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Local delivery to biomaterials scaffold

局部递送至生物材料支架

基本信息

批准号：
17500312
负责人：
TACHIBANA Akira
金额：
$ 1.6万
依托单位：
Osaka City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17500312/
关键词：
local delivery FGF2 BMP2 chitin binding domain cellulose binding domain バイオマテリアル Chitin binding domain Cellulose binding domain

项目摘要

The seats of chitin and cellulose were constructed by casting of chitin and cellulose solution in dimethylactamide containing LiCl. The expression vector plasmids for expression of fusion proteins, chitin binding domain-FGF2, cellulose binding domain-FGF2 and FGF2, were constructed by the new method, sequential SmaI cloning. The fusion proteins were expressed in Escherichia coli and purified by the glutathione affinity column chromatography. The Chitin-binding domain-FGF2 protein was specifically bound on chitin biomaterials, that was detected by immunological techniques using anti-FGF2 antibody. The Chitin-binding domain-FGF2 protein was also bound on acetylated area of chitosan biomaterials. The cellulose-binding domain-FGF2 protein was specifically bound on cellulose biomaterials, that was detected by immunological techniques using anti-FGF2 antibody. On the cellulose biomaterial scaffold bound cellulose binding domain-FGF2, osteoblastic cells MC3T3-E1 were grown without serum. The fusion protein, chitin binding domain-BMP2 also expressed osteoblastic differentiation activity to osteoblastic cells MC3T3-E1 on the chitin biomaterials, chitin seat and acetylated chitosan plate. The fusion proteins, chitin/ cellulose binding domain-FGF2/ BMP2, were locally delivered to specific biomaterials.

通过将甲壳素和纤维素溶液浇铸在含有LiCl的二甲基乙酰胺中，构建了甲壳素和纤维素的座。采用SmaI序列克隆法构建了几丁质结合结构域-FGF 2、纤维素结合结构域-FGF 2和FGF 2融合蛋白的表达载体质粒。在大肠杆菌中表达融合蛋白，并通过谷胱甘肽亲和柱层析进行纯化。利用免疫学技术检测到FGF 2蛋白与甲壳素生物材料特异性结合。几丁质结合域-FGF 2蛋白也结合在壳聚糖生物材料的乙酰化区域上。纤维素结合域-FGF 2蛋白特异性结合于纤维素生物材料上，使用抗FGF 2抗体通过免疫学技术检测。在结合纤维素结合结构域-FGF 2的纤维素生物材料支架上，成骨细胞MC 3 T3-E1在无血清的情况下生长。在几丁质生物材料、几丁质座和乙酰化壳聚糖板上，几丁质结合结构域-BMP 2融合蛋白对成骨细胞MC 3 T3-E1也具有成骨分化活性。融合蛋白，甲壳素/纤维素结合域-FGF 2/BMP 2，局部递送到特定的生物材料。