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Indian J Clin Biochem. 2009 January; 24(1): 42–48.
Published online 2009 May 8. doi:  10.1007/s12291-009-0007-0
PMCID: PMC3453469

Multiple matrix metalloproteinases in type II collagen induced arthritis


To study the relative contribution of various matrix degrading enzymes in the pathogenesis of arthritis, changes in the levels of various matrix metalloprtoteinases (MMPs) during the progression of collagen induced arthritis was studied in experimental animals. Arthritis was induced in male wistar rats by injecting an emulsion containing collagen type II and Freund’s complete adjuvant. The duration of the experiment was 35 days. Synovial effusate was collected at regular intervals after induction. At the end of the experiment serum and cartilage were collected and analysed. Synovial fluid of osteoarthritic patients was also analyzed. Levels of MMP-2, MMP-3, MMP-9 and MT1-MMP were found to be high in synovial effusate and cartilage of experimental animals. In synovial effusate of arthritic animals the expression of MMP-3 was found to be high during the early stages while increase in MMP-2 and MMP-9 occurred at later stages. Synovial fluid of osteoarthritic patients also showed elevated levels of MMP-2, MMP-3 and MMP-9. Our results indicated that sequential action of MMPs such as MMP-3, MMP-2 and MMP-9 can cause degradation of articular cartilage extracellular matrix.

Key Words: Matrix metalloproteinases, Osteoarthritis, Synovial effusate, Synovial fluid

