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Toxicity assessment of two-dimensional nanomaterials molybdenum disulfide in Gallus gallus domesticus

Recently two-dimensional nanomaterials, such as graphene and molybdenum disulfide (MoS2), have received much attention as adsorbent materials for the effective removal of organic contaminants. MoS2 is attracting attention, not only for its chemical-physical properties, but also for its wide availability in nature as a constituent of molybdenite. The aim of this investigation was to assess the effects of different MoS2 concentrations (5 × 10(-1), 5 × 10(-2) and 5 × 10(-3) mg/ml) on the embryonated eggs of Gallus gallus domesticus, according to Beck method. We evaluated the toxic effect of the MoS2 powder purchased at Sigma-Aldrich indicated as "received" and MoS2 powder treated via mechanical milling indicated as "ball mille". Subsequently, the embryos were sacrificed at different times of embryonic development (11th, 15(th) and 19(th) day after incubation) in order to evaluate their embryotoxic and teratogenic effects. The alterations of the embryonic development were studied by morphological and immunohistochemical analysis of the tissues. The results obtained have shown the toxicity of both powders of MoS2 with a high percentage of deaths and growth delays. Moreover, the immunohistochemical analysis performed on several tissue sections showed a strong positivity to the anti-metallothionein1 antibody only for the erythrocytes.

E. M. Scalisi, 2A. Salvaggio, F. Antoci, 2A. Messina, R. Pecoraro, M. Cantarella, G. Gorrasi, G. Impellizzeri, and M. V. Brundo,Toxicity assessment of two-dimensional nanomaterials molybdenum disulfide in Gallus gallus domesticus, Ecotoxicol Environ Saf, 2020, 200, 110772.



The association between metal exposure and semen quality in Chinese males: The mediating effect of androgens

As a crucial factor in male reproduction, androgens may represent an intermediate biological mechanism linking metal exposure with effects on semen quality. This study aimed to investigate the association between metal exposure and semen quality, and to assess the mediating role of seminal androgens between metal exposure and semen quality. We investigated the presence of 10 metals in semen and assessed their effect on semen quality in 1136 men recruited from a hospital in Shenzhen, China. Of these, 464 subjects were randomly selected for 4 androgens detection in semen. Cross-sectional associations between single/multiple metals, androgen levels and semen quality were explored by multivariable linear regressions. Mediation analysis was performed to detect the role of seminal androgens on the association between metal exposure and semen quality. Seminal selenium and iron were positively associated with both sperm concentration and total sperm count. Negative associations were observed between both manganese and zinc and sperm concentration, molybdenum and total sperm count, copper and sperm motility. Furthermore, we found significant dose-dependent relationships between both iron and selenium levels and dihydrotestosterone (DHT), arsenic levels and testosterone, as well as zinc and dehydroepiandrosterone. Mediation analysis indicated that higher seminal iron and selenium were associated with an increasing sperm concentration after controlling for DHT, with 10.32% and 12.89% of these associations were mediated by DHT, respectively. 2A similar mediation effect of DHT was observed in the associations between iron and selenium levels and total sperm count (13.39% and 21.57% mediation, respectively). Our findings suggested that the presence of selenium and iron in semen was beneficial to sperm concentration and total count. Seminal manganese, zinc, molybdenum and copper may be associated with reduced semen quality. The associations between seminal selenium and iron and sperm concentration and total count were partially explained by the concomitant variation of seminal DHT.

P. Liu, G. Yuan, Q. Zhou, Y. Liu, X. He, 1H. Zhang, Y. Guo, Y. Wen, S. Huang, Y. Ke, and J. Chen,The association between metal exposure and semen quality in Chinese males: The mediating effect of androgens, Environ Pollut, 2020, 264, 113975.



Exposure to metals, including essential and nonessential elements, is widespread and may be associated with altered semen quality. This study aimed to examine the association between urinary metal concentrations and semen quality in a Chinese population. We measured semen quality parameters (sperm concentration, count, motility, normal morphology, and abnormal head) and 13 metals [arsenic (As), cadmium (Cd), Cobalt (CO), chromium (Cr), copper (Cu), iron (Fe), lead (Pb), manganese (Mn), molybdenum (Mo), mercury (Ho, nickel (Ni), selenium (Se), and zinc (Zn)] in the mine of 394 men front an infertility clinic. Multivariable logistic and linear regressions were used to assess the relationship between the creatinine-adjusted urinary metal concentrations and semen quality parameters. We found a significant trend for decreased odds ratios (ORs) for below-reference sperm count with increasing Se quartiles (p for trend = 0.04) and a significant trend for increased sperm percent abnormal head with increasing Ni quartiles (p for trend = 0.03). These associations persisted; even when considering multiple metals. Our results suggest that Ni exposure may be associated with deteriorated sperm morphology and that Se exposure may be associated with better semen quality. However, our findings warrant further studies in a larger and general population.

Zeng, Q., Feng, W., Zhou, B., Wang, Y. X., He, X. S., Yang, P., You, L., Yue, J., Li, Y. F., and Lu, W. Q.,Urinary Metal Concentrations in Relation to Semen Quality: A Cross-Sectional Study in China, Environmental Science & Technology, 2015, 49, 5052.


This study investigates the effects of tungsten disulfide nanotubes (WSNTs) and molybdenum disulfide nanoplatelets (MSNPs) on fibroblasts (NIH-3T3) and mesenchymal stem cells (MSCs) to determine safe dosages for potential biomedical applications.


Cytotoxicity of MSNPs and WSNTs (5-300 mug/ml) on NIH-3T3 and MSCs was assessed at 6, 12 or 24 h. MSC differentiation to adipocytes and osteoblasts was assessed following treatment for 24 h.


