University of Ljubljana, Faculty of Electrical Engineering, Trzaska 25, SI-1000 Ljubljana, Slovenia.
Institute of Oncology Ljubljana, Department of Experimental Oncology, Zaloska 2, SI-1000 Ljubljana, Slovenia; University of Ljubljana, Faculty of Medicine, Vrazov trg 2, SI-1000 Ljubljana, Slovenia.
Bioelectrochemistry. 2021 Oct;141:107847. doi: 10.1016/j.bioelechem.2021.107847. Epub 2021 May 24.
High-Intensity Pulsed Electromagnetic Fields (HI-PEMF) treatment is an emerging noninvasive and contactless alternative to conventional electroporation, since the electric field inside the tissue is induced remotely by external pulsed magnetic field. Recently, HI-PEMF was applied for delivering siRNA molecules to silence enhanced green fluorescent protein (EGFP) in tumors in vivo. Still, delivered siRNA molecules were 21 base pairs long, which is 200-times smaller compared to nucleic acids such as plasmid DNA (pDNA) that are delivered in gene therapies to various targets to generate therapeutic effect. In our study, we demonstrate the use HI-PEMF treatment as a feasible noninvasive approach to achieve in vivo transfection by enabling the transport of larger molecules such as pDNA encoding EGFP into muscle and skin. We obtained a long-term expression of EGFP in the muscle and skin after HI-PEMF, in some mice even up to 230 days and up to 190 days, respectively. Histological analysis showed significantly less infiltration of inflammatory mononuclear cells in muscle tissue after the delivery of pEGFP using HI-PEMF compared to conventional gene electrotransfer. Furthermore, the antitumor effectiveness using HI-PEMF for electrotransfer of therapeutic plasmid, i.e., silencing MCAM was demonstrated. In conclusion, feasibility of HI-PEMF was demonstrated for transfection of different tissues (muscle, skin, tumor) and could have great potential in gene therapy and in DNA vaccination.
高强度脉冲电磁场（HI-PEMF）治疗是一种新兴的非侵入性和非接触式替代传统电穿孔的方法，因为组织内的电场是通过外部脉冲磁场远程感应产生的。最近，HI-PEMF 被应用于将 siRNA 分子递送到体内肿瘤中沉默增强型绿色荧光蛋白（EGFP）。然而，递送到体内的 siRNA 分子只有 21 个碱基对长，与质粒 DNA（pDNA）等在基因治疗中递送到各种靶标以产生治疗效果的核酸相比，小了 200 倍。在我们的研究中，我们证明了 HI-PEMF 治疗作为一种可行的非侵入性方法，通过允许将更大的分子如 pDNA 编码 EGFP 转染到肌肉和皮肤中，来实现体内转染。我们在 HI-PEMF 后获得了肌肉和皮肤中 EGFP 的长期表达，在一些小鼠中甚至长达 230 天和 190 天。组织学分析显示，与传统的基因电转移相比，使用 HI-PEMF 递送 pEGFP 后肌肉组织中炎症单核细胞的浸润明显减少。此外，还证明了 HI-PEMF 用于转染治疗性质粒（即沉默 MCAM）的抗肿瘤效果。总之，已经证明 HI-PEMF 用于转染不同组织（肌肉、皮肤、肿瘤）的可行性，并且在基因治疗和 DNA 疫苗接种方面具有很大的潜力。
