An efficient calcium phosphate nanoparticle-based nonviral vector for gene delivery
An efficient calcium phosphate nanoparticle-based nonviral vector for gene delivery
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DOI：
10.2147/ijn.s17096
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发表时间：
2011-01-01
影响因子：
8
通讯作者：
Zhang, Jian
中科院分区：
文献类型：
--
作者：
Liu, Yachun;Wang, Tao;Zhang, Jian
背景：较小的纳米颗粒有助于通过内吞作用将DNA递送到细胞中并提高转染效率。方法：以pEGFP-C1绿色荧光蛋白作为转染效率的指示剂，观察硫酸鱼精蛋白包被的磷酸钙（PS-CaP）是否具有稳定粒径和提高转染效率的作用。原子力显微镜用于评估粒子的形态和大小，并引入MTT法来检测细胞活力和抑制。结果：原子力显微镜图像显示，PS-CaP颗粒比经典的磷酸钙颗粒小得多。在293 FT、HEK 293和NIH 3 T3细胞中，PS-CaP的转染效率高于经典的磷酸钙颗粒。效率的差异意味着较小的纳米颗粒可以促进DNA通过内吞作用递送到细胞中，并且可以提高转染效率。此外，PS-CaP可用于在4 ℃下储存一周后转染HEK 293细胞，与经典磷酸钙相比，效率损失程度较小，表明硫酸鱼精蛋白可增加磷酸钙纳米颗粒的稳定性。细胞活力抑制实验表明，两种纳米粒的细胞毒性相似。结论：PS-CaP可作为一种较好的非病毒转染载体。
Background: Smaller nanoparticles facilitate the delivery of DNA into cells through endocytosis and improve transfection efficiency. The aim of this study was to determine whether protamine sulfate-coated calcium phosphate (PS-CaP) could stabilize particle size and enhance transfection efficiency.Methods: pEGFP-C1 green fluorescence protein was employed as an indicator of transfection efficiency. Atomic force microscopy was used to evaluate the morphology and the size of the particles, and an MTT assay was introduced to detect cell viability and inhibition. The classical calcium phosphate method was used as the control.Results: Atomic force microscopy images showed that the PS-CaP were much smaller than classical calcium phosphate particles. In 293 FT, HEK 293, and NIH 3T3 cells, the transfection efficiency of PS-CaP was higher than for the classical calcium phosphate particles. The difference in efficiencies implies that the smaller nanoparticles may promote the delivery of DNA into cells through endocytosis and could improve transfection efficiency. In addition, PS-CaP could be used to transfect HEK 293 cells after one week of storage at 4 C with a lesser extent of efficiency loss compared with classical calcium phosphate, indicating that protamine sulfate may increase the stability of calcium phosphate nanoparticles. The cell viability inhibition assay indicated that both nanoparticles show similar low cell toxicity.Conclusion: PS-CaP can be used as a better nonviral transfection vector compared with classical calcium phosphate.