Preparation and Characterization of Nanoparticles as Carriers for Gene Delivery

Abstract

In the first part of this thesis, a comprehensive characterization of polyethylene glycol (PEG) modified AuNPs designed for imaging or diagnostic purposed was carried out to investigate the effect of the size, PEG layer conformation and grafting density on the cellular uptake, toxicity and cell cycle phases against prostate (PC3), colon (CaCo2) cancer cell lines and 3T3 Swiss fibroblast cells. It was noticed that the cellular uptake and toxicity profiles of the particles varied depending on the size, surface properties and cell type. The particles were found to show alterations in cell cycle phases by causing DNA damage without apoptotic behavior at certain doses. In the second part of this thesis, efficient multilayer small interfering RNA (siRNA) delivery systems based on gold nanoparticles (AuNPs), cationic pentablock copolymers or fusogenic peptides were developed using cleavable disulfide bonds and electrostatic interactions. siRNA/Polymer (polyplexes) and siRNA/Peptide (peptideplexes) complexes formed by direct electrostatic complexation between siRNA and the cationic pentablock copolymers or peptides were used as controls, respectively. In addition, a conjugate siRNA delivery system based on the cleavable disulfide bonds between siRNA and fusogenic peptide was also proposed as an alternative system. The siRNA activity, toxicity, cellular uptake and intracellular distribution of the developed systems were investigated against luciferase-expressing SKOV3 ovarian cancer cell line. The use of cationic block copolymers or fusogenic peptides in AuNP based multilayer systems and complex systems, provided efficient siRNA condensation and protection from nuclease enzyme and serum protein degradation, in addition to cellular uptake, endosomal escape and siRNA activity in the cytoplasm.