Abstract
The application of silver nanoparticles (AgNPs) in consumer products has been increasing rapidly over the past decades. Therefore, in vitro models capable of accurately predicting the toxicity of AgNPs are much needed. Hepatocyte-like cells (HLCs) derived from human induced pluripotent stem cells (iPSCs) represent an attractive alternative in vitro hepatotoxicity model. Yet, the use of iPSC-derived HLCs (iPSC-HLCs) for the study of nanoparticle toxicity has not been reported so far. In the present study, transcriptomic changes induced by varying concentrations (5–25 μg/ml) of AgNPs were characterized in iPSC-HLCs after 24-h exposure. AgNPs caused concentration-dependent gene expression changes in iPSC-HLCs. At all the concentrations, members of the metallothionein (MT) and the heat shock protein (HSP) families were the dominating upregulated genes, suggesting that exposure to AgNPs induced oxidative stresses in iPSC-HLCs and as a result elicited cellular protective responses in the cells. Functional analysis showed that the differentially expressed genes (DEGs) were majorly involved in the biological processes of metabolism, response to stress, and cell organization and biogenesis. Ingenuity Pathway Analysis revealed that cancer was at the top of diseases and disorders associated with the DEGs at all concentrations. These results were in accordance with those reported previously on hepatoma cell lines and primary hepatocytes. Considering the advantages iPSC-HLCs have over other liver cell models in terms of unlimited supply, consistency in quality, sustainability of function in long-term culture, and, more importantly, affordability of donor specificity, the results of the current study suggest that iPSC-HLCs may serve as a better in vitro model for liver nanotoxicology.
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Data availability
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- AgNPs:
-
Silver nanoparticles
- ANOVA:
-
Analysis of variance
- BP:
-
Biological process
- cDNA:
-
Complimentary DNA
- cRNA:
-
Complimentary RNA
- DAVID:
-
Database for Annotation, Visualization, and Integrated Discovery
- DEGs:
-
Differentially expressed genes
- DLS:
-
Dynamic light scattering
- FC:
-
Fold change
- FDR:
-
False discovery rate
- GO:
-
Gene ontology
- HCA:
-
Hierarchical cluster analysis
- HLCs:
-
Hepatocyte-like cells
- HSP:
-
Heat shock protein
- IPA:
-
Ingenuity Pathway Analysis
- iPSCs:
-
Induced pluripotent stem cells
- IVT:
-
In vitro transcription
- MT:
-
Metallothionein
- PCA:
-
Principal component analysis
- PHHs:
-
Primary human hepatocytes
- RMA:
-
Multi-array average
- ROS:
-
Reactive oxygen species
- SCENIHR:
-
Scientific Committee on Emerging and Newly Identified Health Risks
- TAC:
-
Transcriptome Analysis Console
- TEM:
-
Transmission electron microscopy
- UV-Vis:
-
Ultraviolet-visible
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Acknowledgments
The authors thank Drs. Marianne Miliotis-Solomotis and Mary E. Torrence for critical review and approval of the manuscript as well as for their support of this work. The authors also thank Dr. Michael A. Adams in the Office of Food Additive Safety, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration for critical review and approval of the manuscript.
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This work was supported by internal funds of the U.S. Food and Drug Administration.
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XG conceived and designed the study, carried out the experimental work, analyzed the data, and wrote the manuscript. RL participated in all stages of the experiments. RLS and JJY contributed to the conception of the study, critically reviewed the manuscript, and gave approval of the final version to be published. All authors read and approved the final manuscript.
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Gao, X., Li, R., Sprando, R.L. et al. Concentration-dependent toxicogenomic changes of silver nanoparticles in hepatocyte-like cells derived from human induced pluripotent stem cells. Cell Biol Toxicol 37, 245–259 (2021). https://doi.org/10.1007/s10565-020-09529-1
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DOI: https://doi.org/10.1007/s10565-020-09529-1