Chitosan-Mediated siRNA Delivery <Emphasis Type="Italic">In Vitro</Emphasis>: Effect of Polymer Molecular Weight,Concentration and Salt Forms |
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Authors: | Sunee Techaarpornkul Sirirat Wongkupasert Praneet Opanasopit Auayporn Apirakaramwong Jurairat Nunthanid Uracha Ruktanonchai |
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Institution: | (1) Faculty of Pharmacy, Silpakorn University, Nakhon Pathom, 73000, Thailand;(2) National Nanotechnology Center, Thailand Science Park, Pathumthani, 12120, Thailand; |
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Abstract: | The aim of this study was to investigate chitosan/siRNA complexes formulated with various chitosan salts (CS) including chitosan
aspartate (CS-Asp), chitosan glutamate (CS-Glu), chitosan acetate (CS-Ac), and chitosan hydrochloride (CS-HCl) for in vitro siRNA delivery into stable and constitutive enhanced green fluorescent protein (EGFP)-expressing HeLa cells. The CS/siRNA
complexes were characterized by 2% agarose gel electrophoresis and investigated for their transfection efficiency in stable
and constitutive EGFP-expressing HeLa cells. The cytotoxicity of the complexes was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl
tetrazolium bromide (MTT) assay. The formation of complexes CS/siRNA is mainly dependent on the weight ratio, whereas salt
form and molecular weight has less effect. The particle sizes of the complete complexes were in the range of 270–373 nm except
the complete complex of CS-Ac, with a slightly positive charge of less than 2 mV. The ability of CS to transfer functionally
active siRNA into cell culture is mainly dependent on the weight ratio and molecular weight of CS whereas salt form of CS
has less effect. The high gene-silencing efficiency was observed with low MW of CS (20 kDa) and high weight ratio of 32. Over
80% average cell viabilities were observed for CS/siRNA complexes in all weight ratios comparison to untreated cells. This
study suggests CS salts have the potential to be used as safe siRNA delivery vectors. |
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