Lai Wing-Fu, Tang Gu-Ping, Wang Xin, Li Guo, Yao Hong, Shen Zan, Lu Gang, Poon Wai Sang, Kung Hsiang-Fu, Lin Marie C M
Department of Chemistry, The University of Hong Kong, Hong Kong SAR, China.
Bionanoscience. 2011 Sep;1(3):89-96. doi: 10.1007/s12668-011-0010-9. Epub 2011 Jun 18.
This study aims to modify a cyclodextrin-PEI-based polymer, PEI-β-CyD, with the TAT peptide for plasmid DNA delivery to placenta mesenchymal stem cells (PMSCs). By using the disulfide exchange between the SPDP-activated PEI-β-CyD and TAT peptide, the TAT-PEI-β-CyD polymer was fabricated and the success of this was confirmed by the presence of characteristic peaks for PEI (at δ 2.8-3.2 ppm), CyD (at δ 5.2, 3.8-4.0 and 3.4-3.6 ppm) and TAT (at δ 1.6-1.9 and 6.8-7.2 ppm) in the (1)H NMR spectrum of TAT-PEI-β-CyD. The polymer-plasmid-DNA polyplex could condense DNA at an N/P ratio of 7.0-8.0, and form nanoparticles with the size of 150.6±5.6 nm at its optimal N/P ratio (20/1). By examining the transfection efficiency and cytotoxicity of TAT-PEI-β-CyD, conjugation of the TAT peptide onto PEI-β-CyD was demonstrated to improve the transfection efficiency of PEI-β-CyD in PMSCs after 48 and 96 hours of post-transfection incubation. The viability of PEI-β-CyD-treated PMSCs was shown to be over 80% after 5 h of treatment and 24 h of post-treatment incubation. In summary, this study showed that the TAT-PEI-β-CyD polymer as a vector for plasmid DNA delivery to PMSCs and other cells warrants further investigations. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s12668-011-0010-9) contains supplementary material, which is available to authorized users.
本研究旨在将基于环糊精-聚乙烯亚胺的聚合物PEI-β-CyD与TAT肽进行修饰，用于将质粒DNA递送至胎盘间充质干细胞（PMSC）。通过SPDP活化的PEI-β-CyD与TAT肽之间的二硫键交换，制备了TAT-PEI-β-CyD聚合物，并且通过TAT-PEI-β-CyD的（1）H NMR谱中PEI（δ 2.8 - 3.2 ppm）、环糊精（δ 5.2、3.8 - 4.0和3.4 - 3.6 ppm）以及TAT（δ 1.6 - 1.9和6.8 - 7.2 ppm）的特征峰证实了这一成功。聚合物-质粒-DNA复合物能够在N/P比为7.0 - 8.0时凝聚DNA，并在其最佳N/P比（20/1）下形成尺寸为150.6±5.6 nm的纳米颗粒。通过检测TAT-PEI-β-CyD的转染效率和细胞毒性，结果表明将TAT肽缀合到PEI-β-CyD上可提高转染后48小时和96小时PEI-β-CyD在PMSC中的转染效率。经PEI-β-CyD处理的PMSC在处理5小时和处理后孵育24小时后的活力显示超过80%。总之，本研究表明TAT-PEI-β-CyD聚合物作为将质粒DNA递送至PMSC和其他细胞的载体值得进一步研究。电子补充材料：本文的在线版本（doi:10.1007/s12668-011-0010-9）包含补充材料，授权用户可获取。
