The Enhanced Effect of Static Magnetic Fields on Ultrasound-Microbubble Medicated Gene Delivery In Vitro

• Main Authors: Wei-Wen Chien, 簡瑋文
• Other Authors: Chih-Cheng Lu
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/qj4ttd

碩士 === 國立臺北科技大學 === 機電整合研究所 === 98 === The purpose of this study is to envestigate the enhanced impact of different ultrasound duty cycles (DC) combined with static magnetic fields (SMFs) upon the ultrasound-microbubble mediated transfection on gene delivery. By using microbubbles that can generate cavitation to increase cell membrane permeability, and promote the pGL3 luciferase report gene transported into cells for improved gene transfer efficiency, an ultrasound-assisted system was established for in vitro gene delivery experiments. The target cells were exposed to the static magnetic fields produced by Nd-Fe-B permanent magnets in an incubator for 48 hours. Experiments employing the pGL3 luciferase report gene to be delivered to HAEC (human aortic endothelial cells) for gene expression and cell viability were carried out with specific parametric sets of ultrasound duty cycle and acoustic intensity and Artison® microbubbles. Our experimental results conclude that the gene expression is not significantly increased when using ultrasound alone, but it is significantly increased (p<0.001) when using ultrasound-mediated microbubble compared to the control set. Moreover, the gene transfer efficiency will accordingly improve as the ultrasound duty cycle was increased. Most importantly, the intensity of gene expression is proven to increase when using ultrasound-mediated microbubble with SMFs compared to ultrasound-mediated microbubbles; especially while ultrasound acoustic intensity is between 0.3 and 0.5 MI, the gene expression is increased 2.2 and 1.8 times, respectively. The experimental results also demonstrate that when cells are exposed to SMFs, it really helps to improve the gene transfer efficiency. In addition, the cell viability decreases about 15-25 % when using ultrasound-mediated microbubble with or without SMFs compared to the control set, which means that ultrasonic assisted microbubbles can generate significant cavitation and cause cell casualty.