The role of CTGF as a novel tissue-regenerating factor, regenerin, and its application for medical and dental tissue engineering

CTGF作为新型组织再生因子再生素的作用及其在医学和牙科组织工程中的应用

基本信息

批准号：
15109010
负责人：
TAKIGAWA Masaharu
金额：
$ 66.48万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (S)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2006
项目状态：
已结题

项目摘要

1. Using wild type and/or mutant animals, we found that in addition to endochondral ossification in growth plate, CTGF/CCN2 was involved in secondary ossification center formation, intramembranous ossification, formation of periodontal ligament and articular and auricular cartilages, distraction osteogenesis and repair of tooth extraction socket. In vivo administration of CTGF/CCN2 with gelatin hydrogel into the artificial defect of articular cartilage and bone resulted in repair of these tissues, respectively. Taken together with the finding that platelets contained much CTGF/CCN2, these findings indicate that CTGF/CCN2 functions as a regeneration factor "regenerin".2. We developed CTGF/CCN2 domain-specific antibodies and domain-specific ELISA systems. The function and signal transduction pathway of each domain was different depending on types of cells, such as chondrocytes and endothelial cells. CT domain of CTGF/CCN2 promoted adhesion of mesenchymal stem cells on hydroxyapatite plates, suggesting a possible application for bone regeneration with a combination of CTGF/CCN2 and hydroxyapatite.3. A cis-element in 3'-untranslation region (3'-UTR) of CTGF/CCN2 mRNA, which was involved in destabilization of its mRNA, and a protein, which bound to the element, were found in chondrocytes. The biding between them changed in reverse relation to the process of chondrocyte differentiation. A hypoxia-inducible protein, which stabilized CTGF/CCN2 mRNA by binding to its 3'-UTR was also detected.4. CTGF/CCN2 bound to perlecan, aggrecan and fibronectin, indicating its retention in extracellular matrix. CTGF/CCN2 had collaborative action with M-CSF on cartilage. Low density lipoprotein-related protein I was one of the receptors for CTGF/CCN2 in chondrocytes. Concerning signal transduction pathway of CTGF/CCN2 in chondrocytes, PKC was found as an upstream mediator of ERK and p38MAPK. JNK was involved in cell proliferation. PI3K and PKB were found to be involved in calcification.

1.使用野生型和/或突变型动物，我们发现除了生长板中的软骨内骨化外，CTGF/CCN2还参与次级骨化中心的形成、膜内骨化、牙周膜和关节软骨和耳廓软骨的形成、牵张成骨和拔牙槽的修复。将 CTGF/CCN2 与明胶水凝胶体内施用到关节软骨和骨的人工缺损中，分别导致这些组织的修复。结合血小板含有大量CTGF/CCN2的发现，这些发现表明CTGF/CCN2作为再生因子“regenerin”起作用。2．我们开发了 CTGF/CCN2 域特异性抗体和域特异性 ELISA 系统。每个结构域的功能和信号转导途径根据细胞类型（例如软骨细胞和内皮细胞）而不同。 CTGF/CCN2的CT结构域促进间充质干细胞在羟基磷灰石板上的粘附，提示CTGF/CCN2与羟基磷灰石联合用于骨再生的可能应用。 3．在软骨细胞中发现了 CTGF/CCN2 mRNA 3'-非翻译区 (3'-UTR) 中的顺式元件（参与其 mRNA 的不稳定）以及与该元件结合的蛋白质。它们之间的结合与软骨细胞分化过程呈反向关系。还检测到一种缺氧诱导蛋白，该蛋白通过与其3'-UTR结合来稳定CTGF/CCN2 mRNA。4． CTGF/CCN2 与基底膜蛋白聚糖、聚集蛋白聚糖和纤连蛋白结合，表明其保留在细胞外基质中。 CTGF/CCN2 与 M-CSF 对软骨有协同作用。低密度脂蛋白相关蛋白 I 是软骨细胞中 CTGF/CCN2 的受体之一。关于软骨细胞中CTGF/CCN2的信号转导途径，PKC被发现是ERK和p38MAPK的上游介质。 JNK参与细胞增殖。发现 PI3K 和 PKB 参与钙化。