| 张云燕,李有强,骆书美,钟晓波,王六,王志刚.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[J].中国医学影像技术,2009,25(5):745~748 |
| 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 |
| Ultrasound-mediated lipid microbubble destruction enhances pEGFP-N1 expression in human periodontal ligament fibroblasts cell |
| 投稿时间：2008-10-09 修订日期：2008-12-23 |
| DOI： |
| 中文关键词: 超声检查 微泡 人牙周膜成纤维细胞 基因转染 |
| 英文关键词:Ultrasonography Microbubble Human periodontal ligament fibroblasts Gene transfection |
| 基金项目: |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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. |
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