改變磁性奈米粒子與靜磁場環境對磁轉染效率之影響研究

本研究目的在研究利用靜磁場輔助磁性奈米粒子於基因傳遞(Gene Delivery)之效能，並採取不同靜磁場形式於細胞轉染過程中進行曝照，探討基因傳遞表現量所呈現的趨勢。實驗過程以磁性奈米粒子混合DNA，利用磁場引導性特質使粒子混合體移至細胞附近，藉由內吞作用(endocytosis)來增加DNA進入細胞的效率。本研究首先建構一套適用於體外實驗(in vitro)的磁轉染基因傳遞系統，使用Nd-Fe-B強力磁鐵產生均勻磁場強度，並應用三種不同的磁性奈米粒子(Fe3O4、CombiMAG、PolyMAG)，本研究在基因傳遞過程中將細胞置於磁鐵上方曝照靜磁場，於實驗中利用不同的靜磁場形式與加入Alfa Aesar®及Chemicell®商用磁性奈米粒子，將pGL3 Luciferase報告基因傳遞至人類主動脈內皮細胞(human aortic endothelial cells)與人類胚胎腎細胞(HEK-293 cell line)，檢視對報導基因(reporter gene) 表現量與細胞存活率的影響。實驗結果顯示，單純施加DNA所得到的基因表現量並無明顯增加，但藉由靜磁場-磁性奈米粒子交互作用，報導基因傳遞效率呈現有意義提高，而加入的粒子濃度對於基因表現量及細胞存活率會有一定之影響，且高磁場強度及移動式靜磁場能有效提高基因傳遞效率，與SMF相較增加了45 %。

關鍵字

磁轉染；基因傳遞；靜磁場；磁性奈米粒子；報導基因；人類主動脈內皮細胞；人類胚胎腎細胞

並列摘要

The purpose of this study is to investigate the potential trend of gene expression by manipulating static magnetic fields (SMFs) and employing various magnetic nanoparticles in an incubator for reporter gene delivery. By using the magnetic force that can lead DNA-magnetic nanoparticle complex closer to cells and promote the efficiency of DNA delivery by endocytosis, this study established a suitable in-vitro megnetofetion system including Nd-Fe-B magnets enabling uniform magnetic fields, and application of three different magnetic nanoparticles (Fe3O4、CombiMAG、PolyMAG). In this study, human aortic endothelial cells (HAEC) and HEK-293 cell line were exposed to the SMFs to provoke the delivery of pGL3 Luciferase reporter gene, and evaluated by the reporter gene expression and cell viability. The results indicate that gene expression is not significantly increased by simply DNA. But, with the interaction of SMFs and magnetic nanoparticles, the gene expression is thus significantly increased. It is shown that the concentration of magnetic nanoparticles can increase gene expression but reduce cell viability, and high magnetic flux strength as well as moving magnetic fields (MMFs) manipulation is able to enhance the efficiency of gene delivery up to 45 % compared to the best results of SMFs manipulation .

並列關鍵字

magnetofection ； gene delivery ； static magnetic fields ； magnetic nanoparticles ； reporter gene ； HAEC ； HEK293

參考文獻

[1] R. W. Herzog and V. R. Arruda, “Update on gene therapy for hereditary hematological disorders,” Expert Rev Cardiovasc Ther, vol. 1, no. 2, 2003, pp. 215-232.

[2] F. Wang, D. K. Patel, J. M. Antonello, M. W. Washabaugh, D. C. Kaslow, J. W. Shiver and N. Chirmule, “Development of an adenovirus-shedding assay for the detection of adenoviral vector-based vaccine and gene therapy products in clinical specimens,” Human Gene Therapy, vol. 14, no. 1, 2003, pp. 25-36.

[3] C. Siatskas, J. Chan, J. Field, K. Murphy, Z. Nasa, B. H. Toh and F. Alderuccio, “Gene therapy strategies towards immune tolerance to treat the autoimmune diseases,” Current Gene Therapy, vol. 6, no. 1, 2006, pp. 45-58.

[4] C. R. Mayer and R. Bekeredjian, “Ultrasonic gene and drug delivery to the cardiovascular system,” Advanced Drug Delivery Reviews, vol. 60, no. 10, 2008, pp. 1177-1192.

[5] G. Vassaux, N and R. Lemoine, “Gene therapy for carcinoma of the breast genetic toxins,” Breast Cancer Research, vol. 2, no. 1, 2000, pp. 22-27.

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改變磁性奈米粒子與靜磁場環境對磁轉染效率之影響研究

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