CAJ | 학술논문

目的探讨超声靶向微泡破碎(UTMD)的物理靶向和纳米粒的生物靶向联合的双靶向方法细胞内递送siRNAs的效果.方法制备RGD和非RGD生物可降解纳米粒;L16(45)正交设计筛选超声、微泡、纳米粒的优化参数;实验分成优化参数组、RGD纳米粒组、非RGD纳米粒组和空白对照组;倒置荧光显微镜和流式细胞仪观察并检测分子探针Cy3标记的siRNAs在细胞内的摄取.结果 RGD纳米粒组的细胞摄取率和荧光强度分别为(93.49±1.37)%和34.28±2.06,优化参数组的细胞摄取率和荧光强度分别为(88.33±1.24)%和30.59±3.93,两组比较差异均无统计学意义(P＞0.05).非RGD纳米粒组的细胞摄取率和荧光强度分别为(71.24±2.80)%和18.39±0.90,优化参数组的细胞摄取率和荧光强度分别为(84.78±2.13)%和27.18±0.91,两组比较差异均有统计学意义(P＜0.05).结论 UTMD物理靶向和纳米粒生物靶向联合的双靶向方法不能提高siRNAs的细胞内递送.
목적 탐토초성파향미포파쇄(UTMD)적물리파향화납미립적생물파향연합적쌍파향방법세포내체송siRNAs적효과.방법 제비RGD화비RGD생물가강해납미립;L16(45)정교설계사선초성、미포、납미립적우화삼수;실험분성우화삼수조、RGD납미립조、비RGD납미립조화공백대조조;도치형광현미경화류식세포의관찰병검측분자탐침Cy3표기적siRNAs재세포내적섭취.결과 RGD납미립조적세포섭취솔화형광강도분별위(93.49±1.37)%화34.28±2.06,우화삼수조적세포섭취솔화형광강도분별위(88.33±1.24)%화30.59±3.93,량조비교차이균무통계학의의(P＞0.05).비RGD납미립조적세포섭취솔화형광강도분별위(71.24±2.80)%화18.39±0.90,우화삼수조적세포섭취솔화형광강도분별위(84.78±2.13)%화27.18±0.91,량조비교차이균유통계학의의(P＜0.05).결론 UTMD물리파향화납미립생물파향연합적쌍파향방법불능제고siRNAs적세포내체송.
Objective To investigate the intracellular delivery of siRNAs through the applications of ultrasound targeted microbubbles destruction(UTMD)and biodegradable nanoparticles carriers.Methods Preparation of nanoparticles with and without RGD sequences,parameters optimization via L16(45)orthogonal design,control experiments in groups of optimization,RGD targeted nanoparticles,non-RGD nanoparticles and blank control, and determinations by inverted fluorescence microscope and flow cytometry were performed.Results The uptake and fluorescence intensity of PC-3 cells in group of RGD targeted nanoparticle was (93.49±1.37)% and 34.28±2.06 respectively,and that in group of optimization was (88.33±1.24)% and 30.59±3.93 respectively(P＞0.05).Whereas the uptake and fluorescence intensity of PC-3 cells in group of non-RGD nanoparticles was(71.24±2.80)% and 18.39±0.90 respectively,and that in group of optimization was (84.78±2.13)% and 27.18±0.91 respectively(P＜0.05).ConclusionsThe applications of UTMD with RGD targted nanoparticles cannot increase the intracellular delivery of siRNAs.