Intracellular protein delivery and gene transfection by electroporation using a microneedle electrode array.
Intracellular protein delivery and gene transfection by electroporation using a microneedle electrode array.
复制标题
DOI：
10.1002/smll.201101747
复制
发表时间：
2012-04-10
期刊：
影响因子：
13.3
通讯作者：
Allen, Mark G.
中科院分区：
文献类型：
--
作者：
Choi, Seong-O;Kim, Yeu-Chun;Lee, Jeong Woo;Park, Jung-Hwan;Prausnitz, Mark R.;Allen, Mark G.
关键词：
许多生物药物，包括蛋白质和基因为基础的治疗方法的影响，一直受到限制，需要更好的方法来传递到组织内的细胞。在这里，我们提出了细胞内传递的分子和转染质粒DNA的电穿孔，使用一种新的微针电极阵列设计的皮肤和其他组织表面的靶向治疗。微针阵列由聚乳酸模制而成。电穿孔所需的电极和电路被施加到微针阵列表面的一种新的金属转移微成型方法。微针阵列在插入猪尸体皮肤后保持机械完整性，并且能够在体外电穿孔人前列腺癌细胞。定量测量表明，增加电穿孔脉冲电压增加钙黄绿素和牛血清白蛋白的摄取效率，而增加脉冲长度在研究范围内的影响较小。在高细胞活力下，高达50%的细胞吸收分子，并用绿色荧光蛋白基因转染12%的细胞。我们的结论是，微针电极阵列是能够电穿孔细胞，导致在细胞内摄取的分子，并具有潜在的应用，以改善细胞内的蛋白质，DNA和其他生物药物的交付。
The impact of many biopharmaceuticals, including protein- and gene-based therapies, has been limited by the need for better methods of delivery into cells within tissues. Here, we present intracellular delivery of molecules and transfection with plasmid DNA by electroporation using a novel microneedle electrode array designed for targeted treatment of skin and other tissue surfaces. The microneedle array is molded out of polylactic acid. Electrodes and circuitry required for electroporation are applied to the microneedle array surface by a new metal-transfer micromolding method. The microneedle array maintains mechanical integrity after insertion into pig cadaver skin and is able to electroporate human prostate cancer cells in vitro. Quantitative measurements show that increasing electroporation pulse voltage increases uptake efficiency of calcein and bovine serum albumin, whereas increasing pulse length has lesser effects over the range studied. Uptake of molecules by up to 50 % of cells and transfection of 12 % of cells with a gene for green fluorescent protein is demonstrated at high cell viability. We conclude that the microneedle electrode array is able to electroporate cells, resulting in intracellular uptake of molecules, and has potential applications to improve intracellular delivery of proteins, DNA and other biopharmaceuticals.
登录
查看更多内容
DOI：
10.1038/nrg2432
发表时间：
2008-10
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
--
影响因子：
10.8
作者：
Park, JH;Allen, MG;Prausnitz, MR
通讯作者：
Prausnitz, MR
影响因子：
3.4
作者：
Canatella, PJ;Karr, JF;Prausnitz, MR
通讯作者：
Prausnitz, MR
影响因子：
2.7
作者：
Rajaraman, Swaminathan;Choi, Seong-O;Allen, Mark G.
通讯作者：
Allen, Mark G.
影响因子：
5.5
作者：
Luxembourg, A;Hannaman, D;Bernard, R
通讯作者：
Bernard, R