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使用储库型电极通过电穿孔将质粒DNA导入皮肤。

Plasmid DNA Delivery into the Skin via Electroporation with a Depot-Type Electrode.

作者信息

Yoshida Yuya, Aoki Manami, Nagase Kalin, Marubashi Koichi, Kojima Hiroyuki, Itakura Shoko, Komatsu Syuuhei, Sugibayashi Kenji, Todo Hiroaki

机构信息

Faculty of Pharmacy and Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado 350-0295, Saitama, Japan.

Pharmaceutical Research and Technology Labs., Astellas Pharma Inc., 180 Ozumi, Yaizu 425-0072, Shizuoka, Japan.

出版信息

Pharmaceutics. 2024 Sep 18;16(9):1219. doi: 10.3390/pharmaceutics16091219.

Abstract

Non-viral mediated plasmid DNA transfection by electroporation (EP) is an established method for gene transfection. In this study, the usefulness of direct EP at an intradermal () site () with implanted electrodes to achieve a high protein expression level was investigated. In addition, application with various intervals with a low application voltage was also evaluated to confirm its effect on protein expression. : Green fluorescent protein (GFP)- and luciferase-encoding DNA were administrated, and GFP and luciferase were evaluated. : A higher protein expression level was observed after green fluorescent protein (GFP)- and luciferase-encoding DNA were delivered by injection followed by application. When luciferase expression was observed with an in vivo imaging system, continuous expression was confirmed over 21 days after injection followed by at 100 V. This approach provided increased gene expression levels compared with conventional EP methods via the stratum corneum layer. In addition, the effect of application voltage on luciferase expression was investigated; two-time applications (repeated ) at 20 V with 5 min intervals showed similar luciferase expression level to single application with 100 V. Histological observations showed the skin became thicker after a single at 100 V, whereas no apparent thickness changes were confirmed after repeated at 20 V with 5 min intervals. : This study revealed that direct voltage application achieved high protein expression levels even at low voltages. Skin is a promising administration site for DNA vaccines, so this approach may be effective for DNA vaccine delivery into skin tissue.

摘要

通过电穿孔(EP)进行的非病毒介导的质粒DNA转染是一种成熟的基因转染方法。在本研究中,研究了使用植入电极在皮内()部位进行直接电穿孔以实现高蛋白表达水平的有效性。此外,还评估了以低施加电压在不同间隔下的应用,以确认其对蛋白质表达的影响。:施用了编码绿色荧光蛋白(GFP)和荧光素酶的DNA,并对GFP和荧光素酶进行了评估。:在通过注射然后进行电穿孔递送编码绿色荧光蛋白(GFP)和荧光素酶的DNA后,观察到更高的蛋白质表达水平。当使用体内成像系统观察荧光素酶表达时,在注射然后在100V下进行电穿孔后21天内确认了持续表达。与通过角质层的传统电穿孔方法相比,这种方法提高了基因表达水平。此外,研究了施加电压对荧光素酶表达的影响;在20V下以5分钟间隔进行两次施加(重复电穿孔)显示出与100V单次电穿孔相似的荧光素酶表达水平。组织学观察表明,在100V下单次电穿孔后皮肤变厚,而在20V下以5分钟间隔重复电穿孔后未确认明显的厚度变化。:本研究表明,即使在低电压下,直接施加电压也能实现高蛋白表达水平。皮肤是DNA疫苗的一个有前景的给药部位,因此这种方法可能对将DNA疫苗递送至皮肤组织有效。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d47e/11435037/6c8bb6cf980f/pharmaceutics-16-01219-g001.jpg

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