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

白介素(IL)-1β是颞下颌关节（TMJ）囊内病理状况的关键介质，如内部紊乱（ID）和骨关节炎。利用寡核苷酸芯片分析IL-1β刺激滑膜成纤维细胞的基因表达谱，以阐明与TMJ囊内病理状况相关的候选基因。我们从5例接受TMJ关节镜检查的ID患者身上获得了人类滑膜组织。患者对手术和研究中使用他们的组织给予了完全的知情同意。从5例IL-1β治疗后的滑膜成纤维细胞中分离RNA。用基因芯片进行基因表达谱分析。通过比较il -1β处理细胞和未处理细胞来确定基因表达的变化。IL-1β处理的滑膜成纤维细胞中IL-8、MCP-1、COX2、IL-1β、IL-6和M-CSF的基因表达增加。此外，我们检测了IL-1β被注射到大鼠TMJ腔内。单次关节内注射IL-1β 24小时后，大鼠TMJs出现滑膜样变化，如滑膜内膜厚度增加，血管扩张，滑膜下结缔组织单核细胞浸润。而对照组颞下颌关节滑膜（注射生理盐水）无炎症变化。免疫组化在实验大鼠颞下颌关节滑膜组织的衬里和亚衬里细胞中检测到MCP-1、IL-6和COX2阳性细胞。对照组未见阳性细胞。这些数据应提供有关TMJ病理状况的有用信息，特别是在支持TMJ的诊断和治疗方法方面。实验依据机构审查委员会（ECO3-003）和日本大学松户牙科学院动物护理指南（05-003）制定的指南进行。

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.