Only NIH-3T3 cells treated with MSNPs showed dose or time dependent increase in cytotoxicity. Differentiation markers of MSCs in treated groups were unaffected compared with untreated controls.


MSNPs and WSNTs at concentrations less than 50 microg/ml are potentially safe for treatment of fibroblasts or MSCs for up to 24 h.

Rashkow, J. T., Talukdar, Y., Lalwani, G., and Sitharaman, B.,Interactions of 1D- and 2D-layered inorganic nanoparticles with fibroblasts and human mesenchymal stem cells, Nanomedicine (Lond), 2015, 10, 1693.

Genetic Toxicology

Molybdenum trioxide was not mutagenic in tests with Bacillus subtillis , [Kada et al., 1980], Salmonella typhimurium, [Zeiger et al., 1992; Milvy and Kay, 1978], and Escherichia coli , [Venitt and Levy, 1974].
Kada, T., Hirano, K. and Shirasu, Y., in : Chemical Mutagens: Principles and Methods for their Detection.
Zeiger, E., Anderson, B., Haworth, S., Lawlor, T. and Mortelmans, K., Environ. Mol. Mutagen, 1992, 19 (Suppl. 21), 2.
Milvy, P. and Kay, K., J. Tox. Environ. Health, 1978, 4, 31.
Venitt, S. and Levy, L.S., Nature,1974, 250, 493.

Increased frequencies of chromosomal aberrations were reported in peripheral blood lymphocytes of experimental animals and workers exposed to molybdenum, molybdenite, and molybdenum trioxide [Babaian et al., 1980].

Babaian, E.A., Bagramian, S.B., and Pogosian, A.S., Gig. Tr. Prof. Zabol., 1980, 9, 33.

Cytotoxicity of molybdenum trioxide nanoparticles
There is a lack of information about the impact of nanomaterials on human health and the environment. The purpose of the study was to assess a mouse spermatogonial stem cell line (C18—4from type A spermatogonia isolated from 6-day-old mouse testes) as a model to assess nanotoxicity in the male germline in vitro. The effects of different types of nanoparticles on these cells were evaluated. MoO3 nanoparticles (30 microm) were made in a pulsed plasma reactor which forms the particles in a gas-phase process. There was a concentration-dependent toxicity for all types of particle (MoO3 , CdO, Ag, Al) tested, whereas the corresponding soluble salts had no significant effect. Molybdenum trioxide MoO3 nanoparticles were the least toxic. This cell line provides a valuable model with which to assess the cytotoxicity of nanoparticles in the germ line in vitro. With respect to their mitochondrial function MoO3 nanoparticles had a toxic effect of cellulsar metabolic activity at concentrations of 50 microg/l and above: EC50 90 microg/l. By contrast sodium molybdate in aqueous solution at the same concentrations was hardly toxic (EC50 322 microg/ml). In the LDH leakage test sodium molybdate did not affect the integrity of the plasma membrane whereas a significant increase of LDH leakage was observed with molybdenum nanoparticles: EC50 5 microg/l.

Braydich-Stolle, L., Hussain, S., Schlager, J. J., and Hofmann, M. C., In vitro cytotoxicity of nanoparticles in mammalian germline stem cells, Toxicological Sciences, 2005, 88, 412-419.

The toxicity of (inter alia) MoO3 nanoparticles (30 and 150 nm) was investigated in an in vitro model derived from rat liver cells BRL 3A. The interest was the cellular response to nanosized particle exposure. For toxicity evaluations cellular morphology, mitochondrial function (MTTassay), membrane leakage of lactate dehydrogenase (LDH assay), reduced glutathione (GSH) levels, reactive oxygen species (ROS),and mitochondrial membrane potential (MMP) were assessed under control and exposed conditions (24 h exposure) and at MoO3 concentrations from 0 to 250 ug/l. For MoO3 exposure at concentrations up to 100 microg/l the results of the LDH leakage and MTT assays showedno cytotoxicity but showed a significant effect at 250 microg/l.. The results were summarised as EC50 values (i.e. the concentration of nanoparticles that increased LDH leakage to 50% or decreased MTT reduction by 50%: MTT: 172 ± 25 (30 nm),175 ± 26 (150 nm); LDH: 210 ± 30 (30 nm)and 250 ± 17 (150 nm). MoO3 was less toxic than Ag and CdO but more so than, Fe3O4, Al, MnO2 and W particles.

Hussain, S.M., Hess, K. L., Gearhart, J. M., Geiss, K. T., and Schlager, J. J., In vitro toxicity of nanoparticles in BRL 3A rat liver cells, Toxicology in Vitro, 2005, 19, 975-983.

The mutagenic effect of an industrial enterprise (tungsten and molybdenum factory) was studied in three stages. At the first stage, the putative impact of the industrial sewage of the factory was studied using three plant test systems: Crepis capillaris L., Tradescantia sp. clone 02, and Glycine max (L.) Merill. It was found that the sewage increased the mutation level by a factor of 11-45. At the second stage, the rate of mutation was studied in the native vegetation growing on solid waste piles of the enterprise. It exceeded the corresponding index of uncontaminated areas by a factor of 2.0-4.5. At the third stage, the rates of children with birth defects and miscarriages were studied in the vicinity of the enterprise. The rate of miscarriages proved to be higher than the value averaged over the autonomous republic by a factor of 2.4. No change in the rate of birth defects was detected

Reutova, N.V., Vorobyeva, T. I., and Reutova, T. V., Some approaches to evaluation of the mutagenic effect of industrial waste on the environment, Russian Journal of Genetics, 2005, 41, 608-612.

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