(1. 中南大學(xué) 粉末冶金國(guó)家重點(diǎn)實(shí)驗(yàn)室，長(zhǎng)沙 410083；
2. 中南大學(xué) 湘雅三醫(yī)院，長(zhǎng)沙 410013)
摘 要: 以正硅酸乙酯為原料，通過添加氨基化試劑和釕吡啶配合物水溶液，采用油包水制備硅納米顆粒、表面改性的硅納米顆粒和熒光硅納米顆粒。通過電鏡檢測(cè)到納米顆粒的粒徑約為40 nm；在中性pH條件下，Zeta電位儀檢測(cè)表面改性的硅納米顆粒的凈正電荷約為16 mV；細(xì)胞內(nèi)吞實(shí)驗(yàn)和體外毒性實(shí)驗(yàn)表明，熒光顆粒可被細(xì)胞吞噬，對(duì)細(xì)胞的生長(zhǎng)無(wú)明顯影響；與DNA的結(jié)合試驗(yàn)發(fā)現(xiàn)，氨基化硅納米顆粒能與質(zhì)粒DNA有效結(jié)合，復(fù)合后能有效地抵抗血清或DNase I的降解；細(xì)胞轉(zhuǎn)染實(shí)驗(yàn)表明，顆粒有效地將綠色熒光蛋白(GFP)基因轉(zhuǎn)染到HT1080細(xì)胞和Hela細(xì)胞內(nèi)，并高效表達(dá)。
關(guān)鍵字: 硅納米顆粒；基因載體；細(xì)胞轉(zhuǎn)染；基因治療；表面改性
WANG Guo-hui2, HUANG Dong2, ZHOU Ke-
（1. State Key Laboratory of Powder Metallurgy, Central South University, Changsha 410083, China;
2. The Third Xiangya Hospital, Central South University, Changsha 410013, China）
Abstract:Silica nanoparticles, amino-terminated silica nanoparticles and fluorescent silica nanoparticles were prepared respectively via the formation of hydrolysis of tetraethyl orthosilicate (TEOS) with the synchronous modification of amino functional group in water-in-oil microemulsion. The diameter of the prepared nanoparticles was tested to be 40 nm through TEM analysis, and Zeta potential of the silica nanoparticles surface modified was tested to be 16 mV with zeta potentioneter under the condition of neutral pH value. Agarose gel electrophoresis photos indicate that the amino- terminated silica nanoparticles can bind effectively with DNA molecule, and moreover, nanoparticles-DNA complexes formed could resist digestion of DNase in the serum. The tests of cell by microscopy show that the most of nanoparticles can enter the cells when fluorescence nanoparticles are cultured with live HT1080 cells. Toxicity experiments in vitro indicate that there is no significant influence silica nanoparticles when they are cultured with normal cells. Finally, transfection tests of silica nanoparticles in the cultured cells reveal that plasmid DNA (pEGFP-N1_green fluorescence protein) can obtain expression with certain efficiency when the complexes formed with silica nanoparticles modified and DNA (pEGFP-N1) are cultured with Hela cells.
Key words: silica nanoparticle; gene vector; cell transfection; gene therapy; surface modification
