Department of Electrical and Computer Engineering, Old Dominion University, Norfolk VA, United States.
Department of Medical Engineering, University of South Florida, Tampa, FL, United States.
Bioelectrochemistry. 2021 Aug;140:107814. doi: 10.1016/j.bioelechem.2021.107814. Epub 2021 Apr 17.
A novel monopolar electroporation system and methodologies were developed for in vivo electroporation intended for potential clinical applications such as gene therapy. We hypothesized that an asymmetric anode/cathode electrode applicator geometry could produce favorable electric fields for electroporation, without the typical drawback associated with traditional needle and parallel plate geometries. Three monopolar electrode applicator prototypes were built and tested for gene delivery of reporter genes to the skin in a guinea pig model. Gene expression was evaluated in terms of kinetics over time and expression distribution within the treatment site. Different pulsing parameters, including pulse amplitude, pulse duration, and pulse number were evaluated. Monopolar gene electrotransfer significantly enhanced gene expression compared to controls over the course of 21 days. Gene expression distribution was observed throughout the full thickness of the epidermis, as well as notable expression in the deeper layers of the skin, including the dermis, and the underlying striated muscle without any damage at the treatment site, which is a substantial improvement over previously reported expression confined to the epidermis only. Expression distribution observed is consistent with the electric field distribution model, indicating that our novel electrode geometry results in targeted electroporation and gene transfer. This is important, as it may facilitate translation of many electroporation-based clinical therapies including gene therapies, IRE, and ECT.
开发了一种新型的单极电穿孔系统和方法，用于体内电穿孔，适用于基因治疗等潜在的临床应用。我们假设不对称的阳极/阴极电极敷贴器几何形状可以产生有利于电穿孔的电场，而没有传统的针和平行板几何形状的典型缺点。构建了三种单极电极敷贴器原型，并在豚鼠模型中测试了它们用于将报告基因递送到皮肤的基因传递。根据时间上的动力学和处理部位内的表达分布来评估基因表达。评估了不同的脉冲参数，包括脉冲幅度、脉冲持续时间和脉冲数量。与对照组相比，单极基因电转移在 21 天的过程中显著增强了基因表达。在整个表皮的全厚度以及皮肤的深层（包括真皮）都观察到了基因表达，而在处理部位没有任何损伤，这是对以前仅局限于表皮表达的显著改善。观察到的表达分布与电场分布模型一致，表明我们的新型电极几何形状导致了靶向电穿孔和基因转移。这很重要，因为它可能促进许多基于电穿孔的临床疗法的转化，包括基因治疗、IRE 和 ECT。
