Anatomy Department, School of Medicine, Zanjan University of Medical Science, Zanjan, Iran.
Department of Anatomical Sciences, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran.
Cryobiology. 2024 Dec;117:104978. doi: 10.1016/j.cryobiol.2024.104978. Epub 2024 Oct 21.
Male fertility preservation is still challenged by cell damage induced during sperm cryopreservation and impaired sperm structure and function. Sperm ultra-rapid freezing, despite a higher protective effect compared to conventional freezing method, is still associated with suboptimal sperm cryosurvival and needs to be modified to increase its efficiency in sperm protection. Sperm freezing media supplemented with antioxidants can improve sperm parameters following freezing-warming process. In this study, we aimed to investigate the effect of employing ultra-rapid freezing and myo-inositol on sperm cryosurvival. Thirty semen samples with normal sperm parameters were collected and each one was divided into four portions to cryopreserve by conventional freezing, ultra-rapid freezing, conventional freezing + myo-inositol 2 mg/ml, and ultra-rapid freezing + myo-inositol 2 mg/ml. Sperm samples warmed after at least 24 h of freezing and sperm cryosurvival were analyzed by evaluation of sperm motility, viability, morphology and DNA fragmentation index (DFI). Freezing method had a significant influence on post-thaw sperm DFI and morphology (p < 0.05) and the interaction between freezing method and antioxidant supplementation significantly affected sperm morphology (p < 0.05). The highest percentage of sperm normal morphology and minimal DFI was achieved using ultra-rapid freezing supplemented by myo-inositol antioxidant compared to other groups (P < 0.05). The highest sperm DNA damage after freezing-warming was observed following the conventional freezing method. In conclusion, sperm freezing method was identified as factor strongly influencing sperm DFI and morphology after thawing/warming. Sperm samples can be rapidly frozen using the modified freezing media supplemented by myo-inositol without impacting sperm DNA and morphology.
男性生育力保存仍然受到精子冷冻保存过程中细胞损伤和精子结构与功能受损的挑战。尽管与传统冷冻方法相比，精子超速冷冻具有更高的保护作用，但仍与精子冷冻保存效果不佳有关，需要加以改进以提高其在精子保护方面的效率。在冷冻-解冻过程中添加抗氧化剂的精子冷冻保存液可以改善精子参数。在这项研究中，我们旨在研究采用超速冷冻和肌醇对精子冷冻保存的影响。收集了 30 份具有正常精子参数的精液样本，每个样本分为四部分，分别采用常规冷冻、超速冷冻、常规冷冻+2mg/ml 肌醇和超速冷冻+2mg/ml 肌醇进行冷冻保存。冷冻后至少 24 小时解冻精子样本，并通过评估精子活力、存活率、形态和 DNA 碎片化指数（DFI）来分析精子冷冻保存情况。冷冻方法对解冻后精子 DFI 和形态有显著影响（p<0.05），并且冷冻方法和抗氧化剂补充之间的相互作用显著影响精子形态（p<0.05）。与其他组相比，超速冷冻+抗氧化剂肌醇补充的精子形态正常率最高，DFI 最小（P<0.05）。解冻后观察到常规冷冻方法导致的精子 DNA 损伤最大。结论：冷冻方法是影响解冻后精子 DFI 和形态的重要因素。精子样本可以使用改良的冷冻保存液进行超速冷冻，补充肌醇不会影响精子 DNA 和形态。
