title: Comparison of the degradation of type II collagen and proteoglycan in nasal and articular cartilages induced by interleukin-1 and the selective inhibition of type II collagen cleavage by collagenase.
abstract: OBJECTIVE
  To compare interleukin-1alpha (IL-1alpha)-induced degradation of nasal and articular cartilages in terms of proteoglycan loss and type II collagen cleavage, denaturation, and release; to examine the temporal relationship of these changes; and to investigate the effects of an inhibitor of collagenase 2 and collagenase 3 on these catabolic processes.
  
  
  METHODS
  Discs of mature bovine nasal and articular cartilages were cultured with or without human IL-1alpha (5 ng/ml) with or without RS102,481, a selective synthetic inhibitor of collagenase 2 and collagenase 3 (matrix metalloproteinase 8 [MMP-8] and MMP-13, respectively) but not of collagenase 1 (MMP-1). Immunoassays were used to measure collagenase-generated type II collagen cleavage neoepitope (antibody COL2-3/4C(short)) and denaturation (antibody COL2-3/4m), as well as total type II collagen content (antibody COL2-3/4m) in articular cartilage and culture media. A colorimetric assay was used to measure total proteoglycan concentration (principally of aggrecan) as sulfated glycosaminoglycans (sGAG).
  
  
  RESULTS
  IL-1alpha initially induced a decrease in tissue proteoglycan content in nasal cartilage. A progressive loss of proteoglycan was noted during culture in articular cartilages, irrespective of the presence of IL-1alpha. In both cartilages, proteoglycan loss was followed by IL-1alpha-induced cleavage of type II collagen by collagenase, which was often reflected by increased denaturation. The inhibitor RS102,481 had no clear effect on the reduction in proteoglycan content (measured by sGAG) and collagen denaturation in either cartilage, but at 10 nM it inhibited the enhanced cleavage of type II collagen, partially in nasal cartilage and completely in articular cartilage.
  
  
  CONCLUSIONS
  IL-1alpha-induced cleavage and denaturation of type II collagen is observed in both hyaline cartilages and is secondary to proteoglycan loss. It probably involves different collagenases, since there is no evidence of a rate-limiting role for collagenase 1 in articular cartilage, unlike the case for nasal cartilage. Inhibitors of this kind may be of value in the treatment of cartilage damage in arthritis. Also, the ability to detect the release of type II collagen collagenase-generated fragments from degraded cartilage offers the potential to monitor cartilage collagen damage and its control in vivo.
authors.affiliation: Colorado State University, Fort Collins, USA.
authors.forename: R C
authors.initials: RC
authors.lastname: Billinghurst
authors.forename: W
authors.initials: W
authors.lastname: Wu
authors.forename: M
authors.initials: M
authors.lastname: Ionescu
authors.forename: A
authors.initials: A
authors.lastname: Reiner
authors.forename: L
authors.initials: L
authors.lastname: Dahlberg
authors.forename: J
authors.initials: J
authors.lastname: Chen
authors.forename: H
authors.initials: H
authors.lastname: van Wart
authors.forename: A R
authors.initials: AR
authors.lastname: Poole
mesh_terms: D000818:Animals; D002264:Carboxylic Acids / Q000494:pharmacology*; D002356:Cartilage / Q000187:drug effects / Q000378:metabolism*; D002358:Cartilage, Articular / Q000187:drug effects / Q000378:metabolism*; D002417:Cattle; D003094:Collagen / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D017364:Collagenases / Q000494:pharmacology*; D017077:Culture Media, Conditioned; D046508:Culture Techniques; D006801:Humans; D007211:Indoles / Q000494:pharmacology*; D007375:Interleukin-1 / Q000494:pharmacology*; D053509:Matrix Metalloproteinase 13; D020784:Matrix Metalloproteinase 8 / Q000494:pharmacology; D009300:Nasal Septum; D011489:Protein Denaturation / Q000187:drug effects; D011509:Proteoglycans / Q000378:metabolism*
chemical_list: D002264:Carboxylic Acids; D017077:Culture Media, Conditioned; D007211:Indoles; D007375:Interleukin-1; D011509:Proteoglycans; C406074:RS 102481; D003094:Collagen; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; D020784:Matrix Metalloproteinase 8