Biophysical insight into mechanisms of sonoporation
Biophysical insight into mechanisms of sonoporation
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DOI：
10.1073/pnas.1606915113
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发表时间：
2016-09-06
影响因子：
11.1
通讯作者：
Villanueva, Flordeliza S.
中科院分区：
文献类型：
--
作者：
Helfield, Brandon;Chen, Xucai;Villanueva, Flordeliza S.
这项研究提出了一种独特的方法来理解超声触发细胞膜破裂的生物物理机制（即，声致穿孔）。我们报告超声刺激封装微泡振荡物理和由此产生的细胞膜渗透性之间的直接相关性，通过同时显微镜这两个过程在其固有的物理时间尺度（微秒的微泡动力学和秒到分钟的局部大分子吸收和细胞膜重组）。我们发现，存在一个微泡振荡诱导的剪切应力阈值，在千帕的顺序，超过此内皮细胞膜通透性增加。的剪切应力阈值表现出反平方根的振荡周期数的关系和近似线性依赖于超声频率从0.5到2 MHz。此外，通过实时3D共聚焦显微镜测量，我们的数据提供了声致穿孔事件直接导致立即产生穿过顶端和基底细胞膜层的膜孔的证据，所述膜孔沿其横向区域沿着重新密封（重新密封时间类似于< 2分钟）。最后，我们证明了声致穿孔间接启动相邻融合细胞之间延长的细胞间间隙（类似于> 30-60分钟）的潜力。这种实时显微镜方法提供了深入了解声孔作用的物理，空化为基础的机制和生物物理，细胞膜为基础的机制，微泡声学行为引起急性和持续增强细胞和血管通透性。
This study presents a unique approach to understanding the biophysical mechanisms of ultrasound-triggered cell membrane disruption (i.e., sonoporation). We report direct correlations between ultrasound-stimulated encapsulated microbubble oscillation physics and the resulting cellular membrane permeability by simultaneous microscopy of these two processes over their intrinsic physical timescales (microseconds for microbubble dynamics and seconds to minutes for local macromolecule uptake and cell membrane reorganization). We show that there exists a microbubble oscillation-induced shear-stress threshold, on the order of kilopascals, beyond which endothelial cellular membrane permeability increases. The shear-stress threshold exhibits an inverse squareroot relation to the number of oscillation cycles and an approximately linear dependence on ultrasound frequency from 0.5 to 2 MHz. Further, via real-time 3D confocal microscopy measurements, our data provide evidence that a sonoporation event directly results in the immediate generation of membrane pores through both apical and basal cell membrane layers that reseal along their lateral area (resealing time of similar to< 2 min). Finally, we demonstrate the potential for sonoporation to indirectly initiate prolonged, intercellular gaps between adjacent, confluent cells (similar to> 30-60 min). This real-time microscopic approach has provided insight into both the physical, cavitation-based mechanisms of sonoporation and the biophysical, cell-membrane-based mechanisms by which microbubble acoustic behaviors cause acute and sustained enhancement of cellular and vascular permeability.