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The proteoglycan metabolism, morphology and viability of articular cartilage treated with a synthetic matrix metalloproteinase inhibitor


Matrix metalloproteinases (MMP) are among the key enzymes responsible for the proteolytic destruction of articular cartilage during chronic rheumatic diseases. Articular cartilage is one potential target for drugs designed to inhibit the activity of MMPs in order to stop or to slow down the proteolytic destruction of the extracellular matrix of cartilage. The purpose of this study was to investigate the effect of the synthetic inhibitor of MMPs U-24522 for its ability (1) to inhibit in vitro the activity of MMP-proteoglycanases; (2) to modulate the morphology and viability of cartilage explants; and (3) to modify the biosynthesis and release of proteoglycans from articular cartilage explants. U-24522 dose-dependently inhibited the activity of MMP-proteoglycanases and significantly reduced the release of proteoglycans from interleukin-1 treated bovine articular cartilage explants when tested at concentrations ranging from 10−4 to 10−9 M. This hydroxamic acid derivative proved not to be harmful to chondrocyte viability and cartilage morphology. In addition, U-24522 had no effect on the rate of proteoglycan biosynthesis of interleukin-1 treated cartilage explants and increased the percentage of newly synthesized proteoglycans to form macromolecular aggregates. Thus U-24522 combines direct inhibitory potential on the activity of MMP-proteoglycanases with the inhibition of interleukin-1 stimulated proteoglycan loss from articular cartilage explants without affecting the morphology, viability and biosynthesis of proteoglycans of bovine articular cartilage explants.

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  1. 1.

    Andrews HJ, Plumpton TA, Harper GP, Cawston TE (1992) A synthetic peptide metalloproteinase inhibitor, but not TIMP, prevents the breakdown of proteoglycan with-in articular cartilage in vitro. Agents Actions 37:147–154

  2. 2.

    Azzo W, Woessner JF (1986) Purification and characterization of an acid metalloproteinase from human articular cartilage. J Biol Chem 261:5434–5441

  3. 3.

    Brännström M, Woessner JF Jr, Koos RD, Sear CHJ, LeMaire WJ (1988) Inhibitors of mammalian tissue collagenase and metalloproteinase suppress ovulation in the perfused rat ovary. Endocrinology 122:1715–1721

  4. 4.

    Bunning RAD, Crawford A, Richardson HJ, Opdenakker G, Van Damme J, Russel RGG (1987) Interleukin 1 preferentially stimulates the production of tissue-type plasminogen activator by human articular chondrocytes. Biochim Biophys Acta 924:473–482

  5. 5.

    Burton-Wurster N, Lust G (1990) Fibronectin and proteoglycan synthesis in long term cultures of cartilage explants in Ham’s F-12 supplemented with insulin and calcium: effects of the addition of TGF-?. Arch Biochem Biophys 283:27–33

  6. 6.

    Buttle DJ, Handley CJ, Ilic MZ, Saklatvala J, Murata M, Barrett AJ (1993) Inhibition of cartilage proteoglycan release by a specific inactivator of cathepsin B and an inhibitor of matrix metalloproteinases. Evidence for two converging pathways of chondro-cyte-mediated proteoglycan degradation. Arthritis Rheum 36:1709–1717

  7. 7.

    Campbell IK, Last K, Novak U (1991) Recombinant human interleukin 1 inhibits plasminogen activator inhibitor-1 (PAI-1) production by human articular cartilage and chondrocytes. Biochem Biophys Res Commun 174:251–254

  8. 8.

    Caputo CB, Sygowski LA, Wolanin DJ, Patton SP, Caccese RG, Shaw A, Roberts RA, DiPasquale G (1987) Effect of synthetic metalloprotease inhibitors on cartilage autolysis in vitro. J Pharmacol Exp Ther 240:460–465

  9. 9.

    Caputo CB, Wolanin DJ, Roberts RA, Sygowski LA, Patton SP, Caccese RG, Shaw A, DiPasquale G (1987) Proteoglycan degradation by a chondrocyte metalloprotease. Effects of synthetic protease inhibitors. Biochem Pharmacol 36:995–1002

  10. 10.

    Carney SL (1986) Proteoglycans. In: Chaplin MF, Kennedy JF (eds) Carbohydrate analysis. A practical approach, IRL Press, Oxford, pp 97–142

  11. 11.

    Cawston T, Plempton T, Curry V, Ellis A, Powell L (1994) Role of TIMP and MMP inhibitors in preventing connective tissue breakdown. Ann NY Acad Sci 732:75–83

  12. 12.

