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

纤维软骨表达模式分析以促进颞下颌关节再生医学

• 批准号: 15592098
• 负责人: YODA Tetsuya
• 金额: $ 1.54万
• 依托单位: Saitama Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15592098/
• 关键词: cartilage regeneration; humoral factor; dedifferentiation; temporomandibular joint; statistical analysis; Cdk6; CGK II; cell differentiation; CGK II; 軟骨; 三次元培養; 再生医療

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

颞下颌关节再生医学是口腔颌面外科领域的重要课题之一。为了给颞下颌关节软骨再生的治疗应用提供依据，我们进行了以下几个方面的研究。（1）软骨细胞增殖培养基制备的基础研究：本课题中，我们使用了来自鼻中隔软骨和残余耳廓软骨的软骨细胞。我们的目的是制备软骨细胞增殖培养基，其i）不含胎牛血清（FBS），ii）提供超过1000倍的细胞数量增加，和iii）利用已被证明对临床使用有效的市售生长因子的组合。结果，FGF-2、胰岛素和IGF- 1的组合协同增强了增殖。此外，我们发现BMP-2、胰岛素和PTH的组合对肥大细胞的增殖具有促进作用， ...更多信息 软骨细胞的分化。最后，我们用几种不同的水凝胶对支架的软骨再生性能进行了评价。（2）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的负调控因子的重要功能。因此，CGK II，一种分子开关，通过减弱Sox 9功能，将软骨细胞增殖的停止和肥大分化的开始结合起来。Cdk 6通过Smads介导的下调是BMP-2诱导的成骨细胞分化的关键调节因子，并且过表达Cdk 6的RAW细胞抵抗RANKL诱导的破骨细胞生成;然而，细胞周期调节不受Cdk 6过表达水平的影响。总之，我们已经证明了这些分子在骨组织增殖和分化中的重要性的体外证据。少

项目成果

Cyclic GMP-dependent protein kinase II is a molecular switch from proliferation to hypertrophic differentiation of chondrocytes

• DOI: 10.1101/gad.1224204
• 发表时间: 2004-10-01
• 期刊: GENES & DEVELOPMENT
• 影响因子: 10.5
• 作者: Chikuda, H;Kugimiya, F;Kawaguchi, H
• 通讯作者: Kawaguchi, H

Bone morphogenetic protein 2-induced osteoblast differentiation requires smad-mediated down-regulation of Cdk6

• DOI: 10.1128/mcb.24.15.6560-6568.2004
• 发表时间: 2004-08-01
• 期刊: MOLECULAR AND CELLULAR BIOLOGY
• 影响因子: 5.3
• 作者: Ogasawara, T;Kawaguchi, H;Okayama, H
• 通讯作者: Okayama, H

Mutation in cGMP-dependent protein kinase II causes dwarfism in a rat mutant KML through uncoupling of proliferation and differentiation of chondrocytes.

cGMP 依赖性蛋白激酶 II 的突变通过软骨细胞增殖和分化的解偶联导致大鼠突变 KML 侏儒症。

• 发表时间: 2005
• 期刊: J Bone Miner Metab, 23
• 作者: Hikiji H;Hirotaka chikuda et al.
• 通讯作者: Hirotaka chikuda et al.

Bone morphogenetic protein 2-induced osteoblast differentiation requires Smad-mediated down-regrulation of Cdk6.

骨形态发生蛋白 2 诱导的成骨细胞分化需要 Smad 介导的 Cdk6 下调。

• 发表时间: 2004
• 期刊: Mol Cell Biol 24
• 作者: Toru Ogasawara et al.

Osteoclast differentiation by RANKL requires NF-kappaB-mediated downregulation of cyclin-dependent kinase 6 (Cdk6).

RANKL 的破骨细胞分化需要 NF-kappaB 介导的细胞周期蛋白依赖性激酶 6 (Cdk6) 下调。

• 发表时间: 2004
• 期刊: J. Bone Miner. Res. 19
• 作者: Ogasawara;T.;Katagiri;M.;Yamamoto;A.;Hoshi;K.;Takato;T.;Nakamura;K.;Tanaka;S.;Okayama;H.;Kawaguchi;H.
• 通讯作者: H.