Chitosan-modified poly(D,L-lactide-co-glycolide) nanospheres for plasmid DNA delivery and HBV gene-silencing

Gene silencing using small interfering RNA (siRNA) has several potential therapeutic applications. In the present study, we investigated nanoparticles (NS) formulated using the biodegradable polymer, poly(D,L-lactide-co-glycolide) (PLGA) for plasmid DNA (pDNA) delivery. A cationic polymer, Chitosan (CHS), was incorporated in the PLGA matrix to improve pDNA loading efficiency and cellular uptake ability. PLGA-CHS NS were prepared by a spontaneous emulsion diffusion (SED) method, and various formulation factors were investigated. Spherical nanoparticles with particle size of around 60 nm were obtained under optimum formulation condition. The effectiveness of pDNA-loaded PLGA-CHS nanopartides in expressing the indicative enhanced Green Fluorescent Protein (eGFP) and in slicing Hepatitis B virus (HBV) gene were examined in HepG2.2.15 cells. CHS-modified PLGA NS exhibited much higher loading efficiency than unmodified PLGA NS. CHS-PLGA NS showed a positive zeta potential, while plain-PLGA NS were negatively charged. EGFP expression studies by observation with confocal leaser scanning microscopy (CLSM) indicated that pDNA-loaded CHS-PLGA NS were more effectively taken up by the cells than plain-PLGA NS. The corresponding results showed that the HBV gene-silencing efficiency of CHS-PLGA NS was higher than those of plain-PLGA NS and naked pDNA. Thus, CHS-PLGA NS containing pDNA could provide an effective pDNA delivery system in vitro, showing that such an approach could be useful in the treatment of viral diseases in vivo. (C) 2011 Elsevier B.V. All rights reserved.