Plasma-activated air mediates plasmid DNA delivery in vivo.
Plasma-activated air mediates plasmid DNA delivery in vivo.
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DOI：
10.1038/mtm.2016.28
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Heller R
中科院分区：
文献类型：
--
作者：
Edelblute CM;Heller LC;Malik MA;Bulysheva A;Heller R
等离子体活化空气（PAA）提供了一个非接触的DNA转移平台。在目前的研究中，PAA用于在3D人体皮肤模型以及体内递送质粒DNA。编码荧光素酶的质粒DNA向再细胞化的真皮构建体的递送增强，导致与仅注射相比，荧光素酶表达在120小时内增加四倍（P < 0.05）。在构建体的表皮层中证实了编码绿色荧光蛋白（GFP）的质粒DNA的递送。在BALB/c小鼠中进行体内实验，皮肤作为递送靶点。与单独注射质粒DNA获得的水平相比，PAA暴露显著提高了暴露部位的荧光素酶表达水平460倍（P < 0.001）。当等离子体反应器位于离注射部位更远的位置时，表达水平增强。通过免疫染色证实编码GFP的质粒DNA向小鼠皮肤的递送，其中与GFP表达定位于表皮内的3 mm处的暴露相比，在10 mm距离处的3分钟暴露显示真皮层内深处的递送分布。我们的研究结果表明，PAA介导的交付值得进一步探索作为皮肤靶点的DNA转移的替代方法。
Plasma-activated air (PAA) provides a noncontact DNA transfer platform. In the current study, PAA was used for the delivery of plasmid DNA in a 3D human skin model, as well as in vivo. Delivery of plasmid DNA encoding luciferase to recellularized dermal constructs was enhanced, resulting in a fourfold increase in luciferase expression over 120 hours compared to injection only (P < 0.05). Delivery of plasmid DNA encoding green fluorescent protein (GFP) was confirmed in the epidermal layers of the construct. In vivo experiments were performed in BALB/c mice, with skin as the delivery target. PAA exposure significantly enhanced luciferase expression levels 460-fold in exposed sites compared to levels obtained from the injection of plasmid DNA alone (P < 0.001). Expression levels were enhanced when the plasma reactor was positioned more distant from the injection site. Delivery of plasmid DNA encoding GFP to mouse skin was confirmed by immunostaining, where a 3-minute exposure at a 10 mm distance displayed delivery distribution deep within the dermal layers compared to an exposure at 3 mm where GFP expression was localized within the epidermis. Our findings suggest PAA-mediated delivery warrants further exploration as an alternative approach for DNA transfer for skin targets.