    Chubinskaya S, Huch K, Mikecz K, Cs-Szabo G, Hasty KA, Kuettner KE, Cole AA (1996) Chondrocyte matrix metalloproteinase-8: upregulation of neutrophil collagenase by inter-leukin-1 beta in human cartilage from knee and ankle joints. Lab Invest 74:232–235

  13. 13.

    Cole AA, Chubinskaya S, Schumacher B, Huch K, Szabo G, Yao J, Mikecz K, Hasty KA, Kuettner KE (1996) Chondrocyte matrix metalloproteinase-8. Human articular chondrocytes express neutrophil collagenase. J Biol Chem 271:11023–11026

  14. 14.

    Cole AA, Kuettner KE (1995) MMP-8 mRNA and aggrecanase cleavage products are present in normal and osteoarthritic human articular cartilage. Acta Orthop Scand 266:98–102

  15. 15.

    DiMartino MJ, High W, Galloway WA, Crimmin MJ (1994) Preclinical antiarthritic activity of matrix metalloproteinase inhibitors. Ann NY Acad Sci 732:411–413

  16. 16.

    DiPasquale G, Caccese R, Pasternak R, Conaty J, Hubbs S, Perry K (1986) proteoglycan- and collagen-degrading enzymes from human interleukin-1 stimulated chondrocytes from several species: proteoglycanase and collagenase inhibitors as potentially new disease-modifiying antiarthritic agents. Proc Soc Exp Biol Med 183:262–267

  17. 17.

    Dingle JT, Page Thomas DP, King B, Bard DR (1987) In vivo studies of articular tissue damage mediated by catabolin/interleukin 1. Ann Rheum Dis 46:527–533

  18. 18.

    Ellis AJ, Curry VA, Powell EK, Cawston, TE (1994) The prevention of collagen break-down in bovine nasal cartilage by TIMP, TIMP-2 and a low molecular weight synthetic inhibitor. Biochem Biophys Res Commun 201:94–101

  19. 19.

    Farndale RW, Buttle DJ, Barrett AJ (1986) Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta 883:173–177

  20. 20.

    Farnum CE, Turgai J, Wilsman NJ (1990) Visualization of living terminal hypertrophic chondrocytes of growth plate cartilage in situ by differential interference contrast microscopy and time-lapse cinematography. J Orthop Res 8:750–763

  21. 21.

    Fosang AJ, Last K, Knäuper V, Neame PJ, Murphy G, Hardingham TE, Tschesche H, Hamilton JA (1993) Fibroblast and neutrophil collagenases cleave at two sites in the cartilage aggrecan interglobular domain. Biochem J 295:273–276

  22. 22.

    Fosang AJ, Neame PJ, Last K, Hardingham TE, Murphy G, Hamilton JA (1992) Inter-globular domain of cartilage aggrecan is cleaved by PUMP, gelatinases, and cathepsin B. J Biol Chem 267:19470–19474

  23. 23.

    Galardy RE, Grobelny D, Kortylewiez ZP, Poncz L (1992) Inhibition of human skin fibroblast collagenase by phosphorous-containing peptides. In: Birkedal-Hansen Z, Werb Z, Welgus HG, Van Wart HE (eds) Matrix metalloproteinases and inhibitors (Matrix spec. suppl. no. 1). Gustav Fischer, Stuttgart, pp 259–262

  24. 24.

    Gray RD, Miller RB, Spatola AF (1986) Inhibition of mammalian collagenases by thiolcontaining peptides. J Cell Biochem 32:71–77

  25. 25.

    Harris RB, Strong PD, Wilson IB (1983) Dipeptide-hydroxamates are good inhibitors of the angiotensin I-converting enzyme. Biochem Biophys Res Commun 116:394–399

  26. 26.

    Hasty KA, Reife RA, Kang AH, Stuart JM (1990) The role of stromelysin in the cartilage destruction that accompanies inflammatory arthritis. Arthritis Rheum 33:388–397

  27. 27.

    Heinegard D, Sommarin Y (1987) Isolation and characterization of proteoglycans. Methods Enzymol 144:319–372

  28. 28.

    Henderson B, Davies DE (1991) The design of inhibitors of cartilage breakdown. In: Russell RGG, Dieppe PA (eds) Osteoarthritis: current research and prospectives for pharmacological intervention. IBC Technical Services, London, pp 385–396

  29. 29.

    Johnson WH, Roberts NA, Barkakoti N (1987) Collagenase inhibitors: their design and potential therapeutic use. J Enzyme Inhib 2:1–22

  30. 30.

    Mankin HJ, Dorfman H, Lippiello L and Zarins A (1971) Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. J Bone Joint Surg Am 53:523–537

  31. 31.

