Search Results (121)

Previous studies have shown mouse antigenotoxic and chemopreventive potential with the administration of Citrus paradisi juice (GJ). To evaluate another activity, the aim of the present report was to determine the beneficial effect of GJ on male mouse reproductive damage induced by cadmium chloride (CC). Seven groups of mice were intragastrically (IG) administered for 11 days. A control group was administered purified water daily, three groups were administered GJ daily (4.1, 16.6, and 33.2 µL/g), plus a single administration of CC (3 mg/kg) on the fifth day of the assay, another group was treated daily with 33.2 µL/g GJ, and a positive control group was treated with 3 mg/kg of CC on day 5 of the experiment. The results with the high GJ dose on the CC-treated mice showed a mean reduction of 88% in sperm quality endpoints (viability, motility, malformations) and a 94% sperm concentration increase. With the same dose, we also determined an 81% reduction in the DNA breaking potential and in the number of micronuclei in the spermatids. We also found an 87% decrease in lipoperoxidation and a 68% decrease in protein oxidation with respect to the CC damage, and a strong DPPH scavenging ability. Our results suggest the potential involvement of the GJ antioxidant in the observed effect. Full article

In pathology laboratories, several volatile organic compounds (VOCs) are used, such as formaldehyde, ethanol, and xylene. These substances are recognized as genotoxic and cytotoxic, which is why their handling poses risks to human health. The buccal micronucleus (MN) cytome assay is a non-invasive, useful, and simple method to detect these effects in exposed individuals. The aim of the study was to evaluate the risk of genotoxicity and cytotoxicity of VOCs in pathology professionals of S. Miguel Island, Azores, Portugal. The study comprised two groups: exposed workers (n = 21) from the three laboratories of S. Miguel, and a reference group (n = 50), randomly chosen from other hospital services without known exposure to VOCs. The exfoliated buccal cells were auto-sampled by all the participants using a cytobrush. The samples were processed in ThinPrep^®, stained with modified Feulgen with Fast Green, and visualized for MN and other nuclear anomalies (ONAs), such as karyorrhexis, pyknotic, and karyolytic cells. Results showed that VOCs have a predictive significance for MN frequency, leading to the conclusion that their exposure is an increased risk factor for the health of these professionals, approximately four times greater than in the control group. Full article

Background: The primary objective of this study was to evaluate the cytotoxic and genotoxic effects of calcium silicate-based sealers (BioRoot RCS and MTA Fillapex) compared to an epoxy-based sealer (AH Plus). Materials and methods: The study was conducted in vitro with the cell lines HepG2 and V79 to evaluate cytotoxicity and genotoxicity using the comet and micronucleus assays. Eluates of the materials were tested at two different concentrations (3 cm²/mL and 0.5 cm²/mL) after an exposure time of 72 h. Data were analyzed using the Mann–Whitney and Kruskal–Wallis tests (p < 0.05). Results: At lower concentrations in both cell lines, MTA Fillapex showed no significant difference in the measured comet assay parameters compared to the negative control (p > 0.05). In addition, it showed significantly lower genotoxic effects compared to AH Plus for all comet assay parameters, concentrations, and cell lines (p ≤ 0.001). BioRoot RCS showed lower primary DNA damage (p ≤ 0.001) than AH Plus, only at higher concentrations and in the HepG2 cell line. Concerning the two tested bioceramic sealers, BioRoot RCS showed higher tail intensity values compared to MTA Fillapex (p < 0.05). In contrast to the results of the comet assay, BioRoot RCS significantly reduced the number of nuclear buds and nucleoplasmic bridges in the HepG2 cell line compared to MTA Fillapex, whereas reduction in the V79 cell line was only observed for nuclear buds (p < 0.05). Both materials increased the number of apoptotic cells compared to the negative control (p < 0.05). In comparison to AH Plus, BioRoot RCS and MTA Fillapex significantly reduced the number of cells with micronuclei and increased the number of cells with undamaged chromatin (p < 0.05). Conclusions: The findings suggest that MTA Fillapex and BioRoot RCS exhibit superior biocompatibility over AH Plus, as evidenced by their lower cytotoxic and genotoxic effects in vitro. These results support the use of calcium silicate-based sealers in clinical practice, highlighting the need for further studies to evaluate their performance in vivo and their implications for patient safety. Full article

