超声破坏微泡法介导增强型绿色荧光蛋白质粒转染兔股骨头组织的安全性及可行性(英文)

背景:超声微泡转染系统已尝试于体内多部位的基因转染,但尚未见有用于骨部位基因转染的报告。目的:观察超声破坏微泡法介导增强型绿色荧光蛋白质粒转染兔股骨头组织的效率及可行性。方法:将日本大耳白兔按随机均分为裸转染组,预照射+裸转染组,超声定位转染组,预照射+超声定位转染,重复定位转染组。其中前2组不给予超声定向转染照射,后3组利用超声微泡破裂法介导增强型绿色荧光蛋白质粒,定向基因转染兔股骨头。各组转染1周后,于荧光显微镜下观察增强型绿色荧光蛋白在股骨头组织中的表达情况。结果与结论:超声定位转染组,预照射+超声定位转染,重复定位转染组均有增强型绿色荧光蛋白表达,且复定位转染组增强型绿色荧光蛋白质粒在兔股骨头内的转染效率明显高于其他组(P<0.01)。超声定位转染组,预照射+超声定位转染,重复定位转染组兔超声照射部位软组织和骨组织切片未观察到明显损伤病灶。结果证实,超声微泡破裂法能安全、有效实现增强型绿色荧光蛋白质粒在兔股骨头组织的转染。

Safety and feasibility of ultrasound-targeted microbubble destruction applied for transfection of enhanced green fluorescent protein plasmid into the femoral head of rabbits

BACKGROUND: In recent years, ultrasound microbubble gene transfer system has been applied for gene transfection in many parts of the body, but it has been seldom reported to be used for gene transfection in bone parts. OBJECTIVE: To investigate the efficiency and feasibility of ultrasound-targeted microbubble destruction applied for transfection of enhanced green fluorescent protein plasmid into the femoral head of rabbits. METHODS: Japanese big-ear rabbits were randomly divided into five groups: bare transfection, pre-irradiation + bare transfection, ultrasound transfection, pre-irradiation+ultrasound transfection, and repeatable transfection. In the first two groups, ultrasound-targeted gene transfection and irradiation was not used, but in the latter three groups, ultrasound-targeted microbubble destruction was used to transfect enhanced green fluorescent protein (EGFP) plasmid into the femoral head of rabbits. At 1 week after transfection, EGFP expression in femoral head was observed under the fluorescence microscope. RESULTS AND CONCLUSION: EGFP expression appeared in the ultrasound transfection, pre-irradiation + ultrasound transfection and repeatable transfection. The transfection efficiency of EGFP plasmid was significantly higher in the repeatable transfection group than in the other groups (P < 0.01). Obvious injury loci were not observed in the soft tissue and bone tissue slices of ultrasonic irradiation parts in the ultrasound transfection, pre-irradiation + ultrasound transfection and repeatable transfection groups. These results confirm that ultrasound-targeted microbubble destruction is a safe and effective method to transfect EGFP plasmid into the femoral head of rabbits.