Millions of humans are exposed occupationally and environmentally to lead, mercury and cadmium compounds. Mercury compounds are less abundant but some of them belong to the most toxic chemicals which ...are known. We evaluated the literature to find out if these metals act in humans as genotoxic carcinogens and if their health effects can be predicted by use of micronucleus (MN) assays with lymphocytes and/or with other genotoxicity tests. Numerous studies showed that lead and mercury induce cancer in humans and also in animals, in vitro experiments with cultured cells indicate that they cause DNA damage via different molecular mechanisms including release of reactive oxygen species and interactions with DNA repair processes. Also in most human studies, positive results were obtained in MN tests with lymphocytes (all 15 occupational studies with lead yielded positive results, with mercury 6 out of 7 investigations were positive). For cadmium, there is clear evidence that it causes cancer in humans; however, induction of chromosomal damage was only seen in high dose experiments with mammalian cells while results of animal and human studies yielded conflicting results (only in 2 of 5MN trials with humans positive findings were reported). Possibly, non-genotoxic mechanisms such as inhibition of apoptosis and interaction with signaling pathways account for the carcinogenic properties of cadmium species. The findings of MN studies with lead and mercury are in excellent agreement with results which were obtained with other endpoints (e.g. chromosomal aberrations and comet formations) and it is evident that this approach can be used for occupational and environmental monitoring of exposed individuals. Important future tasks will be the realization of larger studies with a uniform standardized protocol, the additional evaluation of anomalies other than MN (nuclear buds and bridges) and the combination of such trials with investigations which allow to define the molecular mechanisms relevant for exposed humans.
1,2-unsaturated pyrrolizidine alkaloids (PAs) represent a large group of secondary plant metabolites exhibiting hepatotoxic, genotoxic, and carcinogenic properties upon bioactivation.
To examine how ...the degree of esterification affects the genotoxic profile of PA we investigated cytotoxicity, histone H2AX phosphorylation, DNA strand break induction, cell cycle perturbation, micronuclei formation, and aneugenic effects in different cell models.
Analysis of cytotoxicity and phosphorylation of histone H2AX was structure- and concentration-dependent: diester-type PAs (except monocrotaline) showed more pronounced effects than monoester-type PAs. Cell cycle analysis identified that diester-type PAs induced a S-phase arrest and a decrease in the occurrence of cells in the G1-phase. The same structure-dependency was observed by flow-cytometric analysis of PA-induced micronuclei in CYP3A4-overexpressing V79 cells. Analysis of centromeres induced by lasiocarpine in the micronuclei by fluorescence in situ hybridization indicated an aneugenic effect in V79h3A4 cells. Comet assays revealed no significant induction of DNA strand breaks for all investigated PAs.
Overall, diester-type PAs induced more pronounced effects than monoester-type PAs. Furthermore, our results indicate aneugenic effects upon exposure towards lasiocarpine in vitro. These data improve our understanding how structural features of PA influence the genotoxic profile. Especially, the monoester-type PAs seem to induce less severe effects than other PAs.
•Pyrrolizidine alkaloids induce structure-dependently cytotoxic & genotoxic effects.•Pyrrolizidine alkaloids induce a cell cycle arrest in S-phase.•Lasiocarpine induced micronuclei show centromeres indicating an aneugenic mechanism.
•Non-thermal physical plasma is an emerging tool in biomedical application.•We investigated the potential genotoxicity of non-thermal plasma.•The micronucleus assay, the HRPT1 assay, and a ...clonogenicity assay were used.•The assays indicated the absence of genotoxic events for the investigated plasma source.
Cold atmospheric pressure plasmas represent a favorable option for the treatment of heat sensitive materials and human or animal tissue. Beneficial effects have been documented in a variety of medical conditions, e.g., in the treatment of chronic wounds. It is assumed that the main mechanism of the plasma’s efficacy is mediated by a stimulating dissipation of energy via radiation and/or chemical energy. Although no evidence on undesired side effects of a plasma treatment has yet been presented, skepticism toward the safety of the exposure to plasma is present. However, only little data regarding the mutagenic potential of this new treatment option is available. Accordingly, we investigated the mutagenic potential of an argon plasma jet (kinpen) using different testing systems in accordance with ISO norms and multiple cell lines: a HPRT1 mutation assay, a micronucleus formation assay, and a colony formation assay. Moderate plasma treatment up to 180s did not increase genotoxicity in any assay or cell type investigated. We conclude that treatment with the argon plasma jet kinpen did not display a mutagenic potential under the test conditions applied and may from this perspective be regarded as safe for the use in biomedical applications.
