Kawano Takahito, Yamagata Masato, Takahashi Hironobu, Niidome Yasuro, Yamada Sunao, Katayama Yoshiki, Niidome Takuro
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.
J Control Release. 2006 Apr 10;111(3):382-9. doi: 10.1016/j.jconrel.2005.12.022. Epub 2006 Feb 17.
This study aimed to investigate the benefits of combining the use of PEG-modified cationic gold nanoparticles with electroporation for in vivo gene delivery. PEG-modified cationic gold nanoparticles were prepared by NaBH(4) reduction of HAuCl(4) in the presence of 2-aminoethanethiol and mPEG-SH. Zeta-potential of the particles was nearly neutral (+0.1 mV). After forming complexes with plasmid DNA at a w/w ratio of 8.4, nanoparticle complexes were 90 nm for at least 60 min and showed a negative zeta-potential. After intravenous injection of DNA-nanoparticle complexes, 20% of gold were detected in blood at 120 min after injection and 5% of DNA were observed in blood after 5 min, suggesting that PEG-modified nanoparticles were stably circulating in the blood flow, but some of the DNA bound to particles degraded during circulation. When electroporation was applied to a lobe of the liver following injection of DNA-nanoparticle complexes, significant gene expression was specifically observed in the pulsed lobe. We concluded that PEG-modified nanoparticles maintained DNA more stably in the blood flow than in the case of naked DNA and electroporation assisted in restricted gene expression of circulating DNA in limited areas of the liver.
本研究旨在探讨聚乙二醇（PEG）修饰的阳离子金纳米颗粒与电穿孔相结合用于体内基因递送的益处。通过在2-氨基乙硫醇和甲氧基聚乙二醇硫醇（mPEG-SH）存在下用硼氢化钠（NaBH₄）还原氯金酸（HAuCl₄）制备PEG修饰的阳离子金纳米颗粒。颗粒的zeta电位接近中性（+0.1 mV）。在以8.4的重量比与质粒DNA形成复合物后，纳米颗粒复合物至少60分钟内为90纳米，并显示出负的zeta电位。静脉注射DNA-纳米颗粒复合物后，注射后120分钟在血液中检测到20%的金，5分钟后在血液中观察到5%的DNA，这表明PEG修饰的纳米颗粒在血流中稳定循环，但一些与颗粒结合的DNA在循环过程中降解。当在注射DNA-纳米颗粒复合物后对肝脏的一个叶施加电穿孔时，在脉冲叶中特异性地观察到显著的基因表达。我们得出结论，与裸DNA相比，PEG修饰的纳米颗粒在血流中更稳定地保持DNA，并且电穿孔有助于在肝脏有限区域内限制循环DNA的基因表达。
