Wang Haixia, Li Qian, Yang Jing, Guo Jintang, Ren Xiangkui, Feng Yakai, Zhang Wencheng
School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China.
J Mater Chem B. 2017 Feb 21;5(7):1408-1422. doi: 10.1039/c6tb02379g. Epub 2017 Jan 30.
If plasmid complexes prepared from targeting carriers have endothelial cell selectivity and high transfection efficiency, they can specifically enhance rapid endothelialization by delivering the corresponding gene plasmids. A high content of functional groups in the carriers benefits high selectivity efficiency. Herein, we have synthesized a comb-shaped polymer bearing several Arg-Glu-Asp-Val (REDV) peptides and poly(ethylene glycol) as a pEGFP-ZNF580 gene carrier with cell-type recognition of human endothelial cells. An amphiphilic block copolymer of poly(2-hydroxyethyl methacrylate)-block-poly(ε-caprolactone)-graft-poly(ethylene glycol)-graft-poly(ethyleneimine) conjugated with REDV peptide (PHEMA-b-PCL-g-PEG-g-PEI-REDV) is synthesized, and nanoparticles of it are prepared by polymer self-assembly. This polycationic PHEMA-b-PCL-g-PEG-g-PEI-REDV carrier effectively condenses pEGFP-ZNF580 plasmid to form REDV-targeted complexes and protects pEGFP-ZNF580 integrity from enzymatic hydrolysis. These REDV-targeted complexes exhibit low cytotoxicity but high transfection efficiency to EA.hy926 cells compared with non-targeted complexes (PHEMA-b-PCL-g-PEG-g-PEI/pEGFP-ZNF580) as demonstrated by MTT assay, fluorescence-activated cell sorting analysis and fluorescence analysis. Furthermore, the relative protein level of endothelial cells transfected by REDV-targeted complexes is higher than that by non-targeted complexes. Therefore, REDV-bearing carriers may have potential as effective and targeting transfer carriers for pEGFP-ZNF580 plasmid, and their complexes can be used in the endothelialization of artificial vascular grafts.
如果从靶向载体制备的质粒复合物具有内皮细胞选择性和高转染效率，它们可以通过递送相应的基因质粒特异性地促进快速内皮化。载体中高含量的官能团有利于提高选择性效率。在此，我们合成了一种带有多个精氨酸-谷氨酸-天冬氨酸-缬氨酸（REDV）肽和聚乙二醇的梳状聚合物，作为具有人内皮细胞类型识别功能的pEGFP-ZNF580基因载体。合成了与REDV肽共轭的聚（甲基丙烯酸2-羟乙酯）-嵌段-聚（ε-己内酯）-接枝-聚乙二醇-接枝-聚乙烯亚胺两亲性嵌段共聚物（PHEMA-b-PCL-g-PEG-g-PEI-REDV），并通过聚合物自组装制备了其纳米颗粒。这种聚阳离子PHEMA-b-PCL-g-PEG-g-PEI-REDV载体有效地浓缩pEGFP-ZNF580质粒以形成REDV靶向复合物，并保护pEGFP-ZNF580的完整性免受酶水解。与非靶向复合物（PHEMA-b-PCL-g-PEG-g-PEI/pEGFP-ZNF580）相比，这些REDV靶向复合物对EA.hy926细胞表现出低细胞毒性但高转染效率，MTT分析、荧光激活细胞分选分析和荧光分析证明了这一点。此外，REDV靶向复合物转染的内皮细胞的相对蛋白水平高于非靶向复合物。因此，带有REDV的载体可能具有作为pEGFP-ZNF580质粒有效靶向转移载体的潜力，并且它们的复合物可用于人工血管移植物的内皮化。
