BACKGROUND: Cryopreservation is a common method for assisted reproductive technology that causes oxidative stress on sperm, which is the main reason of sperm disruption. Cryoprotective media are commonly supplemented with antioxidants that to improve sperm quality post thawing. OBJECTIVE: This study investigated to assess the effects of Sildenafil Citrate (SC) and Alpha- Lipoic Acid (ALA) supplementation on the oxidative stress during cryopreservation in the asthenozoospermic infertile men. METHODS: 30 semen samples were collected from the asthenozoospermic patients. Each semen sample was divided into five groups: Control (fresh), Freeze (treated with cryo-protectant alone), Freeze+SC (treated with cryo-protectant+ 0.67µM SC), Freeze+ALA (treated with cryo- protectant+ 0.5mM ALA), and Freeze+SC+ALA. In each sample, sperm Tumor necrosis factor alpha (TNF-α), Malondialdehyde (MDA) level, and antioxidant enzymes, including Catalase (CAT), Glutathione (GSH), and Superoxide dismutase (SOD) levels were evaluated by Enzyme- linked immunosorbent assay (ELISA). RESULTS: Our results showed that the freeze group had a significant reduction in antioxidant enzymes levels and a significant increase in sperm TNF-α, and MDA level compared to the control group (p<0.05). In the Freeze+SC, Freeze+SC+ALA and Freeze+ALA groups compared to freeze group, had a significant increase in antioxidant enzymes levels and a significant decrease in TNF-α and MDA level (p<0.05). Antioxidant enzymes levels was significantly higher, but TNF-α and MDA level was significantly lower in the Freeze+ALA group compared to Freeze+SC and Freeze+SC+ALA groups (p˂0.05). Antioxidant enzymes levels in the Freeze+SC+ALA group had significantly increased compared to Freeze+SC group (p˂0.05). CONCLUSION: Our findings revealed that the utilization of ALA and SC along with cryoprotective medium caused improvement the biochemical parameters of sperm against cryodamage.