Platinum-based radiochemotherapy is associated with hematologic side effects, impacting patient outcomes. However, the clinical mechanisms of cisplatin and its interaction with ionizing radiation (IR), including in biodosimetry for radiotherapy, have not yet been fully clarified. For this purpose, healthy donors’ peripheral blood lymphocytes (PBLs) were pretreated with cisplatin in a pulse (1–4 h) or continuous (24 h) regimen followed by X-rays. DNA damage was assessed as DNA double-strand breaks using repair foci of γH2AX and 53BP1 after 0.5 h and 24 h in G1 PBLs and a proliferation-based cytokinesis-block micronucleus assay. Additionally, cell death and proliferation activity were measured. Unlike a 1 h pulse, a 24 h cisplatin pretreatment caused a concentration-dependent increase in cisplatin-induced foci while decreasing IR-induced foci, especially 24 h after irradiation. This was accompanied by increased apoptosis, with cisplatin and IR having additive effects. Both genotoxins alone caused a dose-dependent increase in micronuclei, while cisplatin significantly reduced binuclear cells, especially after the 24 h treatment, leading to lower micronuclei frequencies post-irradiation. Our results show that prolonged cisplatin exposure, even at low concentrations, impacts the vitality and division activity of PBLs, with significantly stronger effects post-irradiation. This has major implications and must be considered for the detection of DNA damage-associated biomarkers in PBLs used in clinical prediction or biodosimetry during radiotherapy. Full article

Objective: This study aims to evaluate cell proliferation capacity and micronuclei incidence in the presence of nickel–chromium (Ni-Cr)-based dental alloys, with and without the addition of beryllium (Be). The use of these alloys in dental prosthetics is widespread; however, the potential risks associated with their genotoxicity and cytotoxicity require further investigation. The study seeks to provide insight into the safety of these materials and their long-term impact on the health of both patients and dental professionals. Methods: The study was conducted through a comparative analysis of genotoxicity and cytotoxicity using human lymphocyte cultures exposed to two types of Ni-Cr-based dental alloys, one containing beryllium and the other without beryllium. The evaluations were performed according to the OECD Test No. 487 guideline, employing the micronucleus assay and cell proliferation assay. Lymphocytes were exposed to three different alloy concentrations (5 mg/mL, 10 mg/mL, and 20 mg/mL), and the effects on genetic material were analyzed microscopically. Descriptive statistics (mean, standard deviation, and variance) were calculated, and one-way ANOVA was used to assess statistical significance between groups, with a significance threshold of p < 0.05. Results: A significant increase in cytotoxicity and micronuclei incidence was observed in the samples containing beryllium compared to those without beryllium. Statistical analysis revealed significant differences (p < 0.001) between the test and control groups and between different concentrations. Additionally, a direct proportional relationship was noted between alloy concentration and the intensity of genotoxic effects. Microscopic analysis confirmed genetic material damage, indicating a potentially increased risk associated with the use of this type of dental material. Conclusions: The data obtained suggest that Ni-Cr-based dental alloys containing beryllium may present a significant risk of genotoxicity and cytotoxicity. Therefore, the selection of materials used in dental prosthetics should be based on solid scientific evidence, and the use of these alloys should be approached with caution. The study highlights the need for further research to better understand the long-term impact of these materials on human health. Full article

Fludioxonil is a widely used fungicide that is frequently used to combat fungal plant diseases. Consequently, excessive concentrations of fludioxonil may enter and accumulate over time in aquatic systems, harming (micro) organisms in several ways. Thus, it is of great importance to evaluate the potential toxic effects of fludioxonil using bioassays. In the present study, various in vitro assays were used to assess the possible effects of fludioxonil in human cells and aquatic microorganisms. For the investigation of the toxic effects of fludioxonil on freshwater microalgae, Scenedesmus rubescens and Dunaliella tertiolecta were exposed to various environmentally relevant concentrations of the fungicide for a period of 96 h. Fludioxonil at 50–200 μg L⁻¹ significantly limited the growth of both microalgae, especially in the first 24 h of the exposure, where inhibitions up to 82.34% were calculated. The toxicity of fludioxonil was further evaluated via the Microtox test, and the studied fungicide was found to be less toxic for the bacteria Aliivibrio fischeri. Regarding human cells, the fludioxonil’s toxic and cyto-genotoxic effects were assessed using the Trypan blue exclusion test and the Cytokinesis Block MicroNucleus (CBMN) assay. Cell viability in all fludioxonil-treated concentrations was similar to control values according to the results of the Trypan blue exclusion test. However, the CBMN assay was used and revealed that fludioxonil had genotoxic potential in higher concentrations and exerted cytotoxic activity against human lymphocytes. Specifically, only the highest dose of fludioxonil, i.e., 10 μg mL⁻¹, exerted genotoxic effects against human lymphocytes, whereas treatment with 0.5, 1, and 5 μg mL⁻¹ did not lead to statistically significant induction of micronuclei (MN) frequencies compared with the control culture. However, fludioxonil-mediated cytotoxicity was statistically significant, which was demonstrated by the decreased CBPI (cytokinesis block proliferation index) values in all cases except for the lowest dose, i.e., 0.5 μg mL⁻¹. Full article

