Liu Liang, Yang Zhaojun, Liu Chaobing, Wang Mengying, Chen Xin
School of Life Science and Technology, Wuhan Polytechnic University, Wuhan, China.
Biotechnol Appl Biochem. 2023 Apr;70(2):824-834. doi: 10.1002/bab.2402. Epub 2022 Sep 19.
Achieving efficient and safe gene delivery is of great significance to promote the development of gene therapy. In this work, a polydopamine (PDA) layer was coated on the surface of Fe O nanoparticles (NPs) by dopamine (DA) self-polymerization, and then magnetic Fe O NPs were prepared by the Michael addition between amino groups in polyethyleneimine (PEI) and PDA. The prepared Fe O NPs (named Fe O @PDA@PEI) were characterized by Fourier transform infrared (FTIR), atomic force microscopy (AFM), and scanning electron microscopy (SEM). As an efficient and safe gene carrier, the potential of Fe O @PDA@PEI was evaluated by agarose gel electrophoresis, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, fluorescence microscopy, and flow cytometry. The results show that the Fe O @PDA@PEI NPs are stable hydrophilic NPs with a particle size of 50-150 nm. It can efficiently condense DNA at low N/P ratios and protect it from nuclease degradation. In addition, the Fe O @PDA@PEI NPs have higher safety than PEI. Further, the Fe O @PDA@PEI/DNA polyplexes could be effectively absorbed by cells and successfully transfected and exhibit higher cellular uptake and gene transfection efficiency than PEI/DNA polyplexes. The findings indicate that the Fe O @PDA@PEI NPs have the potential to be developed into a novel gene vector.
实现高效安全的基因递送对于促进基因治疗的发展具有重要意义。在本研究中,通过多巴胺(DA)自聚合在FeO纳米颗粒(NPs)表面包覆聚多巴胺(PDA)层,然后通过聚乙烯亚胺(PEI)中的氨基与PDA之间的迈克尔加成反应制备磁性FeO NPs。采用傅里叶变换红外光谱(FTIR)、原子力显微镜(AFM)和扫描电子显微镜(SEM)对制备的FeO NPs(命名为FeO@PDA@PEI)进行表征。作为一种高效安全的基因载体,通过琼脂糖凝胶电泳、3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法、荧光显微镜和流式细胞术对FeO@PDA@PEI的潜力进行评估。结果表明,FeO@PDA@PEI NPs是粒径为50-150 nm的稳定亲水性NPs。它能在低N/P比下有效地凝聚DNA并保护其免受核酸酶降解。此外,FeO@PDA@PEI NPs比PEI具有更高的安全性。进一步研究发现,FeO@PDA@PEI/DNA复合物能被细胞有效吸收并成功转染,且比PEI/DNA复合物表现出更高的细胞摄取和基因转染效率。这些研究结果表明,FeO@PDA@PEI NPs有潜力被开发成为一种新型基因载体。