1. A harmonized and standardized in vitro approach produces reliable results on silver nanoparticles toxicity in different cell lines
- Author
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Ettore Meccia, Maria Condello, Cristina Andreoli, Cinzia Butteroni, Gabriella Di Felice, Isabella De Angelis, Bianca Barletta, Marios G. Krokidis, Annalisa Masi, Stefania Meschini, Flavia Barone, Silvia Corinti, Valentina Prota, Chryssostomos Chatgilialoglu, Emiliano Facchini, and Andrea Zijno
- Subjects
silver nanoparticles ,Silver ,ultrastructural analysis ,Metal Nanoparticles ,010501 environmental sciences ,In Vitro Techniques ,Toxicology ,medicine.disease_cause ,01 natural sciences ,Silver nanoparticle ,oxidative DNA damage ,Cell Line ,03 medical and health sciences ,Dynamic light scattering ,Toxicity Tests ,medicine ,Cytotoxicity ,030304 developmental biology ,0105 earth and related environmental sciences ,0303 health sciences ,immunotoxicity ,Micronucleus Tests ,Chemistry ,genotoxicity ,Comet assay ,Cell culture ,Toxicity ,Biophysics ,cytotoxicity ,Comet Assay ,Micronucleus ,characterization in biological media ,Genotoxicity - Abstract
Despite the widespread use of silver nanoparticles (AgNPs) in different fields and the amount of investigations available, to date, there are many contradictory results on their potential toxicity. In the present study, extensively characterized 20-nm AgNPs were investigated using optimized protocols and standardized methods to test several toxicological endpoints in different cell lines. The agglomeration/aggregation state of AgNPs in culture media was measured by dynamic light scattering (DLS). DNA and chromosomal damage on BEAS-2B and RAW 264.7 cells were evaluated by comet and micronucleus assays, while oxidative DNA damage by modified comet assay and 8-oxodG/8-oxodA detection. We also investigated immunotoxicity and immunomodulation by cytokine release and NO production in RAW 264.7 and MH-S cells, with or without lipopolysaccharide (LPS) stimulus. Transmission electron microscope (TEM) analysis was used to analyze cellular uptake of AgNPs. Our results indicate different values of AgNPs hydrodynamic diameter depending on the medium, some genotoxic effect just on BEAS-2B and no or slight effects on function of RAW 264.7 and MH-S in absence or presence of LPS stimulus. This study highlights the relevance of using optimized protocols and multiple endpoints to analyze the potential toxicity of AgNPs and to obtain reliable and comparable results.
- Published
- 2021