Preparation and Physicochemical Characterization of Chitosan-βCyclodextrin-Plasmid DNA Nanoparticles Encoding Interleukin-12

Abstract

Background and Objectives: Interleukin-12 (IL-12) as a cytokine has been proved to possess antitumor effects via stimulating the immune system. Non-viral gene delivery systems offer several advantages, including easiness in production, low cost, safety, low immunogenicity and can carry higher amounts of genetic material without limitation on their sizes.The main goal of the present study was to investigate the potential of a new generation of hybrid polysaccharide nanocarriers, composed of chitosan (CS) and cyclodextrins (CD), for gene delivery. To this end, chitosan/β-cyclodextrin-plasmid DNA (pUMVC3-mIL12) nanoparticles encoding interleukin-12 were prepared and its physicochemical characteristic and cytotoxicity against the CT-26 colon carcinoma cell line were evaluated. Materials and Methods: pUMVC3-mIL12 loaded chitosan/β-cyclodextrin nanoparticles were prepared using ionotropic gelation method and characterized in terms of size, zeta potential, polydispersity index, morphology, loading efficiency and cytotoxicity against the CT-26 colon carcinoma cell line. Results: pUMVC3-mIL12 loaded chitosan/β-cyclodextrin nanoparticles were prepared via ionotropic gelation method. Complex formation between polymers and DNA was demonstrated using agarose gel electrophoresis. All prepared particles were spherical in shape and nanometric in size (171.3±2.165 nm, PdI: 0.231±0.014) and exhibited a positive zeta potential (34.3±1.55). The nanoparticles demonstrated good DNA loading efficiencies (83.315% ± 2.067). Prepared pUMVC3-mIL12 loaded chitosan/β-cyclodextrin nanoparticles showed no acute cell toxicity in murine CT26 colon carcinoma cells. Conclusion: Chitosan-cyclodextrin-plasmid DNA nanoparticles encoding interleukin 12 prepared using ionotropic gelation method with no toxic effect on test cells can potentially be applied to enhance the expression of IL-12 in target cells.