%0 Journal Article
%T Perturbation of adhesion molecule-mediated chondrocyte-matrix interactions by 4-hydroxynonenal binding: implication in osteoarthritis pathogenesis
%A Rana El-Bikai
%A M¨¦lanie Welman
%A Yoran Margaron
%A Jean-Fran£¿ois C£¿t¨¦
%A Luke Macqueen
%A Michael D Buschmann
%A Hassan Fahmi
%A Qin Shi
%A Karim Maghni
%A Julio C Fernandes
%A Mohamed Benderdour
%J Arthritis Research & Therapy
%D 2010
%I BioMed Central
%R 10.1186/ar3173
%X Human Col II was treated with HNE at different molar ratios (MR) (1:20 to 1:200; Col II:HNE). Articular chondrocytes were seeded in HNE/Col II adduct-coated plates and incubated for 48 hours. Cell morphology was studied by phase-contrast and confocal microscopy. Adhesion molecules such as intercellular adhesion molecule-1 (ICAM-1) and ¦Á1¦Â1 integrin at protein and mRNA levels were quantified by Western blotting, flow cytometry and real-time reverse transcription-polymerase chain reaction. Cell death, caspases activity, prostaglandin E2 (PGE2), metalloproteinase-13 (MMP-13), mitogen-activated protein kinases (MAPKs) and nuclear factor-kappa B (NF-¦ÊB) were assessed by commercial kits. Col II, cyclooxygenase-2 (COX-2), MAPK, NF-¦ÊB-p65 levels were analyzed by Western blotting. The formation of ¦Á1¦Â1 integrin-focal adhesion kinase (FAK) complex was revealed by immunoprecipitation.Col II modification by HNE at MR approximately 1:20, strongly induced ICAM-1, ¦Á1¦Â1 integrin and MMP-13 expression as well as extracellular signal-regulated kinases 1 and 2 (ERK1/2) and NF-¦ÊB-p65 phosphorylation without impacting cell adhesion and viability or Col II expression. However, Col II modification with HNE at MR approximately 1:200, altered chondrocyte adhesion by evoking cell death and caspase-3 activity. It inhibited ¦Á1¦Â1 integrin and Col II expression as well as ERK1/2 and NF-¦ÊB-p65 phosphorylation, but, in contrast, markedly elicited PGE2 release, COX-2 expression and p38 MAPK phosphorylation. Immunoprecipitation assay revealed the involvement of FAK in cell-matrix interactions through the formation of ¦Á1¦Â1 integrin-FAK complex. Moreover, the modification of Col II by HNE at a 1:20 or approximately 1:200 MR affects parameters of the cell shape. All these effects were prevented by CAR, an HNE-trapping drug.Our novel findings indicate that HNE-binding to Col II results in multiple abnormalities of chondrocyte phenotype and function, suggesting its contribution in osteoarthritis developm
%U http://arthritis-research.com/content/12/5/R201