Characterization of synovial angiogenesis in osteoarthritis patients and its modulation by chondroitin sulfate

Introduction: This work aimed at comparing the production of inflammatory and pro-and anti-angiogenic factors by normal/reactive (N/R) or inflammatory (I) areas of the osteoarthritic synovial membrane. The effects of interleukin (IL)-1 beta and chondroitin sulfate (CS) on the expression of pro-and anti-angiogenic factors by synovial fibroblasts cells (SFC) were also studied. Methods: Biopsies from N/R or from I areas of osteoarthritic synovial membrane were collected at the time of surgery. The inflammatory status of the synovial membrane was characterized by the surgeon according to macroscopic criteria, including the synovial vascularization, the villi formation and the hypertrophic aspect of the tissue. We assessed the expression of CD45, von Willebrand factor and vascular endothelial growth factor (VEGF) antigen by immunohistochemistry in both N/R and I biopsies. The production of IL-6, -8, VEGF and thrombospondin (TSP)-1 by N/R or I synovial cells was quantified by ELISA. SFC were cultured in the absence or in the presence of IL-1 beta (1 ng/ml) and with or without CS (10, 50, 200 mu g/ml). Gene expression of pro-angiogenic factors (VEGF, basic fibroblast growth factor (bFGF), nerve growth factor (NGF), matrix metalloproteinase (MMP)-2 and angiopoietin (ang)-1) and anti-angiogenic factors (vascular endothelial growth inhibitor (VEGI), TSP-1 and -2) were determined by real time RT-PCR. Production of VEGI and TSP-1 was also estimated by ELISA. Results: Immunohistochemistry showed the increase of lymphocyte infiltration, vascular density and VEGF expression in I compared to N/R synovial biopsies. Synovial cells from I areas produced more IL-6, IL-8 and VEGF but less TSP-1 than cells isolated from N/R synovial biopsies. The expression of pro-angiogenic factors by SFC was stimulated by IL-1 beta. A time dependent regulation of the expression of anti-angiogenic factor genes was observed. IL-1 beta stimulated the expression of anti-angiogenic factor genes but inhibited it after 24 h. CS reversed the inhibitory effect of IL-1 beta on anti-angiogenic factors, VEGI and TSP-1. Conclusions: We demonstrated that synovial biopsies from I areas expressed a pro-angiogenic phenotype. IL-1 beta induced an imbalance between pro-and anti-angiogenic factors in SFC and CS tended to normalize this IL-1 beta-induced imbalance, providing a new possible mechanism of action of this drug.