The evaluation of the frequency of micronuclei (MN) is a broadly utilised approach in in vitro toxicity testing. Nevertheless, the specific properties of nanomaterials (NMs) give rise to concerns regarding the optimal methodological variants of the MN assay. In bronchial epithelial cells (BEAS-2B), we tested the genotoxicity of five types of NMs (TiO₂: NM101, NM103; SiO₂: NM200; Ag: NM300K, NM302) using four variants of MN protocols, differing in the time of exposure and the application of cytochalasin-B combined with the simultaneous and delayed co-treatment with NMs. Using transmission electron microscopy, we evaluated the impact of cytochalasin-B on the transport of NMs into the cells. To assess the behaviour of NMs in a culture media for individual testing conditions, we used dynamic light scattering measurement. The presence of NMs in the cells, their intracellular aggregation and dispersion properties were comparable when tests with or without cytochalasin-B were performed. The genotoxic potential of various TiO₂ and Ag particles differed (NM101 < NM103 and NM302 < NM300K, respectively). The application of cytochalasin-B tended to increase the percentage of aberrant cells. In conclusion, the comparison of the testing strategies revealed that the level of DNA damage induced by NMs is affected by the selected methodological approach. This fact should be considered in the interpretation of the results of genotoxicity tests.

Olfactory function, orchestrated by the cells of the olfactory mucosa at the rooftop of the nasal cavity, is disturbed early in the pathogenesis of Alzheimer’s disease (AD). Biometals including zinc and calcium are known to be important for sense of smell and to be altered in the brains of AD patients. Little is known about elemental homeostasis in the AD patient olfactory mucosa. Here we aimed to assess whether the disease-related alterations to biometal homeostasis observed in the brain are also reflected in the olfactory mucosa. We applied RNA sequencing to discover gene expression changes related to metals in olfactory mucosal cells of cognitively healthy controls, individuals with mild cognitive impairment and AD patients, and performed analysis of the elemental content to determine metal levels. Results demonstrate that the levels of zinc, calcium and sodium are increased in the AD olfactory mucosa concomitantly with alterations to 17 genes related to metal-ion binding or metal-related function of the protein product. A significant elevation in alpha-2-macroglobulin, a known metal-binding biomarker correlated with brain disease burden, was observed on the gene and protein levels in the olfactory mucosa cells of AD patients. These data demonstrate that the olfactory mucosa cells derived from AD patients recapitulate certain impairments of biometal homeostasis observed in the brains of patients.

Efficient and safe carriers of genetic material are crucial for advancing gene therapy. Three new series of cationic dendritic nanocarriers based on a carbosilane scaffold, differentiated by peripheral modifications: saccharide (CS-glyco), amine (CS-N), and phosphonium dendrimers (CS-P) were designed for binding, protecting, and releasing polyanionic compounds like therapeutic siRNA. Besides introducing synthetic methodology, this study brings a unique direct interstructural comparison of 16 dendritic nanovector's characteristics, addressing a gap in typical research that focuses on uniform structural types. The study evaluates the dendrimer's in vitro cytotoxicity, biophysical properties, and complexation capabilities in comparison with widely used PAMAM dendrimers. CS-glyco and PAMAMs were significantly less toxic to MCF-7 and THP-1 cell lines than were CS-N and CS-P, despite having the same peripheral charge density. Notably, CS-glyco maintained biocompatibility comparable to analogous neutral CS glycodendrimers, underscoring the exceptional capability of sugar coating to reduce toxicity. Dendriplexes formed from these nanocarriers protected siRNA from RNase degradation and facilitated its release in the presence of heparin, highlighting its potential in gene delivery applications. The study provides a background for future in-depth investigations into the introduced dendritic nanocarriers, which show significant potential for advancing drug delivery.

Emissions from modern gasoline engines represent an environmental and health risk. In this study, we aimed to compare the toxicity of organic compound mixtures extracted from particulate matter (PM extracts) produced by neat gasoline (E0) and a blend containing 15% ethanol (E15), which is offered as an alternative to non-renewable fossil fuels. Human lung BEAS-2B cells were exposed to PM extracts, and biomarkers of genotoxicity, such as DNA damage evaluated by comet assay, micronuclei formation, levels of phosphorylated histone H2AX, the expression of genes relevant to the DNA damage response, and exposure to polycyclic aromatic hydrocarbons (PAHs), were determined. Results showed that both PM extracts significantly increased the level of oxidized DNA lesions. The E0 extract exhibited a more pronounced effect, possibly due to the higher content of nitrated PAHs. Other endpoints were not substantially affected by any of the PM extracts. Gene expression analysis revealed mild but coordinated induction of genes related to DNA damage response, and a strong induction of PAH-inducible genes, indicating activation of the aryl hydrocarbon receptor (AhR). Our data suggest that the addition of ethanol into the gasoline diminished the oxidative DNA damage, but no effect on other genotoxicity biomarkers was observed. Activated AhR may play an important role in the toxicity of gasoline PM emissions.

Solid fuels are still widely used in household heating in Europe and North America. Emissions from boilers are released in proximity to people. Therefore, there is a need to minimise the toxicity of emissions affecting human health to the greatest extent possible. This study compares the genotoxic potential of the emissions of four boilers of modern and old design (automatic, gasification, down-draft, over-fire) operating at reduced output to simulate the real-life combustion fed by various fossil and renewable solid fuels (hard coal, brown coal, brown coal briquettes, wood pellets, wet and dry spruce). Organic emissions were tested for genotoxic potential by analysing bulky DNA adducts and 8-oxo-dG adduct induction. There was no consistent genotoxic pattern among the fuels used within the boilers. Genotoxicity was strongly correlated with polycyclic aromatic hydrocarbon (PAH) content, and even stronger correlation was observed with particulate matter (PM). In all measured variables (PM, PAHs, genotoxicity), the technology of the boilers was a more important factor in determining the genotoxic potential than the fuels burned. The highest levels of both bulky and 8-oxo-dG DNA adducts were induced by organics originating from the over-fire boiler, while the automatic boiler exhibited genotoxic potential that was ~1000- and 100-fold lower, respectively.