For the first time, this study investigates in vivo the potential of Na-modified natural clinoptilolite to mitigate cadmium toxicity in ICR mice, a model relevant to human health. We enhanced natural clinoptilolite to improve its cadmium (Cd) exchange capacity. Mice were exposed to environmentally realistic cadmium nitrate Cd(NO₃)₂ doses in their drinking water. The detoxification efficacy of the mineral was evaluated over 45 days in four groups: control (no supplementation), Cd(NO₃)₂ only, clinoptilolite only, and a combination of Cd(NO₃)₂ and clinoptilolite. We assessed Cd bioaccumulation in the liver and kidneys, genotoxicity (micronucleus assay), hematological parameters, and oxidative stress markers. Cd exposure resulted in significant bioaccumulation, reduced growth, changes in erythrograms, DNA damage, and oxidative stress. Mice receiving clinoptilolite alone showed a significant increase in body mass. Modified clinoptilolite led to a nearly 48% reduction in Cd accumulation and a 30% increase in Cd excretion in the Cd-plus-clinoptilolite group compared to the Cd-only group. Erythrogram and leukogram parameters returned to near-normal levels, with reductions in malondialdehyde (MDA) and increases in glutathione (GSH) observed by the end of the experiment. No elevated levels of micronuclei were found following clinoptilolite supplementation. These results suggest that modified clinoptilolite may be a cost-effective detoxifier in Cd-polluted regions. Full article

Staphylea pinnata L., (S. pinnata), has long been recognized in Europe as both a wild food source and a traditional medicinal. This study aimed to characterize the metabolomic profile of the leaf extract of S. pinnata and assess its cytotoxic, genotoxic, and antigenotoxic effects in vitro for the first time. The methanolic extract of the leaves was analyzed using Ultra-Performance Liquid Chromatography–High-Resolution Mass Spectrometry (UPLC-HRMS). To evaluate its cytotoxic, genotoxic, and antigenotoxic properties, the cytokinesis block micronucleus assay was performed on Chinese hamster ovarian K1 cells. The analysis revealed a wide variety of metabolites in the extract, with B-type procyanidins and prodelphinidins being the most abundant. The genotoxicity of the extract varied depending on its concentration; at the lowest concentration (75 μg/mL), it showed no genotoxic effects and exhibited antigenotoxic properties by reducing the frequency of micronuclei induced by mitomycin C. However, at the highest concentration (300 μg/mL), the extract demonstrated genotoxic effects. In conclusion, the S. pinnata extract displayed both genotoxic and antigenotoxic properties, which may be attributed to its phytochemical composition. These findings highlight the complex nature of the plant’s bioactive compounds, suggesting potential therapeutic applications with careful consideration of dosage. Additional research is necessary to understand the mechanisms underlying these properties. Full article

