Acoustic Cavitation-Mediated Delivery of Small Interfering Ribonucleic Acids with Phase-Shift Nano-Emulsions.
Acoustic Cavitation-Mediated Delivery of Small Interfering Ribonucleic Acids with Phase-Shift Nano-Emulsions.
复制标题
DOI：
10.1016/j.ultrasmedbio.2015.04.002
复制
发表时间：
2015-08
影响因子：
2.9
通讯作者：
Porter TM
中科院分区：
文献类型：
--
作者：
Burgess MT;Porter TM
小干扰核糖核酸（siRNA）的局部靶向递送一直是使用siRNA治疗各种疾病的最大障碍。在siRNA的合成方面已经取得了重大进展，这导致了更大的靶信使RNA（mRNA）沉默和在生理条件下的稳定性。尽管许多递送策略已显示出前景，但全身施用的siRNA的靶向递送和释放的选择仍然有限。在该体外研究中，探索了相移纳米乳液（PSNE）作为空化核以促进游离siRNA经由声孔递送至癌细胞。将含有不同量的PSNE和siRNA的细胞悬浮液暴露于固定设置（6.2MPa峰值负压、5个循环脉冲、250Hz脉冲重复频率和100秒的总暴露持续时间）的5MHz脉冲超声。使用被动空化检测器在整个暴露过程中检测到惯性空化排放。成功的siRNA递送（即> 50%细胞摄取）以高活力（> 80%活力）实现。具有siRNA摄取的细胞的百分比与蒸发的PSNE产生的惯性空化活性的量相关。siRNA在递送后保持功能性，显著降低稳定转染细胞系中绿色荧光蛋白（GFP）的表达。这些结果表明，蒸发的PSNE可以促进siRNA进入大多数超声处理的细胞的胞质溶胶中，并且可以为siRNA递送提供非内体途径。
Localized, targeted delivery of small interfering ribonucleic acid (siRNA) has been the foremost hurdle in the use of siRNA for the treatment of various diseases. Major advances have been achieved in the synthesis of siRNA, which has led to greater target messenger RNA (mRNA) silencing and stability in physiological conditions. Although numerous delivery strategies have shown promise, there are still limited options for targeted delivery and release of siRNA administered systemically. In this in vitro study, phase-shift nanoemulsions (PSNE) were explored as cavitation nuclei to facilitate free siRNA delivery to cancer cells via sonoporation. A cell suspension containing varying amounts of PSNE and siRNA was exposed to 5 MHz pulsed ultrasound at fixed settings (6.2 MPa peak negative pressure, 5 cycle pulses, 250 Hz pulse repetition frequency, and total exposure duration of 100 seconds). Inertial cavitation emissions were detected throughout the exposure using a passive cavitation detector. Successful siRNA delivery was achieved (i.e. > 50% cell uptake) with high viability (> 80% viability). The percentage of cells with siRNA uptake was correlated with the amount of inertial cavitation activity generated from vaporized PSNE. The siRNA remained functional after delivery, significantly reducing expression of green fluorescent protein (GFP) in a stably transfected cell line. These results show that vaporized PSNE can facilitate siRNA entry into the cytosol of a majority of sonicated cells and may provide a non-endosomal route for siRNA delivery.