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1

No evidence of the long-term in vitro toxicity of Aeroxide P25 TiO2 nanoparticles in three mammalian cell lines despite the initial reduction of cellular mitochondrial activity

Męczyńska-Wielgosz Sylwia, Bartłomiejczyk Teresa, Grądzka Iwona, Sommer Sylwester, Węgierek-Ciuk Aneta, Lankoff Anna, Sikorska Katarzyna, Wojewódzka Maria, Dobrzyńska Małgorzata, Kruszewski Marcin

Nukleonika

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2024

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Vol. 69, No. 1

13--22

EN

We studied the effects of Aeroxide P25 titanium dioxide nanoparticles (TiO2 NPs) with a diameter of 21 nm on induction of DNA damage and long-term survival of three human cell lines: hepatocellular liver carcinoma HepG2, colorectal adenocarcinoma HT29 and lung carcinoma A549. The endpoints examined were DNA breakage estimated by the comet assay and oxidative base damage recognized by formamide-pyrimidine glycosylase (FPG) estimated with the FPG+ comet assay, frequencies of histone H2AX foci and micronuclei, apoptosis, cell metabolic activity measured by mitochondrial activity (MTT) assay and long-term survival measured by colony-forming ability. Each cell line had a different pattern of DNA breakage and base damage vs. nanoparticle (NP) concentration and treatment time. There was no increase in the frequencies of histone H2AX foci and micronuclei as compared to those in the untreated cells. In parallel with these results, no induction of apoptosis has been found in none of the cell lines tested. The reported experiments provided no evidence of the long-term in vitro toxicity of Aeroxide P25 TiO2 NPs, despite a slight decrease in mitochondrial activity and cell survival during the first 72 h.

2

Wpływ biopaliw na częstość występowania mikrojąder w komórkach CHO-9 i A5491

Skowroń Jolanta, Zapór Lidia, Miranowicz-Dzierżawska Katarzyna

Podstawy i Metody Oceny Środowiska Pracy

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2023

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Nr 1 (115)

27--43

PL

Biopaliwa mają wiele zalet, które czynią je atrakcyjnym źródłem energii, jednak ich wpływ na organizm człowieka nie został jeszcze w pełni poznany. W artykule przedstawiono wyniki badań przeprowadzonych różnymi metodami w warunkach in vitro nad działaniem genotoksycznym czterech biopaliw otrzymanych w procesie transestryfikacji tłuszczów odpadowych. Badania uszkodzeń DNA (badanie mikrojąder) powodowanych przez biopaliwa przeprowadzono na komórkach nabłonka płuc pochodzenia nowotworowego (A549) oraz komórkach jajnika chomika chińskiego (CHO-9). Badane biopaliwa powodowały statystycznie istotny wzrost częstości występowania mikrojąder w komórkach CHO-9 (p < 0,05) w zależności od zastosowanych stężeń. Nie powodowały one jednak statystycznie znaczącego wzrostu częstości występowania mikrojąder w komórkach A549. Wyniki przeglądu baz danych (głównie MEDLINE i EMBASE) pozwoliły wskazać cztery główne źródła zagrożeń dla zdrowia ludzkiego, które są związane ze stosowaniem biopaliw: ryzyko zawodowe, zanieczyszczenie wody/gleby, zanieczyszczenie powietrza związane z produkcją i stosowaniem biopaliw oraz wpływ na ceny żywności. Wyniki przedstawionych badań stanowią jedynie etap oceny toksykologicznej biopaliw, których wpływ na komórki zależy od ich składu chemicznego i od rodzaju komórek stosowanych do badań. Biopaliwo II, otrzymywane z tłuszczu zwierzęcego i zawierające największe stężenie estrów metylowych kwasów tłuszczowych, wykazało działanie genotoksyczne (częstość występowania mikrojąder) w komórkach jajnika chomika chińskiego CHO-9. Przedstawione wyniki badań pozwolą producentom i użytkownikom biopaliw zapoznać się z ryzykiem związanym z ich produkcją i stosowaniem. Zakres tematyczny artykułu obejmuje zagadnienia zdrowia oraz bezpieczeństwa i higieny środowiska pracy będące przedmiotem badań z zakresu nauk o zdrowiu i inżynierii środowiska.

EN

Biofuels have a number of advantages that make them an attractive source of energy. However, their effect on the human body has not been fully understood. The article presents the results of studies on the genotoxic effect of four biofuels obtained in the process of transesterification of waste fats with in vitro methods. DNA damage tests (micronucleus test) of biofuels were carried out on the cells of: neoplastic lung epithelium (A549) and Chinese hamster ovary (CHO-9). The tested biofuels caused a statistically significant increase in the frequency of micronuclei in CHO-9 cells (p < 0.05), depending on the concentrations used. However, they did not induce a statistically significant increase in the frequency of micronuclei in A549 cells. The results of the database review (mainly MEDLINE and EMBASE) identified four main sources of human health risks from biofuels: occupational hazards, water / soil contamination, air pollution from biofuel production and use, and the impact on food prices. The results of the presented studies are only a step in the toxicological assessment of biofuels, the effect of which on cells depends on their chemical composition and the type of cells used for the tests. Biofuel II, obtained from animal fat, containing the highest concentration of fatty acid methyl esters showed the strongest genotoxic effect (induced frequency of micronuclei) on CHO-9 Chinese hamster ovary cells. The presented research results could familiarize the producers and users of biofuels with the risks associated with their use. This article discusses the problems of occupational safety and health, which are covered by health sciences and environmental engineering.

