The Effect of Zinc Selenide Nanospheres on Porcine Semen Cryopreservation

Objective: This study aimed to investigate the protective effects of zinc selenide (ZnSe) nanospheres on boar semen during cryopreservation and to determine the optimal supplementation concentration for improving post-thaw sperm quality and antioxidant status.Methods: ZnSe nanospheres were synthesized via a hydrothermal method using NaSeO and Zn(CHCOO) as precursors.Ejaculates with motility 70% were collected from six healthy boars, pooled to minimize individual variation, and the entire experiment was independently repeated three times.Samples were mixed with cryopreservation extenders containing 0 (control), 0.1, 1, or 10 mmol/L ZnSe nanospheres.Post-freezing and thawing, sperm motility parameters, acrosome integrity, plasma membrane integrity, antioxidant enzyme activities, and intracellular reactive oxygen species (ROS) levels were evaluated.Results: Supplementation with 1 mmol/L ZnSe significantly improved post-thaw sperm motility, acrosome integrity, and plasma membrane integrity (P < 0.05).Moreover, 1 mmol/L ZnSe markedly enhanced total antioxidant capacity, catalase, and superoxide dismutase activities, while reducing malondialdehyde concentrations and intracellular reactive oxygen species (ROS) levels (P < 0.05). Conclusion:Supplementation with 1 mmol/L ZnSe in the cryopreservation diluent effectively improves post-thaw sperm quality and antioxidant capacity, offering a promising strategy for optimizing porcine semen cryopreservation.