In this study, the release of cisplatin, loaded on chitosan nanoparticles applied with the appropriate dose and duration in HeLa cancer cell line and its apoptotic, necrotic effects and the expression of glutathione-S-transferase pi isoenzymes in HeLa cancer cells were investigated. Chitosan nanoparticles were synthesized with low molecular weight chitosan by treating with cross-linking, TPP (tripolifosfat). Chitosan nanoparticles size were found to be between 100-200 nm with Scanning Electron Microscope (SEM) and Atomic Force Microscope (AFM). The release of cisplatin loaded on nanoparticles was detected by Graphite Furnace-Atomic Adsorption Spectrometer (GF-AAS). Chitosan nanoparticles loaded with cisplatin (CDDP) and chitosan nanoparticles were incubated with HeLa cancer cells. Images of cells fine structure and their interaction with nanoparticles were observed with Scanning Electron Microscope (SEM). The cytotoxic effects of cisplatin, chitosan nanoparticles and chitosan nanoparticles loaded with cisplatin to HeLa cancer cells were detected by WST-1 assay method. It was observed that chitosan nanoparticles were not toxic, but its toxicity was increased by increasing dose of cisplatin. After the cisplatin and nanoparticles loaded with cisplatin administration into HeLa cells, apoptotic index by the caspase-3 immunocytochemical and double stainings, necrotic index were examined. It was found that their indexes obtained by both staining were parallel, 60±5%, 53±4% in high cisplatin dose respectively, and 64.2±3%, 60.3±2% in necrotic index, respectively. The increasing apoptotic effect were determined. The relationship of GST pi expression and its substrate cisplatin doses were studied in HeLa cancer cells by immunocytochemical staining. GST pi expression was found as 63±2%. |