The cytotoxicity of Au/Ag nanoparticles (NPs) on human spermatozoa was investigated in vitro. Semen from donors were incubated (37 °C, 60'-120') with 30, 60, 125, 250 and 500 μM Au/Ag-NPs. Sperm motility was evaluated following WHO guidelines; sperm viability was assessed with eosin Y test. Au-NPs were characterised and localised with field emission gun-based scanning transmission electron microscope/energy dispersive spectroscopy and transmission electron microscopy. Both tested NPs exerted a significant dose-dependent effect on motility and viability of human spermatozoa (P < 0.001). Ag-NPs seem to show a slightly elevated toxicity although not significant (P > 0.05). Au-NPs were localised in spermatozoa, whereas Ag-NPs were undetectable. In conclusion, Au-NPs and Ag-NPs do not appear to be harmful for human spermatozoa up to high concentrations (250-500 μM) that are probably difficult to reach in vivo. It is mandatory to explore the genotoxic effect of NPs in germ cells.
Aquaporins (AQPs) are a family of 13 small hydrophobic transmembrane proteins expressed in numerous tissues and cells. Some AQPs work as strict water channels, others are permeable to a range of substances, including glycerol. In the male reproductive system their localization in testis, efferent ducts, epididymis, and spermatozoa has been described. We studied the distribution of AQP7 in ejaculated human sperm and the relationship between AQP7 labeling and sperm characteristics. Semen samples from 33 men were examined by light and transmission electron microscopy (TEM). TEM data were quantified using a mathematical formula that calculates a fertility index (FI) and the percentages of sperm apoptosis, immaturity, and necrosis. Immunocytochemistry with a polyclonal antibody anti-AQP7 was performed on the sperm samples. Normal sperm were labeled in the pericentriolar area, midpiece, equatorial segment, and weakly in the tail (grade 1). Abnormal sperm showed a diffuse low intensity of fluorescence evident in the cytoplasmic residues, coiled tails, in the entire head, and acrosome (grade 2). A high number of motile sperm obtained by swim up were labeled in a dotted manner in the mitochondria. A significant positive correlation was found between the spermatozoa with AQP7 grade 1 labeling and the percentage of normal form (P < 0.008), progressive motility and FI (P < 0.005); a negative correlation was noted with the percentages of cytoplasmic residues (P < 0.010) and immaturity (P < 0.006) and coiled tails (P < 0.012). The link between AQP7 distribution and sperm morphology and the particular dotted labeling in swim up selected motile sperm are novel and deserve additional studies.
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