Although the risk of pregnancy with Down syndrome (DS) increases with age, conceptions with trisomy 21 can occur in mothers aged 35 or less. The micronucleus test on peripheral blood lymphocytes is a ...well-recognized method for studying chromosomal instability. The aim of this study was to evaluate the application of the micronucleus assay and fluorescence in situ hybridization (FISH) for estimation of chromosome instability and occurrence of trisomy 21 in young parents having pregnancy or a child with the regular form of Down syndrome. The study included 54 parents (27 couples) who had previous pregnancy with trisomy 21 at age 35 or less. The control group consisted of 30 couples with two healthy children and no previous spontaneous abortions. Parents with trisomy 21 pregnancy had significantly higher frequencies of micronuclei in binucleated cells. There was no statistically significant difference between the study and control groups in the frequencies of micronuclei in mononuclear cells, nuclear buds, or nucleoplasmic bridges. FISH analysis showed higher percentages of micronuclei containing whole chromosomes as well as statistically significant higher numbers of micronuclei containing chromosome 21 in the peripheral blood of DS parents. There was no statistically significant difference between the two groups in the responses of peripheral blood lymphocytes to treatment with the mutagen mitomycin C. Our results suggest that young parents with a history of the regular form of Down syndrome have a higher susceptibility to chromosome nondisjunction in peripheral blood lymphocytes. The micronucleus assay showed high specificity, but moderate sensitivity, for risk assessment of trisomy 21 pregnancy.
•Young parents of Down syndrome have higher susceptibility to nondisjunction.•The micronucleus assay have high specificity for risk assessment of T21.•No significant difference was found in the response to mitomycin C treatment.
There is concern about human exposure to nanoplastics from intentional use or degradation of plastics in the environment. This review assesses genotoxic effects of nanoplastics, defined as particles ...with a primary size of less than 1000 nm. The majority of results on genotoxicity come from studies on polystyrene (PS) particles in mammalian cell cultures. Most studies have measured DNA strand breaks (standard comet assay), oxidatively damaged DNA (Fpg-modified comet assay) and micronuclei. Twenty-nine out of 60 results have shown statistically significant genotoxic effects by PS exposure in cell cultures. A statistical analysis indicates that especially modified PS particles are genotoxic (odds ratio = 8.6, 95 % CI: 1.6, 46) and immune cells seems to be more sensitive to genotoxicity than other cell types such as epithelial cells (odds ratio = 8.0, 95 % CI: 1.6, 39). On the contrary, there is not a clear association between statistically significant effects in genotoxicity tests and the primary size of PS particles, (i.e. smaller versus larger than 100 nm) or between the type of genotoxic endpoint (i.e. repairable versus permanent DNA lesions). Three studies of PS particle exposure in animals have shown increased level of DNA strand breaks in leukocytes and prefrontal cortex cells. Nanoplastics from polyethylene, propylene, polyvinyl chloride and polyethylene terephthalate have been investigated in very few studies and it is currently not possible to draw conclusion about their genotoxic hazard. In summary, there is some evidence suggesting that PS particles may be genotoxic in mammalian cells.
Titanium dioxide nanoparticles (TiO2 NPs, size <100 nm) find applications in a wide range of products including food and cosmetics. Studies have found that exposure to TiO2 NPs can cause ...inflammation, cytotoxicity, genotoxicity and cell apoptosis. In this article, we have reviewed the recent literature on the potential of TiO2 NPs to cause genotoxicity and summarized the results of two standard genotoxicity assays, the comet and micronucleus (MN) assays. Analysis of these peer-reviewed publications shows that the comet assay is the most common genotoxicity test, followed by MN, Ames, and chromosome aberration tests. These assays have reported positive as well as negative results, although there is inconsistency in some results that need to be confirmed further by well-designed experiments. We also discuss the possible mechanisms of TiO2 NP genotoxicity and point out areas that warrant further research.
