Edelblute Chelsea M, Heller Loree C, Malik Muhammad A, Heller Richard
Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, Virginia.
Department of Biology, College of Science, Old Dominion University, Norfolk, Virginia.
Biotechnol Bioeng. 2015 Dec;112(12):2583-90. doi: 10.1002/bit.25660. Epub 2015 Jul 14.
Cold plasma is emerging as a potential method for medical applications. The current study assessed the efficacy of a novel cold plasma reactor based on shielded sliding discharge producing cathode-directed streamers generated in ambient air for the delivery of plasmid DNA. Experiments were performed with mouse melanoma cells (B16F10) and human keratinocyte cells (HaCaT) inoculated with plasmid DNA encoding luciferase. Quantitative results measured over a 72-h period displayed luciferase expression levels as high as 5-fold greater in cells exposed to plasma-activated air (PAA) than levels obtained from the inoculation of plasmid DNA alone (P < 0.05, P < 0.01). No effect on cell viability was observed. Delivery of plasmid encoding GFP to HaCaT cells seeded on polycaprolactone (PCL) scaffolds was confirmed by immunostaining. The use of cold plasma for DNA delivery is attractive as it provides a non-viral, non-invasive method where the electrode or the plasma itself never directly contacts the exposed site. The current device design provides localized DNA transfer using a novel technology. Our report suggests PAA warrants further exploration as an alternative or supplemental approach for DNA transfer.
冷等离子体正成为一种具有医学应用潜力的方法。当前研究评估了一种基于屏蔽滑动放电的新型冷等离子体反应器的功效，该反应器在环境空气中产生指向阴极的流光以递送质粒DNA。实验使用接种了编码荧光素酶的质粒DNA的小鼠黑色素瘤细胞（B16F10）和人角质形成细胞（HaCaT）进行。在72小时内测量的定量结果显示，暴露于等离子体活化空气（PAA）的细胞中荧光素酶表达水平比仅接种质粒DNA所获得的水平高5倍（P < 0.05，P < 0.01）。未观察到对细胞活力的影响。通过免疫染色证实了将编码GFP的质粒递送至接种在聚己内酯（PCL）支架上的HaCaT细胞。冷等离子体用于DNA递送具有吸引力，因为它提供了一种非病毒、非侵入性的方法，其中电极或等离子体本身从不直接接触暴露部位。当前的设备设计使用一种新技术实现了局部DNA转移。我们的报告表明，PAA作为DNA转移的替代或补充方法值得进一步探索。
