Faculty of Pharmacy, Silpakorn University, Nakhon Pathom 73000, Thailand.
Int J Pharm. 2011 May 30;410(1-2):161-8. doi: 10.1016/j.ijpharm.2011.03.008. Epub 2011 Mar 22.
The aim of this research was to investigate the effect of cationic polypeptides mixed with chitosan (CS) on in vitro transfection efficiency and cytotoxicity in human cervical carcinoma cells (HeLa cells). The polypeptides/DNA complexes and ternary complexes (CS, polypeptides and DNA) at varying weight ratios were formulated and characterized by using gel electrophoresis. Their particle sizes and charge were evaluated. The effect of the type and molecular weight (MW) of polypeptides, the weight ratio, order of mixing, the pH and serum on transfection efficiency and cytotoxicity were evaluated in HeLa cells. Three types of polypeptides (poly-L-lysine; PLL, poly-L-arginine; PLA and poly-L-ornithine; PLO) were able to form complete complex with DNA at weight ratio above 0.1. The PLA MW >70 kDa showed the highest transfection efficiency. The order of mixing between CS, PLA and DNA affected the transfection efficiency. The highest transfection efficiency was observed in ternary complexes of PLA/DNA/CS (2:1:4) equal to PEI/DNA complex. For cytotoxicity studies, over 80% the average cell viabilities of the complexes were observed by MTT assay. This study suggests that the addition of CS to PLA/DNA is easy to prepare, safe and exhibits significantly improved DNA delivery potential in vitro.
本研究旨在探讨阳离子多肽与壳聚糖（CS）混合对人宫颈癌（HeLa 细胞）体外转染效率和细胞毒性的影响。采用凝胶电泳法对不同重量比的多肽/DNA 复合物和三元复合物（CS、多肽和 DNA）进行了配方和特性研究。评估了它们的粒径和电荷。在 HeLa 细胞中，评估了多肽类型和分子量（MW）、重量比、混合顺序、pH 值和血清对转染效率和细胞毒性的影响。三种类型的多肽（聚-L-赖氨酸；PLL、聚-L-精氨酸；PLA 和聚-L-鸟氨酸；PLO）在重量比高于 0.1 时能够与 DNA 形成完整复合物。MW>70 kDa 的 PLA 显示出最高的转染效率。CS、PLA 和 DNA 之间的混合顺序影响转染效率。在 PLA/DNA/CS（2:1:4）三元复合物中观察到最高的转染效率，与 PEI/DNA 复合物相当。对于细胞毒性研究，MTT 测定法观察到复合物的平均细胞活力超过 80%。本研究表明，将 CS 添加到 PLA/DNA 中易于制备、安全，并在体外显著提高 DNA 传递潜力。
