Effect of gold nanoparticles on application for drug release and stem cell differentiation

• Main Authors: Chih-Hsuan Chang, 張智鉉
• Other Authors: Huey-Shan Hung
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/jm54n3

碩士 === 中國醫藥大學 === 基礎醫學研究所碩士班 === 102 === Chitosan is natural polymer macromolecules which is composed of beta-1,4 D-glucosamine and N-vinyl D glucosamine. It is widely used in the field of physical medicine and biotechnology because it is not toxic and has property of anti-bacteria. Now the reason of anti-bacteria is known that chitosan in solution produce hydrophobic effect and carry a positive charge cause bacteria can not grow. Nanogold particles is materials widely used in biomedical, which generally have two kinds of process to produce nanogold particles. One is use of chemical processes tetrachloroauric acid to synthesis chemical nanogold particles, another is use of physical way to make physical nanogold particles. The chemical nanogold particles more likely have chemical residues caused cytotoxicity, so this study is use chitosan mixed with two different nanogold particles in different concentrations(25,50,100 ppm) to form Chitosan nanogold(Chi-AuNP), and then to observe biocompatibility using mesenchymal stem cells(MSCs) from human umbilical in vitro. The surface of the nano substrate analyzed by UV / Visible spectrophotometer (UV-Vis) and Fourier Transform Infrared Spectrometer (FTIR). In order to define the biocompatibility of Chi-AuNP with different nanogold particles, we test platelet adhesion, monocytes transfer to macrophages by CD68 staining to observe inflammation, and ROS test against oxidative stress as a material compatibility testing experiments. Next, to investigate the differentiation of mesenchymal stem cells induced by Chi-AuNP nano substrate, we use the dynamic real-time polymerase chain reaction test to observe RNA expression, and use neural class primer GFAP, beta-tubulin, Nestin, vascular endothelial cells class primer CD31, vWF, adipocyte cell class primer PPAR, and bone cell class primer RunX 2 to observe differentiation ability of mesenchymal stem cells by Chi-AuNP induced. Then, to determine whether mesenchymal stem cells differentiate successful, we used immunofluorescence staining to determine and see the protein levels. In the neural class we used GFAP, beta-tubulin, Nestin as marker, and in the blood vessels endothelial growth class, we use CD31, vWF as marker. We also use ORO (Oil red-O Staining) as osteoblasts marker, and ARS (Alizarin Red S Staining) as adipocytes marker to determine.