Development of pH-responsive nanocarriers using trimethylchitosans and methacrylic acid copolymer for siRNA delivery
Development of pH-responsive nanocarriers using trimethylchitosans and methacrylic acid copolymer for siRNA delivery
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DOI：
10.1016/j.biomaterials.2009.11.028
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发表时间：
2010-03-01
期刊：
影响因子：
14
通讯作者：
Evrard, B.
中科院分区：
文献类型：
--
作者：
Dehousse, V.;Garbacki, N.;Evrard, B.
基于RNA干扰的治疗依赖于细胞内siRNA的传递。SiRNA的释放可能是转导过程中的一个限速步骤。本研究的目的是制备三甲基壳聚糖(TMC)的pH响应型纳米载体。为此，在TMC-siRNA配方中添加了对pH敏感的甲基丙烯酸(MAA)共聚物。四种不同的TMC与MAA相关或不与MAA相关作为siRNA载体进行评估。从纳米粒子的尺寸、表面电荷、形貌以及与siRNA的相互作用等方面对其进行了表征。实验中观察到了由于pH降低而引起的溶胀行为，并发现溶胀行为依赖于络合物中MAA的含量。体外实验旨在评估纳米载体在L929细胞中的siRNA转染能力如何受MAA含量的影响。共聚焦显微镜实验表明，与不含这种pH敏感聚合物的荧光粒子相比，含有MAA的荧光粒子在细胞内呈现出不同的分布模式。用抑制RhoA基因表达的方法检测细胞的转染率。MAA-TMC-siRNA复合体对L929细胞的转染率高于相应的TMC-siRNA复合体。这项研究深入了解了pH敏感的纳米载体用于siRNA传递的机会。这些制剂可能是改善siRNA内体逃逸的一种有吸引力的策略。(C)2009爱思唯尔有限公司。保留所有权利。
RNA interference-based therapies are dependent on intracellular delivery of siRNA. The release of siRNA from the endosomal compartment may be a rate limiting step in the transfection process. The purpose of this study was to produce pH-responsive nanocarriers made of trimethylchitosan (TMC). To this end, pH-sensitive methacrylic acid (MAA) copolymer was added to TMC-siRNA formulations. Four different TMCs associated or not with MAA were evaluated as siRNA carriers. Nanoparticles were characterized in terms of size, surface charge, morphology and interaction with siRNA. A swelling behaviour due to a decrease in pH was observed and was found to be dependent on MAA content in the complexes. In vitro experiments aimed at evaluating how the capacity of the nanocarriers to transfect siRNA in L929 cells was affected by MAA content. Confocal microscopy experiments showed that fluorescent MAA-containing particles exhibit a different distribution pattern inside the cells comparing to their counterpart without this pH-sensitive polymer. Transfection efficiency was investigated by RhoA mRNA expression inhibition. MAA-TMC-siRNA complexes were able to transfect L929 cells with greater efficiency than corresponding TMC-siRNA complexes. This study gives an insight into the opportunity of pH-sensitive nanocarriers for siRNA delivery. Such formulations may represent an attractive strategy to improve endosomal escape of siRNA. (C) 2009 Elsevier Ltd. All rights reserved.