Hard Tissue Regeneration using Gene Activated Matrix

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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.