关键词: 超声, 微泡, 人骨形态发生蛋白- 2, 基因治疗

Abstract:

Objective To investigate the transfection efficiency of the recombinant human bone morphogenetic protein- 2（hBMP- 2） gene in targeted cells by ultrasound- mediated microbubble destruction. Methods NIH3T3 cells′ anabiosis was completed and went down to the 3rd or 4th generation, and cultured in 6 well plates. The cells were divided into 2 groups: Plasmid DNA and LipofectamineTM 2000 group（liposome group）, plasmid DNA and ultrasound and microbubble group（ultrasound- mediated microbubble destruction group）. Plasmid DNA was transfected into cells with liposome or ultrasound and microbubble. 24- 48 hours later, the transfection efficiency and the concentrations of hBMP- 2 were measured with fluoresence microscope and enzyme - linked immunosorbent assay（ELISA） respectively. The data were analyzed by curve fitting and t- test of SPSS 11.5. Results The transfection efficiency rate was（7.30± 1.58）% in liposome group, compared with （11.77±3.16）% in ultrasound- mediated microbubble destruction group（P< 0.05）. The concentration of hBMP - 2 after transfection was （1 164.35 ±724.67）pg/mL in liposome group, versus （2 932.70±656.27）pg/mL in ultrasound- mediated microbubble destruction group（P<0.05）. Conclusion Ultrasoundmediated microbubble destruction could significantly improve the transfection efficiency and expression of hBMP- 2 gene in NIH3T3 cells. It may provide a new and effective gene delivery system for gene therapy in periodontal regeneration.

Key words: ultrasound, microbubble, human bone morphogenetic protein- 2, gene therapy