Characterization of cultured synovial cells from human TMJ

本研究的目的是首先从人TMJ滑膜组织中分离培养滑膜细胞并对其进行鉴定，然后研究滑膜细胞中细胞因子和酶的产生。滑膜细胞用免疫细胞化学方法检测细胞特异性标志物。在第二组中，关节滑膜细胞用IL-1β或肿瘤坏死因子-α处理，其在关节紊乱患者的滑液中增加。用酶法检测与细胞外基质降解有关的纤溶酶原激活物(PA)活性。用双抗体夹心法测定IL-6、IL-8和巨噬细胞吸引蛋白(MCP)-1的含量。用RT-PCR方法检测这些细胞因子的基因表达。滑膜细胞表达成纤维细胞特异性标志物和波形蛋白。巨噬细胞标志物、人类白细胞抗原II类和树突状细胞标志物阴性。然后，用或不加IL-1β处理滑膜细胞。IL-1β以时间和剂量依赖的方式增加滑膜细胞产生IL-6、IL-8和单核细胞趋化蛋白-1。IL-1β刺激上述细胞因子的基因表达。IL-1β还可增强滑膜细胞PA活性。接下来，用肿瘤坏死因子-α处理细胞。肿瘤坏死因子-α促进IL-8和MCP-1的产生和基因表达。结论培养的滑膜细胞可能为研究TMJ的细胞和分子机制提供重要的优势。此外，当关节紊乱患者滑液中IL-1β和肿瘤坏死因子-α水平升高时，关节滑膜细胞的PA活性和IL-6、IL-8及单核细胞趋化蛋白-1的产生也可能增加。

The aims of the present study were, firstly, to isolate and to characterize culture synovial cells obtained from human TMJ specimens, and secondly, to investigate the productions of cytokines and enzymes in those cells.<Methods> Synovial cells were isolated from TMJ synovial tissue using an out growth method and then cultured in Ham F12 containing 20% or 10% fetal bovine serum. Synovial cells were examined cell-specific markers using immunocytochemistry. In the next, synovial cells were treated with IL-1β or TNF-α, which was increased in synovial fluids from patients with TMJ disorders. Activity of plasminogen activator (PA), which is associated with extracellular matrix degradation, was measured by enzyme assay. Amounts of IL-6, IL-8 and macrophage attractant protein (MCP)-1 were measured by ELISA. Gene expressions of these cytokines were also investigated using RTPCR method.<Rersults> Synovial cells were positive for fibroblast-specific marker and vimentin. In contrast, the cells were negative for macrophage marker, HLA class II and dendritic cell marker. Then, synovial cells were treated with or without IL-1β. IL-1β increased IL-6, IL-8 and MCP-1 production in synovial cells in a time- and dose-dependent manner. IL-1β stimulated gene expression of these cytokines in them. IL-1β also enhanced PA activity in synovial cells. In the next, the cells were treated with TNF-α. TNF-α enhanced productions and gene expressions of IL-8 and MCP-1.<Conclusions> We found that cultured synovial cells might provide important advantages for studies of the cellular and molecular mechanisms in TMJ. In addition, PA activity and IL-6, IL-8, and MCP-1 productions may be increased in synovial cells when IL-1β and TNF-α levels are increased in synovial fluids from patients with TMJ disorders.