DNA nano-carriers from biodegradable cationic branched polyesters are formed by a modified solvent displacement method
DNA nano-carriers from biodegradable cationic branched polyesters are formed by a modified solvent displacement method
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DOI：
10.1016/j.jconrel.2005.12.004
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发表时间：
2006-04-10
影响因子：
10.8
通讯作者：
Kissel, T
中科院分区：
文献类型：
--
作者：
Oster, CG;Wittmar, M;Kissel, T
基于马兰戈尼效应，DNA纳米载体是由胺改性聚乙烯醇（PVAL）骨架接枝PLGA组成的可生物降解聚酯制成的，从而避免了有害的剪切力或超声波力。这些胺修饰的高分子量可生物降解聚酯结合了特定的特性，如DNA和阳离子支链聚酯之间的静电相互作用，促进了NP与DNA的负载。所得到的含有NP的DNA显示出175-285 nm的流体动力学直径和高度正的xi电位，这取决于用于颗粒形成的氮与磷酸盐（N/P）比。原子力显微镜（AFM）显示了明确的球形颗粒形态。DNA在PBS缓冲液中以完整的超卷曲形式从NP中释放出来。琼脂糖凝胶电泳表明，NP内的DNA不受酶降解的保护。通过四种细胞系的体外转染实验，证明了这种纳米载体的DNA传递的生物效率。胺修饰聚合物的报告基因传递量高于裸DNA（对照），并随着胺取代程度的增加而增加。此外，胺的类型和正电荷与主链的距离也起着重要的作用。此外，在相同的N/P比下，PEI为25 kDa/DNA多聚体的pCMV-Luc质粒和接枝PLGA链长为10个单体的胺修饰聚酯制备的NP的荧光素酶表达也显示了这一特征。在N/P比为9的情况下，P(68)-10负载NP的DNA转染效率比PEI 25 kDa高8倍。这些新的DNA纳米载体值得进一步研究，特别是在体内条件下的DNA疫苗接种。(c) 2005 Elsevier B.V.版权所有
DNA nano-carriers were formulated relying on biodegradable polyesters consisting of amine-modified poly(vinyl alcohol) (PVAL) backbones grafted with PLGA, based on the Marangoni effect thus avoiding detrimental shear or ultrasonic forces. These amine modified high molecular weight biodegradable polyesters combine specific characteristics, such as electrostatic interactions between DNA and cationic branched polyesters facilitating loading of NP with DNA. The resulting DNA containing NP showed hydrodynamic diameters in the range of 175-285 nm and highly positive xi-potentials, depending on the nitrogen to phosphate (N/P) ratio used for the particle formation. Atomic force microscopy (AFM) demonstrated well-defined spherical particle morphologies. DNA was released from NP upon incubation in PBS buffer in its intact supercoiled form. Agarose gel electrophoresis demonstrated that DNA within the NP was protected from enzyme degradation.The biological efficiency of the DNA delivery by this nano-carrier was demonstrated by an in vitro transfection assay using four cell lines. Reporter gene delivery of the amine-modified polymers was higher than naked DNA (Control) and raised with increasing degree of amine substitution. Also type of amine and distance of cationic charge from the backbone play an important role. Further, this feature was shown by Luciferase expression of the pCMV-Luc plasmid with PEI 25 kDa/DNA polyplexes and NP prepared with amine modified polyesters with a grafted PLGA chain length of 10 monomers compared at equal N/P ratios. DNA loaded NP from P(68)-10 showed 8x higher transfection efficiencies than the PEI 25 kDa at an N/P ratio of 9 for both preparations.These novel DNA nano-carriers merit further investigations in particular for DNA vaccination under in vivo conditions. (c) 2005 Elsevier B.V. All rights reserved.