Soil contamination by petroleum derivatives is a growing environmental issue that affects ecosystems and human health, since the hydrocarbons present in them are persistent and toxic, compromising soil quality and biodiversity. This study investigated the potential of a biosurfactant from Candida bombicola URM 3718, to be applied to remove oils from contaminated soils. After isolation, its main surface-active characteristics were evaluated. The biomolecule was then characterized by NMR and FTIR spectroscopy analyses, and its ability to remove motor oil adsorbed on soils with different particle sizes and its genotoxicity profile were determined. Tests to determine surfactant activities revealed a reduction in water surface tension to 30 mN/m with a critical micelle concentration (CMC) of 0.03 g/L. The surfactant was shown to have a glycolipid nature. The removal of burned engine oil sorbed on various kinds of soil was investigated in both static and kinetic assays using the biosurfactant at different concentrations, namely, ½ CMC (0.015 g/L), CMC (0.03 g/L), and 2 × CMC (0.06 g/L). In the static tests, the maximum removal percentage was 65.32% for burned engine oil adsorbed on sandy soil, 59.04% on silty soil, and 57.42% on clayey soil, while in the kinetic tests, this parameter reached 98.60%, 93.22%, and 92.55% for sandy, silty, and clayey soils, respectively. The genotoxicity profile evaluated in Allium cepa roots did not reveal necrosis or the occurrence of micronuclei in the plant root cap cells, demonstrating that the biomolecule thus produced is not toxic. Such findings, when taken together, indicate that the C. bombicola URM 3718 biosurfactant was effective in removing oils and could, therefore, be used as an alternative agent for remediating hydrocarbon-polluted soil. Full article

Alkenylbenzenes occur as natural constituents in a variety of edible plants, in particular those herbs and spices used to give a distinctive flavor to a range of food and feed items. Some alkenylbenzenes with relevance for food, such as estragole and methyleugenol, are known to be genotoxic and carcinogenic in rodents. However, the genotoxic and carcinogenic potential of other structurally related alkenylbenzenes, such as myristicin and elemicin, is still under scientific discussion. Here, we investigated the potential of myristicin and elemicin to induce micronuclei (MN) in V79 cells in comparison to that of estragole and methyleugenol. In addition, we determined the impact of these alkenylbenzenes on cell viability and on the induction of apoptosis and necrosis. All tested alkenylbenzenes affected cell viability in a concentration-dependent manner, albeit to varying degrees. Regarding MN formation, elemicin induced a weak but statistically significant response at 100 µM and 500 µM in the absence of an exogenous metabolizing system (S9 mix). Negative results were obtained for estragole and myristicin at the highest tested non-cytotoxic concentration of 10 µM and 100 µM, respectively. For methyleugenol, the MN assay results were considered equivocal, since the observed change in MN induction was rather small and not supported by a concentration-related trend. These findings indicate that traditional in vitro test systems utilizing exogenous metabolizing systems have limited explanatory power with regard to the genotoxic potential of alkenylbenzenes. Full article

In Europe, there is a growing concern for animal welfare, encompassing both their rights and health. Consequently, identifying biomarkers that predict serious pathological conditions has become crucial in veterinary medicine. The Buccal Micronucleus Cytome (BMCyt) assay is a minimally invasive method that uses biomarkers to evaluate DNA damage and chromosomal instability, using exfoliated buccal cells. A rising frequency of anomalies, such as micronuclei formation, strongly indicates an elevated risk of cancer, neurodegenerative diseases, or accelerated aging, potentially originating from exposure to genotoxins and cytotoxins. This method has been validated in humans, but very little research has been conducted on animals. This work aims to provide a detailed description of an optimized method for collecting buccal exfoliated cells in dogs and to characterize a biomarker related to genomic damage using optical and fluorescent microscopy. Samples from dogs in breeding kennels, including pregnant animals, were tested for chromosomal instability. By following procedures similar to those used in humans, we were able to detect and count major nuclear abnormalities. The percentage of micronuclei was higher compared to other studies. Technical aspects, such as avoiding artifacts and ensuring prior training of the operator, must be taken into account. This work validated the BMCyt method for collecting and processing samples in dogs, potentially enhancing the understanding of micronuclei as biomarkers for pre-pathological states in canines. Full article

The micronucleus test is one of the most popular genotoxicity assays. In order to avoid underestimation of micronuclei frequencies by counting non-replicating cells, the cytokinesis-blocked micronucleus test has been developed. In this technique, only binucleated cells are scored. One underestimated problem is the potential difficulty in discriminating binucleated from mononucleated cells when using DAPI staining, i.e., the possibility that two neighboring mononucleated cells could be mistaken for a binucleated one. The new protocol presented here comprises the addition of acridine orange in order to stain the cytoplasm (in addition to DAPI to stain nuclei and micronuclei). This new technique can increase the sensitivity of the cytokinesis-blocked micronucleus test and avoid underestimation of micronuclei frequencies, an important issue when high doses are employed. Full article