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Selected References

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1. Lotz M., Hashimoto S., Kuhn K. Mechanisms of chondrocyte apoptosis. Osteoarthritis Cartilage. 1999;7:389–391. doi: 10.1053/joca.1998.0220. [PubMed] [Cross Ref]
2. Buttle D.J., Bramwell H., Hollander A.P. Proteolytic mechanisms of cartilage breakdown: a target for arthritis therapy? Clin Pathol Mol Pathol. 1995;48:167–177. doi: 10.1136/mp.48.4.M167. [PMC free article] [PubMed] [Cross Ref]
3. Murphy G., Reynolds J.J. Current views of collagen degradation. Progress towards understanding the resorption of connective tissues. Bio Essays. 1985;2:55–60.
4. Kozaci L.D., Buttle D.J., Hollander A.P. Degradation of type II collagen, but not proteoglycan, correlates with matrix metalloproteinase activity in cartilage explant cultures. Arthritis Rheum. 1997;40:164–174. doi: 10.1002/art.1780400121. [PubMed] [Cross Ref]
5. Makowski G.S., Ramsby M.L. Zymographic analysis of latent and activated forms of matrix metalloproteinase-2 and -9 in synovial fluid: correlation to polymorphonuclear leukocyte infiltration and in response to infection. Clin Chim Acta. 2003;329:77–81. doi: 10.1016/S0009-8981(03)00015-9. [PubMed] [Cross Ref]
6. Matrisian L.M. The matrix-degrading metalloproteinases. Bio Essays. 1992;14:455–463. [PubMed]
7. Murphy G., Hembry R.M., Hughes C.E., Fosang A.J., Hardingham T.E. Role and regulation of metalloproteinases in connective tissue turnover. Biochem Soc Trans. 1990;18:812–815. [PubMed]
8. Murphy G., Reynolds J.J., Hembry R.M. Metalloproteinases and cancer invasion and metastasis. Int J Cancer. 1989;44:757–760. doi: 10.1002/ijc.2910440434. [PubMed] [Cross Ref]
9. Nagase H., Woessner J.F., Jr. Matrix metalloproteinases. J Biol Chem. 1999;274:21491–21494. doi: 10.1074/jbc.274.31.21491. [PubMed] [Cross Ref]
10. Cawston T. Matrix metalloproteinases and TIMPs: properties and implications for the rheumatic diseases. Mol Med Today. 1998;4:130–137. doi: 10.1016/S1357-4310(97)01192-1. [PubMed] [Cross Ref]
11. Brew K., Dinakarpandian D., Nagase H. Tissue inhibitors of metalloproteinases: evolution, structure and function. Biochim Biophys Acta. 2000;1477:267–283. [PubMed]
12. Clark I.M., Rowan A.D., Cawston T.E. Matrix metalloproteinase inhibitors in the treatment of arthritis. Curr Opin Antiinflammatory Immunomodulatory Drugs. 2000;2:16–25.
13. Firestein G.S., Paine M.M., Littman B.H. Gene expression (collagenase, tissue inhibitor of metalloproteinases, complement and HLA-DR) in rheumatoid arthritis and osteoarthritis synovium, Quantitative analysis and effect of intraarticular corticosteroids. Arhtritis Rheum. 1991;34:1094–1105. doi: 10.1002/art.1780340905. [PubMed] [Cross Ref]
14. Gravallese E.M., Darling J.M., Ladd A.L., Katz J.M., Glimcher L.H. In situ hybridization studies of stromelysin and collagenase messenger RNA expression on rheumatoid synovium. Arhtritis Rheum. 1991;34:1076–1084. doi: 10.1002/art.1780340903. [PubMed] [Cross Ref]
15. McCachren S.S. Expression of metalloproteinases and metalloproteinase inhibitor in human arthritic synovium. Arhtritis Rheum. 1991;34:1085–1093. doi: 10.1002/art.1780340904. [PubMed] [Cross Ref]
16. Freije J.M., Diez-Itza I., Balbin M., Sanchez L.M., Blasco R., Tolivia J., et al. Molecular cloning and expression of collagenase-3, a novel human matrix metalloproteinase produced by breast carcinomas. J Biol Chem. 1999;269:16766–16773. [PubMed]
17. Makowski G.S., Ramsby M.L. Autoactivation profiles of calcium-dependent matrix metalloproteinase-2 and -9 in inflammatory synovial fluid: effect of pyrophosphate and bisphosphonates. Clin Chim Acta. 2005;358(1–2):182–191. doi: 10.1016/j.cccn.2005.03.012. [PubMed] [Cross Ref]
18. Balbin M., Fueyo A., Knauper V., Lopez J.M., Alvarez J., Sanchez L.M., et al. Identification and enzymatic characterization of two diverging murine counterparts of human interstitial collagenase (MMP-1) expressed at sites of embryo implantation. J Biol Chem. 2001;276:10253–10262. doi: 10.1074/jbc.M009586200. [PubMed] [Cross Ref]
