Summary
During 5 days of culture, explants of normal rabbit synovium produced no active collagenase, negligible latent collagenase, but significant levels of free collagenase inhibitor. Synovium from joints exhibiting a proliferative arthritis produced greatly elevated levels of collagenase; the appearance of active enzyme in the medium during the second day of culture was associated with the disappearance of free inhibitor. Enzyme levels in the media correlated well with the arthritic status of joints, when explants were prepared up to 10 weeks after the induction of the model arthritis. Synovium from the contralateral joints of rabbits with unilaterally induced arthritis produced no active collagenase, but approximately one-third as much latent collagenase as found with arthritic joints. Enzymatic activities against gelatin and cartilage proteoglycan substrates were demonstrated in synovial culture media in addition to collagenolytic activity. Gel filtration showed that these activities were not due to a single enzyme, and further characterisation confirmed that the enzymes were metalloproteinases. The results are considered in the light of published data, and the involvement of metalloproteinases and their specific inhibitor in the development of arthritic lesions is discussed.
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References
Harris ED Jr, Krane SM (1974) Collagenases. N Engl J Med 291:557–563
Woolley DE, Glanville RW, Crossley MJ, Evanson JM (1975) Purification of rheumatoid synovial collagenase and its action on soluble and insoluble collagen. Eur J Biochem 54:611–622
Sellers A, Cartwright E, Murphy G, Reynolds JJ (1977) Evidence that latent collagenases are enzyme-inhibitor complexes. Biochem J 163:303–307
Murphy G, Cartwright EC, Sellers A, Reynolds JJ (1977) The detection and characterisation of collagenase inhibitors from rabbit tissues in culture. Biochim Biophys Acta 483:493–498
Sellers A, Reynolds JJ, Meikle MC (1978) Neutral metalloproteinases of rabbit bone. Separation in latent forms of distinct enzymes that when activated degrade collagen, gelatin and proteoglycans. Biochem J 171:493–496
Reynolds JJ, Murphy G, Sellers A, Cartwright E (1977) A new factor that may control collagen resorption. Lancet II:333–335
Sellers A, Murphy G, Meikle MC, Reynolds JJ (1979) Rabbit bone collagenase inhibitor blocks the activity of other neutral metalloproteinases. Biochem Biophys Res Commun 87:581–587
Page-Thomas DP (1977a) Aspects of synovial biodegradation. In: Bayer-Symposium VI: Experimental models of chronic inflammatory diseases. Springer, Berlin Heidelberg New York, pp 353–365
Page-Thomas DP (1977b) Aspects of experimental inflammatory arthritis. In: Gordon JL, Hazleman BI (eds) Rheumatoid arthritis. Elsevier/North-Holland Biomedical Press, Amsterdam
Phillips NC, Page-Thomas DP (1979) Monitoring of experimental arthritis in rabbits. Ann Rheum Dis 38:89–93
Phillips NC, Page-Thomas DP, Knight CG, Dingle JT (1979) Liposome-incorporated corticosteroids. II. Therapeutic activity in experimental arthritis. Ann Rheum Dis 38:553–557
Shaw IH, Knight CG, Page-Thomas DP, Phillips NC, and Dingle JT (1979) Liposome-incorporated corticosteroids: I. The interaction of liposomal cortisol palmitate with inflammatory synovial membrane. Br J Exp Pathol 60:142–150
Laemmli UK, Favre M (1973) Maturation of the head of bacteriophage T4: I. DNA packaging events. J Mol Biol 80: 575–599
Sellers A, Reynolds JJ (1977) Identification and partial characterisation of an inhibitor of collagenase from rabbit bone. Biochem J 167:353–360
Harris ED Jr, Krane SM (1972) An endopeptidase from rheumatoid synovial tissue culture. Biochim Biophys Acta 258:566–576
Dingle JT, Blow AMJ, Barrett AJ, Martin PEN (1977) Proteoglycan-degrading enzymes: a radiochemical assay method and the detection of a new enzyme, cathepsin F. Biochem J 167:775–785
Werb Z, Reynolds JJ (1974) Stimulation by endocytosis of the secretion of collagenase and neutral proteinase from rabbit synovial fibroblasts. J Exp Med 140:1482–1497
Lazarus GS, Decker JL, Oliver CH, Daniels JR, Multz CV, Fullmer HM (1968) Collagenolytic activity of synovium in rheumatoid arthritis. N Engl J Med 279:914–919
Woolley DE, Crossley MJ, Evanson JM (1977) Collagenase at sites of cartilage erosion in the rheumatoid joint. Arthritis Rheum 20:1231–1239
Ishikawa H, Ziff M (1976) Electron microscopic observations of immunoreactive cells in the rheumatoid synovial membrane. Arthritis Rheum 19:1–14
Kobayashi I, Ziff M (1975) Electron microscopic studies of the cartilage-pannus junction in rheumatoid arthritis. Arthritis Rheum 18:475–483
Dayer J-M, Krane SM, Russell RGG, Robinson DR (1976) Production of collagenase and prostaglandins by isolated adherent rheumatoid synovial cells. Proc Natl Acad Sci USA 73:945–949
Dayer J-M, Russell RGG, Krane SM (1977) Collagenase production by rheumatoid synovial cells: stimulation by a human lymphocyte factor. Science 195:181–183
Dayer J-M, Goldring SR, Robinson DR, Krane SM (1979) Effects of human mononuclear cell factor on cultured rheumatoid synovial cells: interactions of prostaglandin E2 and cyclic adenosine 3′,5′-monophosphate. Biochim Biophys Acta 586:87–105
Woolley DE, Brinkerhoff CE, Mainardi CL, Vater CA, Evanson JM, Harris ED Jr (1979) Collagenase production by rheumatoid synovial cells: morphological and immunohistochemical studies of the dendritic cell. Ann Rheum Dis 38:262–270
Vater CA, Mainardi CL, Harris ED Jr. (1978) Activation in vitro of rheumatoid synovial collagenase from cell cultures. J Clin Invest 62:987–992
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Cambray, G.J., Murphy, G., Page-Thomas, D.P. et al. The production in culture of metalloproteinases and an inhibitor by joint tissues from normal rabbits, and from rabbits with a model arthritis. Rheumatol Int 1, 11–16 (1981). https://doi.org/10.1007/BF00541217
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DOI: https://doi.org/10.1007/BF00541217