Abstract
Cultures of cartilage explants have long been used to study the effects of modulators of extracellular matrix degradation. We present a simple and rapid assay system, based on culture of rabbit cartilage explants, which permits study of the effects of protease inhibitors on proteoglycan degradation (caused by either aggrecanases or matrix metalloproteinases [MMPs]), and on collagen degradation. The assay is based on the ability of interleukin-1 to stimulate both aggrecanase activity and synthesis of inactive MMPs, which are then activated by p-aminophenylmercuric acetate for the study of MMP-mediated proteoglycan degradation or by plasmin for the study of collagen degradation. Proteoglycan degradation is quantified as percent release of radioactivity from cartilage explants previously labeled with 35SO4 2−. Collagen degradation is calculated as percent release of collagen, measured by colorimetric assay of hydroxyproline.
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References
Goldring, M. B. (2000) The role of chondrocyte in osteoarthritis. Arthritis Rheum. 43, 1916–1926.
Hascall, V. C. and Kimura, J. H. (1992) Proteoglycans: isolation and characterization. Methods Enzymol. 82, 769–800.
Tortorella, M. D., Burn, T. C., Pratta, M. A., et al. (1999) Purification and cloning of aggrecanase-1: a member of the ADAMTS family of proteins. Science 284, 1664–1666.
Abbaszade, I, Liu, R. Q., Yang, F., et al. (1999) Cloning and characterization of ADAMTS11, an aggrecanase from the ADAMTS family. J. Biol. Chem. 274, 23,443–23,450.
Caterson, B., Flannery, C. R., Hughes, C. E., and Little, C. B. (2000) Mechanisms involved in cartilage proteoglycan catabolism. Matrix Biol. 19, 333–344.
Sabatini, M., Bardiot, A., Lesur, C., et al. (2002) Effects of agonists of peroxisome proliferator-activated receptor γ on proteoglycan degradation and matrix metalloproteinase production in rat cartilage in vitro. Osteoarthritis Cartilage 10, 673–679.
Sorbera, L. A. and Castaner, J. (2000) Prinomastat. Drugs Fut. 25, 150–158.
Sugimoto, K., Takahashi, M., Yamamoto, Y., Shimada, K., and Tanzawa, K. (1999) Identification of aggrecanase activity in medium of cartilage culture. J. Biochem. 126, 449–455.
Cremer, M. A., Rosloniec, E. F., and Kang, H. A. (1998) The cartilage collagens; a review of their structure, organization, and role in the pathogenesis of experimental arthritis in animals and in human disease. J. Mol. Med. 76, 275–288.
Billinghurst, R. C., Dahlberg, L., Iionescu, M., et al. (1997) Enhanced cleavage of type II collagen by collagenases in osteoarthritic articular cartilage. J. Clin. Invest. 99, 1534–1545.
Saito, S., Katoh, M., Masumoto, M., Matsumoto S.-I., and Masuho Y. (1997) Collagen degradation induced by the combination of IL-1α and plasminogen in rabbit articular cartilage explant culture. J. Biochem. 122, 49–54.
Kummer, J. A., Abbink, J. J., Deboer, J. P., et al. (1992) Analysis of intraarticular fibrinolytic pathways in patients with inflammatory and non inflammatory joint diseases. Arthritis Rheum. 36, 884–893.
Grant, R. A. (1964) Estimation of OH-proline by the autoanalyser. J. Clinical Pathol. 17, 685–686.
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© 2004 Humana Press Inc., Totowa, NJ
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Lesur, C., Sabatini, M. (2004). Assays of Proteoglycan and Collagen Degradation in Cultures of Rabbit Cartilage Explants. In: Sabatini, M., Pastoureau, P., De Ceuninck, F. (eds) Cartilage and Osteoarthritis. Methods in Molecular Medicine™, vol 100. Humana Press. https://doi.org/10.1385/1-59259-810-2:219
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DOI: https://doi.org/10.1385/1-59259-810-2:219
Publisher Name: Humana Press
Print ISBN: 978-1-58829-247-6
Online ISBN: 978-1-59259-810-6
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