Study On The Compatibility Of Bone Guide Chitosan Memberane Combined With BMP-2 Plasmid DNA And Chinese Medicine

Objective:To investigate the application of bone guide chitosan membrane as a carrier of Chinese medicine-Bingpengsan and BMP-2 plasmid-DNA. In this research, the cellular compatibility of combined chitosan membranes were detected in vitro and the oesteogenetic ability of these new guide bone induced memebranes were evaluated in vivo.Methods:We have divided this study into three parts:1. Chitosan were prepared the guide bone regeneration memebranes. Expression vector encoding bone morphogenetic protain gene was prepared and plasmide DNA were condensed. Chinese medicine Bingpensan and BMP-2 plasmide DNA were combined with chitosan membranes.2. MG63 osteoblast cell were seeded on the complexed chitosan membrane, the cell were observed by SEM and MTT at different time points in vitro. The cellular compatibility of these complexed membrane were estimated.3.48 sprague-Dawley rats were used in this research, the critical size defects of rat calvarium (0.5 mm diameter) were prepared, 10ug BMP-2 plasmide DNA/chitosan membranes and Bingpensan/chitosan membranes were used as guided bone regeneration membrane, pure chitosan and no membrane as control. X-ray, histological inspection were evaluated in vivo.Results:1.The 20.8-41.6 mg/ml Bingpensan showed better ability of bacteriostasis in vitro.The morphologic characteristic of MG63 osteoblast on the chitosan memember of lOug BMP-2 plasmid DNA and 60mg/mltraditional chinese medicine-Bingpensan were evaluated under SEM showed normal appearance. The membranes combined Bingpengsan were observed better ability of earlier period of proliferation of osteoblast. In 24h, the chitosan groups obtained the less number cell adhesive than control group with no chitosan memdrane. In 96h, the ability on proliferation in each chitosan groups showed no difference compared with control.2.The result confirmed the better bone guide regeneration ability of chitosan memebranes of lOug BMP-2-DNA in vivo, due to the new bone formed in 8 week more than other groups. Chitosan memebranes of Bingpengsan were observed antibacterial function and bone guide regeneration ability.Conclusion:1,The chitosan membranes prepared as carrier of medicine are convenient when combined with 60 mg/ml Chinese Medicine Bingpengsan.2,The application of chitosan as the carrier of 10ug BMP-2 plasmid DNA membranes were feasible in this experiment. Moreover, the 10ug plasmid DNA of BMP-2 complexed with chitosan membrane can control the releasing of BMP-2 for guide the regeneration of new bone in vivo.