Mai Kaijin, Lin Jiantao, Zhuang Baoxiong, Li Xiaojun, Zhang Li-Ming
PCFM Lab and GDHPPC Lab, Institute of Polymer Science, School of Chemistry and Chemical Engineering, Sun Yat-sen University, Guangzhou 510275, China.
PCFM Lab and GDHPPC Lab, Institute of Polymer Science, School of Chemistry and Chemical Engineering, Sun Yat-sen University, Guangzhou 510275, China; Guangdong Medical College, Dongguan 523808, China.
Int J Biol Macromol. 2015 Aug;79:209-16. doi: 10.1016/j.ijbiomac.2015.04.064. Epub 2015 May 2.
For the development of effective and safe gene carrier based on starch, the amylose from potato starch was azidized by reacting with 3-azidopropylamine in the presence of N, N'-carbonyldiimidazole and then conjugated with propargyl focal point poly(amidoamine) (PAMAM) dendrons by a Cu(I)-catalyzed azide-alkyne cycloaddition. Such a cationic dendronization was verified by Fourier transform infrared spectroscopy and proton nuclear magnetic resonance analyses. For the resultant amylose conjugates with various contents and generations of PAMAM dendron, their buffering capacity, binding ability with plasmid DNA and in vitro cytotoxicity were investigated. These amylose conjugates were found to exhibit good buffering capacity and biocompatibility. In particular, they could condense effectively plasmid DNA into the nanocomplexes, as confirmed by agarose gel electrophoresis, zeta potential, and particle size analyses as well as transmission electron microscopy observation. For their nanocomplexes with plasmid DNA, the in vitro transfection properties in human embryonic kidney 293T cells were studied by fluorescence microscopy and flow cytometry. It was found that the transfection efficiency could be optimized by the dendronization extent of amylose and the complexation extent of dendronized amylose with plasmid DNA.
为了开发基于淀粉的有效且安全的基因载体，在N，N'-羰基二咪唑存在下，将马铃薯淀粉中的直链淀粉与3-叠氮基丙胺反应进行叠氮化，然后通过铜（I）催化的叠氮化物-炔烃环加成反应与炔丙基焦点聚（酰胺胺）（PAMAM）树枝状分子共轭。通过傅里叶变换红外光谱和质子核磁共振分析验证了这种阳离子树枝状化。对于具有不同含量和代数PAMAM树枝状分子的所得直链淀粉共轭物，研究了它们的缓冲能力、与质粒DNA的结合能力和体外细胞毒性。发现这些直链淀粉共轭物表现出良好的缓冲能力和生物相容性。特别是，通过琼脂糖凝胶电泳、zeta电位、粒径分析以及透射电子显微镜观察证实，它们可以有效地将质粒DNA浓缩成纳米复合物。对于它们与质粒DNA的纳米复合物，通过荧光显微镜和流式细胞术研究了在人胚肾293T细胞中的体外转染特性。发现转染效率可以通过直链淀粉的树枝状化程度以及树枝状化直链淀粉与质粒DNA的络合程度来优化。
