Establishment of synvoial cells and analysis of interactions between the cells from synovial tissue

白细胞介素1(IL-1)β是关节囊内病变的关键介质，如关节内结构紊乱和骨关节炎。应用寡核苷酸芯片技术研究IL-1β刺激的滑膜成纤维细胞基因表达谱，以阐明与关节囊内病变相关的候选基因。滑膜组织取自5例接受TMJ关节镜检查的ID患者。患者对手术和在研究中使用他们的组织表示完全知情同意。从5例患者经IL-1β处理后的滑膜成纤维细胞中提取RNA。用基因芯片进行基因表达谱分析。通过比较IL-1β处理的细胞和未处理的细胞来确定基因表达的变化。滑膜成纤维细胞经IL-1β作用后，IL-8、单核细胞趋化蛋白-1、COX2、IL-1β、IL-6、M-CSF5基因表达增加。此外，我们还观察了IL-1β在大鼠颞下颌关节腔内的注射情况。单次关节内注射IL-1β后24小时，大鼠关节滑膜出现滑膜炎样改变，滑膜衬里增厚，血管扩张，滑膜下结缔组织内有单个核细胞浸润。而对照组(注射生理盐水)的滑膜表现为非炎性改变。免疫组织化学显示，MCP-1、IL-6和COX2在实验性大鼠关节滑膜组织的衬里和衬里下层细胞中均有表达。对照组未见阳性细胞。这些数据应提供有关TMJ病理状态的有用信息，特别是支持TMJ的诊断和治疗方法。这些实验是根据机构审查委员会(Eco3-003)和日本大学松岛牙科学院动物护理指南(05-003)制定的指南进行的。

Interleukin (IL)-1β is a key mediator of intracapsular pathologic conditions of the temporomandibular joint (TMJ), such as internal derangement (ID) and osteoarthritis. Gene expression profile of synovial fibroblasts stimulated with IL-1β was studied by oligonucleotide microarray analysis to elucidate candidate genes associated with intracapsular pathologic conditions of TMJ. Human synovial tissue was obtained from five patients with ID who underwent arthroscopy of the TMJ. The patients gave complete informed consent for the surgery and for the use of their tissue in research. RNA was isolated from synovial fibroblasts from five patients after IL-1β treatment. Gene expression profiling was performed with a GeneChip. Changes in gene expression were determined by comparing IL-1β-treated cells with untreated cells. Gene expressions of IL-8, MCP-1, COX2, IL-1β, IL-6, and M-CSF were increased in synovial fibroblasts treated with IL-1β. Furthermore, we examined that IL-1β was injected into the cavity of the rat TMJ. Twenty-four hours after a single intraarticular injection of IL-1β in rat TMJs, the TMJs showed synovitis-like changes such as an increase in the thickness of the synovial lining, vascular dilation, and infiltration of mononuclear cells in the subsynovial connective tissue. However, the synovial membranes of control TMJs (injected with saline solution) showed non-inflammatory changes. In immunohistochemistry, positive cells of MCP-1, IL-6, and COX2 were detected in the lining and the sublining cells of the synovial tissue of experimental rat TMJs. No positive cells appeared in the control. These data should provided useful information about the pathologic conditions of TMJ, especially in support of diagnosis and therapeutic approaches to TMJ. These experiments were performed according to the guidelines established by the Institutional Review Board (ECO3-003) and the Guidelines for Animal Care (05-003) of Nihon University School of Dentistry at Matsudo.