Abstract
Zinc oxide nanoparticles (ZnO-NPs) are commonly used for industrial applications. Consequently, there is increasing exposure of humans to them. The in vitro analysis of cytotoxicity and genotoxicity is commonly performed under standard cell culture conditions. Thus, the question arises of how the results of genotoxicity and cytotoxicity experiments would alter if human plasma was used instead of cell culture medium containing of fetal calf serum (FCS). Human mesenchymal stem cells (hMSCs) were cultured in human plasma and exposed to ZnO-NPs. A cultivation in expansion medium made of DMEM consisting 10% FCS (DMEM-EM) served as control. Genotoxic and cytotoxic effects were evaluated with the comet and MTT assay, respectively. hMSC differentiation capacity and ZnO-NP disposition were evaluated by histology and transmission electron microscopy (TEM). The protein concentration and the amount of soluble Zn2+ were measured. The cultivation of hMSCs in plasma leads to an attenuation of genotoxic and cytotoxic effects of ZnO-NPs compared to control. The differentiation capacity of hMSCs was not altered. The TEM showed ZnO-NP persistence in cytoplasm in both groups. The concentrations of protein and Zn2+ were higher in plasma than in DMEM-EM. In conclusion, the cultivation of hMSCs in plasma compared to DMEM-EM leads to an attenuation of cytotoxicity and genotoxicity in vitro.
Highlights
The use of nanotechnology has been increasing in industry, biomedicine and research during the last years
The effects of Zinc oxide nanoparticles (ZnO-NPs) on Human mesenchymal stem cells (hMSCs) were analyzed under the presence of standard cell culture medium consisting of Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal calf serum (FCS) (Linaris, Wertheim-Bettingen, Germany), 1% penicillin and streptomycin (Sigma-Aldrich, Schnelldorf, Germany)
Single zinc oxide (ZnO)-NPs were visible by transmission electron microscopy (TEM) within the cytoplasm sparing the nucleus in both groups, DMEM-EM and plasma
Summary
The use of nanotechnology has been increasing in industry, biomedicine and research during the last years. NPs which can be oxidized, reduced or dissolved are often cytotoxic and genotoxic to cellular organisms [7] Concerning this topic, we were able to show a correlation between ion levels and genotoxic reactions of ZnO-NPs in vitro [8]. The in vitro analysis of cytotoxicity and genotoxicity of NPs is commonly performed under standard cell culture conditions. The media of these systems are routinely supplemented with animal serum like 10% fetal calf serum (FCS). The question arises of how the results of genotoxicity and cytotoxicity assays would alter if 100% of human plasma was used instead of cell culture medium containing of 10% FCS. The current study aimed to investigate differences in cytotoxic and genotoxic effects of ZnO-NPs in primary human cells. Our group favors the use of human mesenchymal stem cells (hMSCs) for nanotoxicological evaluations, since they are a very stable primary cell system with a high regenerative ability after toxic events and the possibility to perform long-term cultivation
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