Department of Chemistry, Shahid Bahonar University of Kerman, Kerman, Iran.
Department of Biochemistry, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran.
Mater Sci Eng C Mater Biol Appl. 2019 Jan 1;94:234-246. doi: 10.1016/j.msec.2018.09.026. Epub 2018 Sep 8.
A low transfection efficiency and failure to deliver therapeutic genes to target organs limit the use of vesicular systems in gene therapy. In this study, magnetic niosomes were used to improve transfection efficiency and overcome limitations. In this light, Tween 60 and Span 60 molecules were employed as the bilayer component and ergosterol and/or cholesterol as membrane-stabilizing agents. We studied the structural and dynamical properties of cholesterol-containing niosomes (ST60/Chol) and ergosterol-containing vesicles (ST60/Ergo) using the molecular dynamics (MD) simulation technique. In in vitro experiments, the protamine-condensed DNA along with magnetic nanoparticles were prepared and incorporated into the niosome to form magnetic niosome-entrapped protamine-condensed DNA (M-NPD). The MD simulation comparison of two bilayers showed that the ST60/Ergo vesicles have better properties for gene delivery. Our in vitro results confirmed the in silico results and revealed that Ergo-niosomes have smaller size, better polydispersity, and slower release of plasmid than Chol-niosome. Moreover, M-NPD-Ergo showed higher cellular uptake and gene expresssion in HEK-293T cell line compared to M-NPD-Chol vesicles.
转染效率低，无法将治疗基因递送到靶器官，限制了囊泡系统在基因治疗中的应用。在这项研究中，使用磁性脂质体来提高转染效率并克服这些限制。为此，我们使用 Tween 60 和 Span 60 分子作为双层成分，并使用甾醇和/或胆固醇作为膜稳定剂。我们使用分子动力学 (MD) 模拟技术研究了含有胆固醇的脂质体 (ST60/Chol) 和含有麦角固醇的囊泡 (ST60/Ergo) 的结构和动力学特性。在体外实验中，将与磁纳米颗粒结合的鱼精蛋白-缩合 DNA 制备并掺入脂质体中，形成磁性脂质体包封的鱼精蛋白-缩合 DNA (M-NPD)。对两种双层膜的 MD 模拟比较表明，ST60/Ergo 囊泡具有更好的基因传递性能。我们的体外实验结果证实了计算结果，并表明与 Chol-niosome 相比，Ergo-niosomes 具有更小的粒径、更好的多分散性和更慢的质粒释放。此外，与 M-NPD-Chol 囊泡相比，M-NPD-Ergo 在 HEK-293T 细胞系中表现出更高的细胞摄取和基因表达。
