| Summary: | 碩士 === 中原大學 === 醫學工程研究所 === 96 === The goal of the study was to explore the effects of biocompatibility on the of human bone marrow mesenchymal stem cells ( SV40T-hTERT-hBMSC，hBMSCs ) and human hepatoma cells ( HuH-7 ) using water-soluble gold nanoparticles with different sizes and concentrations. The effect of cytotoxicity was analyzed with MTT assay at different time points after co-culturing with gold nanoparticles. The results showed that low concentration of gold nanoparticles exhibited more than 80% cell survival in two different types of cells. However, cell survival decreased after incubation with high concentration of gold nanoparticles for a longer period of time. From the results of Annexin V and propidium iodide ( PI ) staining, the percentage of PI positive cells increased with the concentration of gold nanoparticles, indicating that high concentration led to cell death via necrosis. The level of reactive oxygen species ( ROS ) increased after co-culturing with gold nanoparticles, suggesting that the elevation of ROS level might be the reason why necrosis occurred after gold nanoparticles were endocytosed by cells. The results from osteogenic hBMSCs showed that the activity of alkaline phosphatase and the level of calcification were both inhibited by addition of gold nanoparticles. Similar results were observed with adipogenic hBMSCs, in which the accumulation of triacylglycerides was repressed by addition of gold nanoparticles. Gold nanoparticles were found to localize within the vesicles and nucleus after endocytosis from TEM images. Besides, gold nanoparticles conjugated with FITC ( GNP-FITC ) were prepared in this study. The diameter of GNP-FITC was around 50 nm by TEM and DLS analyses. The feasibility of the method to prepare GNP-FITC was demonstrated by agarose gel electrophoresis and FTIR analysis, in which gold nanoparticles with positive surface charge were conjugated with FITC with negative charge. The study has not only accomplished the biocompatibility study of water-soluble gold nanoparticles but also prepared fluorescent GNP-FITC successfully. GNP-FITC will be applied in cell imaging as well as cell tracing in the field of cell biology.
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