评价N,N,N-三甲基壳聚糖盐酸盐(TMC)及其纳米粒子对质粒DNA(pDNA)的负载及保护能力,考察了其纳米复合物对人类肝癌细胞株(HepG 2细胞)的转染能力.通过复凝聚法制备TMC/pDNA纳米粒子,并采用透射电镜及原子力显微镜表征粒子形态和粒径;采用凝胶阻滞分析观察其对pDNA的保护情况;采用四噻氮唑盐(MTT)比色法测定细胞毒性;以Lipofectamine 2000转染试剂作为阳性对照,检测其对HepG 2细胞的转染活性;采用荧光倒置显微镜观察转染情况.结果表明负载pDNA的TMC纳米粒子多呈球形,粒径为100~300 nm,能有效地包裹和保护基因不被DNaseⅠ酶消化;当TMC与pDNA的质量比为10∶1时,在48 h达到最高的转染效率.
参考文献
[1] | Mao S et al.Chitosan-based formulations for delivery of DNA and siRNA[J].Advanced Drug Delivery Reviews,2009,62(01):12. |
[2] | Elsie Khai-Woon Toh;Hsing-Yin Chen;Yu-Lun Lo et al.Succinated chitosan as a gene carrier for improved chitosan solubility and gene transfection[J].Nanomedicine:Nanotechn Biol Med,2011,2(07):174. |
[3] | Lavertu M;Methot S;Tran-Khanh N;Buschmann MD .High efficiency gene transfer using chitosan/DNA nanoparticles with specific combinations of molecular weight and degree of deacetylation[J].Biomaterials,2006(27):4815-4824. |
[4] | Prabaharan M;Mano JF .Chitosan-based particles as controlled drug delivery systems.[J].Drug delivery,2005(1):41-57. |
[5] | 刘璠娜,饶华新,张子勇.季铵盐壳聚糖纳米荧光探针的制备及细胞渗入研究[J].功能材料,2008(05):857-860,863. |
[6] | 刘瑶娜,张子勇,饶华新.一种新型季胺盐壳聚糖纳米载药体系的制备与性能[J].功能高分子学报,2008(01):50-54,75. |
[7] | Mao ZW;Ma L;Jiang Y;Yan M;Gao CY;Shen JC .N,N,N-Trimethylchitosan chloride as a gene vector: Synthesis and application[J].Macromolecular bioscience,2007(6):855-863. |
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