Perturbation of adhesion molecule-mediated chondrocyte-matrix interactions by 4-hydroxynonenal binding: implication in osteoarthritis pathogenesis

Abstract

IntroductionObjectives were to investigate whether interactions between human osteoarthritic chondrocytes and 4-hydroxynonenal (HNE)-modified type II collagen (Col II) affect cell phenotype and functions and to determine the protective role of carnosine (CAR) treatment in preventing these effects.MethodsHuman 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 α1β1 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-κB) were assessed by commercial kits. Col II, cyclooxygenase-2 (COX-2), MAPK, NF-κB-p65 levels were analyzed by Western blotting. The formation of α1β1 integrin-focal adhesion kinase (FAK) complex was revealed by immunoprecipitation.ResultsCol II modification by HNE at MR approximately 1:20, strongly induced ICAM-1, α1β1 integrin and MMP-13 expression as well as extracellular signal-regulated kinases 1 and 2 (ERK1/2) and NF-κB-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 α1β1 integrin and Col II expression as well as ERK1/2 and NF-κB-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 α1β1 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.ConclusionsOur novel findings indicate that HNE-binding to Col II results in multiple abnormalities of chondrocyte phenotype and function, suggesting its contribution in osteoarthritis development. CAR was shown to be an efficient HNE-snaring agent capable of counteracting these outcomes.

Matrix-Embedded Cytokines to Simulate Osteoarthritis-Like Cartilage Microenvironments

“…41 El-Bikai et al supported this concept by showing that ICAM-1, integrin a 1 b 1 , MMP13, extracellular signal-regulated kinases 1 and 2, and COX-2 were strongly upregulated in chondrocytes cultured over 4-hydroxynonenal anchored collagen 2 fibrils. 42 These findings indicate that the intimate interaction between receptors of chondrocytes with inflammatory mediators anchored to the underlying matrix or modulated matrix composition initiates cascades of signaling pathways triggering early features of OA pathogenesis.…”

Integrin signaling in skeletal development and function

“…In the presence of α1 or β1 blocking antibodies the induction of MMP‐13 expression was inhibited (Ronziere et al, ). Furthermore, a recent report has suggested that collagen II fibrils modified by the lipid peroxidation end‐product 4‐hydroxynonenal, which is upregulated in synovial fluid of OA patients, induce cell death and catabolic and inflammatory responses of human AC chondrocytes, presumably through perturbation of α1β1 signaling (El‐Bikai et al, ).…”

Transforming dietary peptides in promising lead compounds: the case of bioavailable carnosine analogs

“…As an example, HNE binding to type II collagen and the consequent ALE product results in signal transduction, inducing multiple abnormalities of chondrocytes phenotype and function, suggesting its contribution in osteoarthritis development (El-Bikai et al 2010). AGEs can also induce vascular smooth muscle proliferation via a prolonged agonist-induced Ca 2?…”