Assali Mohyeddin, Kittana Naim, Badran Ismail, Omari Safa
Department of Pharmacy, Faculty of Medicine and Health Sciences, An-Najah National University Nablus Palestine
Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, An-Najah National University Nablus Palestine
RSC Adv. 2023 Mar 2;13(10):7000-7008. doi: 10.1039/d3ra00727h. eCollection 2023 Feb 21.
Several approaches, including plasmid transfection and viral vectors, were used to deliver genes into cells for therapeutic and experimental purposes. However, due to the limited efficacy and questionable safety issues, researchers are looking for better new approaches. Over the past decade, graphene has attracted tremendous attention in versatile medical applications, including gene delivery, which could be safer than the traditional viral vectors. This work aims to covalently functionalize pristine graphene sheets with a polyamine to allow the loading of plasmid DNA (pDNA) and enhance its delivery into cells. Graphene sheets were successfully covalently functionalized with a derivative of tetraethylene glycol connected to polyamine groups to improve their water dispersibility and capacity to interact with the pDNA. The improved dispersibility of the graphene sheets was demonstrated visually and by transmission electron microscopy. Also, it was shown by thermogravimetric analysis that the degree of functionalization was about 58%. Moreover, the surface charge of the functionalized graphene was +29 mV as confirmed by zeta potential analysis. The complexion of f-graphene with pDNA was achieved at a relatively low mass ratio (10 : 1). The incubation of HeLa cells with f-graphene loaded with pDNA that encodes enhanced green fluorescence protein (eGFP) resulted in the detection of fluorescence signal in the cells within one hour. f-Graphene showed no toxic effect . Density functional theory (DFT) and quantum theory of atoms in molecules (QTAIM) calculations revealed strong binding with Δ = 74.9 kJ mol. QTAIM between the f-graphene and a simplified model of pDNA. Taken together, the developed functionalized graphene could be used for the development of a new non-viral gene delivery system.
包括质粒转染和病毒载体在内的几种方法被用于将基因导入细胞以用于治疗和实验目的。然而，由于疗效有限和安全性问题存疑，研究人员正在寻找更好的新方法。在过去十年中，石墨烯在包括基因递送在内的多种医学应用中引起了极大关注，其可能比传统病毒载体更安全。这项工作旨在用多胺对原始石墨烯片进行共价功能化，以实现质粒DNA（pDNA）的负载并增强其向细胞内的递送。石墨烯片成功地与连接有多胺基团的四甘醇衍生物进行了共价功能化，以改善其水分散性和与pDNA相互作用的能力。通过肉眼观察和透射电子显微镜证明了石墨烯片分散性的改善。此外，热重分析表明功能化程度约为58%。此外，通过zeta电位分析证实功能化石墨烯的表面电荷为+29 mV。功能化石墨烯与pDNA在相对较低的质量比（10∶1）下实现了络合。用负载有编码增强型绿色荧光蛋白（eGFP）的pDNA的功能化石墨烯孵育HeLa细胞，一小时内即可在细胞中检测到荧光信号。功能化石墨烯未显示出毒性作用。密度泛函理论（DFT）和分子中原子的量子理论（QTAIM）计算表明，功能化石墨烯与pDNA的简化模型之间具有很强的结合力，Δ = 74.9 kJ/mol。综上所述，所开发的功能化石墨烯可用于开发一种新型非病毒基因递送系统。