Cistus monspeliensis L. (C. monspeliensis) is used in Italian folk medicine. This study was performed to determine genotoxic and antigenotoxic effects of C. monspeliensis leaf extract against mitomycin C (MMC) using an in vitro cytokinesis-block micronucleus assay (CBMN) in the Chinese Hamster Ovarian K1 (CHO-K1) cell line. The phytochemical composition of C. monspeliensis extract was evaluated using an untargeted metabolomic approach by employing UPLC-PDA-ESI/MS. The automated in vitro CBMN assay was carried out using image analysis systems with a widefield fluorescence microscope and the ImageStreamX imaging flow cytometer. The phytochemical profile of C. monspeliensis extract showed, as the most abundant metabolites, punicalagin, myricetin, gallocathechin, and a labdane-type diterpene. C. monspeliensis, at the tested concentrations of 50, 100, and 200 μg/mL, did not induce significant micronuclei frequency, thus indicating the absence of a genotoxic potential. When testing the C. monspeliensis extract for antigenotoxicity in the presence of MMC, we observed a hormetic concentration-dependent effect, where low concentrations resulted in a significant protective effect against MMC-induced micronuclei frequency, and higher concentrations resulted in no effect. In conclusion, our findings demonstrate that C. monspeliensis extract is not genotoxic and, at low concentration, exhibits an antigenotoxic effect. In relation to this final point, C. monspeliensis may act as a potential chemo-preventive against genotoxic agents. Full article

This study evaluates the cytotoxic and genotoxic effects of PM_2.5 collected from an open-cast coal mining area in northern Colombia. Cyclohexane (CH), dichloromethane (DCM), and acetone (ACE) extracts were obtained using Soxhlet extraction to isolate compounds of different polarities. Human lymphocytes were exposed to the extracted compounds, and cytotoxicity and genotoxicity were assessed using the cytokinesis block micronucleus (CBMN) and comet assays, incorporating FPG and ENDO III enzymes to detect oxidative DNA damage. Chemical analysis revealed that the organic fractions consisted mainly of modified hydrocarbons and volatile organic compounds. The CBMN assay showed a significant increase in micronuclei in binucleated (MNBN) and mononucleated (MNMONO) cells and nucleoplasmic bridges (NPB) in exposed lymphocytes. The comet assay revealed substantial oxidative DNA damage, particularly with the ACE extract, which significantly increased oxidized purines and pyrimidines. DCM induced similar effects, while CH showed moderate effects. CREST immunostaining revealed aneugenic activity, particularly in cells exposed to ACE and DCM extracts. These results suggest that polar fractions of PM_2.5, likely containing metals and modified PAHs, contribute to DNA damage and chromosomal instability. The study highlights the need to monitor the composition of PM_2.5 in mining regions to implement stricter environmental policies to reduce exposure and health risks. Full article

The widespread use of silver nanoparticles in many industries is increasing every year. Along with this use, there is growing concern about the potential unintentional exposure of human and animal organisms to these nanomaterials. It has been shown that AgNPs have the ability to penetrate organisms and can have harmful effects on cells and organs in the body. In order to reduce the effects of AgNPs on living organisms, newer solutions are being investigated, such as particle stabilization or other methods of synthesizing these particles. The physical synthesis of AgNPs using high-voltage arc discharge (HVAD) may be one of these alternatives. To determine the effect of silver nanoparticles obtained by this method, cytogenetic analysis was performed on domestic dog somatic cells using a cytokinesis-blocking micronucleus assay. In the experiments performed, peripheral blood cells of the domestic dog were exposed in vitro for 3 and 24 h to three tested colloidal silver compounds (unstable AgNP-HVAD, sodium citrate-stabilized silver nanoparticles—AgNP+C, and silver nitrate). The toxicity of these compounds was evaluated at concentrations of 5, 10, and 20 µg/L, and the presence of the following cellular abnormalities was analyzed: micronuclei, nuclear buds, nucleoplasmic bridges, or multinucleated cells. The study showed a significant increase in the number of micronuclei compared to the control sample, as well as the presence of nuclear buds and nucleoplasmic bridges in somatic cells of the domestic dog, confirming the genotoxic nature of the particles. However, there was no cytotoxic effect due to the lower number of multinucleated cells and the absence of apoptotic or necrotic cells in the samples analyzed. Further studies are needed to better understand the mechanisms of toxicity of AgNPs produced by the HVAD method and the extent of their effects on mammalian somatic cells. Full article