超声微泡介导pEGFP-N1质粒转染人牙周膜成纤维细胞的实验研究

目的 探讨超声微泡造影剂在一定能量的超声波辐照下,介导质粒pEGFP-N1转染人牙周膜成纤维细胞(HPDLFs)的效率及安全性.方法 体外原代培养HPDLFs,以EGFP基因为报告基因,脂质微泡造影剂为载体,用超声辐照介导质粒pEGFP-N1转染HPDLFs.实验组超声+微泡+质粒组根据转染条件不同分成不同亚组,对照组为质粒组、微泡+质粒组、超声+质粒组和脂质体+质粒组.转染48 h后在倒置荧光显微镜下观察绿色荧光蛋白GFP表达.同时用MTT法检测HPDLFs的活力.结果 超声微泡介导的质粒对HPDLFs的转染效率与脂质体介导的质粒转染效率相似,明显高于其他对照组.超声+微泡+质粒组中HPDLFs的活力明显高于脂质体+质粒组.结论 在一定条件下,超声微泡能安全、有效地介导外源基因的转染与表达.

Ultrasound-mediated lipid microbubble destruction enhances pEGFP-N1 expression in human periodontal ligament fibroblasts cell

Objective To investigate whether ultrasound-mediated microbubble destruction can safely and effectively deliver pEGFP-N1 plasmid to human periodontal ligament fibroblasts. Methods The primary cultured human periodontal ligament fibroblasts were divided into five groups. Experiment groups were ultrasound+microbubble+plasmid. Based on different conditions of transfection, experiment groups were further divided into sub groups. Control groups included naked plasmid, ultrasound+plasmid, microbubble+plasmid and liposome+plasmid. After 48 h, GFP expression in the human periodontal ligament fibroblasts was detected using phase-contrast fluorescence microscopy. MTT was adopted to measure the cells vitality of every group. Results The transfection efficiency of ultrasound+microbubble+plasmid group was similar to that of the liposome+plasmid group, which were both higher than that of the other control groups, but the cells vitality of the ultrasound+microbubble+plasmid group was higher than that of the liposome+plasmid group. Conclusion Under some specific conditions, ultrasound mediated microbubble destruction method can safely and effectively enhance the reporter gene transfection and expression.