The in vitro micronucleus (MN) assay is a component of most test batteries used in assessing potential genotoxicity. Our previous study adapted metabolically competent HepaRG cells to the ...high-throughput (HT) flow-cytometry-based MN assay for genotoxicity assessment (Guo et al. in J Toxicol Environ Health A 83:702–717, 2020b,
https://doi.org/10.1080/15287394.2020.1822972
). We also demonstrated that, compared to HepaRG cells grown as two-dimensional (2D) cultures, 3D HepaRG spheroids have increased metabolic capacity and improved sensitivity in detecting DNA damage induced by genotoxicants using the comet assay (Seo et al. in ALTEX 39:583–604, 2022,
https://doi.org/10.14573/altex.22011212022
). In the present study, we have compared the performance of the HT flow-cytometry-based MN assay in HepaRG spheroids and 2D HepaRG cells by testing 34 compounds, including 19 genotoxicants or carcinogens and 15 compounds that show different genotoxic responses in vitro and in vivo. 2D HepaRG cells and spheroids were exposed to the test compounds for 24 h, followed by an additional 3- or 6-day incubation with human epidermal growth factor to stimulate cell division. The results demonstrated that HepaRG spheroids showed generally higher sensitivity in detecting several indirect-acting genotoxicants (require metabolic activation) compared to 2D cultures, with 7,12-dimethylbenzanthracene and
N
-nitrosodimethylamine inducing higher % MN formation along with having significantly lower benchmark dose values for MN induction in 3D spheroids. These data suggest that 3D HepaRG spheroids can be adapted to the HT flow-cytometry-based MN assay for genotoxicity testing. Our findings also indicate that integration of the MN and comet assays improved the sensitivity for detecting genotoxicants that require metabolic activation. These results suggest that HepaRG spheroids may contribute to New Approach Methodologies for genotoxicity assessment.
Nanomaterials (NMs) generally display fascinating physical and chemical properties that are not always present in bulk materials; therefore, any modification to their size, shape, or coating tends to ...cause significant changes in their chemical/physical and biological characteristics. The dramatic increase in efforts to use NMs renders the risk assessment of their toxicity highly crucial due to the possible health perils of this relatively uncharted territory. The different sizes and shapes of the nanoparticles are known to have an impact on organisms and an important place in clinical applications. The shape of nanoparticles, namely, whether they are rods, wires, or spheres, is a particularly critical parameter to affect cell uptake and site‐specific drug delivery, representing a significant factor in determining the potency and magnitude of the effect. This review, therefore, intends to offer a picture of research into the toxicity of different shapes (nanorods, nanowires, and nanospheres) of NMs to in vitro and in vivo models, presenting an in‐depth analysis of health risks associated with exposure to such nanostructures and benefits achieved by using certain model organisms in genotoxicity testing. Nanotoxicity experiments use various models and tests, such as cell cultures, cores, shells, and coating materials. This review article also attempts to raise awareness about practical applications of NMs in different shapes in biology, to evaluate their potential genotoxicity, and to suggest approaches to explain underlying mechanisms of their toxicity and genotoxicity depending on nanoparticle shape.
This review, therefore, intends to offer a complete picture of all research into the cytotoxicity and genotoxicity of different shapes of NMs (nanorods NRs, nanowires NWs, and nanospheres NSs) to in vitro and in vivo models. Summary of in vitro and in vivo cytotoxicity and genotoxicity studies with 13 types of NMs (Ag, Al2O3, Au, BNNT, CeO2, CdS, Co, GaP, Fe, Ni, Si, Ti, and Zn) with different shapes including NRs, NWs, and NSs using in vitro and in vivo test systems.
With the aim to investigate the mechanisms of action of nano plastics (nano PS) on plants, seeds of Allium cepa were germinated for 72 h in the presence of polystyrene nano PS (50 nm size, at ...concentrations of 0.01, 0.1 and 1 g L−1) and, subsequently, roots were analysed by a multifaceted approach. No effect was induced by any concentration of nano PS on the percentage of seed germination while root growth was inhibited by 0.1 and 1 g L−1 nano PS. Cytological analysis of the root meristems indicated cytotoxicity (reduction of mitotic index) and genotoxicity (induction of cytogenetic anomalies and micronuclei) starting from the lowest dose. Moreover, the biochemical and histochemical analysis of oxidative stress markers gave evidence of stress induction, especially at the highest doses. Damages reported could be due to mechanical surface contact in root external layers, as evidenced by histological localization, and to the internalization of nano PS in different cellular compartments, observed under TEM. The present research underlines the hazardous nature of nano PS, that for their ability to be internalized into crop plants, can enter into different trophic levels of the food chain.
