<span class="paragraphSection"><div class="boxTitle">Abstract</div>A number of <span style="font-style:italic;">in vitro</span> methodologies have been used to assess the genotoxicity of different nanomaterials, including titanium dioxide nanoparticles (TiO<sub>2</sub> NPs) and silver nanoparticles (AgNPs). The <span style="font-style:italic;">in vitro</span> micronucleus assay is one of the most commonly used test methods for genotoxicity evaluation of nanomaterials. However, due to the novel features of nanomaterials, such as high adsorption capacity and fluorescence properties, there are unexpected interactions with experimental components and detection systems. In this study, we evaluate the interference by two nanoparticles, AgNPs and TiO<sub>2</sub> NPs, with the <span style="font-style:italic;">in vitro</span> micronucleus assay system and possible confounding factors affecting cytotoxicity and genotoxicity assessment of the nanomaterials including cell lines with different <span style="font-style:italic;">p</span>53 status, nanoparticle coatings and fluorescence, cytochalasin B, fetal bovine serum in cell treatment medium and different measurement methodologies for detecting micronuclei. Our results showed that micronucleus induction by AgNPs was similar when evaluated using flow cytometry or microscope, whereas the induction by TiO<sub>2</sub> NPs was different using the two methods due to TiO<sub>2</sub>’s fluorescence interference with the cytometry equipment. Cells with the mutated <span style="font-style:italic;">p</span>53 gene were more sensitive to micronucleus induction by AgNPs than the <span style="font-style:italic;">p</span>53 wild-type cells. The presence of serum during treatment increased the toxicity of AgNPs. The coatings of nanoparticles played an important role in the genotoxicity of AgNPs. These collective data highlight the importance of considering the unique properties of nanoparticles in assessing their genotoxicity using the <span style="font-style:italic;">in vitro</span> micronucleus assay.</span>
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