Department of Biosciences and Informatics, Keio University, Yokohama, Kanagawa, Japan.
Biomaterials. 2012 Oct;33(29):7251-60. doi: 10.1016/j.biomaterials.2012.06.040. Epub 2012 Jul 13.
pDNA/chitosan complexes have been investigated as promising non-viral vectors for gene delivery. However, an increase in transfection efficiency and enhancement of physicochemical stability are required for their practical use. In this study, chondroitin sulfate (CS) was employed as a coating agent to increase the stability and transfection efficiency of a pDNA/chitosan complex. The pDNA/chitosan/CS ternary complexes formed with six kinds of CSs having different limiting viscosities (0.2-1.6) and sulfation degrees (5.0-7.0%) showed considerable differences in particle size, surface charge, and morphology. Among them, CS having a medium limiting viscosity (0.5-0.6) and a high sulfation degree (6.9%) showed significant enhancements in cell transfection efficiency. Analyses of cellular uptake and intracellular trafficking revealed that increased cellular uptake via macropinocytosis, together with reduced entry into lysosomes, may explain the promotion of transfection efficiency of ternary complexes.
pDNA/壳聚糖复合物已被研究作为有前途的非病毒基因载体。然而，为了实际应用，需要提高转染效率和增强物理化学稳定性。在本研究中，硫酸软骨素（CS）被用作涂层剂，以提高 pDNA/壳聚糖复合物的稳定性和转染效率。用六种具有不同极限粘度（0.2-1.6）和硫酸化度（5.0-7.0%）的 CS 制备的 pDNA/壳聚糖/CS 三元复合物在粒径、表面电荷和形态上表现出明显的差异。其中，具有中等极限粘度（0.5-0.6）和高硫酸化度（6.9%）的 CS 显著提高了细胞转染效率。细胞摄取和细胞内转运分析表明，通过巨胞饮作用增加细胞摄取，同时减少进入溶酶体，可能解释了三元复合物转染效率的提高。
