Biomaterial having active sites of collagen-binding adhesion molecules (CMP and RGD-CAP)

具有胶原蛋白结合粘附分子活性位点的生物材料（CMP 和 RGD-CAP）

• 批准号: 09557146
• 负责人: KATO Yukio
• 金额: $ 7.81万
• 依托单位: Hiroshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09557146/
• 关键词: CMP; RGD-CAP; chondrocyte; integrin; collagen; biomatenal; 生体材料; 線維芽細胞

The CMP coating enhanced the adhesion and spreading of various chondrocytes and fibroblasts. The effect of CMP on the cell spreading was synergistically increased by type 11 collagen. We also showed that 1251-CMP selectively binds to alphalbetal integrin directly using monoclonal antibodies to various integrin subunits. In addition, we cloned cDNAS for rabbit matrilin1 (CMP) and rabbit matrilin3 to identify active sites for the stimulation of cell adhesion.The RGD-CAP coating also enhanced the adhesion and spreading of various chondrocytes and fibroblasts. The RGD sequence near the C-terminal was not essential for the adhesion activity. Studies with monoclonal antibodies to various integrin subunits showed that RGD-CAP is a novel ligand for alpha1beta1 integrin that does not bind to the RGD motif. We have shown that at least two repeated structures within the RGD-CAP molecule are required for the stimulation of cell adhesion. These findings will be useful for clinical application of CMP and RGD-CAP, as well as their fragments, as biomaterials in tissue engineering.

化学机械抛光涂层增强了各种软骨细胞和成纤维细胞的黏附和铺展。11型胶原协同增强了cMP对细胞铺展的作用。我们还发现，1251-CMP利用针对各种整合素亚基的单抗，选择性地直接与α-β整合素结合。此外，我们还克隆了兔Matrilin1和兔matrilin3的cDNA，以确定刺激细胞黏附的活性部位。RGD-CAP涂层还增强了各种软骨细胞和成纤维细胞的黏附和铺展。C末端附近的RGD序列对黏附活性不是必需的。对不同整合素亚基的单抗研究表明，RGD-CAP是一种不与RGD基序结合的新的α1β1整合素配体。我们已经证明，RGD-CAP分子中至少有两个重复结构是刺激细胞黏附所必需的。这些发现将有助于将CMP和RGD-CAP及其片段作为生物材料在组织工程中的临床应用。

项目成果

Okimura, A.Y.Okada, S.Makihira, H.Pan, L Yu,K.Tanne K Iamai,H. Yamada T, Kawamoto, M.Nosiro, W Yan, and Y,Kato.: "Enhancement of cartilage matrix protein synthesis in arthritic cartilage." Arthritis and Rheumatism.40. 1029-1036 (1997)

冲村，A.Y.Okada，S.Makihira，H.Pan，L Yu，K.Tanne K Iamai，H.

Kawashima-Ohya,Y.,et al.: "Retinol-binding protein is produced by rabbit chondrocytes and responds to parathyroid hormone(PTH)/PTH-related peptide-cyclic adenosine monophophate pathway"Endocrinology. 140. 1075-1081 (1999)

Kawashima-Ohya, Y., et al.：“视黄醇结合蛋白由兔软骨细胞产生，并对甲状旁腺激素 (PTH)/PTH 相关肽-环磷酸腺苷途径作出反应”内分泌学。

Makihira,S.,et al.: "Enhancement of cell adhesion and spreading by a cartilage-speciffic noncollagenous protein,cartilage matrix protein (CMP/Matrilin-1),via integrin α1β1."J.Biol.Chem.. 274. 11417-11423 (1999)

Makihira, S. 等人：“通过整合素 α1β1，通过软骨特异性非胶原蛋白、软骨基质蛋白 (CMP/Matrilin-1) 增强细胞粘附和扩散。J.Biol.Chem.. 274. 11417” -11423 (1999)

Ohno,S.,et al.: "RGD CAP(βin-h3) enhances the spreading of chondrocyten and fibroblastes via integrin α1β1"Biochimica et Biophysica Acta. 1451. 196-205 (1999)

Ohno, S., 等人：“RGD CAP(βin-h3) 通过整合素 α1β1 增强软骨细胞和成纤维细胞的扩散”Biochimica et Biophysical Acta. 1451. 196-205 (1999)

Shimazu,A.,et al.: "Expression of syndecan-2,-4,and fibroblast growth factor receptor type I in periodontal ligament fibroblasts and down-regulation of these membrance proteins during differentiation"Journal of Dental Research. 78. 1791-1799 (1999)

Shimazu, A., et al.：“牙周膜成纤维细胞中 Syndecan-2、-4 和成纤维细胞生长因子受体 I 型的表达以及分化过程中这些膜蛋白的下调”《牙科研究杂志》。