacent joint tissue. The present study was undertaken to study the combined effects of compression injury, tumor necrosis factor alpha (TNFalpha), and interleukin-6 (IL-6) and its soluble receptor (sIL-6R) on immature bovine and adult human knee and ankle cartilage, using an in vitro model, and to test the hypothesis that endogenous IL-6 plays a role in proteoglycan loss caused by a combination of injury and TNFalpha. METHODS Injured or uninjured cartilage disks were incubated with or without TNFalpha and/or IL-6/sIL-6R. Additional samples were preincubated with an IL-6-blocking antibody Fab fragment and subjected to injury and TNFalpha treatment. Treatment effects were assessed by histologic analysis, measurement of glycosaminoglycan (GAG) loss, Western blot to determine proteoglycan degradation, zymography, radiolabeling to determine chondrocyte biosynthesis, and Western blot and enzyme-linked immunosorbent assay to determine chondrocyte production of IL-6. RESULTS In bovine cartilage samples, injury combined with TNFalpha and IL-6/sIL-6R exposure caused the most severe GAG loss. Findings in human knee and ankle cartilage were strikingly similar to those in bovine samples, although in human ankle tissue, the GAG loss was less severe than that observed in human knee tissue. Without exogenous IL-6/sIL-6R, injury plus TNFalpha exposure up-regulated chondrocyte production of IL-6, but incubation with the IL-6-blocking Fab significantly reduced proteoglycan degradation. CONCLUSION Our findings indicate that mechanical injury potentiates the catabolic effects of TNFalpha and IL-6/sIL-6R in causing proteoglycan degradation in human and bovine cartilage. The temporal and spatial evolution of degradation suggests the importance of transport of biomolecules, which may be altered by overload injury. The catabolic effects of injury plus TNFalpha appeared partly due to endogenous IL-6, since GAG loss was partially abrogated by an IL-6-blocking Fab.

[1]  H. Iwaso,et al.  Intraarticular inflammatory cytokines in acute anterior cruciate ligament injured knee. , 2003, The Knee.

[2]  P. Lipsky.  Interleukin-6 and rheumatic diseases , 2006, Arthritis research & therapy.

[3]  K. Kuettner,et al.  Prevalence of degenerative morphological changes in the joints of the lower extremity. , 1997, Osteoarthritis and cartilage.

[4]  J. Kingma,et al.  Injuries Due to School Sports Accidents in 4 to 13-Yr.-Old Children , 2000, Perceptual and motor skills.

[5]  D. Eyre,et al.  The release of crosslinked peptides from type II collagen into human synovial fluid is increased soon after joint injury and in osteoarthritis. , 2003, Arthritis and rheumatism.

[6]  A. Grodzinsky,et al.  Biosynthetic response of cartilage explants to dynamic compression , 1989, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[7]  S. Connolly,et al.  Acute injury of the articular cartilage and subchondral bone: a common but unrecognized lesion in the immature knee. , 2004, AJR. American journal of roentgenology.

[8]  J. Saklatvala,et al.  FGF-2 is bound to perlecan in the pericellular matrix of articular cartilage, where it acts as a chondrocyte mechanotransducer. , 2007, Osteoarthritis and cartilage.

[9]  J. Bertram,et al.  Swelling and fibronectin accumulation in articular cartilage explants after cyclical impact , 1996, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[10]  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.

[11]  Alan J Grodzinsky,et al.  Mechanisms and kinetics of glycosaminoglycan release following in vitro cartilage injury. , 2004, Arthritis and rheumatism.

[12]  C. Little,et al.  IL-6 and its soluble receptor augment aggrecanase-mediated proteoglycan catabolism in articular cartilage. , 2000, Matrix biology : journal of the International Society for Matrix Biology.

[13]  H. Roos,et al.  Temporal patterns of stromelysin‐1, tissue inhibitor, and proteoglycan fragments in human knee joint fluid after injury to the cruciate ligament or meniscus , 1994, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[14]  P. Patwari,et al.  Proteoglycan degradation after injurious compression of bovine and human articular cartilage in vitro: interaction with exogenous cytokines. , 2003, Arthritis and rheumatism.

[15]  A. Grodzinsky,et al.  Mechanical injury of cartilage explants causes specific time-dependent changes in chondrocyte gene expression. , 2005, Arthritis and rheumatism.

[16]  C. Gabay,et al.  Interleukin-6 and chronic inflammation , 2006, Arthritis research & therapy.

[17]  A. Cole,et al.  Human knee and ankle cartilage explants: Catabolic differences , 2002, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

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

[19]  M E Levenston,et al.  A versatile shear and compression apparatus for mechanical stimulation of tissue culture explants. , 2000, Journal of biomechanics.

[20]  G. Lust,et al.  Chondrocyte necrosis and apoptosis in impact damaged articular cartilage , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[21]  A. L. Stevens,et al.  Nitric oxide enhances aggrecan degradation by aggrecanase in response to TNF-alpha but not IL-1beta treatment at a post-transcriptional level in bovine cartilage explants. , 2008, Osteoarthritis and cartilage.

