Fibroblast growth factor 2 inhibits induction of aggrecanase activity in human articular cartilage

Abstract

Articular chondrocytes are surrounded by an extracellular pool of fibroblast growth factor 2 (FGF-2). We undertook this study to investigate the possible role of FGF-2 in aggrecan catabolism by aggrecanase in human articular cartilage. Aggrecan catabolism was induced by interleukin-1alpha (IL-1alpha) in normal human articular cartilage and assessed by measuring the release of glycosaminoglycan (GAG) and aggrecanase-dependent fragments by Western blotting with antibodies against neoepitopes. ADAMTS-4 and ADAMTS-5 messenger RNA (mRNA) expression was measured by quantitative real-time reverse transcriptase-polymerase chain reaction. Production of matrix metalloproteinases (MMPs) 1, 3, and 13 and tissue inhibitors of metalloproteinases (TIMPs) 1 and 3 was measured by Western blotting. IL-6 and IL-8 were measured by enzyme-linked immunosorbent assay. Proteoglycan synthesis was monitored by 35S-sulfate incorporation. IL-1alpha caused cleavage of aggrecan in cultured human articular cartilage explants, with release of GAG and aggrecan fragments containing ARGS and AGEG neoepitopes. This was inhibited by FGF-2 (1-100 ng/ml). Tumor necrosis factor alpha and retinoic acid also stimulated release of neoepitope, and this was also suppressed by FGF-2. IL-1alpha induced ADAMTS-4 and ADAMTS-5 mRNA in primary human chondrocytes, and this was inhibited by FGF-2. IL-1alpha-induced aggrecan breakdown was inhibited by TIMP-1 or by the N-terminal portion of TIMP-3, although FGF-2 did not affect production of the inhibitors TIMP-1 and TIMP-3 when IL-1alpha was present. FGF-2 did not prevent IL-1alpha suppression of proteoglycan synthesis and did not negate its ability to stimulate the production of IL-6, IL-8, and MMPs 1, 3, and 13. Our findings suggest that FGF-2 may play a chondroprotective role in human articular cartilage by controlling the expression and activity of the aggrecanases ADAMTS-4 and ADAMTS-5.