Objective: The aim of this study was to determine the effects of melatonin and selenium in freezing extenders on frozen-thawed rat sperm. Methods: Semen samples were collected from 20 adult male Wistar albino rats. Following dilution, the samples were divided into six groups: four cryopreserved groups with 1 mM and 0.5 mM melatonin and selenium supplements, and two fresh and cryopreserved control groups. The rapid freezing technique was used to freeze the samples. Flow cytometry was used to assess plasma membrane integrity, mitochondrial membrane potential, and DNA damage, while computer-assisted sperm analysis was used to assess motility. Results: Total motility was higher in the 1 mM melatonin supplementation group than in the cryopreserved control group (mean±standard error of the mean, 69.89±3.05 vs. 59.21±1.31; p≤0.05). The group with 1 mM selenium had the highest plasma membrane integrity (42.35%±1.01%). The cryopreserved group with 0.5 mM selenium had the highest mitochondrial membrane potential, whereas the cryopreserved control group had the lowest (45.92%±4.53% and 39.45%±3.52%, respectively). Conclusion Cryopreservation of rat semen supplemented with 1 mM melatonin increased sperm motility after freeze-thawing, while supplementation with 0.5 mM selenium increased mitochondrial activity.

Objective: Given the destructive effects of oxidative stress on sperm structure, this study was conducted to investigate the antioxidant effects of different concentrations of Ceratonia siliqua plant extract on human sperm parameters after the freezing-thawing process. Methods: A total of 20 normozoospermic samples were frozen. Each sample was divided into two control groups (fresh and cryopreservation) and three cryopreservation experimental groups (containing C. siliqua extract at concentrations of 20, 30, and 40 μg/mL in the freezing extender). Motility, intracellular levels of reactive oxygen species (ROS), plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), viability, and acrosome reaction parameters were evaluated. Results: Statistical analysis showed that the highest motility, viability, and PMI were associated with the 20 μg/mL concentration of C. siliqua extract. At all concentrations, intracellular ROS levels were significantly lower and the levels of MMP and the acrosome reaction were significantly higher than in the cryopreservation control group (p≤0.05). Conclusion C. siliqua extract supplements at concentrations of 20, 30, and 40 μg/mL improved sperm motility, viability, PMI, MMP, intracellular ROS, and the acrosome reaction.