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Hard Tissue Regeneration using Gene Activated Matrix

使用基因激活基质进行硬组织再生

基本信息

批准号：
13470429
负责人：
ASAHINA Izumi
金额：
$ 8.96万
依托单位：
The University of Tokyo (2003)Tokyo Medical and Dental University (2001-2002)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

1)Bone Formation by the cells transfected with BMP cDNAhBMP-2 and 4 cDNAs were cloned from human dental pulp derived cDNA library. pcDNA3 transfected with hBMP-2 or 4 cDNA was transfected into HEK293 cells by lipofectin method. The cells with Three types of scaffolds, collagen sponge, gelatin spoge, and honey-combed collage sponge, were implanted into subcutaneous tissue of mice. The implants were harvested at 2, 4, 8 weeks after operation, and the bone formation were evaluated. As the results, the cells on honey-combed collage sponge formed new bone, whereas the cells on the other scaffolds did not formed bone.2)Bone Regeneration using Gene Activated MatrixcDNAs were not transfected effectively into cells when collagen was used as matrix of gene activated matrix. However, the addition of CaP granules to the matrix improved the in vivo gene transfection efficiency. The combination of the plasmid expressing rhBMP-2, collagen and Cap grafted into 5mm bone defect created in rat tibia. The bone defects were regenerated completely with new bone 6 weeks after the implantation. Only 10 ug of plasmid DNA, which was 11100 quantity compared with the conventional way, could regenerate bone.3)Gene Activated Matrix using PGLACollagen is useful biomaterial, but is derived from animal source which has risk of transmitting disease. We found that poly glicoic acid, which is synthetic polymer, could be used as the matrix instead of collagen.

1)从人牙髓cDNA文库中克隆了hBMP-2和4个cDNA。用脂质体法将hBMP-2和hBMP-4基因转染HEK 293细胞。以胶原海绵、明胶海绵和蜂窝状胶原海绵为支架材料，将细胞植入小鼠皮下。分别于术后2、4、8周取种植体，评价种植体成骨情况。结果表明，蜂窝状胶原海绵上的细胞能形成新骨，而其他支架上的细胞不能形成新骨。2）基因活化基质骨再生实验：胶原作为基因活化基质的基质时，cDNA不能有效转染细胞。然而，向基质中添加CaP颗粒提高了体内基因转染效率。将表达rhBMP-2的质粒、胶原和Cap复合移植于大鼠胫骨5 mm缺损处。植入后6周，骨缺损完全再生，有新骨形成。仅需10 μ g质粒DNA，与常规方法相比，其量为11100。3）使用PGLA的基因活化基质胶原是有用的生物材料，但来自动物源，具有传播疾病的风险。我们发现，聚葡萄糖酸，这是一种合成的聚合物，可以作为基质，而不是胶原蛋白。