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Nanomaterials 2020, 10, 2255 9 of 11 AgNPs may be due to the high amount of released ionic Ag inducing cell toxicity. The toxicity of aged Citrate AgNPs may be relevant to the aggregations of Citrate AgNPs. Our results are consistent with others showing the impact of aging on NP properties resulting in influences on their toxicity [24–27]. For example, Kittler et al. (2010) showed that silver NPs are increasingly toxic to human mesenchymal stem cells due to the increased release of silver ions [34]. 5. Conclusions Our study showed that media had a significant impact on the physicochemical properties of Citrate AgNPs. The electrolyte solution containing positive ions (e.g., Na+) can increase the ionic strength in media, which induced significant aggregation of Citrate AgNPs in PBS. However, Citrate AgNPs in saline did not show aggregation, which indicated that the kinetics of aggregation depends on the ionic composition of media. The decreased zeta potential of Citrate AgNPs in serum demonstrates that the diversity of the proteins forming the protein corona may be affected, reducing the aggregation of AgNPs. The aging of AgNPs with different coatings shows that the observed changes in AgNPs’ toxicity are related to different processes such as dissolution, agglomeration, and capping agent degradation. Our study confirms the strong impact of aging and media properties on increased toxicity of AgNPs. These processes need to be explicitly modeled to predict the fate of silver in the environment and humans. Author Contributions: Conceptualization, methodology, writing—Original draft preparation, C.P.; review and editing, P.Z., Y.M., J.R., and B.Z.; project administration, C.P. and B.Z.; funding acquisition, C.P. and B.Z. All authors have read and agreed to the published version of the manuscript. Funding: This research was funded by the National Natural Science Foundation of China (No. 21407162) and partly supported by VILLUM FONDEN-VILLUM Experiment of Denmark. Conflicts of Interest: The authors declare no conflict of interest. References 1. DTU Environment. The Nanodatabase. 2013. Available online: http://nanodb.dk/products (accessed on 17 December 2013). 2. Rejeski, D. Nanotechnology and Consumer Products. 2009. Available online: http://www.nanotechproject. org/process/assets/files/8278/pen_submission_cpsc.pdf (accessed on 18 September 2013). 3. Kaegi, R.; Voegelin, A.; Sinnet, B.; Zuleeg, S.; Hagendorfer, H.; Burkhardt, M.; Siegrist, H. Behavior of metallic silver nanoparticles in a pilot wastewater treatment plant. Environ. Sci. Technol. 2011, 45, 3902–3908. [CrossRef] [PubMed] 4. Lynch, I.; Dawson, K.A. Protein-nanoparticle interactions. Nano Today 2008, 3, 40–47. [CrossRef] 5. Stegemeier, J.P.; Avellan, A.; Lowry, G.V. Effect of initial speciation of copper-and silver-based nanoparticles on their long-term fate and phytoavailability in freshwater wetland mesocosms. Environ. Sci. Technol. 2017, 51, 12114–12122. [CrossRef] [PubMed] 6. Siemer, S.; Westmeier, D.; Barz, M.; Eckrich, J.; Wünsch, D.; Seckert, C.; Thyssen, C.; Schilling, O.; Hasenberg, M.; Pang, E.; et al. Biomolecule-corona formation confers resistance of bacteria to nanoparticle- induced killing: Implications for the design of improved nanoantibiotics. Biomaterials 2019, 192, 551–559. [CrossRef] 7. Wang, L.; Li, J.; Pan, J.; Jiang, X.; Ji, Y.; Li, Y.; Qu, Y.; Zhao, Y.; Wu, X.; Chen, C. Revealing the binding structure of the protein corona on gold nanorods using synchrotron radiation-based techniques: Understanding the reduced damage in cell membranes. J. Am. Chem. Soc. 2013, 135, 17359–17368. [CrossRef] 8. Nguyen, K.C.; Richards, L.; Rippstein, P.; Massarsky, A.; Moon, T.W.; Tayabali, A.F. Toxicological evaluation of representative silver nanoparticles in macrophages and epithelial cells. Toxicol. In Vitro 2016, 33, 163–173. [CrossRef] 9. Gliga, A.R.; Skoglund, S.; Wallinder, I.O.; Fadeel, B.; Karlsson, H.L. Size-dependent cytotoxicity of silver nanoparticles in human lung cells: The role of cellular uptake, agglomeration and Ag release. Part. Fibre Toxicol. 2014, 11, 11. [CrossRef]PDF Image | Silver Nanoparticles Undergoing Long-Term Aging
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