超声微泡-阳离子纳米脂质体复合体制备的实验研究

摘要

Objective To combine ultrasound microbubbles with cationic nanoliposomes by biotin-avidin system and prepare a new type of gene vector. Methods Biotinylated ultrasound microbubbles (Bio-MB) were prepared by mechanical shaking. Biotinylated cationic nanoliposomes (Bio-CNLP) were prepared by film dispersion method-film extrusion method. They were combined by Biotin-avidin system. With nonbiotinylated ultrasound microbubbles (MB) and nonbiotinylated cationic nanoliposomes (CNLP) served as the control group,the morphology and connect effect of complex were observed by laser scanning confocal microscope. Results The mean size of Bio-CNLP was 291.7 nm,and the mean surface potential was (21.6±2. 1) mV. In Bio-MB+Bio-CNLP group,it showed that the green Bio-MB wall was not smooth and a number of red small round Bio-CNLP gathered around Bio-MB under confocal imaging. In MB+ CNLP group,the MB wall was smooth and no conjugation of CNLP was observed. Conclusions Bio-MB and Bio-CNLP can be combined firmly and successfuly by avidin-biotin system. It is potential to provide a new means of gene therapy and a new train of thought on probing gene vectors and transfection methods.%目的 利用生物素亲和素系统连接超声微泡与阳离子纳米脂质体,制备一种新型的基因载体.方法 机械振荡法制备生物素化超声微泡(Bio-MB).薄膜分散-膜挤压法制备生物素化阳离子纳米脂质体(Bio-CNLP).采用生物素亲和素系统偶联Bio-MB与Bio-CNLP.以非生物素化超声微泡(MB)加非生物素化阳离子纳米脂质体(CNLP)组作为对照,行激光共聚焦显微镜观察复合物形态与连接效果.结果 Bio-CNLP的平均粒径约291.7 nm,平均表面电荷约(21.6±2.1)mV.激光共聚焦显微镜下Bio-MB+Bio-CNLP组可见绿色Bio-MB壁因连接Bio-CNLP而不光滑,周围可见红色小圆形点状Bio-CNLP呈花环状聚集,而MB+CNLP组MB壁光滑,周围未见CNLP聚集.结论 生物素亲和素系统可成功将Bio-CNLP与Bio-MB牢固连接,有望为肿瘤基因治疗提供一种新的手段,为探索基因载体和转染方法提供一种新的思路.