Uptake and intracellular distribution of silver nanoparticles in human mesenchymal stem cells

被引:308
作者
Greulich, C. [1 ]
Diendorf, J. [2 ]
Simon, T. [3 ]
Eggeler, G. [3 ]
Epple, M. [2 ]
Koeller, M. [1 ]
机构
[1] Ruhr Univ Bochum, Bergmannsheil Univ Hosp Surg Res, D-44789 Bochum, Germany
[2] Univ Duisburg Essen, Ctr Narointegrat Duisburg Essen, D-45117 Essen, Germany
[3] Ruhr Univ Bochum, Fac Mech Engn, Inst Mat, D-44789 Bochum, Germany
关键词
Silver nanoparticles; Human mesenchymal stem cells; Cellular uptake; Focused ion beam; Fluorescence microscopy; GOLD NANOPARTICLES; UPTAKE MECHANISM; PATHWAYS; RELEASE; FATE;
D O I
10.1016/j.actbio.2010.08.003
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
Silver nanoparticles (Ag-NP) are widely used due to their well-known antibacterial effects. In medicine Ag-NP have found applications as wound dressings, surgical instruments and bone substitute biomaterials, e.g. silver-containing calcium phosphate cements. Depending on the coating technique, during resorption of a biomaterial Ag-NP may come into close contact with body tissues, including human mesenchymal stem cells (hMSC). Despite the widespread uses of Ag-NP, there is a serious lack of information concerning their biological effects on human cells. In this study the uptake of Ag-NP into hMSC has been analyzed and the intracellular distribution of Ag-NP after exposure determined. Non-agglomerated (dispersed) Ag-NP from the cell culture medium were detected as agglomerates of nanoparticles within the hMSC by combined focused ion beam/scanning electron microscopy. The silver agglomerates were typically located in the perinuclear region, as determined by light microscopy. Specific staining of cellular structures (endo-lysosomes, nuclei, Golgi complex and endoplasmatic reticulum) using fluorescent probes showed that the silver nanoparticles occurred mainly within endo-lysosomal structures, not in the cell nucleus, endoplasmic reticulum or Golgi complex. Quantitative determination of the uptake of Ag-NP by flow cytometry (scattergram analysis) revealed a concentration-dependent uptake of the particles which was significantly inhibited by chlorpromazine and wortmannin but not by nystatin, indicating clathrin-dependent endocytosis and macropinocytosis as the primary uptake mechanisms. (C) 2010 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码:347 / 354
页数:8
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