Self-assembled biodegradable micellar nanoparticles of amphiphilic and cationic block copolymer for siRNA delivery
Self-assembled biodegradable micellar nanoparticles of amphiphilic and cationic block copolymer for siRNA delivery
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DOI：
10.1016/j.biomaterials.2008.07.036
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发表时间：
2008-11-01
期刊：
影响因子：
14
通讯作者：
Wang, Jun
中科院分区：
文献类型：
--
作者：
Sun, Tian-Meng;Du, Jin-Zhi;Wang, Jun
设计并合成了由单甲氧基聚（乙二醇）、聚（ε-己内酯）（PCL）和聚（2-氨基乙基亚乙基磷酸酯）组成的新型两亲性阳离子三嵌段共聚物，表示为mPEG（45）-b-PCL 100-b-PPEEA（12），用于siRNA递送。用核磁共振氢谱和凝胶渗透色谱对共聚物进行了表征。用动态光散射、透射电镜和荧光技术研究了该两亲性共聚物在水溶液中胶束纳米粒子的形成。MNP呈均匀的球形形态，zeta电位约为45 mV，并通过PCL核中的疏水-疏水相互作用稳定，表现出临界胶束浓度为2.7 x 10（-3）mg/mL。这样的MNP允许在纳米颗粒形成后siRNA负载而不改变均匀性。根据N/P比，siRNA结合后纳米颗粒的平均直径范围为98至125 nm。装载siRNA的纳米颗粒可以有效地内化并随后在HEK 293细胞中释放siRNA，导致显著的基因敲除活性，这通过递送靶向绿色荧光蛋白（GFP）的两种siRNA来证明。它有效地沉默了40- 70%GFP表达的HEK 293细胞中的GFP表达，并且观察到较高的N/P比导致更有效的沉默，这可能是由于在较高的N/P比下更好的细胞内化。MTT法表明，无论是MNP本身还是siRNA负载的MNP，即使在高浓度下也不显示细胞毒性。由生物相容性和可生物降解的聚合物制成的这种阳离子MNP有希望用于siRNA递送。(C)2008爱思唯尔有限公司保留所有权利。
A novel amphiphilic and cationic triblock copolymer consisting of monomethoxy poly(ethylene glycol), poly(epsilon-caprolactone) (PCL) and poly(2-aminoethyl ethylene phosphate) denoted as mPEG(45)-b-PCL100-b-PPEEA(12) was designed and synthesized for siRNA delivery. The copolymers were well characterized by H-1 NMR spectroscopy and gel permeation chromatography. Micelle nanoparticles'(MNPs) formation of this amphiphilic copolymer in aqueous solution was studied by dynamic light scattering, transmission electron microscopy and fluorescence technique. MNPs took uniform spherical morphology with zeta potential of around 45 mV and were stabilized by hydrophobic-hydrophobic interaction in the PCL core, exhibiting the critical micelle concentration at 2.7 x 10(-3) mg/mL. Such MNPs allowed siRNA loading post nanoparticle formation without change in uniformity. The average diameter of nanoparticles after siRNA binding ranged from 98 to 125 nm depending on N/P ratios. The siRNA loaded nanoparticles can be effectively internalized and subsequently release siRNA in HEK293 cells, resulting in significant gene knockdown activities, which was demonstrated by delivering two siRNAs targeting green fluorescence protein (GFP). It effectively silenced GFP expression in 40-70% GFP-expressed HEK293 cells and it was observed that higher N/P ratio resulted in more effective silence which was likely due to better cell internalization at higher N/P ratio. MTT assay demonstrated that neither MNPs themselves nor siRNA loaded MNPs showed cytotoxicity even at high concentrations. Such cationic MNPs made from biocompatible and biodegradable polymers are promising for siRNA delivery. (C) 2008 Elsevier Ltd. All rights reserved.