A Study of the durability of sperm cell motility and deoxyribo-nucleic acid damage during cryopreservation

Semen cryopreservation is an essential technique in artificial insemination (AI) and assisted reproductive technology (ART). The addition of antioxidants to the freezing medium is a promising strategy to reduce cryo-induced damage to sperm cells. While sperm freezing has been extensively studied, this research aimed to improve the post-thaw durability and functionality of sperm. A total of 110 semen samples 55 from fertile and 55 from infertile individuals were collected and processed. After centrifugation, each sample was mixed with a cryoprotectant medium at a 1:1 (v/v) ratio. Three different cryoprotectant formulations (S1, S2, and S3), each containing varying concentrations of protective agents, were evaluated. The samples were then frozen in liquid nitrogen at -196°C. Post-thaw analyses included assessments of sperm count, motility, vitality, morphology, DNA fragmentation, and reactive oxygen species (ROS) levels representing a major strength of the study due to the comprehensive evaluation of sperm quality. The results showed significant differences in DNA fragmentation between fresh and cryopreserved samples. Among the tested formulations, S3 supplemented with citric acid anhydrous and taurine produced the best outcomes. It significantly improved sperm motility and vitality, while effectively reducing oxidative stress and cryodamage. Statistical analysis using one-way ANOVA and Tukey's HSD test revealed significant differences in sperm morphology between fresh and post-thaw S3-treated samples (p < 0.05), with a 95% confidence level. A limitation of the study was the long interval between the use of fresh control samples and the analysis of cryopreserved samples, which may influence comparative accuracy. Despite this limitation, the findings suggest that the S3 formulation is highly effective in preserving sperm quality. These results support its potential integration into fertility preservation protocols in ART. Furthermore, the outcomes may guide clinical practice and support the development of national policies aimed at improving access to advanced cryopreservation techniques as part of comprehensive reproductive healthcare services.