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通过多巴胺自聚合制备聚乙烯亚胺修饰的纳米颗粒用于高效DNA递送

Preparation of PEI-modified nanoparticles by dopamine self-polymerization for efficient DNA delivery.

作者信息

Liu Liang, Yang Zhaojun, Liu Chaobing, Wang Mengying, Chen Xin

机构信息

School of Life Science and Technology, Wuhan Polytechnic University, Wuhan, China.

出版信息

Biotechnol Appl Biochem. 2023 Apr;70(2):824-834. doi: 10.1002/bab.2402. Epub 2022 Sep 19.

Abstract

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.

摘要

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。

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