Insights into the impact of silver nanoparticles on human keratinocytes metabolism through NMR metabolomics.
Due to their antimicrobial properties, silver nanoparticles (AgNPs) are increasingly incorporated into consumer goods and medical products. Their potential toxicity to human cells is however a major concern, and there is a need for improved understanding of their effects on cell metabolism and function. Here, Nuclear Magnetic Resonance (NMR) metabolomics was used to investigate the metabolic profile of human epidermis keratinocytes (HaCaT cell line) exposed for 48 h to 30 nm citrate-stabilized spherical AgNPs (10 and 40 μg/mL). Intracellular aqueous extracts, organic extracts and extracellular culture medium were analysed to provide an integrated view of the cellular metabolic response. The specific metabolite variations, highlighted through multivariate analysis and confirmed by spectral integration, suggested that HaCaT cells exposed to AgNPs displayed upregulated glutathione-based antioxidant protection, increased glutaminolysis, downregulated tricarboxylic acid (TCA) cycle activity, energy depletion and cell membrane modification. Importantly, most metabolic changes were apparent in cells exposed to a concentration of AgNPs which did not affect cell viability at significant levels, thus underlying the sensitivity of NMR metabolomics to detect early biochemical events, even in the absence of a clear cytotoxic response. It can be concluded that NMR metabolomics is an important new tool in the field of in vitro nanotoxicology.
CICECO - Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal.
Joana
J
Carrola
CESAM & Laboratory of Biotechnology and Cytomics, Department of Biology, University of Aveiro, 3810-193 Aveiro, Portugal.
Verónica
V
Bastos
CESAM & Laboratory of Biotechnology and Cytomics, Department of Biology, University of Aveiro, 3810-193 Aveiro, Portugal.
José Miguel P
JM
Ferreira de Oliveira
CESAM & Laboratory of Biotechnology and Cytomics, Department of Biology, University of Aveiro, 3810-193 Aveiro, Portugal.
Helena
H
Oliveira
CESAM & Laboratory of Biotechnology and Cytomics, Department of Biology, University of Aveiro, 3810-193 Aveiro, Portugal.
Conceição
C
Santos
CICECO - Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal.
Ana M
AM
Gil
CICECO - Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal. Electronic address: ioladuarte@ua.pt.
Iola F
IF
Duarte
D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002951:Citrates / Q000737:chemistry; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D055432:Metabolomics; D053768:Metal Nanoparticles / Q000633:toxicity*; D008928:Mitochondria / Q000187:drug effects; D012834:Silver / Q000737:chemistry* / Q000633:toxicity*; D018675:Toxicity Tests
D002951:Citrates; D012834:Silver
Keratinocytes; Metabolomics; NMR spectroscopy; Nanotoxicology; Silver nanoparticles