CQM-Centro de Química da Madeira, MMRG, Universidade da Madeira, Campus da Penteada, 9000-390, Funchal, Portugal.
J Mater Sci Mater Med. 2012 Dec;23(12):2967-80. doi: 10.1007/s10856-012-4753-9. Epub 2012 Sep 4.
In the present study, the effect of N,N-dimethylaminoethyl methacrylate (DMAEMA) conjugation onto branched poly(ethylenimine) (PEI) with different grafting degree was examined for gene delivery applications. The DMAEMA-grafted-PEI conjugates were characterized and complexed with plasmid DNA (pDNA) at various concentrations, and the physicochemical properties, cell viability, and in vitro transfection efficiency of the complexes were evaluated in HEK 293T cells. Computational techniques were used to analyze the interaction energies and possible binding modes between DNA and conjugates at different grafting degrees. The cytotoxicity analysis and in vitro transfection efficiency of the conjugate/pDNA complexes exhibited a beneficial effect of DMAEMA conjugation when compared to PEI alone. The computational results revealed that the DNA/vector interaction energy decreases with increasing grafting degree, which can be associated to an enhanced release of the pDNA from the carrier once inside cells. The results indicate the significance of DMAEMA conjugation onto PEI as a promising approach for gene delivery applications.
在本研究中,研究了不同接枝度的支化聚(亚乙基亚胺)(PEI)接枝 N,N-二甲基氨基乙基甲基丙烯酸酯(DMAEMA)对基因传递应用的影响。对 DMAEMA 接枝的 PEI 缀合物进行了表征,并在不同浓度下与质粒 DNA(pDNA)复合,在 HEK 293T 细胞中评估了复合物的物理化学性质、细胞活力和体外转染效率。计算技术用于分析不同接枝度下 DNA 和缀合物之间的相互作用能和可能的结合模式。与单独的 PEI 相比,缀合物 / pDNA 复合物的细胞毒性分析和体外转染效率显示出 DMAEMA 接枝的有益效果。计算结果表明,DNA/载体相互作用能随着接枝度的增加而降低,这与载体进入细胞后 pDNA 的释放增强有关。结果表明,PEI 接枝 DMAEMA 作为基因传递应用的一种有前途的方法具有重要意义。