3

Cytotoxicity and genotoxicity of GO-Fe3O4 hybrid in cultured mammalian cells

Jedrzejczak-Silicka M.

Polish Journal of Chemical Technology

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2017

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Vol. 19, nr 1

27--33

EN

The study was aimed at investigating the effect of the Fe3O4 hybrid deposited on graphene oxide (GO- Fe3O4) on the relative viability and DNA integrity. The properties of the GO-Fe3O4 hybrid were analyzed using a transmission electron microscopy (TEM), X-ray diffraction technique (XRD) and thermal gravimetric method (TGA), while the efficiency of graphene oxide covalent functionalization with iron oxide nanospheres was determined by Fourier transform infrared spectroscopy (FT-IR). L929 and MCF-7 cell lines were selected to analyze the biocompatibility of GO-Fe3O4 nanoparticles. The hybrid was tested using WST-1 and LDH leakage assays. DNA integrity was analyzed by agarose gel electrophoresis and micronucleus assay was performed to examine chromosomal damage in the exposed cell lines. The tested GO-Fe3O4 hybrid did not significantly reduce cell metabolism of L929 cells. GO-Fe3O4 hybrid particles only slightly affected the integrity of cell membranes. DNA integrity and micronucleus assays did not indicate genotoxicity of the hybrid.

4

Test mikrojądrowy w retrospektywnej dozymetrii biologicznej

Rawojć K., Miszczyk J.

Technical Issues

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2016

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nr 4

78--82

EN

Rapid retrospective biological dosimetry allows absorbed dose evaluation post exposure to ionizing radiation. One of the main tools of biodosimetry is based on the analysis of the effects resulting from the impact of ionizing radiation on the cell. Various cytogenetic tests give possibility of the accurate dose estimation. To investigate cell response to radiation one performs the analysis of biomarkers approved by International Atomic Energy Agency e.g. the analysis of dicentric chromosomes or micronuclei frequency. Micronucleus test is relatively a faster and therefore more effective method to study changes in the genetic material, induced by various genotoxic agents. This study confirms that micronulei frequency and nuclear division index analysis allows for appropriate absorbed dose estimation when it comes to ionizing radiation. In order to further optimize and facilitate the micronucleus assay and other cytogenetic tests in rapid retrospective biological dosimetry, the research are still ongoing.

5

Micronucleus assay in epithelial cells from the oral cavity and urinary tract in female smokers and non-smokers

Błaszczyk E., Mielzynska-Svach D.

Environmental Biotechnology

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2014

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Vol. 10, No. 2

60--65

EN

The aim of the work was to investigate the possible use of epithelial cells from the oral cavity and urinary tract in identifying smoking-related effects in women. Epithelial cells from the oral cavity and urinary tract were collected from 9 smoking and 9 non-smoking women and subjected to micronucleus assay. The DNA damage (cells with micronuclei and nuclear buds), cytokinetic defects (binucleated cells) and cell death (condensed chromatin, karyorrhexis, pyknosis and karyolysis) were observed after DNA specific staining. In pooled analysis of the frequency of binucleated cells and condensed chromatin cells in 18 studied women, statistically significant differences were noted only in epithelial cells from the oral cavity in comparison to those of the urinary tract. Non pooled results demonstrated no differences in damage frequency in cells collected from the oral cavity and isolated from the urine. The lack of differences in the observed frequencies of micronuclei in buccal and urothelial cells could be an effect of the small size of the sampled group, smoking pattern of the women and the number of cigarettes smoked per day.

6

Induction of Micronuclei in Mice Bone Marrow Cells by Cobalt and Copper Chlorides

Goc Rasgele P., Kekecoglu M., Gokalp Muranli F. D.

Archives of Environmental Protection

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2013

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Vol. 39, no. 1

75-82

EN

The aim of our research was to investigate the genotoxic effects of cobalt chloride and copper chloride in mouse bone marrow cells using the micronucleus (MN) assay. The three different concentrations of cobalt chloride (11.2, 22.5 and 45 mg kg-1) and copper chloride (1.17, 2.35 and 4.70 mg kg-1) were injected intraperitoneally to mice for 24 and 48 hours. It was observed that both of these heavy metals induced a signifi cant increase in frequency of micronucleated polychromatic erythrocytes (MNPCE) at different concentrations in mice for 24 and 48 hours when compared with the control. Furthermore, the signifi cant reduction for the polychromatic erythrocyte/normochromatic erythrocyte (PCE/NCE) ratio which is indicative of bone marrow cytotoxicity was observed in bone marrow cells which were treated with copper chloride at all concentrations for 24 and 48 hours. No reduction of the PCE/NCE ratio was observed both 24 and 48 hours after all the doses of cobalt chloride tested as compared to the negative control. These results lead us to the conclusion that copper chloride may have genotoxic and cytotoxic properties due to induction in the frequency of MN and a reduction in PCE/NCE ratio in bone marrow cells of mice, whereas cobalt chloride induced only genotoxic effect in mice bone marrow.