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Real-time impedance feedback to enhance cutaneous gene electrotransfer in a murine skin model
Bioelectrochemistry ( IF 4.8 ) Pub Date : 2021-07-13 , DOI: 10.1016/j.bioelechem.2021.107885
Reginald M Atkins 1 , Timothy J Fawcett 2 , Richard Gilbert 1 , Andrew M Hoff 3 , Richard Connolly 1 , Douglas W Brown 4 , Mark J Jaroszeski 5
Affiliation  

Electric field mediated gene delivery methods have the ability to efficiently transfect cells in vivo with an excellent safety profile. The method has historically used a fixed number of electric pulses with identical characteristics in induce delivery. Electrical treatment does not typically compensate for subject-to-subject variation and other differences. This study was designed to investigate if delivery/expression could be increased using a novel electropulsation method that compensated for variation using real-time electrical impedance measurements. The method involved delivering plasmid DNA encoding luciferase to murine skin. Tissue impedance in a 1–3 KHz range was measured before electric pulses were applied. Impedance was also measured after each successive pulse. Pulsation was stopped when impedance values were reduced by either 80% or 95% relative to prepulse values. Standard/fixed pulsing parameters were also used for comparison. The results indicated that up to 15-fold increases in luciferase expression could be obtained when electrical treatment was ceased based upon impedance reductions. Furthermore, peak expression levels of all treatment groups pulsed using the novel pulsing method were statistically higher than those that employed standard pulsing. These results strongly suggest that applying pulses until a defined impedance-based endpoint results in higher expression.



中文翻译:

实时阻抗反馈增强小鼠皮肤模型中的皮肤基因电转移

电场介导的基因传递方法能够在体内有效地转染细胞具有出色的安全性。该方法历来在诱导递送中使用固定数量的具有相同特性的电脉冲。电疗通常不会补偿受试者之间的差异和其他差异。本研究旨在研究是否可以使用一种新颖的电脉冲方法来增加递送/表达,该方法使用实时电阻抗测量来补偿变化。该方法涉及将编码荧光素酶的质粒 DNA 递送至小鼠皮肤。在施加电脉冲之前测量 1–3 KHz 范围内的组织阻抗。还在每个连续脉冲后测量阻抗。当阻抗值相对于预脉冲值降低 80% 或 95% 时,脉动停止。标准/固定脉冲参数也用于比较。结果表明,当基于阻抗降低而停止电疗时,荧光素酶表达最多可增加 15 倍。此外,使用新型脉冲方法脉冲的所有治疗组的峰值表达水平在统计学上高于采用标准脉冲的那些。这些结果强烈表明,施加脉冲直到定义的基于阻抗的终点导致更高的表达。

更新日期:2021-07-22
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