Choi Joon Sig, Nam Kihoon, Park Jong-Yeun, Kim Jung-Bin, Lee Ja-Kyeong, Park Jong-Sang
Department of Biochemistry, Chungnam National University, Gung-dong 220, Yuseong-gu, Daejeon 305-764, South Korea.
J Control Release. 2004 Oct 19;99(3):445-56. doi: 10.1016/j.jconrel.2004.07.027.
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-arginine-grafted-PAMAM dendrimer (PAMAM-Arg). For comparison, an L-lysine-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.
我们设计了一种新型的富含精氨酸的树枝状聚合物，其结构基于定义明确的树枝状聚合物聚酰胺-胺树枝状聚合物（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树枝状聚合物（PAMAM-Arg）。为了进行比较，还生成了L-赖氨酸接枝的PAMAM树枝状聚合物（PAMAM-Lys）并作为对照试剂进行比较。发现这些聚合物与质粒DNA通过静电作用自组装，形成纳米级复合物。通过动态光散射实验观察到，多聚体的平均直径约为200nm。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的复合物形成。与天然PAMAM树枝状聚合物和PAMAM-Lys相比，由PAMAM-Arg/DNA组成的复合物显示出更高的基因传递能力。通过与聚乙烯亚胺（PEI）和PAMAM比较，测定了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显示出大大提高的转染效率。这些发现表明，L-精氨酸接枝的PAMAM树枝状聚合物具有成为用于基因治疗的新型基因传递载体的潜力。
