为了制备理想的非病毒基因载体,合成了甲壳三糖-g-壳聚糖(TCS),并与钙基累托石(REC)插层复合制备纳米复合材料,采用X射线衍射和透射电镜方法对产物进行了表征,考察了其细胞相容性,将REC、TCS和纳米复合材料分别与质粒DNA复合形成复合物,并考察了其体外基因转染效果.结果表明:TCS、REC及纳米复合材料的细胞相容性良好,当N/P比为3时,TCS/DNA复合物粒径大小都在75 nm左右,REC加入后,其粒径不同程度地增大,最大达到191 nm;相对分子质量高的TCS所得DNA复合物较相对分子质量低的TCS所得DNA复合物稳定,而REC的加入降低了TCS/DNA复合物的稳定性;TCS/REC-DNA复合物能介入肝癌细胞中并表达荧光.说明该纳米复合材料可作为潜在的非病毒基因载体.
In order to obtain ideal non-viral gene carriers,trisaccharide-g-chitosan(TCS) was synthesized and was intercalated into rectorite(REC) to prepare novel nanocomposite.The product was then characterized by means of XRD and TEM,with its cellular compatibility being also analyzed.Moreover,REC,TCS and the prepared nanocomposite were respectively combined with plasmid DNA to form novel complexes,whose gene transfection effects in vitro were also evaluated.The results indicate that REC,TCS and the prepared nanocomposite all have good cellular compatibility,that the TCS/DNA complex is about 75nm in size at a N/P ratio of 3,that the addition of REC may increase the size up to 191nm,that the stability of TCS/DNA complex is positively correlated to the relative molecular mass of TCS and may degrade due to the addition of REC,and that TCS/REC-DNA complex can enter into hepatoma cells with fluorescence expression.It is thus concluded that TCS/REC nanocomposite is a potential non-viral gene carrier.