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KMID : 1033620220490020117
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Clinical and Experimental Reproductive Medicine
2022 Volume.49 No. 2 p.117 ~ p.126
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The relationship between reactive oxygen species, DNA fragmentation, and sperm parameters in human sperm using simplified sucrose vitrification with or without triple antioxidant supplementation
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Juanpanich Theesit
Suttirojpattana Tayita
Parnpai Rangsun
Vutyavanich Teraporn
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Abstract
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Objective: This study examined whether the addition of triple antioxidants (3A)?10 μM acetyl-L-carnitine, 10 μM N-acetyl-L-cysteine, and 5 μM α-lipoic acid?in freezing-thawing medium during human sperm cryopreservation using the sucrose vitrification (SuV) and liquid nitrogen vapor (Vapor) techniques could improve post-thaw survival of spermatozoa.
Methods: We analyzed 30 samples from healthy human sperm donors. Each sample was allocated into one of five groups: fresh control, SuV, SuV+3A, Vapor, and Vapor+3A. The sperm motility, morphology, viability, intracellular and extracellular reactive oxygen species (ROS) levels, and sperm DNA fragmentation (SDF) were evaluated.
Results: The cryopreserved spermatozoa had significantly reduced percentages of motility (p<0.05) and viability (p<0.05). Antioxidant supplementation non-significantly improved these parameters (p>0.05). No significant differences were found in sperm morphology between the fresh and frozen-thawed groups (p>0.05). After freezing, the extracellular ROS levels in the frozen-thawed groups were significantly higher (p<0.05) than in the fresh group. However, we did not find any differences in intracellular ROS parameters among these groups (p>0.05). The SDF was higher in the SuV and Vapor groups than in the fresh group, but without statistical significance (p=0.075 and p=0.077, respectively).
Conclusion: Cryopreservation had detrimental effects on sperm motility, viability, and extracellular ROS levels, without changing the morphology or intracellular ROS levels. Antioxidant supplementation was slightly effective in preventing SDF in frozen-thawed spermatozoa.
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KEYWORD
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Antioxidants, DNA fragmentation, Human, Spermatozoa, Sucrose, Vitrification
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