Department of Polymer Science and Engineering, Sunchon National University, 255 Jungang-ro, Suncheon, Jeonnam 540-742, Republic of Korea.
Department of Polymer Science and Engineering, Sunchon National University, 255 Jungang-ro, Suncheon, Jeonnam 540-742, Republic of Korea.
Carbohydr Polym. 2017 Dec 15;178:322-330. doi: 10.1016/j.carbpol.2017.09.048. Epub 2017 Sep 14.
For efficient gene delivery, non-viral vectors should have high cellular uptake, excellent endosomal escape, and the ability to rapidly release the gene into the cytoplasm. Here, we developed a disulfide bond-conjugated bioreducible LMWSC-g-bPEI (LCP) composed of low molecular-weight water soluble chitosan (LMWSC), bPEI, and cystamine (Cys). The developed LCP had advantages such as low toxicity, great endosomal escape, and rapid release of pDNA into the cytoplasm. The polyplexes with LCP showed higher uptake into the nucleus and greater transfection efficiency than that without disulfide bond. Moreover, LCP polymer and polyplexes with LCP indicated lower cytotoxicity than bPEI 25kDa. In addition, a gel retardation assay and particle size were analyzed to demonstrate the reduction-sensitive gene delivery system. Besides, intracellular uptake pathway of polyplexes was investigated by various endocytosis inhibitor and confirmed to internalization into cell via macropinocytosis. These results suggest that bioreducible LCP is a superb non-viral vector for efficient gene delivery.
为了实现高效的基因传递，非病毒载体应该具有高细胞摄取能力、出色的内体逃逸能力以及快速将基因释放到细胞质中的能力。在这里，我们开发了一种由低分子量水溶性壳聚糖（LMWSC）、bPEI 和半胱胺（Cys）组成的二硫键偶联的生物还原型 LMWSC-g-bPEI（LCP）。所开发的 LCP 具有低毒性、出色的内体逃逸和快速将 pDNA 释放到细胞质中等优点。与没有二硫键的 LCP 相比，具有 LCP 的聚阳离子复合物具有更高的核摄取和更高的转染效率。此外，LCP 聚合物和具有 LCP 的聚阳离子复合物的细胞毒性低于 bPEI 25kDa。此外，通过凝胶阻滞实验和粒径分析来证明还原敏感的基因传递系统。此外，通过各种内吞作用抑制剂研究了多聚物的细胞内摄取途径，并通过巨胞饮作用证实了多聚物通过巨胞饮作用进入细胞内。这些结果表明，生物还原型 LCP 是一种高效的基因传递的优秀非病毒载体。
