Effects of Crocin on Human Sperm Viability, Motility, Morphology, DNA Fragmentation and Reactive Oxygen Species Levels after Freezing and Thawing
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Abstract
Objectives: To investigate the effects of crocin supplementation during solid surface vitrification (SSV) and liquid nitrogen vapor freezing (LNF) of human spermatozoa on post-thaw sperm parameters.
Materials and Methods: Thirty-six normozoospermic semen samples were used in the study. Post prepared semen samples were divided into five aliquots: one served as non-cryopreserved control; two were vitrified, with or without crocin supplementation (SSV ± 10 µg/ml crocin); and the last two aliquots were frozen in liquid nitrogen vapor, with or without crocin supplementation (LNF ± 10 µg/ml crocin).
Results: After cryopreservation, sperm motility (93.54 ± 3.55%, 70.10 ± 10.94% and 53.58 ± 12.06% in controls, SSV and LNF groups, respectively, p < 0.001) and sperm viability (79.82 ± 18.86%, 57.74 ± 21.99% and 49.69 ± 22.25%, p < 0.001) decreased significantly in both methods. However, the SSV groups had a significantly higher sperm motility (70.10 ± 10.94% and 53.58 ± 12.06%, p < 0.001) and viability (57.74 ± 21.99% and 49.69 ± 22.25%, p < 0.001) than the LNF groups. Supplementation with crocin 10 µg/ml in cryoprotective agent did not improve sperm motility and viability in both cryopreservation methods. Also, no effect was noted on sperm morphology, sperm deoxyribonucleic acid (DNA) integrity, and both the intracellular and extracellular reactive oxygen species (ROS) levels.
Conclusion: Crocin supplementation during vitrification and liquid nitrogen vapor freezing did not improve the outcome of post-thaw sperm.
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