SHORT CHAIN nanoparticle formulation CHITOSAN-pEGFP-C1 transfection AND CAPABILITY IN CELL CULTURE Cervical cancer BY IN VITRO
ABSTRACT: The use of interferon combined with chemotherapy agents on cancer therapy could damage not only cancer cells but also normal cells. Nanoparticles delivery system using DNA and polycationic polymers, like chitosan, is one of the promising delivery systems to deliver non-viral genes. The purposes of this study were to make nanoparticles using short chain chitosan conjugated with plasmid Enhanced Green Fluorescence Protein (pEGFP-C1) on pH 4.0 and pH 5.0, to see their cytotoxicity and transfection ability on Hela Cell culture. Electrophoresis was used to see the forming of short chain chitosan- pEGFP-C1 nanoparticles complex. The size and the size distribution of nanoparticles were determined using Particle Size Analyzer (PSA), and their morphologies were determined using Transmission Electron Microscopy (TEM). The stability studies were done to see the stability of the nanoparticles complex in DNAse and blood serum. The cytotoxicity and transfection abilities of nanoparticles complex were determined on Hela cell. This study produces short chain chitosan- pEGFP-C1 nanoparticles with average size diameter 190.5-623.3 nm and 0.988 polydispersity index. It has a spherical morphology. The nanoparticles system could improve the stability of pEGFP-C1 in DNAse and blood serum. The results show that pEGFP-C1 nanoparticles has relatively low toxicity to Hela cell culture with viability more than 70% and the nanoparticles could be transfected in to the Hela cell culture.
