Optimization of a plasma facilitated DNA delivery method
Optimization of a plasma facilitated DNA delivery method
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DOI：
10.1016/j.bioelechem.2014.09.003
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发表时间：
2015-06-01
影响因子：
5
通讯作者：
Jaroszeski, Mark J.
中科院分区：
文献类型：
--
作者：
Connolly, Richard J.;Hoff, Andrew M.;Jaroszeski, Mark J.
基于等离子体的方法最近出现了一种增加质粒DNA递送到皮肤的技术。这种输送方式依赖于电离气体分子沉积到目标细胞或组织上以建立电场。据推测，这个电场导致细胞膜的介电击穿，使细胞对外源分子具有渗透性。这项体内研究试图通过调节血浆源和质粒DNA的总暴露量来优化表达质粒DNA的荧光素酶的皮内递送。改变血浆暴露时间从2,5,10和20分钟，可以发现导致荧光素酶最高表达的条件。这些条件与+ 8kv或- 8kv等离子体的10分钟暴露时间相关，当发生器在距离表皮组织表面3cm处以15l /min的氦气流速运行时。将注射的侧腹皮肤暴露10分钟，在+ 8kv产生的等离子体中增加37.3倍，在- 8kv产生的等离子体中增加27.1倍。用50、100、200 μ g的荧光素酶表达质粒处理时，发现100 μ g的荧光素酶表达质粒发光峰值最高。(C) 2014 Elsevier B.V.版权所有
Plasma-based methods have recently emerged as a technique for augmenting plasmid DNA delivery to skin. This delivery modality relies on the deposition of ionized gas molecules on to targeted cells or tissue to establish an electric field. It is hypothesized that this electric field results in the dielectric breakdown of cell membranes, making cells permeable to exogenous molecules. This in vivo investigation sought to optimize the intradermal delivery of a luciferase expressing plasmid DNA by modulating the total exposure to the plasma source and the plasmid DNA dose. Varying the plasma exposure time from 2, 5, 10, and 20 min allowed the conditions resulting in the highest expression of luciferase to be found. These conditions correlated to the 10 minute exposure time for a plasma derived from either +8 kV or -8 kV, when the generator was operated 3 cm from the epidermal tissue surface with a helium flow rate of 15 L/min. Exposing the injected flank skin for 10 min resulted in a rise of 37.3-fold for a plasma created with +8 kV and 27.1-fold for a plasma created with -8 kV. When using this treatment time with 50, 100, or 200 mu g of a luciferase expressing plasmid, it was found that 100 mu g resulted in the highest peak luminescence. (C) 2014 Elsevier B.V. All rights reserved.