Quantitative Assessment of the Comparative Nanoparticle-Uptake Efficiency of a Range of Cell Lines
Quantitative Assessment of the Comparative Nanoparticle-Uptake Efficiency of a Range of Cell Lines
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DOI：
10.1002/smll.201101076
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发表时间：
2011-12-02
期刊：
影响因子：
13.3
通讯作者：
Dawson, Kenneth A.
中科院分区：
文献类型：
--
作者：
dos Santos, Tiago;Varela, Juan;Dawson, Kenneth A.
纳米粒子与细胞相互作用的机制尚不清楚。目前，人们对纳米颗粒进入细胞内并在细胞内定位的相关长度和时间尺度，或纳米颗粒摄取和定位的细胞特异性知之甚少。本文在不同细胞系中研究了粒径对模型荧光羧基修饰聚苯乙烯纳米颗粒体外细胞内摄取的影响。将一系列确定尺寸的微粒子和纳米颗粒（40 nm至2 μ m）与一系列细胞类型（包括HeLa和A549上皮细胞、1321N1星形胶质细胞、HCMEC D3内皮细胞和小鼠RAW 264.7巨噬细胞）一起培养。共聚焦显微镜和流式细胞术等技术被用于研究颗粒摄取和亚细胞定位，为确保半定量方法的可重复性做出了重大努力。结果表明，在所研究的所有细胞系中，（纳米）颗粒的内化高度依赖于大小，并且对同一类型纳米颗粒的摄取动力学在不同的细胞类型中有所不同。有趣的是，即使不是专门用于吞噬的细胞也能够内化较大的纳米颗粒。所有大小颗粒的细胞内摄取在RAW 264.7细胞（一种特殊的吞噬细胞系）中最高，在HeLa细胞中最低。这些结果表明（纳米）颗粒摄取可能不遵循通常定义的摄取过程的尺寸限制，并突出了不同细胞类型中相同材料的摄取动力学的可变性。这些结论对评价纳米材料的安全性和纳米颗粒潜在的生物医学应用具有重要意义。
The mechanism(s) of nanoparticlecell interactions are still not understood. At present there is little knowledge of the relevant length- and timescales for nanoparticle intracellular entry and localization within cells, or the cell-specificity of nanoparticle uptake and localisation. Here, the effect of particle size on the in-vitro intracellular uptake of model fluorescent carboxyl-modified polystyrene nanoparticles is investigated in various cell lines. A range of micro- and nanoparticles of defined sizes (40 nm to 2 mu m) are incubated with a series of cell types, including HeLa and A549 epithelial cells, 1321N1 astrocytes, HCMEC D3 endothelial cells, and murine RAW 264.7 macrophages. Techniques such as confocal microscopy and flow cytometry are used to study particle uptake and subcellular localisation, making significant efforts to ensure reproducibility in a semiquantitative approach. The results indicate that internalization of (nano)particles is highly size-dependent for all cell lines studied, and the kinetics of uptake for the same type of nanoparticle varies in the different cell types. Interestingly, even cells not specialized for phagocytosis are able to internalize the larger nanoparticles. Intracellular uptake of all sizes of particles is observed to be highest in RAW 264.7 cells (a specialized phagocytic cell line) and the lowest in the HeLa cells. These results suggest that (nano)particle uptake might not follow commonly defined size limits for uptake processes, and highlight the variability of uptake kinetics for the same material in different cell types. These conclusions have important implications for the assessment of the safety of nanomaterials and for the potential biomedical applications of nanoparticles.