POTENTIAL OF CATIONIC LIPOSOMES AND CHITOSAN NANOPARTICLES FOR DELIVERY DNA VACCINE MODEL NTC8685-EGFP

Abstract

The development of livestock vaccines has important significance in preventing livestock industry losses due to infectious diseases. DNA vaccines that have the potential as a preventive effort against viral diseases have a weakness in the form of low efficacy due to limitation in the delivery system. This study aims to explore the delivery system in the form of liposomes-DNA and chitosan-DNA nanoparticles for DNA vaccine delivery. The making of chitosan-DNA nanoparticles using complex coarsevation method with the ratio of DNA and chitosan mass is 1: 0.5; 1: 1,0; 1: 2,0; and 1: 4,0. Observation of complex formation with gel retardation assay. Liposome-DNA complex was made using LipofectamineTM 3000. In vitro tests were carried out on HeLa cells with a confluent rate of 70% -90%. Chitosan-DNA nanoparticles and liposome-DNA complex with a total of 8 ?g DNA components were transfected for 4 hours then replaced with new growth media and further incubated for 24 hours and analyzed by eGFP protein glow with confocal microscopy. Chitosan-DNA complex was successfully made and visualized as a band of DNA held in agarose wells. Visualization shows that nanoparticles with a mass ratio of 1: 2 are a minimum ratio of chitosan that can trap the whole plasmid DNA to form polyplex. The delivery is able to show the presence of luminescence under a confocal microscope. Fluorescence on chitosan DNA nanoparticles sample qualitatively tends to be less when compared to luminescence in samples with liposome-DNA treatment. The vaccine vector NTC8685-eGFP through the cationic liposome delivery system and chitosan nanoparticles can enter the HeLa cell (model) so that it can be expressed as eGFP protein. Cationic liposomes qualitatively have a higher effectiveness than chitosan nanoparticles in delivering NTC8685-eGFP plasmids into HeLa cells.