Analysis of expression pattern of fibrocartilage with a view to promoting regenerative medicine of temporomandibular joint

摘要颞下颌关节再生医学是口腔颌面外科领域最重要的学科之一。为了给TMJ软骨再生的治疗应用提供依据，我们进行了以下几个项目的研究：(1)软骨细胞增殖液的基础研究：本项目中，我们使用了来源于鼻中隔软骨的软骨细胞和残留的耳廓软骨细胞。我们的目标是制备一种软骨细胞增殖液，i)不含胎牛血清(FBS)，ii)细胞数量增加1000倍以上，iii)使用已被证明有效的商业生长因子的组合。结果，联合应用成纤维细胞生长因子-2、胰岛素和胰岛素样生长因子-1协同促进了细胞的增殖。此外，我们还发现骨形态发生蛋白-2、胰岛素和甲状旁腺素联合应用对肥大…的增殖有促进作用。软骨细胞分化较多。了解软骨细胞分化的分子机制：我们研究了CGK II在软骨细胞肥大分化中的新作用，以及CDK 6作为成骨细胞、破骨细胞和软骨细胞分化负调控因子的关键作用。因此，作为一种分子开关，CGK II通过抑制SOX-9功能，使软骨细胞停止增殖和开始肥大分化。CDK6是Smads介导的BMP-2诱导成骨细胞分化的关键调节因子，高表达CDK6的RAW细胞可拮抗RANKL诱导的破骨细胞生成，但细胞周期调控不受CDK6过表达水平的影响。总之，我们已经在体外证明了这些分子在骨组织的增殖和分化中的重要性。较少

Regenerative medicine of the temporomandibular joint (TMJ) is one of the most important subjects in the field of oral and maxillofacial surgery. In order to provide a basis of therapeutic application of TMJ cartilage regeneration, we have investigated in the following projects.(1)Basic study to prepare a chondrocyte proliferation medium : In this project, we used cartilage cells derived from the cartilage of the nasal septum and the remnant auricular cartilage. We have aimed to prepare a chondrocyte proliferation medium that i)does not contain fetal bovine serum (FBS), ii)provides more than a 1000-fold increase in cell numbers, and iii)makes use of a combination of commercially available growth factors that has been proven to be effective for clinical use. As a result, a combination of FGF-2, insulin, and IGF- 1 synergistically enhanced the proliferation. Furthermore, we showed that a combination of BMP-2, insulin, and PTH possessed promotional effects on proliferation of hypertrophic … More differentiation of chondrocyte. Finally, we evaluated the property of the scaffold using some kinds of hydro-gel on cartilage regeneration.(2)Understanding the molecular mechanism of chondrocyte differentiation : We have investigated a novel role of CGK II in hypertrophic differentiation of chondrocytes and a critical function of Cdk 6 as a negative regulator of differentiation of osteoblast, osteoclast and chonrocyte. As a result, CGK II, a molecular switch, coupled the cessation of proliferation and the start of hypertrophic differentiation of chondrocytes through attenuation of Sox 9 function. Cdk 6 was a critical regulator of BMP-2-induced osteoblast differentiation by Smads-mediated down-regulation and, RAW cells overexpressing Cdk 6 resisted RANKL-induced osteoclastgenesis ; however, cell cycle regulation was not affected by the levels of Cdk 6 overexpression. In conclusion, we have demonstrated in vitro evidence of the importance of these molecules in proliferation and differentiation of bone tissue. Less