Μεταπτυχιακή εργασία - Σχολή Γεωτεχνικών Επιστημών - Τμήμα Γεωπονίας, 2023 (α/α 14061)
(EL)
Nanotechnology has become one of the most promising fields of research with its significant application in all fields of science. The effects of nanoparticles (NPs) at the cellular and tissue level may be used in assisted reproduction to improve male fertility, and particularly enhance sperm quality under in vivo or in vitro conditions. In recent years, the tin oxide nanoparticles have received enormous attention due to their properties. In this aspect, the object of this study was as a first step to find the non-detrimental concentration of tin nanoparticles (stock solution: ΝPs SnO2 0,25g / 100 mL distilled water) in the qualitative characteristics of cryopreserved bull semen.
Ninety-six (96) straws of cryopreserved bull semen (12 replicates; 4 experimental groups; 2 straws/group) were thawed (37℃; 30 sec) and four experimental groups were processed: control group (no treatment), low concentration NPs (20 ιL stock solution NPsSnO2/ mL semen), medium concentration NPs (40 ιL stock solution NPs SnO2/ mL semen), and high concentration NPs (80 ιL stock solution NPs SnO2/ mL semen). All groups were further extended (1:2; v/v) with pre-warmed (37μ C) phosphate-buffered saline and finally incubated (37℃; 2 hours). For all experimental groups, sperm motility and kinetics (CASA), viability (eosin-nigrosin), morphology (eosin-nigrosin) and functional integrity of sperm plasma membrane (Hypo-Osmotic Swelling Test; HOST) were evaluated at 0 (time of NPs supplementation), 1 and 2 hrs of incubation.
Data analysis was done in the methodological frame of mixed linear models and was based on the randomized completely block (RCBD) split plot design (subgroups within groups). The significance level α of the statistical tests was predetermined at α=0.05 (p<0.05). Analyses were performed at statistical package IBM SPSS Statistics (Version 28).
The results of the study revealed that the low concentration of tin nanoparticles did not affect the examined semen variables after 1 h of incubation, but lower values were noticed for kinematics BCF and VCL after 2 hrs of incubation compared to the control group. The medium tin nanoparticles concentration negatively affected the kinematics VCL, VSL, BCF, and the percentage of hyperactivated spermatozoa. However, it was not affected the total and progressive motility, rapid, medium, slow moving, spermatozoa). Finally, the high concentration of tin was detrimental to almost all sperm parameters.
In conclusion, the study revealed that the tin nanoparticles‟ concentration of 20 ιL/ml\L of bull semen was non cytotoxic and, the safe time of nanoparticles‟ interaction with bull spermatozoa is 1 h. Moreover, it was proved that the medium and high tin nanoparticles‟ concentrations were partly and strong cytotoxic for cryopreserved bull semen, respectively. Therefore, bull semen handling and processing with tin nanoparticles is feasible, when the concentration limits of this study are taken in consideration. The determination of the non-cytotoxic concentration may guarantee further research about the possible antimicrobial action of tin oxide NPs and their potential use as alternative to conventional antibiotics.
(EN)
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