Department of Polymer Science and Engineering, Sunchon National University, Jeonnam 57922, Republic of Korea.
Department of Polymer Science and Engineering, Sunchon National University, Jeonnam 57922, Republic of Korea.
Int J Biol Macromol. 2018 Aug;115:459-468. doi: 10.1016/j.ijbiomac.2018.04.053. Epub 2018 Apr 19.
Branched polyethylenimine (bPEI) was well known as high transfection agent, which has many amine group. However, utilization of bPEI was limited due to high toxicity. To solve these problems, bPEI was introduced to low molecular weight water-soluble chitosan (LMWSC) with coupling agent. In addition, hyaluronic acid (HA), one of natural anion polymer, was introduced to binary complex of pDNA/bPEI-grafted LMWSC (LMPEI) to target the specific cancer cell and impart the serum resistant. Ternary complexes of pDNA/LMPEI/HA were prepared by electrostatic charge interaction and their binding affinity and DNase protection assay were conducted by gel retardation assay. Particle size of ternary complexes showed that had each 482 ± 245.4 (pDNA/LMPEI2%/HA, 1:16:1, w/w/w) and 410 ± 78.5 nm (pDNA/LMPEI4%/HA, 1:16:2, w/w/w). Moreover, to demonstrate serum-resistant effect of ternary complexes, particle size of them was measured according to incubated time (0-10 h) under serum condition. Transfection assay of ternary complexes showed that their transfection efficiency in CD44-receptor overexpressed HCT116 cell was higher than CD44-receptor negative CT26 cell. Additionally, intracellular uptake of ternary complexes with propidium iodide (PI)-labeled pDNA was observed to confirm targeting effect and cellular internalization by fluorescence microscopy. These results suggest that ternary complexes are superb gene carrier with excellent serum-resistant and high gene transfection.
支化聚乙烯亚胺（bPEI）作为一种高转染试剂而广为人知，它具有许多胺基。然而，由于其毒性高，bPEI 的应用受到限制。为了解决这些问题，bPEI 被用偶联剂引入到低分子量水溶性壳聚糖（LMWSC）中。此外，将天然阴离子聚合物之一的透明质酸（HA）引入到 pDNA/bPEI 接枝 LMWSC（LMPEI）的二元复合物中，以靶向特定的癌细胞并赋予血清抗性。通过静电电荷相互作用制备 pDNA/LMPEI/HA 的三元复合物，并通过凝胶阻滞试验进行其结合亲和力和 DNA 酶保护试验。三元复合物的粒径表明，pDNA/LMPEI2%/HA（1:16:1，w/w/w）和 pDNA/LMPEI4%/HA（1:16:2，w/w/w）的粒径分别为 482±245.4nm 和 410±78.5nm。此外，为了证明三元复合物的耐血清效应，根据在血清条件下孵育时间（0-10h）测量它们的粒径。三元复合物的转染实验表明，它们在 CD44 受体过表达的 HCT116 细胞中的转染效率高于 CD44 受体阴性的 CT26 细胞。此外，通过荧光显微镜观察用碘化丙啶（PI）标记的 pDNA 的三元复合物的细胞内摄取，以确认靶向作用和细胞内化。这些结果表明，三元复合物是一种具有出色的耐血清性和高基因转染效率的优异基因载体。
