Transfection efficiency influenced by aggregation of DNA/polyethylenimine max/magnetic nanoparticle complexes
Transfection efficiency influenced by aggregation of DNA/polyethylenimine max/magnetic nanoparticle complexes
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DOI：
10.1007/s11051-013-1653-y
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发表时间：
2013-04
影响因子：
2.5
通讯作者：
S. Ota;Y. Takahashi;A. Tomitaka;Tsutomu Yamada;D. Kami;Masatoshi Watanabe;Y. Takemura
中科院分区：
文献类型：
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作者：
S. Ota;Y. Takahashi;A. Tomitaka;Tsutomu Yamada;D. Kami;Masatoshi Watanabe;Y. Takemura
使用磁性纳米颗粒（MNP）的基因递送被称为磁转染，并且是一种有效的非病毒基因递送系统。制备了粒径为29 nm的γ-Fe 2 O3纳米粒子和粒径为20-30 nm的Fe 3 O 4纳米粒子，并将其与DNA偶联。评估转染效率对MNP重量、HeLa细胞活力和DNA/PEI max/MNP复合物大小的依赖性。转染效率最初随着复合物的重量而增加;然而，随着重量的进一步增加而降低。相反，细胞活力随着重量的进一步增加而增加。细胞毒性试验表明，在较高的重量转染效率的下降是不是归因于DNA/PEI max/MNP复合物的细胞毒性。DNA/PEI max/MNP复合物由于DNA结合和pH与介质的相互作用而聚集。证实了依赖于MNP重量的聚集。复合物的数量估计从大小分布。此外，使用复合物的细胞内吞途径的转染效率对聚集的依赖性进行了评估。复合物通过网格蛋白依赖的内吞作用内化，这是一种大小依赖性途径。这项研究表明，降低转染效率与聚集的程度，这是由高重量的MNPs诱导。
Gene delivery using magnetic nanoparticles (MNPs) is known as magnetofection and is an efficient non-viral gene delivery system. γ-Fe2O3nanoparticles (primary diameter = 29 nm) and Fe3O4nanoparticles (primary diameter = 20–30 nm) coated with deacylated linear polyethylenimine (PEI max) were prepared and conjugated with DNA. The dependency of transfection efficiency on the weight of MNPs, viability of HeLa cells, and size of DNA/PEI max/MNP complexes was evaluated. Transfection efficiency initially increased with the weight of the complexes; however, it decreased with further increase in weight. In contrast, cell viability increased with further increase in weight. Cytotoxicity assay showed that the decline in transfection efficiency at higher weights was not attributable to cytotoxicity of DNA/PEI max/MNP complexes. The DNA/PEI max/MNP complexes aggregated because of DNA binding and pH interaction with the medium. Aggregation depending on the weight of MNPs was confirmed. The number of complexes was estimated from the size distribution. In addition, the dependency of the transfection efficiency on aggregation was assessed with respect to cellular endocytic pathways using the complexes. The complexes were internalized through clathrin-dependent endocytosis, which was a size-dependent pathway. This study reveals that decreased transfection efficiency was associated with the extent of aggregation, which was induced by high weight of MNPs.