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Proteomic and lipidomic analysis of primary mouse hepatocytes exposed tometal and metal oxide nanoparticles
Stockholm University, Faculty of Science, Department of Biochemistry and Biophysics.ORCID iD: 0000-0002-1986-2845
Stockholm University, Faculty of Science, Department of Biochemistry and Biophysics.
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2015 (English)In: Journal of Integrated Omics, ISSN 2182-0287, Vol. 5, no 1Article in journal (Refereed) Published
Abstract [en]

The global analysis of the cellular lipid and protein content upon exposure to metal and metal oxide nanoparticles (NPs) can provide an overview of the possible impact of exposure. Proteomic analysis has been applied to understand the nanoimpact however the relevance of the alteration on the lipidic profile has been underestimated. In our study, primary mouse hepatocytes were treated with ultra-small (US) TiO2-USNPs as well as ZnO-NPs, CuO-NPs and Ag-NPs. The protein extracts were analysed by 2D-DIGE and quantified by imaging software and the selected differentially expressed proteins were identified by nLC-ESI-MS/MS. In parallel, lipidomic analysis of the samples was performed using thin layer chromatography (TLC) and analyzed by imaging software. Our findings show an overall ranking of the nanoimpact at the cellular and molecular level: TiO2-USNPs<ZnO-NPs<Ag-NPs<CuO-NPs. CuO-NPs and Ag-NPs were cytotoxic while ZnO-NPs and CuO-NPs had oxidative capacity. TiO2-USNPs did not have oxidative capacity and were not cytotoxic.  The most common cellular impact of the exposure was the down-regulation of proteins. The proteins identified were involved in urea cycle, lipid metabolism, electron transport chain, metabolism signaling, cellular structure and we could also identify nuclear proteins. CuO-NPs exposure decreased phosphatidylethanolamine and phosphatidylinositol and caused down-regulation of electron transferring protein subunit beta. Ag-NPs exposure caused increased of total lipids and triacylglycerol and decrease of sphingomyelin. TiO2-USNPs also caused decrease of sphingomyelin as well as up-regulation of ATP synthase and electron transferring protein alfa. ZnO-NPs affected the proteome in a concentration-independent manner with down-regulation of RNA helicase.  ZnO-NPs exposure did not affect the cellular lipids. To our knowledge this work represents the first integrated proteomic and lipidomic approach to study the effect of NPs exposure to primary mouse hepatocytes in vitro.

Place, publisher, year, edition, pages
2015. Vol. 5, no 1
Keyword [en]
nanoparticles, hepatocytes, proteomics, lipidomics, mass spectrometry, toxicity
National Category
Biochemistry and Molecular Biology Cell Biology
Research subject
URN: urn:nbn:se:su:diva-117027DOI: 10.5584/jiomics.v5i1.184OAI: diva2:809752
Available from: 2015-05-05 Created: 2015-05-05 Last updated: 2015-05-12Bibliographically approved
In thesis
1. Toxicity and biological impact of metal and metal oxide nanoparticles: Focus on the vascular toxicity of ultra-small titanium dioxide nanoparticles
Open this publication in new window or tab >>Toxicity and biological impact of metal and metal oxide nanoparticles: Focus on the vascular toxicity of ultra-small titanium dioxide nanoparticles
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The application of nanoparticles (NPs) in different technologies has led to tremendous advancement in those fields.  Moreover, there is growing interest in application of ultra-small NPs (USNPs) at 1-3 nm due to their distinct molecule like features. Parallel to these promises, there is a growing concern regarding their safety. The main goal of this thesis was to investigate the toxicity and underlying mechanisms following exposure to different metal and metal oxide NPs as well as USNPs. Their effects were studied on Saccharomyces cerevisiae, on hepatocytes and endothelial cells and finally in vivo on zebrafish embryos (Danio rerio). By selecting the rutile form of titanium dioxide (TiO2-USNPs) without intrinsic or intracellular reactive oxygen species (ROS) production, we could study biological impacts solely due to size and direct interaction with the cells. We showed that TiO2-USNPs were not cytotoxic but induced DNA damage. They had anti-angiogenic effects both in vitro and in vivo. Also, at high concentrations they caused complete mortality in zebrafish embryos exposed in water, while at lower concentrations induced delay in hatching. When injected they caused malformations. They specifically induced the differential overexpression of transcripts involved in lipid and cholesterol metabolism in endothelial cells. In hepatocytes they induced the overexpression of proteins in the electron transport chain and decreased lipids in lipid rafts. At 30 nm, TiO2-NPs, were also not cytotoxic but were genotoxic. They had no effects in vivo or on angiogenesis. However, they induced differential expression of transcripts involved in endoplasmic reticulum (ER) stress and heat shock response as well as cholesterol metabolism. This suggests a more toxic response in the cells compared to TiO2-USNPs.  Single walled carbon nanotubes (SWCNTs) despite having the highest oxidative activity among the NPs studied, were not severely cyto- or genotoxic but induced expression of transcripts involved in early ER stress response. Copper oxide (CuO-NPs) was the most toxic NPs studied due to both ion release and ROS production, affecting lipid metabolism of the cells. Silver (Ag-NPs) were also cytotoxic and caused the disruption of cellular components and lipids. ZnO-NPs were not cytotoxic, did not affect cellular lipids but they increased the size of vacuoles in yeast cells. Finally by using superparamagnetic iron oxide NPs (SPIONs) with different coatings, and using a mathematical model, a nano impact index (INI) was developed as a tool to enable the comparison of nanotoxicology data.

Place, publisher, year, edition, pages
Stockholm: Department of Biochemistry and Biophysics, Stockholm University, 2015. 78 p.
Nanoparticles, Nanotoxicology, Ultra-small nanoparticles
National Category
Biochemistry and Molecular Biology
Research subject
urn:nbn:se:su:diva-116708 (URN)978-91-7649-193-5 (ISBN)
Public defence
2015-06-02, Magnélisalen, Kemiska övningslaboratoriet, Svante Arrhenius väg 16 B, Stockholm, 10:00 (Swedish)

At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 2: Accepted. Paper 3: Manuscript. Paper 4: Manuscript. Paper 5: Manuscript.

Available from: 2015-05-11 Created: 2015-04-23 Last updated: 2015-05-12Bibliographically approved

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