    Matrisian LM (1990) Metalloproteinases and their inhibitors in matrix remodeling. Trends Genet 6:121–125

  32. 32.

    Morales TI, Hascall VC (1989) Factors involved in the regulation of proteoglycan metabolism in articular cartilage. Arthritis Rheum 32:1197–1201

  33. 33.

    Mort JS, Dodge GR, Roughley PJ, Liu J, Finch SJ, DiPasquale G, Poole AR (1993) Direct evidence for active metalloproteinases mediating matrix degradation in inter-leukin 1-stimulated human articular cartilage. Matrix 13:95–102

  34. 34.

    Muldrew K, Novak K, Hurtig MB, Schachar NS, McGann LE (1993) Cryopreservation of articular cartilage using in vitro and in vivo assays. Trans 39th Ann Mtg Orthop Res Soc 18:275

  35. 35.

    Nagase H, Woessner JF (1980) An improved assay for proteases and polysaccharidases employing a cartilage proteoglycan substrate entrapped in polyacrylamide particles. Anal Biochem 107:385–392

  36. 36.

    Netzel-Arnett S, Mallya SK, Nagase H, Birkedal-Hansen Z, Van Wart HE (1991) Continuously recording fluorescent assays optimized for five human matrix metalloproteinases. Anal Biochem 195:86–92

  37. 37.

    Nixon JS, Bottomley KMK, Bradhurst MJ, Brown PA, Johnson WH, Lawton G, Marley J, Sedgwick AD, Wilkinson SE (1991) Potent collagenase inhibitors prevent inter-leukin 1-induced cartilage degradation in vitro. Int J Tissue React 13:237–243

  38. 38.

    Ohlendorf C, Tomford WW, Mankin HJ (1996) Chondrocyte survival in cryopreserved osteochondral articular cartilage. J Orthop Res 14:413–416

  39. 39.

    Pelletier JP, Howell DS (1993) Etiopathogenesis of osteoarthritis. In: McCarthy, Koopman WJ (eds) Arthritis and allied conditions, 12th edn. Lea & Febiger, Philadelphia, pp 1723–1734

  40. 40.

    Pettipher ER, Higgs GA, Henderson B (1986) Interleukin 1 induces leucocyte infiltration and cartilage proteoglycan degradation in the synovial joint. Proc Natl Acad Sci USA 85:8749–8853

  41. 41.

    Sandy JD, Flannery CR, Neame PJ, Lohmander LS (1992) 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. J Clin Invest 89:1512–1516

  42. 42.

    Schwartz MA, Van Wart HE (1992) Synthetic inhibitors of bacterial and mammalian interstitial collagenases. In: Ellis GP, Luscombe DK (eds) Progress in medicinal chemistry, Elsevier, Amsterdam, pp 1124–1131

  43. 43.

    Schwartz MA, Venkataraman S, Ghaffari MA, Libby A, Mookhtiar KA, Mallya SK, Birkedal-Hansen H, Van Wart HE (1991) Inhibition of human collagenases by sulfur-based substrate analogs. Biochem Biophys Res Commun 176:173–179

  44. 44.

    Seed MP, Ismaiel S, Cheung CY, Thomson TA, Gardner CR, Atkins RM, Elson CJ (1993) Inhibition of interleukin 1 beta induced rat and human cartilage degradation in vitro by the metalloproteinase inhibitor U27391. Ann Rheum Dis 52:37–43

  45. 45.

    Williams RJ, Smith RL, Schurman DJ (1991) Purified staphylococcal culture medium stimulates neutral metalloprotease secretion from human articular cartilage. J Orthop Res 9:258–265

  46. 46.

    Williams LM, Gibbons DL, Gearing A, Maini RN, Feldmann M, Brennan FM (1996) Paradoxical effects of a synthetic metalloproteinase inhibitor that blocks both p55 and p75 receptor shedding and TNF alpha processing in RA synovial membrane cell cultures. J Clin Invest 97:2833–2841

  47. 47.

    Woessner JF Jr (1991) Matrix metalloproteinases and their inhibitors in connective tissue remodeling. FASEB J 5:2145–2154

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Correspondence to Jürgen Steinmeyer.

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Steinmeyer, J., Daufeldt, S. & Kalbhen, D.A. The proteoglycan metabolism, morphology and viability of articular cartilage treated with a synthetic matrix metalloproteinase inhibitor. Res. Exp. Med. 197, 63–79 (1997).

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Key words

  • Cartilage
  • Matrix metalloproteinase
  • Synthetic inhibitor
  • Proteoglycan
  • Interleukin-1