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新型非热等离子体源对细胞内质粒 DNA 的传递和表达。

Delivery and expression of plasmid DNA into cells by a novel non-thermal plasma source.

机构信息

Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA, USA.

Department of Chemistry and Biochemistry, Hampton University, Hampton, VA, USA.

出版信息

Bioelectrochemistry. 2021 Aug;140:107816. doi: 10.1016/j.bioelechem.2021.107816. Epub 2021 Apr 5.

Abstract

Medical applications such as plasma assisted gene transfer is a minimally invasive approach that can substantially reduce potential discomfort of treated area. Atmospheric pressure plasma discharge is an effective approach to deliver plasmid DNA for in vitro and in vivo applications. We investigated plasma assisted delivery in vitro in mouse melanoma cells (B16F10) using a novel surface plasma device, which is operated in air. We evaluated the influence of applied voltage and distance between the surface device and cell monolayer. We found no significant effect on the viability of cells. Highest expression following delivery of a plasmid encoding green fluorescent protein was achieved with an applied voltage of 11.25 kV at a 2 mm distance and 5 s exposure time. To better understand the influence of oxidative damages and stress on cells after plasma delivery, a mRNA expression study was performed. Our results indicated that TNFα mRNA was significantly upregulated. The mRNA response may be attributed to the RONS generated by plasma; however, this mRNA upregulation was not adequate to be reflected in a coordinate protein upregulation. From the results reported here, it is clear that this novel plasma device could be used for plasmid delivery.

摘要

医疗应用,如等离子体辅助基因转移是一种微创的方法,可以大大减少治疗区域的潜在不适。大气压等离子体放电是一种有效的方法来提供质粒 DNA 进行体外和体内应用。我们使用一种新型的表面等离子体装置研究了体外等离子体辅助传递,该装置在空气中运行。我们评估了施加电压和表面装置与细胞单层之间的距离的影响。我们发现,施加电压和暴露时间对细胞活力没有显著影响。在用编码绿色荧光蛋白的质粒进行转染时,在 11.25 kV 的施加电压下,在 2 毫米的距离和 5 秒的暴露时间下,可实现最高的表达。为了更好地理解等离子体转染后氧化损伤和应激对细胞的影响,进行了 mRNA 表达研究。我们的结果表明,TNFα mRNA 显著上调。mRNA 的反应可能归因于等离子体产生的 RONS;然而,这种 mRNA 的上调不足以在协调的蛋白质上调中得到反映。从这里报道的结果可以清楚地看出,这种新型等离子体装置可用于质粒传递。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f60a/8187306/26be8e59fa80/nihms-1696833-f0001.jpg

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