Department of Chemistry and Biochemistry, National Chung Cheng University, 168 University Rd., Min-Hsiung Chia-Yi, Taiwan, ROC.
J Mater Sci Mater Med. 2010 May;21(5):1543-51. doi: 10.1007/s10856-010-4004-x. Epub 2010 Feb 5.
To improve the cytotoxicity of PEI25k and the transfection efficiency of poly(beta-amino ester) with DNA, we synthesized a poly(beta-amino ester), PEDP, bearing ester linkages in the backbone and tertiary amines in the backbone and side chain and prepared a binary mixture, PEDP-PEI25k, using physical blending meyhod. Both poly(beta-amino ester) PEDP and binary mixture PEDP-PEI25k, readily self-assembled with plasmid DNA (pCMV-beta gal) in a HEPES buffer, were characterized by dynamic light scattering. The results reveal that PEDP-PEI25k was able to self-assemble plasmid DNA into PEDP-PEI25k/DNA nano-complexes small enough to enter a cell through endocytosis. Titration studies were performed to determine the buffering capacities of PEDP and PEDP-PEI25k. The COS-7 cell viabilities in the presence of PEDP and PEDP-PEI25k were studied. At low mass ratio of PEDP/PEI25k (1/1), it is found that the transfection curve of PEDP-PEI25k/DNA bearing a maximum peak is similar to that of PEI25k/DNA. In addition, the PEDP-PEI25k/DNA complexes were able to transfect COS-7 cells in vitro with a high efficiency comparable to a well-known gene carrier PEI25k/DNA. The results indicate that binary mixture PEDP-PEI25k is an attractive cationic carrier for gene delivery and an interesting candidate for further study.
为了提高 PEI25k 的细胞毒性和聚(β-氨基酸酯)与 DNA 的转染效率,我们合成了一种聚(β-氨基酸酯)PEDP,其主链中含有酯键,主链和侧链中含有叔胺,并采用物理共混法制备了二元混合物 PEDP-PEI25k。聚(β-氨基酸酯)PEDP 和二元混合物 PEDP-PEI25k 都很容易在 HEPES 缓冲液中与质粒 DNA(pCMV-beta gal)自组装,通过动态光散射进行了表征。结果表明,PEDP-PEI25k 能够将质粒 DNA 自组装成 PEDP-PEI25k/DNA 纳米复合物,这些复合物足够小,可以通过内吞作用进入细胞。进行了滴定研究以确定 PEDP 和 PEDP-PEI25k 的缓冲能力。研究了 COS-7 细胞在 PEDP 和 PEDP-PEI25k 存在下的活力。在低质量比的 PEDP/PEI25k(1/1)下,发现具有最大峰值的 PEDP-PEI25k/DNA 的转染曲线与 PEI25k/DNA 的转染曲线相似。此外,PEDP-PEI25k/DNA 复合物能够在体外转染 COS-7 细胞,效率与著名的基因载体 PEI25k/DNA 相当。结果表明,二元混合物 PEDP-PEI25k 是一种有吸引力的阳离子载体,可用于基因传递,是进一步研究的有趣候选物。