Sakai Shinji, Yamada Yusuke, Yamaguchi Tetsu, Ciach Tomasz, Kawakami Koei
Department of Chemical Engineering, Faculty of Engineering, Kyushu University, Fukuoka, Japan.
J Biomed Mater Res A. 2009 Feb;88(2):281-7. doi: 10.1002/jbm.a.31870.
The method of coating electrospun ultrafine poly(L-lactic acid) fibers with DNA, by building up polyelectrolyte layer(s) of poly(ethyleneimine) (PEI) and plasmid DNA using an electrostatic layer-by-layer deposition method, for gene delivery is presented. The pGL3 encoding luciferase was applied as plasmid DNA. The quantity of pGL3 immobilized on individual fibers increased with increasing pGL3 concentration in the immersion solution (0.017-0.870 mg/mL) and increasing bilayer number of PEI/pGL3 (single-triple). With the exception of one specimen prepared under the condition 0.870 mg/mL pGL3 solution and double PEI/pGL3 layers, the transfection efficiency of COS-7 cells, defined by the ratio of fluorescence intensity (resulting from the presence of luciferase) with respect to the quantity of cellular protein on the fibrous mat increased with increasing quantity of pGL3 on the fibers. In addition to the ease of controlling the quality of polyelectrolyte bilayer(s) by simply changing the concentrations of substances and number of immersing cycles, the features of the electrospun fibrous mat such as a very large surface-to-volume ratio and flexibility, could potentially be employed as a strategy for gene therapy combined with tissue engineering technology.
本文介绍了一种通过静电逐层沉积法构建聚(乙烯亚胺)(PEI)和质粒DNA的聚电解质层,从而用DNA包覆静电纺丝超细聚(L-乳酸)纤维以进行基因递送的方法。编码荧光素酶的pGL3用作质粒DNA。固定在单根纤维上的pGL3数量随着浸泡溶液中pGL3浓度(0.017 - 0.870 mg/mL)的增加以及PEI/pGL3双层数量(单层 - 三层)的增加而增加。除了在0.870 mg/mL pGL3溶液和双层PEI/pGL3条件下制备的一个样本外,由纤维垫上荧光强度(由荧光素酶的存在导致)与细胞蛋白量的比值定义的COS - 7细胞转染效率随着纤维上pGL3量的增加而增加。除了通过简单改变物质浓度和浸泡循环次数就能轻松控制聚电解质双层的质量外,静电纺丝纤维垫的特性,如非常大的表面积与体积比和柔韧性,有可能被用作与组织工程技术相结合的基因治疗策略。