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Enhanced siRNA Delivery and Silencing Gold–Chitosan Nanosystem with Surface Charge-Reversal Polymer Assembly and Good Biocompatibility

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CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, National Center for Nanoscience and Technology, Beijing 100190, People’s Republic of China
Laboratory of Molecular Iron Metabolism, College of Life Science, Hebei Normal University, Shijiazhuang 050016, People’s Republic of China
Cite this: ACS Nano 2012, 6, 8, 7340–7351
Publication Date (Web):July 27, 2012
https://doi.org/10.1021/nn3024688
Copyright © 2012 American Chemical Society

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    Abstract

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    A simple nanocarrier coated with chitosan and the pH-responsive charge-reversible polymer, PAH-Cit, was constructed using layer-by-layer assembly to deliver siRNA. Gold nanoparticles (AuNPs) were di-rectly reduced and stabilized by chitosan (CS), forming a positively charged AuNP-CS core. Charge-reversible PAH-Cit and polyethylenimine (PEI) were sequentially deposited onto the surface of AuNP-CS through electrostatic interaction, forming a PEI/PAH-Cit/AuNP-CS shell/core structure. After loading siRNA, the cytotoxicity of siRNA/PEI/PAH-Cit/AuNP-CS against HeLa and MCF-7R cells was negligible. This vehicle completely protected siRNA against enzymatic degradation at vector/RNA mass ratios of 2.5:1 and above. An in vitro release profile demonstrated an efficient siRNA release (79%) from siRNA/PEI/PAH-Cit/AuNP-CS at pH 5.5, suggesting a pH-induced charge-reversing action of PAH-Cit. This mechanism also worked in vivo and facilitated the escape of siRNA from endosomes. Using this carrier, the uptake of cy5-siRNA by HeLa cells was significantly increased compared to PEI, an efficient polycationic transfection reagent. In drug-resistant MCF-7 cells, specific gene silencing effectively reduced expression of MDR1, the gene encoding the drug exporter P-gp, and consequently promoted the uptake of doxorubicin. This simple charge-reversal polymer assembly nanosystem has three essential benefits (protection, efficient uptake, and facilitated escape) and provides a safe strategy with good biocompatibility for enhanced siRNA delivery and silencing.

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