Protective effect of the ethanolic extract of Punica granatum L. during the cooling of bovine ovarian tissue.

INTRODUCTION: Reproductive biotechnologies such as cryopreservation are essential for preserving reproductive potential, particularly in species of high zootechnical value. During ovarian collection and transport, cooling of the tissue is a critical step. In this context, the addition of antioxidant compounds has been investigated to enhance cellular integrity and prevent oxidative and structural damage. Extracts of Punica granatum L. have been widely described for their strong antioxidant properties, promoting cellular protection against free radicals. Therefore, this study aimed to evaluate the effect of different concentrations of pomegranate extract during ovarian tissue preservation. METHODOLOGY: The ethanolic extract of Punica granatum L. peel (EEPG) was produced according to Nascimento et al. (2018). Ovaries (n = 5) were collected, fragmented, and distributed into groups maintained in 0.9% saline solution supplemented with antibiotics (penicillin and streptomycin, 100 μg/mL), in the absence (CTRL) or presence of 10 and 100 μg/mL of EEPG, and stored at 4 °C. Lipid peroxidation was assessed using the thiobarbituric acid reactive substances (TBARS) method. Data were analyzed using two-way ANOVA in GraphPad Prism, with significance set at p < 0.05. RESULTS: The following values were obtained: Fresh control (SEM = 0.107975). At 4 h, SEM values were CTRL = 0.565526, 10 μg/mL = 0.105036, and 100 μg/mL = 0.164488; at 12 h, CTRL = 0.998575, 10 μg/mL = 0.118822, and 100 μg/mL = 0.081847; and at 24 h, CTRL = 0.117856, 10 μg/mL = 0.098463, and 100 μg/mL = 0.06771. No significant differences were observed among the experimental groups or between the treated groups and the fresh or conserved controls at any evaluated time point (p > 0.05). CONCLUSION: At the tested concentrations, the extract appears to be safe for ovarian tissue, maintaining redox balance throughout the storage period. However, further analysis of follicular morphology is required to confirm the effectiveness of EEPG in preserving ovarian tissue quality.