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首页> 外文期刊>Nanotechnology >Synthesis and evaluation of diethylethylamine-chitosan for gene delivery: Composition effects on the in vitro transfection efficiency
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Synthesis and evaluation of diethylethylamine-chitosan for gene delivery: Composition effects on the in vitro transfection efficiency

机译：二乙基乙胺-壳聚糖的合成和基因传递评估：组成对体外转染效率的影响

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摘要

Chitosan has been indicated as a safe and promising polycation vector for gene delivery. However its low transfection efficiency has been a challenging obstacle for its application. To address this limitation, we synthesized chitosan derivatives which had increasing amounts of diethylethylamine groups (DEAE) attached to the chitosan main chain. The plasmid DNA VR1412 (pDNA), encoding the ?-galactosidase (?-gal) reporter gene was used to prepare nanoparticles with the chitosan derivatives, and the transfection studies were performed with HeLa cells. By means of dynamic light scattering and zeta potential measurements, it was shown that diethylethylamine-chitosan derivatives (DEAE_x-CH) were able to condense DNA into small particles having a surface charge depending on the polymer/DNA ratio (N/P ratio). Nanoparticles prepared with derivatives containing 15 and 25% of DEAE groups (DEAE_(15)-CH and DEAE_(25)-CH) exhibited transfection efficiencies ten times higher than that observed with deacetylated chitosan (CH). For derivatives with higher degrees of substitution (DS), transfection efficiency decreased. The most effective carriers showed low cytotoxicity and good transfection activities at low charge ratios (N/P). Vectors with low DS were easily degraded in the presence of lysozyme at physiological conditions in vitro and the nontoxicity displayed by these vectors opens up new opportunities in the design of DEAE-chitosan-based nanoparticles for gene delivery.
机译：壳聚糖已被证明是用于基因递送的安全且有希望的聚阳离子载体。然而，其低转染效率一直是其应用的挑战性障碍。为了解决这一局限性，我们合成了壳聚糖衍生物，该衍生物具有越来越多的连接到壳聚糖主链上的二乙乙基胺基（DEAE）。使用编码β-半乳糖苷酶（β-gal）报告基因的质粒DNA VR1412（pDNA）制备具有壳聚糖衍生物的纳米颗粒，并用HeLa细胞进行转染研究。通过动态光散射和ζ电势测量，表明二乙基乙胺-壳聚糖衍生物（DEAE_x-CH）能够将DNA冷凝成具有表面电荷的小颗粒，具体取决于聚合物/ DNA的比率（N / P比率）。用含有15％和25％的DEAE基团的衍生物（DEAE_（15）-CH和DEAE_（25）-CH）制备的纳米颗粒的转染效率是脱乙酰壳聚糖（CH）的十倍。对于具有较高取代度（DS）的衍生物，转染效率降低。最有效的载体在低电荷比（N / P）下显示出低细胞毒性和良好的转染活性。具有低DS的载体在体外在生理条件下在溶菌酶的存在下很容易降解，这些载体显示的无毒性为基于DEAE-壳聚糖的纳米粒基因递送设计开辟了新的机会。

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