19. Konttinen Y.T., Ainola M., Valleala H., Ma J., Ida H., Mandelin J., et al. Analysis of 16 different matrix metalloproteinases (MMP-1 to MMP-20) in synovial membrane: different profiles in trauma and rheumatoid arthritis. Ann Rheum Dis. 1999;58:691–697. doi: 10.1136/ard.58.11.691. [PMC free article] [PubMed] [Cross Ref]
20. Mattsson L., Larsson P., Erlandsson-Harris H., Klareskog L., Harris R.A. Parasite-mediated down -regulation of collagen induced arthritis (CIA) in DA rats. Clin Exp Immunol. 2000;122:477–483. doi: 10.1046/j.1365-2249.2000.01384.x. [PubMed] [Cross Ref]
21. Kawai Y., Anno K. Mucopolysaccharise-degrading enzymes from the liver of the squid, Ommastrephes solani pacificus. I. Hyaluronidase. Biochim Biophys Acta. 1971;242:428–436. [PubMed]
22. Johnson W.G., Moak G., Brady R.O. In: Methods in Enzymology. Colowick S.P., Kaplan N.O., editors. NY: Academic Press; 1972. pp. 857–861.
23. Ambili M., Jayasree K., Sudhakaran P.R. 60k gelatinase involved in mammary gland involution is regulated by â estradiol. Biochim Biophys Acta. 1998;1403:219–231. doi: 10.1016/S0167-4889(98)00030-5. [PubMed] [Cross Ref]
24. Engvall E., Perlman P. Enzyme-linked immunosorbent assay (ELISA), Quantitative assay of immunoglobulin. Immunochem. 1971;8:871–874. doi: 10.1016/0019-2791(71)90454-X. [PubMed] [Cross Ref]
25. Mastroianni C., Vollov F., Paloetti A.D., Massetti F., Sorice S., Dellia I. Detection of mycobacterial antigens by dotblot assay in cerebrospinal fluids of patients with tubercular meningities. J Infect. 1991;22:106–107. doi: 10.1016/0163-4453(91)91374-7. [PubMed] [Cross Ref]
26. Rao S.K., Mathrubutham M., Karteron A., Sorensen K., Cohen J.R. A versatile Microassay for Elastase using succinylated Elastin. Anal Biochem. 1997;250:222–227. doi: 10.1006/abio.1997.2223. [PubMed] [Cross Ref]
27. Smolian H., Aurer A., Sittinger M., Zacher J., Bernimoulin J.P., Burmester G.R., Kolkenbrock H. Secretion of gelatinases and activation of gelatinase A (MMP-2) by human rheumatoid synovial fibroblasts. Biol Chem. 2001;382:1491–1499. doi: 10.1515/BC.2001.183. [PubMed] [Cross Ref]
28. Buttle D.J., Handley M.Z., Ilic M., Saklatvala J., Murata M., Barrett A.J., et al. Inhibition of cartilage proteoglycan release by a specific inactivator of Cathepsin B and an inhibitor of matrix metalloproteinases. Evidence for two converging pathways of chondrocyte-mediated proteoglycan degradation. Arthritis Rheum. 1993;36:1709–1717. doi: 10.1002/art.1780361210. [PubMed] [Cross Ref]
29. Knauper V.C., Will H., Lopez-Otin C., Smith B., Atkinson S.J., Stanton H., et al. Cellular mechanisms for human procollagenase -3 (MMP-13) activation. Evidence that MT1- MMP (MMP-14) and gelatinase A (MMP-2) are able to generate active enzyme. J Biol Chem. 1996;271:17124–17131. doi: 10.1074/jbc.271.29.17124. [PubMed] [Cross Ref]
30. Gomis-Ruth F.X., Maskos K., Betz M., Bergner A., Huber R., Suzuki K., et al. Mechanism of inhibition of the human metalloproteinase stromelysin-1 by TIMP-1. Nature. 1997;389:77–81. doi: 10.1038/37995. [PubMed] [Cross Ref]
31. McKenna L.A., Liu H., Sansom P.A., Dean M.F. An N-terminal peptide from link protein stimulates proteoglycan synthesis in human articular cartilage in vitro. Arthritis Rheum. 1998;41:157–161. doi: 10.1002/1529-0131(199801)41:1<157::AID-ART19>3.0.CO;2-J. [PubMed] [Cross Ref]
32. Peake N.J., Foster H.E., Khawaja K., Cawston L.E., Rowan A.D. Assessment of the clinical significance of gelatinase activity in patients with juvenile idiopathic arthritis using quantitative protein substrate zymography. Ann Rheum Dis. 2006;65(4):501–517. doi: 10.1136/ard.2005.039032. [PMC free article] [PubMed] [Cross Ref]
33. Salinardi B.J., Roush J.K., Schermerhorn T., Mitchell K.E. Matrix metalloproteinase and tissue inhibitor of metalloproteinase in serum and synovial fluid of osteoarthritic dogs. Vet Comp Orthop Traumatol. 2006;19:49–55. [PubMed]
34. Kanyama M., Kuboki T., Kojima S., Fujisawa T., Hattori T., Takigawa M., Yamashita A. Matrix metalloproteinases and tissue inhibitors of metalloproteinases in synovial fluids of patients with temporomandibular joint osteoarthritis. J Orofac Pain. 2000;14:20–30. [PubMed]

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