Pan Junjie, Chiang Chi-Ling, Wang Xinyu, Bertani Paul, Ma Yifan, Cheng Junao, Talesara Vishank, Lee Ly James, Lu Wu
Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio 43210, USA.
Department of Electrical and Computer Engineering, The Ohio State University, Columbus, Ohio 43210, USA.
Nanoscale. 2023 Feb 23;15(8):4080-4089. doi: 10.1039/d2nr05575a.
Nanochannel electroporation (NEP) is a new technology for cell transfection, which provides superior gene delivery and cell viability to conventional bulk electroporation (BEP). In NEP, the cells laid on a porous substrate are subjected to an asymmetric electric field which induces asymmetric membrane poration. The cell membrane near the channel outlet ('transfection membrane') is porated intensely, allowing direct delivery of genetic materials, while the rest of the cell membrane ('non-transfection membrane') remains much less perturbed for low cellular damage. In this work, the transfection window of NEP for the delivery of different sized molecules is systematically investigated. The results show that small molecules (∼0.6 kDa) can be delivered into cells at a relatively lower voltage without significantly impacting the non-transfection membrane. To deliver larger molecules (∼6 kDa), a higher working voltage is required at the cost of cell viability due to more severe damage of the non-transfection membrane. Through numerical analysis of both transient transmembrane potential (t-TMP) and dynamic transmembrane potential (d-TMP), here we show that the membrane damage on both transfection and non-transfection sides of the cell membrane can be predicted. The agreement between experimental results and numerical analysis provides a comprehensive understanding of cell membrane damage and cargo delivery in NEP.
纳米通道电穿孔（NEP）是一种用于细胞转染的新技术，与传统的大容量电穿孔（BEP）相比，它能提供更优异的基因递送效果和细胞活力。在NEP中，置于多孔基质上的细胞会受到非对称电场作用，该电场会诱导细胞膜出现非对称穿孔。靠近通道出口处的细胞膜（“转染膜”）会被强烈穿孔，从而允许遗传物质直接递送，而细胞膜的其余部分（“非转染膜”）受到的扰动则小得多，因而细胞损伤较低。在这项工作中，系统地研究了NEP递送不同大小分子的转染窗口。结果表明，小分子（约0.6 kDa）可以在相对较低的电压下递送至细胞内，而不会对非转染膜产生显著影响。为了递送更大的分子（约6 kDa），由于非转染膜受到更严重的损伤，需要更高的工作电压，但这是以细胞活力为代价的。通过对瞬态跨膜电位（t-TMP）和动态跨膜电位（d-TMP）的数值分析，我们在此表明，细胞膜转染侧和非转染侧的膜损伤都可以被预测。实验结果与数值分析之间的一致性为全面理解NEP中的细胞膜损伤和货物递送提供了依据。
