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Cationic lipid nanosystems as carriers for nucleic acids

机译:阳离子脂质纳米系统作为核酸载体

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Solid lipid nanoparticles (SLNs) consisting of tristearin or tribehenin, and monoolein aqueous dispersions (MADs) consisting of glyceryl-monoolein have been studied as potential nanocarriers for nucleic acids. The cationic character of nanocarriers was obtained by adding cationic surfactants, such as diisobutylphenoxyethyl-dimethylbenzyl ammonium chloride (DEBDA) or PEG-15 Cocopolyamine (PCPA), to the lipid composition. The products were characterised in terms of size and morphology by Cryo-TEM and PCS. The charge properties were determined by measuring the zeta potential. Our experimental protocol enabled us to obtain homogeneous and stable cationic nanosystems within 3-6 months of production. Assessment of cytotoxicity on HepG2 cells by MTT assays indicated that MAD preparations were less toxic than SLN, and in general PCPA-containing formulations are less cytotoxic than DEBDA-containing ones. The formation of electrostatic complexes with salmon sperm or plasmid DNA, used as model nucleic acids, was evaluated by electrophoresis on agarose gel. The results confirmed that all the formulations studied are able to form the complex. Finally, we investigated the ability of SLN and MAD to deliver DNA into HepG2 cells, and to this purpose we exploited expression plasmids for green fluorescent protein or firefly luciferase. Although with reduced efficiency, the results showed that the produced nanocarriers are able to convey plasmids into cells. The data obtained encourage further study aimed at improving these new formulations and proposing them as novel in vitro transfection reagents with potential application to in vivo delivery of nucleic acids.
机译:已经研究了由甘油三硬脂精或tribehenin组成的固体脂质纳米颗粒(SLNs)和由甘油基单油精组成的单油精水分散体(MAD)作为核酸的潜在纳米载体。通过向脂质组合物中添加阳离子表面活性剂(例如二异丁基苯氧基乙基-二甲基苄基氯化铵(DEBDA)或PEG-15 Cocopolyamine(PCPA))来获得纳米载体的阳离子特性。通过Cryo-TEM和PCS对产物的尺寸和形态进行表征。通过测量ζ电势来确定电荷性质。我们的实验方案使我们能够在生产3-6个月内获得均质且稳定的阳离子纳米系统。通过MTT分析评估对HepG2细胞的细胞毒性表明,MAD制剂的毒性小于SLN,一般而言,含PCPA的制剂的细胞毒性小于含DEBDA的制剂。通过在琼脂糖凝胶上电泳来评估与鲑鱼精子或质粒DNA用作模型核酸的静电复合物的形成。结果证实,所有研究的制剂都能够形成复合物。最后,我们研究了SLN和MAD的递送DNA染HepG2细胞的能力,并为此目的,我们利用表达质粒为绿色荧光蛋白或萤火虫荧光素酶。尽管效率降低,但结果表明产生的纳米载体能够将质粒转运到细胞中。所获得的数据鼓励了进一步的研究,旨在改进这些新配方并将其作为新型的体外转染试剂提出,并有望应用于核酸的体内递送。

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