Intracellular disposition of chitosan nanoparticles in macrophages: intracellular uptake, exocytosis, and intercellular transport.
Intracellular disposition of chitosan nanoparticles in macrophages: intracellular uptake, exocytosis, and intercellular transport.
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壳聚糖纳米颗粒在巨噬细胞中的细胞内分布：细胞内摄取、胞吐作用和细胞间运输。
DOI：
10.2147/ijn.s142060
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发表时间：
2017
影响因子：
8
通讯作者：
Zheng CL
中科院分区：
文献类型：
--
作者：
Jiang LQ;Wang TY;Webster TJ;Duan HJ;Qiu JY;Zhao ZM;Yin XX;Zheng CL
可生物降解纳米材料已广泛应用于众多医学领域。为了进一步改善这些努力，本研究重点研究了壳聚糖纳米颗粒(CsNPs)在单核巨噬细胞系统(MPS)的主要细胞巨噬细胞中的分布。这种与MPS的相互作用决定了纳米颗粒在体内的滞留时间，因此在其自身的临床安全性中发挥着重要作用。本研究制备了多种染料标记的CsNPs(约250 nm)，并选择小鼠巨噬细胞系RAW-264.7作为模型巨噬细胞。结果表明，CsNPs在巨噬细胞中的掺入有两种机制，即一种主要的网状蛋白介导的内吞途径和一种吞噬作用。在内化后，这些颗粒在细胞中部分解离，表明细胞消化了纳米颗粒。结果表明，细胞内摄取CsNPs后，大部分CsNPs在24 h内排出，细胞内荧光强度下降69%，其余颗粒难以清除。Wortmannin和液泡素-1均能抑制CsNP的胞吐作用，表明CsNP的摄取是通过溶酶体和多囊泡体途径介导的，胞吐后CsNPs被邻近细胞重新摄取的实验进一步证实。因此，本研究阐明了CsNPs在巨噬细胞中的去向，并确定了细胞处置机制，为CsNPs如何被MPS识别提供了基础；这些信息对于CsNPs的众多医学应用是至关重要的。
Biodegradable nanomaterials have been widely used in numerous medical fields. To further improve such efforts, this study focused on the intracellular disposition of chitosan nanoparticles (CsNPs) in macrophages, a primary cell of the mononuclear phagocyte system (MPS). Such interactions with the MPS determine the nanoparticle retention time in the body and consequently play a significant role in their own clinical safety. In this study, various dye-labeled CsNPs (about 250 nm) were prepared, and a murine macrophage cell line (RAW 264.7) was selected as a model macrophage. The results showed two mechanisms of macrophage incorporation of CsNPs, ie, a clathrin-mediated endocytosis pathway (the primary) and phagocytosis. Following internalization, the particles partly dissociated in the cells, indicating cellular digestion of the nanoparticles. It was proved that, after intracellular uptake, a large proportion of CsNPs were exocytosed within 24 h; this excretion induced a decrease in fluorescence intensity in cells by 69%, with the remaining particles possessing difficulty being cleared. Exocytosis could be inhibited by both wortmannin and vacuolin-1, indicating that CsNP uptake was mediated by lysosomal and multivesicular body pathways, and after exocytosis, the reuptake of CsNPs by neighboring cells was verified by further experiments. This study, thus, elucidated the fate of CsNPs in macrophages as well as identified cellular disposition mechanisms, providing the basis for how CsNPs are recognized by the MPS; such information is crucial to numerous medical applications of CsNPs.