Division of Nanobiotechnology Section, Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran.
Division of Nanobiotechnology Section, Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran.
Mater Sci Eng C Mater Biol Appl. 2017 Oct 1;79:770-782. doi: 10.1016/j.msec.2017.05.068. Epub 2017 May 14.
Non-viral vectors have attracted great interest, as they are simple to prepare, easy to modify and relatively safe, compared to viral vectors. Kidney-targeted gene delivery systems depict a promising technology to improve drug efficacy in renal diseases treatments. In order to develop a novel kidney-targeted gene delivery system, we synthesized polyamine-PEI conjugates using polymyxin B as ligand and investigated their potential targeting efficiency. After grafting either PEI kDa or PEI kDa with polymyxin B through amidation reaction, the modified-polymyxin-PEI/DNA-nanoplexes were produced via electrostatic attraction between the cationic polymers and EGFP plasmid. The properties of modified polymers including size, surface charge density, DNA condensation ability, buffering capacity and cytotoxicity were evaluated. Results revealed that the average size of -modified-polymyxin- PEIkDa was about 143-180nm and modified-polymyxin-PEIkDa 115-194nm. The zeta potentials were in the range of 16.4±1.87 to 23.43±1.25mV and 11.3±1.4 to 19.3±2.1mV for conjugates based on PEI and PEI respectively. The AFM images revealed that the complexes were spherical and nano-sized at C/P=4. The buffering capacity of both PEI 10 and 25kDa increased as the percentage of polymyxin B grafting increased. In vitro study demonstrated that modified-polymyxin-PEI conjugates could remarkably improve the gene transfection efficiency to kidney cells. The transfection efficiency of the polyplexes was dependent on the weight ratio of ligand in the formulation (~12 and 8 fold increase for PEI and PEIkDa, respectively) and was significantly higher than that of unmodified PEIs/DNA nanoparticles. These results suggest that modified-polymyxin-PEI /DNA nanoparticles can effectively target megalin-expressing kidney cells and show improved transfection efficiency and low cytotoxicity in In vitro and In vivo studies. Animal studies confirmed the in vivo study. Thus, these conjugates can be considered as a safe and efficient non-viral therapeutic therapy vector for kidney diseases.
非病毒载体由于其制备简单、易于修饰且相对安全，因此受到了极大的关注，与病毒载体相比，非病毒载体具有明显的优势。肾脏靶向基因传递系统是一种很有前途的技术，可以提高肾脏疾病治疗的药物疗效。为了开发一种新型的肾脏靶向基因传递系统，我们合成了聚赖氨酸-PEI 缀合物，使用多粘菌素 B 作为配体，并研究了它们潜在的靶向效率。通过酰胺反应将 PEI kDa 或 PEI kDa 与多粘菌素 B 接枝后，通过阳离子聚合物与 EGFP 质粒之间的静电吸引作用产生修饰后的多粘菌素-PEI/DNA-纳米复合物。对修饰聚合物的性能，包括粒径、表面电荷密度、DNA 缩合能力、缓冲能力和细胞毒性进行了评价。结果表明，-修饰多粘菌素-PEI kDa 的平均粒径约为 143-180nm，修饰多粘菌素-PEI kDa115-194nm。PEI 和 PEI 对应的共轭物的 ζ 电位分别为 16.4±1.87 至 23.43±1.25mV 和 11.3±1.4 至 19.3±2.1mV。原子力显微镜图像显示，复合物在 C/P=4 时为球形且纳米级。随着多粘菌素 B 接枝率的增加，PEI10 和 25kDa 的缓冲能力均增加。体外研究表明，修饰的多粘菌素-PEI 缀合物可以显著提高肾脏细胞的基因转染效率。多聚物的转染效率取决于配方中配体的重量比（PEI 和 PEI kDa 分别约增加 12 倍和 8 倍），显著高于未修饰的 PEIs/DNA 纳米颗粒。这些结果表明，修饰的多粘菌素-PEI/DNA 纳米颗粒可以有效地靶向表达 megalin 的肾脏细胞，并在体内外研究中显示出更高的转染效率和低细胞毒性。动物研究证实了体内研究。因此，这些缀合物可被视为用于肾脏疾病的安全有效的非病毒治疗载体。
