靜磁場與超音波微氣泡轉染對於基因傳遞影響研究

A Study on The Effect of Ultrasonic Microbubble Transfection and Static Magnetic Fields on Gene Delivery In Vitro

白啟宏(Chi-Hung Pai)

指導教授 ： 呂志誠

國立臺北科技大學/機電學院/機電整合研究所/碩士(2009年)

https://doi.org/10.6841/NTUT.2009.00203

摘要

本研究目的在探討利用超音波微氣泡轉染於報導基因傳遞(reporter gene delivery)之效應，並加入靜磁場作用對細胞培養過程中進行曝照後，其基因傳遞表現量所呈現的趨勢。基因傳遞實驗是以超音波微氣泡轉染作為方法。微氣泡破裂會釋放聲壓、光、熱能與自由基，稱空穴效應(cavitation)，當能量傳遞至細胞膜對其作用會產生短暫性的開洞，稱聲孔效應(sonoporaton)，效應會促進DNA進入細胞的效率。在本研究中將建構一套適用於體外實驗(in vitro)的超音波基因傳遞系統，自製的壓電傳感器將超音波透過壓克力導管傳送至細胞培養皿，使細胞有效曝照於超音波場強範圍。在細胞培養過程中將細胞置於磁鐵(不同的磁鐵型式)上方曝照靜磁場，於實驗中利用不同的超音波聲壓強度與加入Artison®商用微氣泡，將pGL3 luciferase報告基因傳遞至HAEC(人類主動脈內皮細胞, Human Aortic Endothelial Cells)，來測試對基因表現量與細胞存活率之影響。實驗結果顯示，單純施加超音波所得到的基因表現量並無明顯增加，但超音波微氣泡轉染則可得到較明顯的luciferase表現量。另外超音波與靜磁場作用對luciferase表現量無明顯差異，但受靜磁場曝照後的細胞存活數有些微增加趨勢；而超音波微氣泡加靜磁場作用與超音波微氣泡的結果相比，luciferase表現量並無顯著增加，此外超音波聲強越高，細胞存活數並無降低趨勢。

關鍵字

微氣泡 ； 基因傳遞 ； 靜磁場 ； 空穴效應 ； 聲孔效應

並列摘要

The purpose of this study was to investigate the trend of luciferase expression on the effect of ultrasound microbubble transfection and exposed static magnetic fields (SMFs) to cell in incubator for report gene delivery. Gene delivery experiment was a method that using ultrasound microbubble. The destruction of microbubble will release pressure, lights, thermal energy and free radicals, also called cavitation effect. When the energy transferred to cell membrane can able to generate permeability of transient pores, also called sonoporation. Those effects promote the efficiency of DNA into cells. An ultrasound system was established for in vitro gene delivery in this study. It is exposing ultrasound acoustic field when the piezoelectric transducer producing ultrasound that can through probe to the well of bottom of culture plate. The culture plate will putted onto magnet (variance magnet type) in culture cell stage. Then experiment for the gene expressions and cell viabilities when the PGL3 luciferase report gene delivered to HAECs after that added difference ultrasound intensity and Artison® microbubble. The results show that the gene expression was not significant increased when using ultrasound alone, but it was significant increased when using ultrasound microbubble transfection. In addition, the luciferase expression was not significant difference when using ultrasound and SMFs, but it was slightly enhanced of the cell viabilities after SMFs exposed. And then the luciferase expression was not significant difference when using ultrasound microbubble with SMFs compared to ultrasound microbubble. However the cell viabilities were not decreased clearly when the ultrasound intensity was increased.

並列關鍵字

microbubble ； gene delivery ； static magnetic fields ； cavitation ； sonoporation

參考文獻

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