Qin Peng, Xu Lin, Han Tao, Du Lianfang, Yu Alfred C H
Department of Instrument Science and Engineering, Shanghai Jiao Tong University, Shanghai, China.
National Laboratory of Plant Molecular Genetics, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
Ultrason Sonochem. 2016 Jul;31:107-15. doi: 10.1016/j.ultsonch.2015.12.001. Epub 2015 Dec 9.
Sonoporation-transient plasma membrane perforation elicited by the interaction of ultrasound waves with microbubbles--has shown great potential for drug delivery and gene therapy. However, the heterogeneity of sonoporation introduces complexities and challenges in the realization of controllable and predictable drug delivery. The aim of this investigation was to understand how non-acoustic parameters (bubble related and bubble-cell interaction parameters) affect sonoporation. Using a customized ultrasound-exposure and fluorescence-imaging platform, we observed sonoporation dynamics at the single-cell level and quantified exogenous molecular uptake levels to characterize the degree of sonoporation. Sonovue microbubbles were introduced to passively regulate microbubble-to-cell distance and number, and bubble size. 1 MHz ultrasound with 10-cycle pulse duration and 0.6 MPa peak negative pressure were applied to trigger the inertial collapse of microbubbles. Our data revealed the impact of non-acoustic parameters on the heterogeneity of sonoporation. (i) The localized collapse of relatively small bubbles (diameter, D<5.5 μm) led to predictable sonoporation, the degree of which depended on the bubble-to-cell distance (d). No sonoporation was observed when d/D>1, whereas reversible sonoporation occurred when d/D<1. (ii) Large bubbles (D>5.5 μm) exhibited translational movement over large distances, resulting in unpredictable sonoporation. Translation towards the cell surface led to variable reversible sonoporation or irreversible sonoporation, and translation away from the cell caused either no or reversible sonoporation. (iii) The number of bubbles correlated positively with the degree of sonoporation when D<5.5 μm and d/D<1. Localized collapse of two to three bubbles mainly resulted in reversible sonoporation, whereas irreversible sonoporation was more likely following the collapse of four or more bubbles. These findings offer useful insight into the relationship between non-acoustic parameters and the degree of sonoporation.
声穿孔——由超声波与微泡相互作用引发的瞬时细胞膜穿孔——在药物递送和基因治疗方面已显示出巨大潜力。然而，声穿孔的异质性给实现可控且可预测的药物递送带来了复杂性和挑战。本研究的目的是了解非声学参数（与气泡相关的参数以及气泡 - 细胞相互作用参数）如何影响声穿孔。使用定制的超声暴露和荧光成像平台，我们在单细胞水平观察了声穿孔动力学，并对外源分子摄取水平进行了量化，以表征声穿孔的程度。引入声诺维微泡来被动调节微泡与细胞的距离、数量以及气泡大小。施加具有10个周期脉冲持续时间和0.6兆帕峰值负压的1兆赫超声波，以触发微泡的惯性空化。我们的数据揭示了非声学参数对声穿孔异质性的影响。（i）相对较小气泡（直径D<5.5μm）的局部空化导致可预测的声穿孔，其程度取决于气泡与细胞的距离（d）。当d/D>1时未观察到声穿孔，而当d/D<1时发生可逆声穿孔。（ii）大气泡（D>5.5μm）表现出远距离平移运动，导致不可预测的声穿孔。向细胞表面平移导致可变的可逆声穿孔或不可逆声穿孔，而远离细胞平移则导致无穿孔或可逆声穿孔。（iii）当D<5.5μm且d/D<1时，气泡数量与声穿孔程度呈正相关。两到三个气泡的局部空化主要导致可逆声穿孔，而四个或更多气泡空化后更有可能发生不可逆声穿孔。这些发现为非声学参数与声穿孔程度之间的关系提供了有用的见解。
