Development and characterisation of chitosan nanoparticles for siRNA delivery


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Abstract

Gene silencing mediated by double-stranded small interfering RNA (siRNA) has been widely investigated as a potential therapeutic approach for diseases with genetic defects. The use of siRNA, however, is hampered by its rapid degradation and poor cellular uptake into cells in vitro or in vivo. Therefore, we have explored chitosan as a siRNA vector due to its advantages such as low toxicity, biodegradability and biocompatibility. Chitosan nanoparticles were prepared by two methods of ionic cross-linking, simple complexation and ionic gelation using sodium tripolyphosphate (TPP). Both methods produced nanosize particles, less than 500 nm depending on type, molecular weight as well as concentration of chitosan. In the case of ionic gelation, two further factors, namely chitosan to TPP weight ratio and pH, affected the particle size. In vitro studies in two types of cells lines, CHO K1 and HEK 293, have revealed that preparation method of siRNA association to the chitosan plays an important role on the silencing effect. Chitosan–TPP nanoparticles with entrapped siRNA are shown to be better vectors as siRNA delivery vehicles compared to chitosan–siRNA complexes possibly due to their high binding capacity and loading efficiency. Therefore, chitosan–TPP nanoparticles show much potential as viable vector candidates for safer and cost-effective siRNA delivery.

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