Enhanced transfection efficiency of PAMAM dendrimer by surface modification with L-arginine
Enhanced transfection efficiency of PAMAM dendrimer by surface modification with L-arginine
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DOI：
10.1016/j.jconrel.2004.07.027
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发表时间：
2004-10-19
影响因子：
10.8
通讯作者：
Park, J
中科院分区：
文献类型：
--
作者：
Choi, JS;Nam, K;Park, J
我们设计了一种新型的富含精氨酸的树状大分子，其结构基于定义良好的树状大分子聚氨基胺树状大分子（PAMAM）。进行了进一步的表征，以证明该聚合物是一种有效的非病毒基因传递载体。将位于PAMAM表面的伯胺与l -精氨酸偶联，生成l -精氨酸接枝的PAMAM树状大分子（PAMAM- arg）。为了进行比较，还生成了l - lysiie接枝的pamam树状大分子（PAMAM-Lys）作为对照试剂进行了比较。这些聚合物可以与质粒DNA静电自组装，形成纳米级的复合物。通过动态光散射实验，观察到多聚体的平均直径约为200 nm。我们使用PicoGreen试剂作为一种有效的探针来检测聚合物与质粒DNA的复杂形成。由PAMAM- arg /DNA组成的复合物与天然PAMAM树状大分子和PAMAM- lys相比，具有更高的基因传递能力。通过PEI和PAMAM的比较，测定293、HepG2和Neuro， 2A细胞的细胞毒性和转染效率。此外，在原代大鼠血管平滑肌细胞中进行了转染实验，PAMAM-Arg的转染效率明显提高。这些发现表明，l -精氨酸接枝的pamam树突状物具有成为基因治疗的新型基因传递载体的潜力。(C) 2004 Elsevier B.V.版权所有
We designed a novel type of arginine-rich dendrimer, with a structure based on the well-defined dendrimer, polyamidoamine dendrimer (PAMAM). Further characterization was performed to prove that the polymer is a potent nonviral gene delivery carrier. The primary amines located on the surface of PAMAM were conjugated with L-arginine to generate an L-argininegrafted-PAMAM dendrimer (PAMAM-Arg). For comparison, an L-lysiiie-grafted-PAMAM dendrimer (PAMAM-Lys) was also generated and compared as a control reagent. The polymers were found to self-assemble electrostatically with plasmid DNA, forming nanometer-scale complexes. From dynamic light scattering experiments, the mean diameter of the polyplexes was observed to be around 200 nm. We used PicoGreen reagent as an efficient probe for assaying complex formation of polymers with plasmid DNA. The complex composed of PAMAM-Arg/DNA showed increased gene delivery potency compared to native PAMAM dendrimer and PAMAM-Lys. The cytotoxicity and transfection efficiencies for 293, HepG2, and Neuro, 2A cells were measured by comparison with PEI and PAMAM. In addition, transfection experiments were performed in primary rat vascular smooth muscle cells, and PAMAM-Arg showed much enhanced transfection efficiency. These findings suggest that the L-arginine-grafted-PAMAM dendrimer possesses the potential to be a novel gene delivery carrier for gene therapy. (C) 2004 Elsevier B.V. All rights reserved.