Enhanced gene transfection efficiency by polyamidoamine (PAMAM) dendrimers modified with ornithine residues.
Enhanced gene transfection efficiency by polyamidoamine (PAMAM) dendrimers modified with ornithine residues.
复制标题
DOI：
10.1016/j.ijpharm.2010.03.054
复制
发表时间：
2010-06
影响因子：
5.8
通讯作者：
Ajay Kumar;V. Yellepeddi;G. Davies;K. Strychar;S. Palakurthi
中科院分区：
文献类型：
--
作者：
Ajay Kumar;V. Yellepeddi;G. Davies;K. Strychar;S. Palakurthi
本研究的目的是制备鸟氨酸偶联的PAMAMG4树枝状大分子，并对其基因转染率和细胞毒性进行评价。采用Fmoc合成法合成了鸟氨酸偶联的PAMAMG4树状大分子。在HEK 293T、GM7373和NCI H157G细胞中进行了PAMAMG4树状分子和表面修饰的树状分子的比较基因转染研究。通过单独实验考察了过量鸟氨酸(100μM)对鸟氨酸偶联PAMAMG4树枝状大分子转染率的影响。用四甲基偶氮唑盐比色法测定了树突状网络对HEK 293T细胞的细胞毒作用。核磁共振氢谱和MALDI-TOF光谱分析表明，约60个鸟氨酸分子(PAMAMG4-ORN60)偶联到PAMAMG4树状大分子上。初步研究表明，电荷比(N/P)为10的树突状网络具有较高的转染率，而血清的存在不影响树状网络的转染率。与HEK 293T细胞相比，PAMAMG4-ORN60树突状网络在癌细胞(NCI H157G)中的转染率略高。过量鸟氨酸的存在降低了PAMAMG4-ORN60树状大分子的转染率，而对亲本PAMAMG4树状大分子没有影响。细胞毒性实验表明，N/P10的PAMAMG4-ORN60树突状网络在≤50μg/mL浓度下是安全的。因此，鸟氨酸偶联的树状大分子有可能成为新型的基因载体。
Aim of the study was to prepare and to evaluate gene transfection efficiency and cytotoxicity of the ornithine-conjugated PAMAMG4 dendrimers. Ornithine-conjugated PAMAMG4 dendrimers were prepared by Fmoc synthesis. A comparative gene transfection study between PAMAMG4 dendrimers and the surface modified dendrimers was conducted in HEK 293T, GM7373 and NCI H157G cell lines. Effect of excess of ornithine (100μM) on transfection efficiency of the ornithine-conjugated PAMAMG4 dendrimers was investigated in separate experiment. Cytotoxicity of the dendriplexes was tested in HEK 293T cells by MTT assay.1H NMR and MALDI-TOF spectral analysis showed that about 60 molecules of ornithine (PAMAMG4-ORN60) were conjugated to a PAMAMG4 dendrimer. Preliminary studies indicated that dendriplexes at charge ratio (N/P 10) show higher transfection efficiency and presence of serum does not affect the transfection efficiency of the dendriplexes. Transfection efficiency of PAMAMG4-ORN60 dendriplexes was slightly higher in cancer cells (NCI H157G) as compared to HEK 293T cells. Transfection efficiency of the PAMAMG4-ORN60 dendrimers decreased in presence of excess of ornithine while there was no effect on the parent PAMAMG4 dendrimers. Cytotoxicity assay has shown that PAMAMG4-ORN60 dendriplexes at N/P 10 were safe at concentrations ≤50μg/mL. It may be concluded that the ornithine-conjugated dendrimers possess the potential to be novel gene carrier.