title: Characterisation of equine matrix metalloproteinase 2 and 9; and identification of the cellular sources of these enzymes in joints.
abstract: The cellular production by resident articular cells and infiltrating inflammatory cells of the gelatinase matrix metalloproteinases (MMP) was investigated by tissue culture methods and analysis of cell supernatants by gelatin zymography. Peripheral blood neutrophils in short term culture produced MMP-9, as did peripheral blood monocytes in culture. Isolated articular chondrocytes in monolayer culture produced both MMP-2 and MMP-9, although articular cartilage maintained as explant culture produced MMP-2 alone. Synovial fibroblasts grown in monolayer culture produced MMP-2 alone, although synovial membrane in explant culture produced both MMP-2 and the active form of MMP-2. Lysis of blood polymorph neutrophils produced large quantities of MMP-9, but lysis of blood monocytes, synovial fibroblasts and articular chondrocytes produced little enzyme indicating that, unlike the other cell types, polymorph neutrophils store MMPs intracellularly. Equine MMP-2 was purified from synovial fibroblast cell culture supernatant, and equine MMP-9 from polymorph neutrophil cell culture supernatant, by gelatin-sepharose affinity chromatography. The 2 enzymes were identified from their molecular weights and by their respective N-terminal amino acid sequences which showed homology with the enzymes from other species. The demonstration that invasive cells and resident articular cells can produce enzymes which are capable of digestion of certain component molecules of the articular cartilage matrix, shows that therapeutic targeting of these enzymes could be a valid proposition in the prevention of cartilage destruction in osteoarthritis.
authors.affiliation: Department of Veterinary Clinical Science and Animal Husbandry, University Veterinary Teaching Hospital, University of Liverpool, Leahurst, UK.
authors.forename: P D
authors.initials: PD
authors.lastname: Clegg
authors.forename: R M
authors.initials: RM
authors.lastname: Burke
authors.forename: A R
authors.initials: AR
authors.lastname: Coughlan
authors.forename: C M
authors.initials: CM
authors.lastname: Riggs
authors.forename: S D
authors.initials: SD
authors.lastname: Carter
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology / Q000201:enzymology; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology; D019902:Chondrocytes / Q000166:cytology / Q000201:enzymology; D002846:Chromatography, Affinity / Q000379:methods / Q000662:veterinary; D017364:Collagenases / Q000096:biosynthesis* / Q000737:chemistry; D046508:Culture Techniques; D004285:Dogs; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods / Q000662:veterinary; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D018093:Gelatinases / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000737:chemistry; D006736:Horses / Q000378:metabolism*; D006801:Humans; D007596:Joints / Q000166:cytology / Q000201:enzymology*; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D061965:Matrix Metalloproteinase Inhibitors; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009000:Monocytes / Q000166:cytology / Q000201:enzymology*; D009504:Neutrophils / Q000166:cytology / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D013583:Synovial Membrane / Q000166:cytology / Q000201:enzymology*
chemical_list: D002614:Chelating Agents; D061965:Matrix Metalloproteinase Inhibitors; D004492:Edetic Acid; D017364:Collagenases; D018093:Gelatinases; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9