Kazemi Oskuee Reza, Mahmoudi Asma, Gholami Leila, Rahmatkhah Alireza, Malaekeh-Nikouei Bizhan
Neurogenic Inflammation Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.; Department of Medical Biotechnology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Adv Pharm Bull. 2016 Dec;6(4):515-520. doi: 10.15171/apb.2016.065. Epub 2016 Dec 22.
Cationic polymers and cationic liposomes have shown to be effective non-viral gene delivery vectors. In this study, we tried to improve the transfection efficiency by employing the advantages of both. For this purpose, modified polyallylamines (PAAs) were synthesized. These modifications were done through the reaction of PAA (15 KDa) with acrylate and 6-bromoalkanoic acid derivatives. Liposomes comprising of these cationic polymers and cationic lipid were prepared and extruded through polycarbonate filters to obtain desired size. Liposome-DNA nanocomplexes were prepared in three carrier to plasmid (C/P) ratios. Size, zeta potential and DNA condensation ability of each complex were characterized separately and finally transfection efficiency and cytotoxicity of prepared vectors were evaluated in Neuro2A cell line. The results showed that mean particle size of all these nanocomplexes was lower than 266 nm with surface charge of 22.0 to 33.9 mV. Almost the same condensation pattern was observed in all vectors and complete condensation was occurred at C/P ratio of 1.5. The lipoplexes containing modified PAA 15 kDa with 10% hexyl acrylate showed the highest transfection efficacy and lowest cytotoxicity in C/P ratio of 0.5. In some cases nanocomplexes consisting of cationic liposome and modified PAA showed better transfection activity and lower cytotoxicity compared to PAA.
阳离子聚合物和阳离子脂质体已被证明是有效的非病毒基因传递载体。在本研究中，我们试图通过利用两者的优势来提高转染效率。为此，合成了改性聚烯丙胺（PAA）。这些改性是通过使15 kDa的PAA与丙烯酸酯和6-溴链烷酸衍生物反应来完成的。制备了由这些阳离子聚合物和阳离子脂质组成的脂质体，并通过聚碳酸酯滤膜挤出以获得所需尺寸。脂质体-DNA纳米复合物以三种载体与质粒（C/P）比例制备。分别对每种复合物的尺寸、zeta电位和DNA凝聚能力进行了表征，最后在Neuro2A细胞系中评估了所制备载体的转染效率和细胞毒性。结果表明，所有这些纳米复合物的平均粒径均低于266 nm，表面电荷为22.0至33.9 mV。在所有载体中观察到几乎相同的凝聚模式，并且在C/P比例为1.5时发生完全凝聚。含有10%丙烯酸己酯的改性15 kDa PAA的脂质体复合物在C/P比例为0.5时显示出最高的转染效率和最低的细胞毒性。在某些情况下，与PAA相比，由阳离子脂质体和改性PAA组成的纳米复合物表现出更好的转染活性和更低的细胞毒性。