[22]  J. Bijlsma,et al.  ROLE OF TNFα, IN RELATION TO IL-1 AND IL-6 IN THE PROTEOGLYCAN TURNOVER OF HUMAN ARTICULAR CARTILAGE , 1991 .

[23]  H. Roos,et al.  Osteoarthritis of the knee after injury to the anterior cruciate ligament or meniscus: the influence of time and age. , 1995, Osteoarthritis and cartilage.

[24]  D A Lauffenburger,et al.  Real-time quantitative measurement of autocrine ligand binding indicates that autocrine loops are spatially localized. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[25]  I. Clark.  Matrix Metalloproteinase Protocols , 2000, Methods in Molecular Biology.

[26]  L. Lohmander,et al.  The structure of aggrecan fragments in human synovial fluid. Evidence for the involvement in osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. , 1992, The Journal of clinical investigation.

[27]  A. Grodzinsky,et al.  Effects of injurious compression on matrix turnover around individual cells in calf articular cartilage explants , 1998, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[28]  R. Stockwell,et al.  Distribution of Acid Glycosaminoglycans in Human Articular Cartilage , 1967, Nature.

[29]  E. Hunziker,et al.  Improved cartilage fixation by ruthenium hexammine trichloride (RHT). A prerequisite for morphometry in growth cartilage. , 1982, Journal of ultrastructure research.

[30]  A. Cole,et al.  Comparison of biomechanical and biochemical properties of cartilage from human knee and ankle pairs , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[31]  D. D’Lima,et al.  Impact of Mechanical Trauma on Matrix and Cells , 2001, Clinical orthopaedics and related research.

[32]  B Kurz,et al.  Biosynthetic response and mechanical properties of articular cartilage after injurious compression , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[33]  Alan J. Grodzinsky,et al.  Analysis of the Relationship between Peak Stress and Proteoglycan Loss following Injurious Compression of Human Post-mortem Knee and Ankle Cartilage , 2007, Biomechanics and modeling in mechanobiology.

[34]  A. Grodzinsky,et al.  Injurious mechanical compression of bovine articular cartilage induces chondrocyte apoptosis. , 2000, Archives of biochemistry and biophysics.

[35]  M. Terauchi,et al.  Changes in biochemical parameters after anterior cruciate ligament injury , 2006, International Orthopaedics.

[36]  P. Chockalingam,et al.  Release of hyaluronan and hyaladherins (aggrecan G1 domain and link proteins) from articular cartilage exposed to ADAMTS-4 (aggrecanase 1) or ADAMTS-5 (aggrecanase 2). , 2004, Arthritis and rheumatism.

[37]  C. Handley,et al.  Coincubation of bovine synovial or capsular tissue with cartilage generates a soluble "Aggrecanase" activity. , 1999, Biochemical and biophysical research communications.

[38]  C. Handley,et al.  "Aggrecanase" activity is implicated in tumour necrosis factor alpha mediated cartilage aggrecan breakdown but is not detected by an in vitro assay. , 1997, Molecular pathology : MP.

[39]  Itai Cohen,et al.  Mapping the depth dependence of shear properties in articular cartilage. , 2007, Journal of biomechanics.

[40]  Modulation of lubricin biosynthesis and tissue surface properties following cartilage mechanical injury. , 2009, Arthritis and rheumatism.

[41]  J. Steinmeyer,et al.  The proteoglycan metabolism of mature bovine articular cartilage explants superimposed to continuously applied cyclic mechanical loading. , 1997, Biochemical and biophysical research communications.

[42]  J. Reginster,et al.  Effects of exogenous IL-1β, TNFα, IL-6, IL-8 and LIF on cytokine production by human articular chondrocytes , 1996 .

[43]  R. Wardale,et al.  Up-regulation of matrix metalloproteinase expression and activation following cyclical compressive loading of articular cartilage in vitro. , 2001, Archives of biochemistry and biophysics.

[44]  J. Heath,et al.  Synergistic effects of glycoprotein 130 binding cytokines in combination with interleukin-1 on cartilage collagen breakdown. , 2001, Arthritis and rheumatism.

[45]  P. Torzilli,et al.  Effect of impact load on articular cartilage: cell metabolism and viability, and matrix water content. , 1999, Journal of biomechanical engineering.

[46]  R. Haut,et al.  The extent of matrix damage and chondrocyte death in mechanically traumatized articular cartilage explants depends on rate of loading , 2001, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[47]  E B Hunziker,et al.  Physical and Biological Regulation of Proteoglycan Turnover around Chondrocytes in Cartilage Explants: Implications for Tissue Degradation and Repair , 1999, Annals of the New York Academy of Sciences.

[48]  G. Lust,et al.  Compositional and metabolic changes in damaged cartilage are peak-stress, stress-rate, and loading-duration dependent. , 2000, Journal of orthopaedic research : official publication of the Orthopaedic Research Society.

[49]  M. Hochberg,et al.  Joint Injury in Young Adults and Risk for Subsequent Knee and Hip Osteoarthritis , 2000, Annals of Internal Medicine.