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

滑膜细胞的建立和滑膜组织细胞之间相互作用的分析

基本信息

批准号：
17592111
负责人：
OGURA Naomi
金额：
$ 2.37万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17592111/
关键词：
temporomandibular joint synovial fibroblasts IL-1β GeneChip analysis chemokines COX2 IL-6 MCP-1 顎関節由来滑膜細胞 Microarray TNF-α 株化テロメアーゼ遺伝子

项目摘要

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.

白细胞介素（IL）-1β是颞下颌关节（TMJ）囊内病理状态如内紊乱（ID）和骨关节炎的关键介质。应用寡核苷酸芯片技术研究IL-1β刺激后滑膜成纤维细胞基因表达谱的变化，以探讨与颞下颌关节囊内病变相关的候选基因。人滑膜组织来自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给予了完全的知情同意。从IL-1β治疗后的5名患者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处理的细胞与未处理的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、COX 2、IL-1β、IL-6和M-CSF的基因表达增加。此外，我们还观察了IL-1β注射到大鼠颞下颌关节腔中。在大鼠TMJ中单次关节内注射IL-1β后24小时，TMJ显示滑膜炎样变化，例如滑膜衬里厚度增加、血管扩张和滑膜下结缔组织中单核细胞浸润。然而，对照TMJ的滑膜（注射盐水溶液）显示非炎症性变化。免疫组化结果显示，实验性大鼠TMJ滑膜组织的内层和下层细胞中均可见MCP-1、IL-6和COX-2的阳性细胞。对照组未见阳性细胞。这些数据将提供有关TMJ病理状况的有用信息，特别是支持TMJ的诊断和治疗方法。根据机构审查委员会（ECO 3 -003）制定的指南和日本大学松户牙科学院动物护理指南（05-003）进行这些实验。