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•Cyto-physiological traits in onion roots treated by nano-polystyrene were studied.•Nano PS reduced root elongation in onion seedlings during germination.•Nano PS induced cyto/genotoxicity on root meristem.•The highest applied concentrations of nano PS triggered oxidative stress.•Nano PS were internalized in different root cellular compartments.
The species Rosmarinus officinalis L. (rosemary) is an herb from the Lamiaceae family, widely used in cooking as a food preservative, seasoning or condiment. It also stands out for its therapeutic ...properties, mainly presenting antioxidant, antibacterial and antitumor activity. The aim of this study was to evaluate the cytotoxic and mutagenic activity of R. officinalis L. essential oil through the Allium cepa bioassay. This test constitutes an excellent plant model routinely used due to its sensitivity, low cost and good correlation with test systems in mammals. The defined concentrations for carrying out the test were 750, 243, 81 and 27 µg mL-1. Five bulbs were used, 4 roots of each, measuring approximately 2 cm, and they were analyzed on two slides. All assays were performed at least in triplicate and compared to the negative control. The statistical test of analysis of variance (ANOVA with a fixed factor) was used, followed by Tukey's multiple comparisons test, for p < 0.05. For this purpose, the GraphPad Prism program (version 6.0) was used. The results showed a cytotoxic and mutagenic effect for all concentrations used of the essential oil of R. officinalis L. However, it is important to conduct further research using other genotoxicological tests with different endpoints and at different concentrations, in order to clarify the interaction of the essential oil of the species R. officinalis L. with the genetic material of the cell and its possible mechanism of action.
Meso-zooplankton plays a vital role in maintaining healthy marine ecosystems, and some of the taxa provide biological indications for the monitoring of environmental and climate change. Recently, ...several newly emerging stressors were shown to impact marine and coastal meso-zooplankton in some ways. Marine organisms' genomic core, tightly packed with high-level integrity, can be damaged by anthropogenic activities in coastal zones worldwide and impact their integrity. Genomic integrity loss leads to a cascade of effects on the destruction of the food chain sequences, from primary producers to higher invertebrates. Therefore, monitoring genomic integrity loss using ecotoxicological approaches that focus on genetic changes appears to be a suitable approach. A literature review shows that different stressors severely impact genomic integrity through DNA damage at different concentrations and exposure times. Contaminated sediments also strongly impact the genomic integrity of estuaries and adjacent coastal meso-zooplankton communities.
•Mechanisms of genotoxicity in the meso-zooplankton evaluated.•Scarcity of genotoxicity assessment in the coastal meso-zooplankton demonstrated.•Several stressors have a significant impact on genomic integrity by causing DNA damage at varying concentrations and exposure periods.•Even contaminated sediments have a strong impact on genomic integrity of coastal meso-zooplankton.
Anthocyanins belong to phenolic pigments and are known to have various pharmacological activities. This study aimed to investigate whether anthocyanins could inhibit hydrogen peroxide (H 2 O 2 ...)-induced oxidative damage in human retinal pigment epithelial ARPE-19 cells. Our results indicated that anthocyanins suppressed H 2 O 2 -induced genotoxicity, while inhibiting reactive oxygen species (ROS) production and preserving diminished glutathione. Anthocyanins also suppressed H 2 O 2 -induced apoptosis by reversing the Bcl-2/Bax ratio and inhibiting caspase-3 activation. Additionally, anthocyanins attenuated the release of cytochrome c into the cytosol, which was achieved by interfering with mitochondrial membrane disruption. Moreover, anthocyanins increased the expression of heme oxygenase-1 (HO-1) as well as its activity, which was correlated with the phosphorylation and nuclear translocation of nuclear factor-erythroid-2 related factor 2 (Nrf2). However, the cytoprotective and anti-apoptotic effects of anthocyanins were significantly attenuated by the HO-1 inhibitor, demonstrating that anthocyanins promoted Nrf2-induced HO-1 activity to prevent ARPE-19 cells from oxidative stress. Therefore, our findings suggest that anthocyanins, as Nrf2 activators, have potent ROS scavenging activity and may have the potential to protect ocular injury caused by oxidative stress.
