Chitosan nanoparticles as non-viral gene delivery vehicles based on atomic force microscopy study
Chitosan (CS), a biocompatible and biodegradable material, can act as a non-viral delivery vehicle with low toxicity. In this study, plasmid DNA (pDNA) and siRNA were encapsulated in CS nanoparticles (NPs) to prepare CS–DNA and CS–siRNA NPs using a complex coacervation process. The CS–DNA particle size was within the range of 180–370 nm with a surface charge ranging from 0 to 18 mV at pH 5.5. The stability of pDNA in CS–DNA was investigated by pDNA release study and DNase I protection assay. The release of pDNA from NPs was studied in pH 7.4 phosphate-buffered saline at 37°C and the CS–DNA NPs could delay the DNA release. Results of DNase I protection assay showed that CS–DNA NPs could protect the encapsulated pDNA from nuclease degradation. In the transfection study, it was found that the transfection efficiency in vitro was dependent on the molecular weight, charge ratio, and DNA concentration of the CS–DNA NP as well as the type of cell transfected. Moreover, the morphology of HeLa cells transfected with CS–siRNA complexes was studied using atomic force microscopy. The results suggest that CS may be more capable than liposome in delivering siRNA to target cells. In summary, our analysis suggests that pDNA and siRNA can be encapsulated in CS NPs without being damaged.