Molecular mechanism of temporomandibular joint osteoarthritis and gene therapy for degraded, cartilage

颞下颌关节骨关节炎的分子机制及退化软骨的基因治疗

• 批准号: 14571843
• 负责人: FUJISAWA Takuo
• 金额: $ 2.56万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571843/
• 关键词: Osteoarthritis; Mechanical Stress; Gene Therapy; Articular Cartilage; Cartilage Degradation; Apoptosis; Nitric Oxide; 軟骨細胞; NO; 細胞増殖; 細胞分化; 結合組織成長因子

The purposes of this research are to clarify the mechanism of cartilage degradation in osteoarthritis (OA) and to investigate the possibility of gene therapy for articular cartlige restoration using connective tissue growth factor (CTGF).First, we investigated the effects of CTGF/Hcs24 transduced by recombinant adenoviruses on the rabbit articular cartilage (RAC) cells in vitro. When RAC cells were infected with adenoviruses containing the CTGF/Hcs24 gene, RAC cells expressed CTGF/Hcs24 mRNA and produced CTGF/Hcs24 protein. RAC cells synthesized more proteoglycan than the control cells. These results suggest that CTGF is useful factor for cartilage repair.Second, we establish a genuine mechanical-stress-induced OA model of the rabbit TMJ. In the experimental rabbits, repetitive forced jaw opening (RFJO) 3 hours/day for 5 days was applied. By histological assessment of the TMJ articular tissues, partial eburnation of the articular cartilage, reactive marginal proliferation of the articular cartilage chondrocytes and nested proliferation of chondrocytes in the subchondral bone area were observed at 7 days after the RFJO period. Furthermore, apoptotic chondrocytes were observed in the cartilage degradation area at 7 days after the RFJO period. And nitrotyrosine, a marker of NO production, and MMP-3, a key factor of cartilage ECM degradation, were observed where chondrocyte apoptosis was evident. These results suggest the RFJO protocol without any surgical intervention can induce evident OA-like lesions in the rabbit TMJ, and cartilage degradation in OA may be induced via chondrocytes apoptosis. This OA model may greatly contribute to the elucidation of the cartilage degradation mechanism in TMJ OA.

本研究的目的是阐明骨关节炎（OA）软骨退化的机制，并探讨利用结缔组织生长因子（CTGF）进行基因治疗修复关节软骨的可能性。首先，我们在体外研究了重组腺病毒转导的CTGF/Hcs24对兔关节软骨（RAC）细胞的影响。当RAC细胞感染含有CTGF/Hcs24基因的腺病毒时，RAC细胞表达CTGF/Hcs24 mRNA并产生CTGF/Hcs24蛋白。 RAC 细胞比对照细胞合成更多的蛋白多糖。这些结果表明CTGF是软骨修复的有用因子。其次，我们建立了真正的机械应力诱导的兔TMJ骨关节炎模型。在实验兔子中，每天进行 3 小时的重复强制张口 (RFJO)，持续 5 天。通过对TMJ关节组织的组织学评估，在RFJO期后7天观察到关节软骨的部分烧伤、关节软骨软骨细胞的反应性边缘增殖和软骨下骨区域的软骨细胞的巢式增殖。此外，RFJO期后7天，在软骨退化区域观察到凋亡的软骨细胞。在软骨细胞凋亡明显的情况下，观察到硝基酪氨酸（NO 产生的标志物）和 MMP-3（软骨 ECM 降解的关键因素）。这些结果表明，无需任何手术干预的 RFJO 方案即可在兔 TMJ 中诱导明显的 OA 样病变，并且 OA 中的软骨退化可能是通过软骨细胞凋亡诱导的。该 OA 模型可能极大地有助于阐明 TMJ OA 中的软骨退化机制。

项目成果

T.Fujisawa, T.Kuboki, T.Kasai, W.Sonoyama, S.Kojima, J.Uehara, C.Komori, H.Yatani, T.Hattori, M.Takigawa: "A repetitive mouth opening induced osteoarthritis-like lesion in rabbit temporomandibular joint."J Dent Res. 82. 731-735 (2003)

T.Fujisawa、T.Kuboki、T.Kasai、W.Sonoyama、S.Kojima、J.Uehara、C.Komori、H.Yatani、T.Hattori、M.Takikawa：“重复张口引起的骨关节炎样病变

Fujisawa et al.: "A repetitive mouth opening induced osteoarthritis-like lesion in rabbit temporomandibular joint."J Dent Res. 82(8). 731-735 (2003)

Fujisawa 等人：“反复张口会导致兔子颞下颌关节出现骨关节炎样病变。”J Dent Res。

T Fujisawa et al.: "A repetitive mouth opening induced osteoarthritis-like lesion in rabbit temporomandibular joint."J Dent Res. 82. 731-735 (2003)

T Fujisawa 等人：“反复张口会导致兔子颞下颌关节出现骨关节炎样病变。”J Dent Res。

T.Nishida et al.: "CTGF/Hcs24, a hypertrophic chondrocytes specific gene product, stimulates proliferation and differentiation but not hypertrophy of cultured articular chondrocytes"Journal of Cellular Physiology. 192. 55-63 (2002)

T.Nishida 等人：“CTGF/Hcs24，一种肥大软骨细胞特异性基因产物，刺激增殖和分化，但不刺激培养的关节软骨细胞肥大”《细胞生理学杂志》。

T.Fujisawa et al.: "NO Production in Mechanical-Stress-Induced OA Cartilage of the Rabbit TMJ"Journal of Dental Research. 81. 379 (2002)

T.Fujisawa 等人：“兔子 TMJ 机械应力诱导的 OA 软骨中的 NO 产生”牙科研究杂志。