Emodin is an anthraquinone secondary metabolite produced by several species of plants and fungi. Emodin is known for its pharmacological versatility, and, in the textile industry, for its good dyeing ...properties. However, its use in the textile industry can result in the formation and disposal of large volumes of wastewater. Emodin mutagenicity has been shown in bacteria and in human cells, but little is known about its possible toxic, genotoxic, or mutagenic effects in aquatic organisms. We have evaluated the eco/genotoxicity of emodin to aquatic organisms. Emodin was toxic to Daphnia similis (EC50 = 130 μg L−1) and zebrafish embryos (LC50 = 25 μg L−1). No toxicity was observed for Raphidocelis subcapitata, Ceriodaphnia dubia, or Parhyale hawaiensis. Additional biochemistry/molecular studies are needed to elucidate the toxic/mutagenic pathways of emodin in aquatic organisms. The PNEC value for emodin was 0.025 μg L−1. In addition to mutagenicity in the Salmonella/microsome assay, emodin was mutagenic in the micronucleus assay in the amphipod P. hawaiensis. Among the anthraquinone dyes tested to date, natural or synthetic, emodin was the most toxic to aquatic species.
•Emodin is toxic to Daphnia similis and Danio rerio embryos.•Among the organisms studied, Danio rerio embryo was the most sensitive to emodin.•Emodin is mutagenic to Parhyale hawaiensis.•Emodin is more toxic than dermorubin and dermocybin to aquatic species.
Furan is a widespread endogenous contaminant in heat-processed foods that can accumulate rapidly in the food chain and has been widely detected in foods, such as wheat, bread, coffee, canned meat ...products, and baby food. Dietary exposure to this chemical may bring health risk. Furan is classified as a possible category 2B human carcinogen by the International Agency for Research on Cancer, with the liver as its primary target organ. Hepatic fibrosis is the most important nontumoral harmful effect of furan and also an important event in the carcinogenesis of furan. Although the specific mechanism of furan-induced liver fibrosis is still unclear, it may involve oxidative stress and genetic toxicity, in which the activation of cytochrome P450 2E1 (CYP2E1) may be the key event. Thus, we conducted a study using an integrating multi-endpoint genotoxicity platform in 120-day in vivo subchronic toxicity test in rats. Results showed that the rats with activated CYP2E1 exhibited DNA double-strand breaks in D4, gene mutations in D60, and increased expression of reactive oxygen species and nuclear factor erythroid 2-related factor 2 in D120. Necrosis, apoptosis, hepatic stellate cell activation, and fibrosis also occurred in the liver, suggesting that furan can independently affect liver fibrosis through oxidative stress and genotoxicity pathways. Point of Departure (PoD) was obtained by benchmark-dose (BMD) method to establish health-based guidance values. The human equivalent dose of PoD derived from BMDL05 was 2.26 μg/kg bw/d. The findings laid a foundation for the safety evaluation and risk assessment of furan and provided data for the further construction and improvement of the adverse outcome pathway network in liver fibrosis.
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•An integrated genotoxicity platform was applied to explore the AOP of furan.•Furan may induce gene mutation in a cumulative way, independent of oxidative stress.•The AOP of CYP2E1 activation leading to liver fibrosis was preliminary established.•BMDLs of key events from AOP are more protective than values from apical endpoints.
Ibuprofen (IBP) is an anti-inflammatory drug found in aquatic environments, potentially toxic for the biota. We exposed the test fish C. decemmaculatus to two environmentally relevant concentrations ...(50 and 100 μg IBP/L) for 4 and 12 d and evaluated the effect on some biomarkers. Micronucleus test, nuclear abnormality test and comet assay indicated cyto-genotoxicity at both concentrations and exposure periods. Oxidative stress and biochemical biomarkers were not affected, excepting muscle AChE activity for 4 d. Muscle metabolic biomarkers showed significant decrease in ETS, lipid and protein content, while carbohydrate content was not affected. The CEA index increased at the lower IBP concentration for 4 and 12 d, possibly due to changes in body energy reserves. A full-factorial GLM performed to assess the effects of IBP and exposure times showed that the metabolic and genotoxicity biomarkers were the most sensitive to IBP toxicity, mainly at 50 μg IBP/L for 4 d.
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•Exposure of Cnesterodon decemmaculatus to IBP was more toxic at 4 than at 12 d.•Exposure of Cnesterodon decemmaculatus was more toxic at 50 than at 100 μg IBP/L.•Metabolic and genotoxicity biomarkers were more sensitive to IBP toxicity than oxidative stress biomarkers.•CEA index was affected by the interaction of IBP concentration and exposure time.•IBP concentration-exposure time interaction may compromise fish energy budget.
