Inflammatory conditions partly impair the mechanically mediated activation of Smad2/3 signaling in articular cartilage

Background: Joint trauma, which is frequently related with mechanical overloading of articular cartilage, is a well-established risk for osteoarthritis (OA) development. Additionally, reports show that trauma leads to synovial joint inflammation. In consequence, after joint trauma, cartilage is influenced by deleterious excessive loading combined with the catabolic activity of proinflammatory mediators. Since the activation of TGF-beta signaling by loading is considered to be a key regulatory pathway for maintaining cartilage homeostasis, we tested the effect of proinflammatory conditions on mechanically mediated activation of TGF-beta/Smad2/3P signaling in cartilage. Methods: Cartilage explants were subjected to dynamic mechanical compression in the presence of interleukin-1 beta (IL-1 beta) or osteoarthritic synovium-conditioned medium (OAS-CM). Subsequently, the activation of the Smad2/3P pathway was monitored with QPCR analysis of reporter genes and additionally the expression of receptors activating the Smad2/3P pathway was analyzed. Finally, the ability for mechanically mediated activation of Smad2/3P was tested in human OA cartilage. Results: IL-1 beta presence during compression did not impair the upregulation of Smad2/3P reporter genes, however the results were affected by IL-1 beta-mediated upregulations in unloaded controls. OAS-CM significantly impaired the compression-mediated upregulation of bSmad7 and Tgbfb1. IL-1 beta suppressed the compression-mediated bAlk5 upregulation where 12 MPa compression applied in the presence of OAS-CM downregulated the bTgfbr2. Mechanically driven upregulation of Smad2/3P reporter genes was present in OA cartilage. Conclusions: Proinflammatory conditions partly impair the mechanically mediated activation of the protective TGF-beta/Smad2/3P pathway. Additionally, the excessive mechanical compression, applied in the presence of proinflammatory conditions diminishes the expression of the type II TGF-beta receptor, a receptor critical for maintenance of articular cartilage.