title: Metabolomics of silver nanoparticles toxicity in HaCaT cells: structure-activity relationships and role of ionic silver and oxidative stress.
abstract: The widespread use of silver nanoparticles (AgNPs) is accompanied by a growing concern regarding their potential risks to human health, thus calling for an increased understanding of their biological effects. The aim of this work was to systematically study the extent to which changes in cellular metabolism were dependent on the properties of AgNPs, using NMR metabolomics. Human skin keratinocytes (HaCaT cells) were exposed to citrate-coated AgNPs of 10, 30 or 60 nm diameter and to 30 nm AgNPs coated either with citrate (CIT), polyethylene glycol (PEG) or bovine serum albumin (BSA), to assess the influence of NP size and surface chemistry. Overall, CIT-coated 60 nm and PEG-coated 30 nm AgNPs had the least impact on cell viability and metabolism. The role of ionic silver and reactive oxygen species (ROS)-mediated effects was also studied, in comparison to CIT-coated 30 nm particles. At concentrations causing an equivalent decrease in cell viability, Ag(+ )ions produced a change in the metabolic profile that was remarkably similar to that seen for AgNPs, the main difference being the lesser impact on the Krebs cycle and energy metabolism. Finally, this study newly reported that while down-regulated glycolysis and disruption of energy production were common to AgNPs and H2O2, the impact on some metabolic pathways (GSH synthesis, glutaminolysis and the Krebs cycle) was independent of ROS-mediated mechanisms. In conclusion, this study shows the ability of NMR metabolomics to define subtle biochemical changes induced by AgNPs and demonstrates the potential of this approach for rapid, untargeted screening of pre-clinical toxicity of nanomaterials in general.
authors.affiliation: a CICECO - Aveiro Institute of Materials, Department of Chemistry , University of Aveiro , Aveiro , Portugal .
authors.forename: Joana
authors.initials: J
authors.lastname: Carrola
authors.affiliation: b CESAM & Laboratory of Biotechnology and Cytomics, Department of Biology, University of Aveiro , Aveiro , Portugal , and.
authors.forename: Verónica
authors.initials: V
authors.lastname: Bastos
authors.affiliation: a CICECO - Aveiro Institute of Materials, Department of Chemistry , University of Aveiro , Aveiro , Portugal .
authors.forename: Ivana
authors.initials: I
authors.lastname: Jarak
authors.affiliation: a CICECO - Aveiro Institute of Materials, Department of Chemistry , University of Aveiro , Aveiro , Portugal .
authors.forename: Rui
authors.initials: R
authors.lastname: Oliveira-Silva
authors.affiliation: a CICECO - Aveiro Institute of Materials, Department of Chemistry , University of Aveiro , Aveiro , Portugal .
authors.forename: Eliana
authors.initials: E
authors.lastname: Malheiro
authors.affiliation: a CICECO - Aveiro Institute of Materials, Department of Chemistry , University of Aveiro , Aveiro , Portugal .
authors.forename: Ana L
authors.initials: AL
authors.lastname: Daniel-da-Silva
authors.affiliation: b CESAM & Laboratory of Biotechnology and Cytomics, Department of Biology, University of Aveiro , Aveiro , Portugal , and.
authors.forename: Helena
authors.initials: H
authors.lastname: Oliveira
authors.affiliation: b CESAM & Laboratory of Biotechnology and Cytomics, Department of Biology, University of Aveiro , Aveiro , Portugal , and.|c Department of Biology , Faculty of Sciences, University of Porto , Porto , Portugal.
authors.forename: Conceição
authors.initials: C
authors.lastname: Santos
authors.affiliation: a CICECO - Aveiro Institute of Materials, Department of Chemistry , University of Aveiro , Aveiro , Portugal .
authors.forename: Ana M
authors.initials: AM
authors.lastname: Gil
authors.affiliation: a CICECO - Aveiro Institute of Materials, Department of Chemistry , University of Aveiro , Aveiro , Portugal .
authors.forename: Iola F
authors.initials: IF
authors.lastname: Duarte
mesh_terms: D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D019343:Citric Acid / Q000737:chemistry / Q000633:toxicity; D006801:Humans; D007477:Ions; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism; D055432:Metabolomics / Q000379:methods*; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000633:toxicity*; D018384:Oxidative Stress / Q000187:drug effects*; D011092:Polyethylene Glycols / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D012834:Silver / Q000737:chemistry* / Q000633:toxicity*; D013329:Structure-Activity Relationship
chemical_list: D007477:Ions; D017382:Reactive Oxygen Species; D019343:Citric Acid; D012834:Silver; D011092:Polyethylene Glycols
keywords: AgNPs; NMR spectroscopy; keratinocytes; metabolism; reactive oxygen species