a School of Medicine , Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University , Shanghai , 200032 , China.
J Biomater Sci Polym Ed. 2013;24(15):1757-66. doi: 10.1080/09205063.2013.801702. Epub 2013 Jun 8.
Calcium phosphate-based transfection method had been used to transfer DNA into living cells. However, it had so far not been studied in detail to what extend siRNA delivery system. In this study, Pluronic F127/calcium phosphate hybrid nanoparticles (F127/CaP) were prepared by a facile room temperature method and employed as carriers to deliver siRNA to silence tumor cell. The morphology of the F127/CaP hybrid nanoparticles was investigated with TEM. In order to determine the ratio of F127 to CaP in the hybrid nanoparticles, TGA (the thermogravimetric analysis) was applied. MTT assays confirmed that the F127/CaP hybrid nanoparticles were quite safe. The hybrid F127/CaP nanoparticles obtained were 120-210 nm in diameter, and they were applied as siRNA carriers for siRNA loading and in vitro transfection. The siRNA encapsulating efficiency was 91.5 wt.% with a loading content of 6.5 wt.%. Compared to traditional CaP transfection method, the siRNA-loaded F127/CaP exhibited higher gene inhibition efficiency, and this was supported by fluorescence microscopy. Quantitative analysis of GFP silencing efficiency of various siRNA formulations was measured by using FACS flow cytometry analysis. Additionally, both custom CaP and F127/CaP are biocompatible and biodegradable, thus the as-prepared F127/CaP hybrid nanoparticles are promising for siRNA delivery.
基于磷酸钙的转染方法已被用于将 DNA 转入活细胞。然而，迄今为止，人们尚未详细研究其在多大程度上可以作为 siRNA 的递送系统。在这项研究中，通过一种简单的室温方法制备了 Pluronic F127/磷酸钙杂化纳米粒子（F127/CaP），并将其用作载体将 siRNA 递送至沉默肿瘤细胞。通过 TEM 研究了 F127/CaP 杂化纳米粒子的形态。为了确定杂化纳米粒子中 F127 与 CaP 的比例，应用了 TGA（热重分析）。MTT 测定证实 F127/CaP 杂化纳米粒子非常安全。获得的杂化 F127/CaP 纳米粒子的直径为 120-210nm，它们被用作 siRNA 载体进行 siRNA 负载和体外转染。siRNA 的包封效率为 91.5wt.%，负载量为 6.5wt.%。与传统的 CaP 转染方法相比，负载 siRNA 的 F127/CaP 表现出更高的基因抑制效率，这一点通过荧光显微镜得到了证实。通过使用 FACS 流式细胞术分析对各种 siRNA 制剂的 GFP 沉默效率进行了定量分析。此外，定制的 CaP 和 F127/CaP 均具有生物相容性和可生物降解性，因此所制备的 F127/CaP 杂化纳米粒子有望用于 siRNA 的递送。
