Roth Caleb C, Barnes Ronald A, Ibey Bennett L, Beier Hope T, Christopher Mimun L, Maswadi Saher M, Shadaram Mehdi, Glickman Randolph D
School of Medicine, Dept. of Radiological Sciences, University of Texas Health Science Center San Antonio, 7703 Floyd Curl Drive, San Antonio, Texas, USA 78229.
Dept. of Electrical Engineering, University of Texas San Antonio, 1 UTSA Circle, San Antonio, Texas, USA 78249.
Sci Rep. 2015 Oct 9;5:15063. doi: 10.1038/srep15063.
The mechanism(s) responsible for the breakdown (nanoporation) of cell plasma membranes after nanosecond pulse (nsEP) exposure remains poorly understood. Current theories focus exclusively on the electrical field, citing electrostriction, water dipole alignment and/or electrodeformation as the primary mechanisms for pore formation. However, the delivery of a high-voltage nsEP to cells by tungsten electrodes creates a multitude of biophysical phenomena, including electrohydraulic cavitation, electrochemical interactions, thermoelastic expansion, and others. To date, very limited research has investigated non-electric phenomena occurring during nsEP exposures and their potential effect on cell nanoporation. Of primary interest is the production of acoustic shock waves during nsEP exposure, as it is known that acoustic shock waves can cause membrane poration (sonoporation). Based on these observations, our group characterized the acoustic pressure transients generated by nsEP and determined if such transients played any role in nanoporation. In this paper, we show that nsEP exposures, equivalent to those used in cellular studies, are capable of generating high-frequency (2.5 MHz), high-intensity (>13 kPa) pressure transients. Using confocal microscopy to measure cell uptake of YO-PRO®-1 (indicator of nanoporation of the plasma membrane) and changing the electrode geometry, we determined that acoustic waves alone are not responsible for poration of the membrane.
纳秒脉冲(nsEP)暴露后导致细胞质膜破裂(纳米孔形成)的机制仍未得到充分理解。目前的理论仅关注电场,将电致伸缩、水偶极排列和/或电极化作为孔形成的主要机制。然而,通过钨电极向细胞施加高压nsEP会产生多种生物物理现象,包括电液压空化、电化学相互作用、热弹性膨胀等。迄今为止,极少有研究调查nsEP暴露期间发生的非电现象及其对细胞纳米孔形成的潜在影响。主要关注的是nsEP暴露期间产生的声冲击波,因为已知声冲击波可导致膜孔形成(声孔效应)。基于这些观察结果,我们团队对nsEP产生的声压瞬变进行了表征,并确定这些瞬变是否在纳米孔形成中起作用。在本文中,我们表明,与细胞研究中使用的等效nsEP暴露能够产生高频(2.5 MHz)、高强度(>13 kPa)的压力瞬变。使用共聚焦显微镜测量YO-PRO®-1(质膜纳米孔形成的指示剂)的细胞摄取并改变电极几何形状,我们确定仅声波并不导致膜孔形成。