title: Selective enhancement of collagenase-mediated cleavage of resident type II collagen in cultured osteoarthritic cartilage and arrest with a synthetic inhibitor that spares collagenase 1 (matrix metalloproteinase 1).
abstract: OBJECTIVE
  To examine whether type II collagen cleavage by collagenase and loss of proteoglycan are excessive in human osteoarthritic (OA) articular cartilage compared with nonarthritic articular cartilage, and whether this can be inhibited by a selective synthetic inhibitor that spares collagenase 1 (matrix metalloproteinase 1 [MMP-1]).
  
  
  METHODS
  Articular cartilage samples were obtained during surgery from 11 patients with OA and at autopsy from 5 adults without arthritis. The articular cartilage samples were cultured in serum-free medium. A collagenase-generated neoepitope, which reflects cleavage of type II collagen, and proteoglycan glycosaminoglycan (GAG), which predominantly reflects aggrecan release, were assayed in culture media. In addition, cultures were performed using either of 2 synthetic MMP inhibitors, both of which inhibited collagenase 2 (MMP-8) and collagenase 3 (MMP-13), but one of which spared collagenase 1. Cultures were also biolabeled with 3H-proline in the presence and absence of these inhibitors to measure collagen synthesis (as tritiated hydroxyproline) and incorporation in articular cartilage.
  
  
  RESULTS
  As a group, cleavage of type II collagen by collagenase was significantly increased in OA cartilage samples. In contrast, proteoglycan (GAG) release was not increased. This release of a collagenase-generated epitope was inhibited by both MMP inhibitors in 2 of 5 nonarthritic samples and in 9 of 11 OA cartilage samples. The inhibitor that spared collagenase 1 was generally more effective and inhibited release from 4 of 5 nonarthritic cartilage samples and the same OA cartilage samples. Group analyses revealed that the inhibition of collagenase neoepitope release by both inhibitors was significant in the OA patient cartilage, but not in the nonarthritic cartilage. Proteoglycan loss was unaffected by either inhibitor. Newly synthesized collagen (predominantly, type II) exhibited increased incorporation in OA cartilage, but only in the presence of the inhibitor that arrested collagenase 1 activity.
  
  
  CONCLUSIONS
  These results further indicate that the digestion of type II collagen by collagenase is selectively increased in OA cartilage, and that this can be inhibited in the majority of cases by a synthetic inhibitor that can inhibit collagenases 2 and 3, but not collagenase 1. The results also suggest that in OA, newly synthesized collagen is digested, but in a different manner than that of resident molecules. Proteoglycan release was not increased in OA cartilage and was unaffected by these inhibitors. Inhibitors of this kind may be of value in preventing damage to type II collagen in human arthritic articular cartilage.
authors.affiliation: University Hospital, Malmö, Sweden.
authors.forename: L
authors.initials: L
authors.lastname: Dahlberg
authors.forename: R C
authors.initials: RC
authors.lastname: Billinghurst
authors.forename: P
authors.initials: P
authors.lastname: Manner
authors.forename: F
authors.initials: F
authors.lastname: Nelson
authors.forename: G
authors.initials: G
authors.lastname: Webb
authors.forename: M
authors.initials: M
authors.lastname: Ionescu
authors.forename: A
authors.initials: A
authors.lastname: Reiner
authors.forename: M
authors.initials: M
authors.lastname: Tanzer
authors.forename: D
authors.initials: D
authors.lastname: Zukor
authors.forename: J
authors.initials: J
authors.lastname: Chen
authors.forename: H E
authors.initials: HE
authors.lastname: van Wart
authors.forename: A R
authors.initials: AR
authors.lastname: Poole
mesh_terms: D000328:Adult; D053674:Aggrecans; D002358:Cartilage, Articular / Q000378:metabolism*; D003094:Collagen / Q000378:metabolism*; D017364:Collagenases / Q000494:pharmacology*; D046508:Culture Techniques; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D016326:Extracellular Matrix Proteins; D005260:Female; D006801:Humans; D037181:Lectins, C-Type; D008297:Male; D053509:Matrix Metalloproteinase 13; D061965:Matrix Metalloproteinase Inhibitors; D008875:Middle Aged; D010003:Osteoarthritis / Q000378:metabolism*; D011489:Protein Denaturation; D011509:Proteoglycans / Q000187:drug effects / Q000378:metabolism
chemical_list: D053674:Aggrecans; D004791:Enzyme Inhibitors; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D061965:Matrix Metalloproteinase Inhibitors; D011509:Proteoglycans; D003094:Collagen; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13