Assays of proteoglycan and collagen degradation in cultures of rabbit cartilage explants.
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 (35)SO4(2-). Collagen degradation is calculated as percent release of collagen, measured by colorimetric assay of hydroxyproline.
Division der Rhumatologie, Institut de Recherches Servier, Suresnes, France.
Christophe
C
Lesur
Massimo
M
Sabatini
D000818:Animals; D001681:Biological Assay; D002356:Cartilage / Q000737:chemistry / Q000378:metabolism*; D020134:Catalytic Domain; D002478:Cells, Cultured; D003094:Collagen / Q000032:analysis / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism; D005341:Fibrinolysin / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D061965:Matrix Metalloproteinase Inhibitors; D009930:Organic Chemicals / Q000494:pharmacology; D010662:Phenylmercuric Acetate / Q000031:analogs & derivatives* / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D011509:Proteoglycans / Q000032:analysis / Q000378:metabolism*; D011817:Rabbits
D007375:Interleukin-1; D061965:Matrix Metalloproteinase Inhibitors; D009930:Organic Chemicals; D011480:Protease Inhibitors; D011509:Proteoglycans; C113282:prinomastat; C010024:4-aminophenylmercuriacetate; D003094:Collagen; D010450:Endopeptidases; D005341:Fibrinolysin; C083641:aggrecanase; D010662:Phenylmercuric Acetate