title: Cannabinoid WIN-55,212-2 mesylate inhibits interleukin-1β induced matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase expression in human chondrocytes.
abstract: OBJECTIVE
  Interleukin-1β (IL-1β) is involved in the up-regulation of matrix metalloproteinases (MMPs) leading to cartilage degradation. Cannabinoids are anti-inflammatory and reduce joint damage in animal models of arthritis. This study aimed to determine a mechanism whereby the synthetic cannabinoid WIN-55,212-2 mesylate (WIN-55) may inhibit cartilage degradation.
  
  
  METHODS
  Effects of WIN-55 were studied on IL-1β stimulated production of MMP-3 and -13 and their inhibitors TIMP-1 and -2 in human chondrocytes. Chondrocytes were obtained from articular cartilage of patients undergoing total knee replacement. Chondrocytes were grown in monolayer and 3D alginate bead cultures. Real-time polymerase chain reaction (PCR) was used to determine the gene expression of MMP-3, -13, TIMP-1 and -2 and Enzyme Linked Immunosorbent Assay (ELISA) to measure the amount of MMP-3 and MMP-13 protein released into media. Immunocytochemistry was used to investigate the expression of cannabinoid receptors in chondrocyte cultures.
  
  
  RESULTS
  Treatment with WIN-55 alone or in combination with IL-1β, decreased or abolished MMP-3, -13, TIMP-1 and -2 gene expression in human chondrocyte monolayer and alginate bead cultures in both a concentration and time dependent manner. WIN-55 treatment alone, and in combination with IL-1β, reduced MMP-3 and -13 protein production by chondrocytes cultured in alginate beads. Immunocytochemistry demonstrated the expression of cannabinoid receptors in chondrocyte cultures.
  
  
  CONCLUSIONS
  Cannabinoid WIN-55 can reduce both basal and IL-1β stimulated gene and protein expression of MMP-3 and -13. However WIN-55 also decreased basal levels of TIMP-1 and -2 mRNA. These actions of WIN-55 suggest a mechanism by which cannabinoids may act to prevent cartilage breakdown in arthritis.
authors.affiliation: Biomedical Research Centre, Faculty of Health and Wellbeing, Sheffield Hallam University, Sheffield S1 1WB, UK. Electronic address: sldunn@my.shu.ac.uk.
authors.forename: S L
authors.initials: SL
authors.lastname: Dunn
authors.affiliation: Academic Unit of Bone Metabolism, Department of Human Metabolism, University of Sheffield, UK. Electronic address: j.m.wilkinson@sheffield.ac.uk.
authors.forename: J M
authors.initials: JM
authors.lastname: Wilkinson
authors.affiliation: Centre for Biomaterials & Tissue Engineering, University of Sheffield, School of Clinical Dentistry, UK. Electronic address: a.crawford@sheffield.ac.uk.
authors.forename: A
authors.initials: A
authors.lastname: Crawford
authors.affiliation: Biomedical Research Centre, Faculty of Health and Wellbeing, Sheffield Hallam University, Sheffield S1 1WB, UK. Electronic address: C.Lemaitre@shu.ac.uk.
authors.forename: C L
authors.initials: CL
authors.lastname: Le Maitre
authors.affiliation: Biomedical Research Centre, Faculty of Health and Wellbeing, Sheffield Hallam University, Sheffield S1 1WB, UK. Electronic address: r.a.bunning@shu.ac.uk.
authors.forename: R A D
authors.initials: RA
authors.lastname: Bunning
mesh_terms: D000464:Alginates; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D048588:Benzoxazines / Q000494:pharmacology*; D002358:Cartilage, Articular / Q000378:metabolism / Q000473:pathology; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects* / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005786:Gene Expression Regulation / Q000187:drug effects; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D053583:Interleukin-1beta / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D053509:Matrix Metalloproteinase 13 / Q000096:biosynthesis / Q000235:genetics; D019278:Matrix Metalloproteinase 3 / Q000096:biosynthesis / Q000235:genetics; D020782:Matrix Metalloproteinases / Q000096:biosynthesis*; D009025:Morpholines / Q000494:pharmacology*; D009281:Naphthalenes / Q000494:pharmacology*; D020370:Osteoarthritis, Knee / Q000378:metabolism / Q000473:pathology; D043882:Receptors, Cannabinoid / Q000378:metabolism; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000096:biosynthesis; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000096:biosynthesis; D019714:Tissue Inhibitor of Metalloproteinases / Q000096:biosynthesis*
chemical_list: D000464:Alginates; D000894:Anti-Inflammatory Agents, Non-Steroidal; D048588:Benzoxazines; D006603:Hexuronic Acids; D053583:Interleukin-1beta; D009025:Morpholines; D009281:Naphthalenes; D043882:Receptors, Cannabinoid; C560822:TIMP1 protein, human; C560823:TIMP2 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D019714:Tissue Inhibitor of Metalloproteinases; D019716:Tissue Inhibitor of Metalloproteinase-2; C070417:(3R)-((2,3-dihydro-5-methyl-3-((4-morpholinyl)methyl)pyrrolo-(1,2,3-de)-1,4-benzoxazin-6-yl)(1-naphthalenyl))methanone; D020723:Glucuronic Acid; D053509:Matrix Metalloproteinase 13; D020782:Matrix Metalloproteinases; D019278:Matrix Metalloproteinase 3
keywords: Cannabinoid; Cartilage degradation; Chondrocytes; Interleukin 1 (IL-1); Matrix metalloproteinases (MMPs); Tissue inhibitors of matrix metalloproteinases (TIMP)