Effect of Pulsed Electric Field on Electroporation Area of Cell Membrane
Wu Xiao1,2, Mi Yan1, Zheng Wei1, Ma Chi1, Zhang Mengnan1
1. State Key Laboratory of Power Transmission Equipment & System Security and New Technology Chongqing University Chongqing 400030 China; 2. Marketing Service Center of State Grid Henan Electric Power Company (Metrology Center) Zhengzhou 450000 China
Abstract：In the field of electrobiology, simulation and experimental methods can be used to study the cell electroporation effect under pulsed electric field. In simulation, parameters such as transmembrane voltage, current density, pore density, pore radius and pore area are usually used to characterize the electroporation effect. In the experiment, the killing effect or the overall situation of the electric field on the cells is usually characterized macroscopically, which is not equivalent to the specific parameters such as the number and size of the pores formed, and it is not easy to establish a close relationship between simulation and experiment. Therefore, the electroporation area of cell membrane, a new characterization parameter to directly measure the electroporation effect, and its experimental measurement method are proposed in this paper. Firstly, the measurement principle of cell electroporation area was analyzed based on the molecular transport dynamics, and it was found that the electroporation area could be measured and calculated by cell fluorescence experiment. Then, the fluorescent molecular staining experiment was carried out based on the above principle, the fluorescence intensity changes of cells under the electric field were captured in real time under the fluorescence microscope. Before carrying out cell electroporation experiments, the cell fluorescence intensity needs to be corrected to obtain the relationship between the cell fluorescence intensity and the intracellular concentration of propidium iodide ions. Next, the maximum cell fluorescence intensity and the pore recovery time constant were extracted from the fluorescence intensity curve under the action of the electric field, from which the electroporation area of cell membrane formed during the pulse could be calculated. Finally, experiments were carried out under different pulsed electric field parameters (field strength and pulse width) to study the relationship and law between cell electroporation area and pulsed electric field parameters, and the obtained laws were analyzed and explained theoretically. Under the pulsed electric field parameters selected in this paper, the electroporation area calculated by the cell fluorescent staining experiment is in the order of 10-15 m2, which is quantitatively close to the value calculated in the literature. Under different pulse electric field parameters, with the increase of pulse electric field parameters (field strength and pulse width), the fluorescence intensity of cells increases, the fluorescence intensity curve moves up, and the electroporation effect is enhanced. Moreover, the calculated electroporation area increases with the increase of pulse field strength, pulse width and pulse electric shock, and all have a linear relationship. The obtained law is analyzed in combination with theory, and the laws obtained in this paper are qualitatively consistent with those in the existing literature. By comparing the electroporation area between this paper and the existing conclusions quantitatively and qualitatively, the accuracy of the method and results of calculating the cell electroporation area by experimental measurements can be evaluated. According to these laws, it can provide guidance for the pulsed electric field treatment. For example, in the electrochemotherapy, the pulsed electric field parameters (field strength and pulse width) are increased linearly, the electroporation area is increased, and more drugs enter the cells, and the treatment is more effective.
吴晓, 米彦, 郑伟, 马驰, 张梦男. 脉冲电场对细胞膜电穿孔面积的影响研究[J]. 电工技术学报, 2023, 38(14): 3779-3788.
Wu Xiao, Mi Yan, Zheng Wei, Ma Chi, Zhang Mengnan. Effect of Pulsed Electric Field on Electroporation Area of Cell Membrane. Transactions of China Electrotechnical Society, 2023, 38(14): 3779-3788.
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2023, Vol. 38 
