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Characterization and Identification of Proteinases and Proteinase Inhibitors Synthesized by Temporomandibular Joint Disc Cells

Division of Orthodontics, Department of Growth and Development, University of California, San Francisco, California 94143-0640

C. Lee

D.W. Richards

Department of Stomatology, School of Dentistry, University of California, San Francisco, California 94143-0640

The adult mammalian temporomandibular joint (TMJ) disc is a fibrocartilaginous tissue that undergoes normal developmental remodeling, requiring removal of the existing extracellular matrix and its replacement by new matrix macromolecules. This remodeling is probably mediated by matrix-degrading enzymes, but to date none has been demonstrated in association with the TMJ disc. We characterized, identified, and determined the regulation of proteinases and proteinase inhibitors (PIs) synthesized by TMJ disc cells in organ and cell cultures. TMJ discs were retrieved from 14-week-old male NZW rabbits and both tissue- and disc-derived cells were cultured in serum-free medium. The conditioned media were retrieved at 12-hour intervals and assayed for proteinases and PIs in gelatin- and casein-impregnated polyacrylamide gels. Three proteinases with gelatinolytic activities at 92 kDa, 72 kDa, and 53/57 kDa and one caseinolytic activity at 51/54 kDa were detected. All were inhibited by 1,10-1 phenanthroline, thus characterizing these enzymes as matrix metalloproteinases (MMPs), most likely 92-kDa gelatinase (proMMP-9), 72-kDa gelatinase (proMMP-2), procollagenase (proMMP-1), and prostromelysin (proMMP-3). The identity of the latter two MMPs was confirmed by Western blots. Two PIs of 30 kDa and 20 kDa, probably tissue inhibitors of metalloproteinase (TIMP) and TIMP-2, were observed on reverse zymograms. TPA, a protein kinase-C agonist, increased the expression of 92-kDa gelatinase and 30-kDa PI by both explanted discs and isolated disc cells. The profile of MMPs constitutively expressed by disc cells is similar to that of synovial fibroblasts but different from that of chondrocytes. These findings implicate MMPs and Pis in the normal remodeling of the TMJ disc, and potentially in the pathologic degradation of the disc during various arthritides.

Key Words: temporomandibular joint disc • matrix metalloproteinases • proteinase inhibitors • tissue inhibitor of metalloproteinases • matrix degradation.

Journal of Dental Research, Vol. 74, No. 6, 1328-1336 (1995)
DOI: 10.1177/00220345950740061301


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