Formulation of Nanoparticles from Ahort Chain Chitosan As Gene Delivery System and Transfection Against T4&D Cell Line

Abstract

Recently numerous prototype DNA-based biopharmecuticals can be used to control disease progression by induction and inhibitin the overexpression of Genes. Since there are poor cellular uptake and rapid in vivo the overexpression of DNA-based therapeutics therefore the use of delivery systems to facilitate cellular internalization and preserve their activity is necessary. Cationic polymers dommonly used as carriers to delivery gene because of easy to form complexes and higher stalibity compared to that lipoplex. Chitosan, a cationic, are polymer most widely used in gene delivery systems because of the low tocity, and blocompatible. The aim pf this study was to formulate nanoparticles of short chain chitosan-pEGPP C1 and short chain chitosan/TPP-pEGFP-C1 by coaservation complex method. Stability test of the formula was performed by incubating the anoparticles complex with DN ase 1 and artificial Intestinal Fluid. Cytotoxicity and transfection studies were evaluated aganst T470 cell line. The diameter of chitosan-pEGFP-C1 chistosan/TPP-PEGFP-C1 nanoparticles were +16.6 mV. Stability studies showed that potential was determined to Be +14.03-+16.6 mV. Stability studies showed that chitosan-PEGP-C1 and chitosan/TPP-pEGFP-C1 nanopaartides were stable, undegradable by Dnase 1 and artificial intestinal fluid. Cytotoxic Assay of Chitosasn-pEEGFP-C1 and chitosan/TPP-pEGFP- C1 nanoparticles (pH 4.0) showed that the vability of cell was > 90% for allformulas. EGFP-C1 plasmid gene delivered by chitosan nanoparticles can be expressed in T47D cell culture. According to these results chitosan and chitosan/TPP nanoparticles had potentially to be used as a non-viral vector