In the last decades, the micronucleus assay has been recognized as a suitable biomarker for monitoring populations exposed to many different occupational factors, lifestyle, environmental conditions, ...radiation exposure, and deleterious effects of pesticides. The objective of this work is to direct the design of future field studies in the assessment of the risk of children exposed to environmental mutagens, radiation, and pesticides. This review sought available information on the analysis of micronuclei in oral cells in children. A literature search for papers investigating DNA damage, genetic damage, oral cells, buccal cells, genotoxicity, mutagenicity and micronucleus was begun in 2000 and is scheduled to be concluded in May, 2022. Briefly, a search of PubMed, MEDLINE, and Google Scholar for a variety of articles was performed. The results showed that there are still few studies that addressed micronuclei of oral cells in children exposed to the most diverse environmental conditions. Only environmental pollution was associated with damage to the genome of oral cells in children. Therefore, researchers need to be calibrated in cell analysis, standardization of field study protocols and the development of new research in the evaluation of children using the micronucleus test as a tool in child biomonitoring.
Micronuclei (MNs) are extranuclear DNA-containing bodies and determining MN frequencies is a measure of genomic instability. An age-related increase in MN frequencies in lymphocytes has been ...quantified, but this effect has not yet been measured in nasal and buccal cells.
We determined the effect of age on the MN frequency distributions in buccal and nasal cells among a sample of a general adult population in Switzerland. To maximize the power to detect an effect of age in our population study, we recruited preferentially younger and older working age adults. We harvested buccal and nasal cells from 32 young (19–36 year) and 33 working age (47–71 years) participants. The collected cells were washed, centrifuged, and stained (Feulgen) before microscopic manual counting in 2000 cells. Based on these results, we developed an age-dependent background MN frequency chart to help interpret an individual's MN frequency score as an early signal for the effect of genotoxic exposure.
MN frequencies were respectively 0.53‰ and 0.47‰ for buccal and nasal among the younger and 0.87‰ and 1.03‰ in the older working age group. This corresponded to a multiplicative slope of 14% and 20% per 10 years of age for buccal and nasal cells, respectively.
Based on our study results, we are able to propose an approach for interpreting an individual's MN screening results.
•An individual's MN results can be interpreted using baseline MN frequencies values.•Age dependent baseline buccal cell MN frequencies were 0.53‰–0.87‰.•Age dependent baseline nasal cell MN frequencies were 0.47‰–1.03‰.
Shipyards are industrial areas where workers are likely exposed to environmental pollutants such as welding fumes, fine organic solvent and dye dust, that render the occupational environment a high ...risk one. Assessing the risk that workers are exposed to is a high critical factor in improving their working conditions. The present study aims to investigate the potential genetic damage to workers exposed to a harsh environment in a Greek shipyard. It is focused on assessing the percentage of induced micronuclei, as well as on changes in the various cell types of shipyard workers’ oral mucosa epithelium by implementing the buccal micronucleus cytome assay. Exposed workers appeared with statistically significant induced micronuclei as compared to office employees. Statistically, significant cell lesions were detected and are related to workers’ exposure to environmental conditions. The workers’ smoking habit contributed as well to the observed buccal epithelial cell alterations. The observed data signify the high-risk workers are exposed; resulting in the shipyard’s management the need to implement measures improving the working environment conditions and to reevaluate the workers’ personal protective equipment requirements.
► We consider current in vitro OECD genotoxicity tests for nanomaterials. ► Ames test does not appear to be suitable for nanomaterial assessment. ► In vitro HPRT and micronucleus assays require ...nanomaterial specific protocols. ► We recommend a strategic in vitro genotoxicity testing strategy for nanomaterials.
There is a pressing requirement to define a hazard identification and risk management strategy for nanomaterials due to the rapid growth in the nanotechnology industry and their promise of life-style revolutions through the development of wide-ranging nano-containing consumer products. Consequently, a battery of well defined and appropriate in vitro assays to assess a number of genotoxicity endpoints is required to minimise extensive and costly in vivo testing. However, the validity of the established protocols in current OECD recognised genotoxicity assays for nanomaterials is currently being questioned. In this report, we therefore consider the in vitro OECD genotoxicity test battery including the Ames, micronucleus and HPRT forward mutation assays, and their potential role in the safety assessment of nanomaterial induced DNA damage in vitro.
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