Cellbiological study for gene expression on the cultured synovial cell in case of internalderangement of the temporomandibular joint

颞下颌关节内部紊乱时培养的滑膜细胞基因表达的细胞生物学研究

• 批准号: 08835011
• 负责人: FUJITA Shigeyuki
• 金额: $ 1.34万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08835011/
• 关键词: Internalderangement; Cellculture; Temporomandibularjoint; Vimentin; inteleukin-6; TGF-β; テネイシン; 顎関節; 細胞外基質; サイトカイン; In-situ hybridization

Many patientens who are suffering from pain and disorder due to the internalderangement on temporomandibular join (TMJ) visited our clinic but the it's etiology still unclear.Especially the fuction of TMJ synoviocytes in the case of internalderangement is not investigated at all. We tried to establish the culture synovial celllines from the internalderangement of TMJ.At first, we tried just the same method as the synovial cell culture method on knee joint by Dayr JC.With this method, in the end of the screening the synovial tissue materials, the number of rest of synoviocytes was very little and it's activity was not so hight as synoviocytes of rheumatism arthritis on knee joints. With this method, we could not get the cultured synovial cellline from our TMJ materials. Therefore we added some arrangement to this method ; after that, we found that DMEM medium with 10% fetal bovine serum (BSA) was best to culture TMJ synovial cell. In this condition, one of cellline could grow after 38 h … More ours later, another of cellline takes more than 70 hours. What was interesting for us the hormer cellline derived from the patient who had much more pain in TMJ than the latter cellline.We also studied the cellline with anti-cytokeratin, anti-vimentin, anti-leukocyte common antigen, and anti-macrophage antigen monoclonal antibodies immunohistochemically. As the result of these study only anti-vimentin monoclonal antibodies reacted strongly a lot of numbers of culture cells, with another antibodies, no positive reaction could be seen.Farther more, with the culture supernatants from synoviocytes, the consentration of trans forming growth facto beta, interleukin-1 beta and interleukin-6 were analized with ELISA method. As the result of these study, only the consentration of interleukin-6 was hight level in every celllines but there ware no capable difference between them. The other consentration of another cytokines was not detectable with ELISA.The other hand, on the hyperplastic synovial tissues in these disease m-RNA production could be recognized with in-situ-hybridization. Less

许多因颞下颌关节（TMJ）内部紊乱而遭受疼痛和紊乱的患者来我们诊所就诊，但其病因仍不清楚。特别是颞下颌关节滑膜细胞在内部紊乱情况下的功能根本没有被研究过。我们试图从TMJ内部紊乱中建立培养滑膜细胞系。首先，我们尝试了与Dayr JC膝关节滑膜细胞培养方法相同的方法。用这种方法，在筛选滑膜组织材料的最后，剩余的滑膜细胞数量很少，其活性也不如膝关节风湿性关节炎滑膜细胞高。通过这种方法，我们无法从我们的颞下颌关节材料中获得培养的滑膜细胞系。因此我们在这个方法上添加了一些安排；之后我们发现含有10%胎牛血清（BSA）的DMEM培养基最适合培养TMJ滑膜细胞。在这种情况下，一种细胞系可以在38小时后生长……而我们的细胞系则需要70多个小时。对我们来说有趣的是来自于颞下颌关节疼痛比后者更严重的患者的hormer细胞系。我们还用免疫组织化学方法研究了具有抗细胞角蛋白、抗波形蛋白、抗白细胞共同抗原和抗巨噬细胞抗原单克隆抗体的细胞系。本研究结果表明，只有抗波形蛋白单克隆抗体对大量培养细胞有强烈反应，与其他抗体未见阳性反应。此外，用ELISA方法分析了滑膜细胞培养上清中转化生长因子β、白细胞介素1β和白细胞介素6的一致性。这些研究的结果是，只有IL-6在各细胞系中的表达水平较高，但它们之间没有显着差异。 ELISA无法检测到另一种细胞因子的其他一致性。另一方面，在这些疾病的增生性滑膜组织上，mRNA的产生可以通过原位杂交来识别。较少的

项目成果

吉田 博昭: "ヒト顎関節内障の滑膜病変にともなうテネシンの特異的発現" 近畿北陸地区歯科医学大会誌. 49. 48-52 (1997)

Hiroaki Yoshida：“与人类颞下颌关节疾病滑膜病变相关的 Tennessine 的特异性表达”近畿北陆地区牙科会议杂志 49. 48-52 (1997)。

Shinya Yasuda, Shigeyuki Fujita, Norihiro Iwashiro, Tadahiko Iizuka: "Establishment of cultured cell lines of synoviocytes from the case with internalderangement of temporomandibular joint." Jpn J Oral Diagnosis. (submitted to print).

Shinya Yasuda、Shigeyuki Fujita、Norihiro Iwashiro、Tadahiko Iizuka：“从颞下颌关节内部紊乱病例中建立滑膜细胞培养细胞系。”

HIROAKI YOSHIDA: "The specific expression of tenascin in the synovial membrane of the temporamandibular joint with internal derangement:An immuhohistachemical study" Histochemistry and Cell Biology. (In press). (1997)

HIROAKI YOSHIDA：“内部紊乱的颞下颌关节滑膜中肌腱蛋白的特异性表达：一项免疫组织化学研究”组织化学和细胞生物学。

Shinya Yasuda, Shigeyuki Fujita, Norihiro Iwashiro, Tadahiko Iizuka: "Production of cytokine from the cultured cell line of synoviocyte in the case of internalderangement of temporomandibular joint" Jpn J Oral Diagnosis. (submitted to print).

Shinya Yasuda、Shigeyuki Fujita、Norihiro Iwashiro、Tadahiko Iizuka：“在颞下颌关节内部紊乱的情况下从滑膜细胞培养细胞系中产生细胞因子”Jpn J Oral Diagnosis。

Shigeyuki Fujita: "Transforming growth factor and tenascin in synovial chonclromatosis of the TMJ" Int.J.Oral Maxillofac Surg. 26. 258-259 (1997)

Shigeyuki Fujita：“TMJ 滑膜软骨瘤病中的转化生长因子和肌腱蛋白”Int.J.Oral Maxillofac Surg。