Cellular uptake mechanism and intracellular fate of hydrophobically modified pullulan nanoparticles.
Cellular uptake mechanism and intracellular fate of hydrophobically modified pullulan nanoparticles.
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DOI：
10.2147/ijn.s44342
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发表时间：
2013
影响因子：
8
通讯作者：
Zhang Q
中科院分区：
文献类型：
--
作者：
Jiang L;Li X;Liu L;Zhang Q
研究了胆固醇修饰普鲁兰多糖（CHSP）自组装纳米粒子（NPs）在人肝癌细胞（HepG 2）中的摄取机制和细胞内命运。与异硫氰酸荧光素（FITC）的共价缀合产生稳定标记的CHSP（FITC-CHSP），其通过透析成功地配制成NP（平均粒径63.0 ± 1.9 nm）。细胞毒性测定清楚地表明CHSP NP在HepG 2细胞中没有显示出显著的毒性。NP浓度，孵育时间和温度对NP的细胞摄取的影响进行了系统的评价，通过荧光测定法，结果表明，细胞摄取的NP的浓度，时间和温度依赖性。内吞抑制剂的体外实验表明，网格蛋白介导的内吞作用和巨胞饮作用参与了CHSP NP的内化。细胞内运输研究表明，CHSP NP在孵育后1小时被截留在溶酶体中;在整个研究过程中未观察到NP与高尔基体或内质网的共定位。这些结果表明，CHSP纳米颗粒可以作为一个通用的载体细胞内输送的治疗剂。
The cellular uptake mechanism and intracellular fate of self-assembled nanoparticles (NPs) of cholesterol-modified pullulan (CHSP) by human hepatocellular carcinoma (HepG2) cells were investigated. Covalent conjugation with fluorescein isothiocyanate (FITC) yielded stably labeled CHSP (FITC-CHSP), which was successfully formulated into NPs (mean particle size 63.0 ± 1.9 nm) by dialysis. A cytotoxicity assay clearly indicated that the CHSP NPs did not show significant toxicity in HepG2 cells. The effects of NP concentration, incubation time, and temperature on the cellular uptake of the NPs were systematically evaluated by fluorometry, and the results suggested that cellular uptake of the NPs was concentration-,time-, and temperature-dependent. In vitro experiments with endocytic inhibitors revealed that clathrin-mediated endocytosis and macropinocytosis were involved in the internalization of CHSP NPs. The intracellular trafficking study demonstrated that CHSP NPs were entrapped in the lysosomes at 1 hour after incubation; colocalization of NPs with either the Golgi apparatus or the endoplasmic reticula was not observed during the entire course of the study. These results suggested that the CHSP NPs may serve as a versatile carrier for intracellular delivery of therapeutic agents.