Mou Qiongyao, Xu Mengli, Deng Jinan, Hu Ning, Yang Jun
Key Laboratory of Biorheological Science and Technology, Ministry of Education and Bioengineering College, Chongqing University, Chongqing 400044, China.
APL Bioeng. 2023 May 8;7(2):026103. doi: 10.1063/5.0147104. eCollection 2023 Jun.
Electroporation shows great potential in biology and biomedical applications. However, there is still a lack of reliable protocol for cell electroporation to achieve a high perforation efficiency due to the unclear influence mechanism of various factors, especially the salt ions in buffer solution. The tiny membrane structure of a cell and the electroporation scale make it difficult to monitor the electroporation process. In this study, we used both molecular dynamics (MD) simulation and experimental methods to explore the influence of salt ions on the electroporation process. Giant unilamellar vesicles (GUVs) were constructed as the model, and sodium chloride (NaCl) was selected as the representative salt ion in this study. The results show that the electroporation process follows lag-burst kinetics, where the lag period first appears after applying the electric field, followed by a rapid pore expansion. For the first time, we find that the salt ion plays opposite roles in different stages of the electroporation process. The accumulation of salt ions near the membrane surface provides an extra potential to promote the pore initiation, while the charge screening effect of the ions within the pore increases the line tension of the pore to induce the instability of the pore and lead to the closure. The GUV electroporation experiments obtain qualitatively consistent results with MD simulations. This work can provide guidance for the selection of parameters for cell electroporation process.
电穿孔在生物学和生物医学应用中显示出巨大潜力。然而,由于各种因素(尤其是缓冲溶液中的盐离子)的影响机制尚不清楚,目前仍缺乏可靠的细胞电穿孔方案来实现高穿孔效率。细胞微小的膜结构和电穿孔规模使得监测电穿孔过程变得困难。在本研究中,我们使用分子动力学(MD)模拟和实验方法来探究盐离子对电穿孔过程的影响。构建了巨型单层囊泡(GUVs)作为模型,并选择氯化钠(NaCl)作为本研究中的代表性盐离子。结果表明,电穿孔过程遵循滞后-爆发动力学,即在施加电场后首先出现滞后阶段,随后是快速的孔扩张。我们首次发现盐离子在电穿孔过程的不同阶段发挥相反作用。膜表面附近盐离子的积累提供了额外的电势以促进孔的起始,而孔内离子的电荷屏蔽效应增加了孔的线张力,从而导致孔的不稳定并使其关闭。GUV电穿孔实验获得了与MD模拟定性一致的结果。这项工作可为细胞电穿孔过程参数的选择提供指导。