Vakilian Haghighat, Andres Rojas Eduardo, Habibi Rezaei Lida, Behmanesh Mehrdad
Nano biotechnology Department, Faculty of Bioscience, Tarbiat Modares University, Tehran, Iran.
Department of Pathology and Molecular Medicine, McMaster Immunology Research Centre, McMaster University, Hamilton, ON, Canada.
Rep Biochem Mol Biol. 2020 Oct;9(3):297-308. doi: 10.29252/rbmb.9.3.297.
One of the major challenges in gene therapy is producing gene carriers that possess high transfection efficiency and low cytotoxicity (1). To achieve this purpose, crystal nanocellulose (CNC) -based nanoparticles grafted with polyethylenimine (PEI) have been developed as an alternative to traditional viral vectors to eliminate potential toxicity and immunogenicity.
In this study, CNC-PEI10kDa (CNCP) nanoparticles were synthetized and their transfection efficiency was evaluated and compared with linear cationic PEI10kDa (PEI) polymer in HEK293T (HEK) cells. Synthetized nanoparticles were characterized with AFM, FTIR, DLS, and gel retardation assays. gene delivery efficiency by nano-complexes and their effects on cell viability were determined with fluorescent microscopy and flow cytometry.
Prepared CNC was oxidized with sodium periodate and its surface cationized with linear PEI. The new CNCP nano-complex showed different transfection efficiencies at different nanoparticle/plasmid ratios, which were greater than those of PEI polymer. CNPC and Lipofectamine were similar in their transfection efficiencies and effect on cell viability after transfection.
CNCP nanoparticles are appropriate candidates for gene delivery. This result highlights CNC as an attractive biomaterial and demonstrates how its different cationized forms may be applied in designing gene delivery systems.
基因治疗的主要挑战之一是制备具有高转染效率和低细胞毒性的基因载体(1)。为实现这一目的，已开发出接枝聚乙烯亚胺(PEI)的基于结晶纳米纤维素(CNC)的纳米颗粒，作为传统病毒载体的替代品，以消除潜在的毒性和免疫原性。
在本研究中，合成了CNC-PEI10kDa(CNCP)纳米颗粒，并在HEK293T(HEK)细胞中评估了其转染效率，并与线性阳离子PEI10kDa(PEI)聚合物进行了比较。用原子力显微镜(AFM)、傅里叶变换红外光谱(FTIR)、动态光散射(DLS)和凝胶阻滞试验对合成的纳米颗粒进行了表征。用荧光显微镜和流式细胞术测定了纳米复合物的基因传递效率及其对细胞活力的影响。
制备的CNC用过碘酸钠氧化，其表面用线性PEI阳离子化。新的CNCP纳米复合物在不同的纳米颗粒/质粒比例下表现出不同的转染效率，高于PEI聚合物。CNPC和脂质体转染效率及其对转染后细胞活力的影响相似。
CNCP纳米颗粒是基因传递的合适候选物。这一结果突出了CNC作为一种有吸引力的生物材料，并展示了其不同的阳离子化形式如何应用于设计基因传递系统。
