Vectorology and Anticancer Therapies, UMR 8203, CNRS, Univ. Paris-Sud, Gustave Roussy, Université Paris-Saclay, 94805 Villejuif, France.
Vectorology and Anticancer Therapies, UMR 8203, CNRS, Univ. Paris-Sud, Gustave Roussy, Université Paris-Saclay, 94805 Villejuif, France.
Biochim Biophys Acta Biomembr. 2018 May;1860(5):1022-1034. doi: 10.1016/j.bbamem.2018.01.018. Epub 2018 Feb 2.
The permeabilization of biological membranes by electric fields, known as electroporation, has been traditionally performed with square electric pulses. These signals distribute the energy applied to cells in a wide frequency band. This paper investigates the use of sine waves, which are narrow band signals, to provoke electropermeabilization and the frequency dependence of this phenomenon. Single bursts of sine waves at different frequencies in the range from 8 kHz-130 kHz were applied to cells in vitro. Electroporation was studied in the plasma membrane and the internal organelles membrane using calcium as a permeabilization marker. Additionally, a double-shell electrical model was simulated to give a theoretical framework to our results. The electroporation efficiency shows a low pass filter frequency dependence for both the plasma membrane and the internal organelles membrane. The mismatch between the theoretical response and the observed behavior for the internal organelles membrane is explained by a two-step permeabilization process: first the permeabilization of the external membrane and afterwards that of the internal membranes. The simulations in the model confirm this two-step hypothesis when a variable plasma membrane conductivity is considered in the analysis. This study demonstrates how the use of narrow-band signals as sine waves is a suitable method to perform electroporation in a controlled manner. We suggest that the use of this type of signals could bring a simplification in the investigations of the very complex phenomenon of electroporation, thus representing an interesting option in future fundamental studies.
电场对生物膜的通透性作用，即电穿孔，传统上是通过方波电脉冲来实现的。这些信号将施加于细胞的能量分布在很宽的频带上。本文研究了使用正弦波（窄带信号）来引发电穿孔以及这种现象的频率依赖性。在 8 kHz-130 kHz 的范围内，对体外细胞施加不同频率的单个正弦波脉冲。使用钙作为通透性标记物，研究了细胞膜和细胞器膜的电穿孔。此外，还模拟了双壳层电模型，为我们的结果提供了理论框架。对于细胞膜和细胞器膜，电穿孔效率均表现出低通滤波器频率依赖性。细胞器膜的理论响应与观察到的行为不匹配，这可以通过两步通透性过程来解释：首先是外部膜的通透性，然后是内部膜的通透性。在分析中考虑可变的细胞膜电导率时，模型中的模拟证实了这种两步假设。本研究表明，使用窄带信号（如正弦波）作为一种合适的方法来以可控的方式进行电穿孔。我们建议，使用这种类型的信号可以简化电穿孔这一非常复杂的现象的研究，因此在未来的基础研究中是一个很有前途的选择。
