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两亲性偶氮苯-新霉素共轭物自组装成纳米结构,并将质粒DNA高效转运到哺乳动物细胞中。

Amphiphilic azobenzene-neomycin conjugate self-assembles into nanostructures and transports plasmid DNA efficiently into the mammalian cells.

作者信息

Yadav Santosh, Deka Smriti Rekha, Jha Diksha, Gautam Hemant Kumar, Sharma Ashwani Kumar

机构信息

Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi-110 007, India; Academy of Scientific and Innovative Research, New Delhi, India.

Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Mall Road, Delhi-110 007, India.

出版信息

Colloids Surf B Biointerfaces. 2016 Dec 1;148:481-486. doi: 10.1016/j.colsurfb.2016.09.008. Epub 2016 Sep 16.

Abstract

The present study demonstrates the use of self-assembled nanostructures of cationic amphiphilic azobenzene-neomycin (a small molecule) conjugate, Azo-Neo, as delivery vector for plasmid DNA. These nanostructures efficiently condensed nucleic acid and formed more compact nanoassemblies. DLS analysis showed size and zeta potential of the resulting Azo-Neo/pDNA nanoassemblies ∼153.7nm and +7.26mV, respectively. The nanoassemblies were characterized by physicochemical techniques and evaluated for its toxicity and ability to deliver nucleic acid therapeutics. The flow cytometry results on MCF-7 and HEK293T cells revealed that Azo-Neo/pDNA nanoassemblies transfected ∼31% and 23% cells, respectively, at a w/w ratio of 250, while the standard transfection reagent, bPEI/pDNA complex, could transfect only ∼21% and 29% cells, respectively, at its best w:w ratio of 2.3. MTT and hemolysis assays showed the non-toxic nature of the projected nanoassemblies and nanostructures, respectively, at various concentrations. Further, Azo-Neo nanostructures showed efficient antibacterial activity against different strains, laboratory strain of Staphylococcus aureus (MTCC 740) as well as MRSA strains (Staphylococcus aureus ATCC 33591, ATCC 43300 and ATCC 700699). These results ensure the great potential of these nanostructures in gene delivery and antimicrobial applications.

摘要

本研究展示了阳离子两亲性偶氮苯 - 新霉素(一种小分子)共轭物Azo - Neo的自组装纳米结构作为质粒DNA递送载体的应用。这些纳米结构有效地凝聚了核酸并形成了更紧凑的纳米组装体。动态光散射(DLS)分析表明,所得Azo - Neo/pDNA纳米组装体的尺寸和zeta电位分别约为153.7nm和 +7.26mV。通过物理化学技术对纳米组装体进行了表征,并评估了其毒性和递送核酸治疗剂的能力。在MCF - 7和HEK293T细胞上的流式细胞术结果显示,在w/w比为250时,Azo - Neo/pDNA纳米组装体分别转染了约31%和23%的细胞,而标准转染试剂bPEI/pDNA复合物在其最佳w:w比为2.3时,分别仅能转染约21%和29%的细胞。MTT和溶血试验表明,在不同浓度下,所预测的纳米组装体和纳米结构分别具有无毒性。此外,Azo - Neo纳米结构对不同菌株,即金黄色葡萄球菌实验室菌株(MTCC 740)以及耐甲氧西林金黄色葡萄球菌菌株(金黄色葡萄球菌ATCC 33591、ATCC 43300和ATCC 700699)表现出高效的抗菌活性。这些结果确保了这些纳米结构在基因递送和抗菌应用方面具有巨大潜力。

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