Background

Despite its routine and frequent application, cryopreservation of human sperm is far from the desired efficacy, as freezing and thawing impair motility, viability, acrosomal unity, and DNA integrity.

Objectives

In this study, the authors aimed to investigate whether adding antioxidants, coenzyme Q10, and curcumin into the freezing medium provide better efficacy in the cryopreservation of human sperm.

Methods

The semen samples from 40 healthy men aged 18–45 were collected in sterile containers by masturbation. Samples within normal reference values for sperm concentration (≥15 million/mL) and motility (progressive motile ≥ 32% and total motility ≥ 40%) were included in the study. Semen samples were equally divided into five groups and evaluated; i) pre-freezing sperm suspension, ii) frozen-thawed control (Ctrl) without any supplementation in freezing medium, iii) frozen-thawed with curcumin supplementation of 0.25 mM (Cur), iv) frozen-thawed coenzyme Q10 supplementation of 25 µM (CoQ10) and v) frozen-thawed curcumin (0.25 mM) plus coenzyme Q10 (25 µM) supplementation (CurCoQ10) into the freezing medium. Liquid nitrogen vapour freezing and rapid thawing were performed in each group (ii-v). Sperm motility, viability, acrosome integrity, and DNA fragmentation rates were compared and ultrastructural evaluations by transmission electron microscopy were undertaken between the groups. Additionally, the total antioxidant capacity/total oxidant capacity values were measured.

Results

According to CASA results, progressive motility was significantly higher in the CoQ10 group (9.4 ± 7.6) when compared with the Ctrl (7.1 ± 6.3), Cur (6.4 ± 4.8) and CurCoQ10 (8.1 ± 7.7) groups (p < 0.05). Flow cytometry results showed no difference in the viability and acrosome integrity values after thawing, but DNA fragmentation was significantly increased in the curcumin-added groups (p < 0.05). Acrosomal changes and sub-acrosomal defects were seen in all groups after thawing at the ultrastructural level. Mitochondrial membrane structure was preserved in CoQ10 and CurCoQ10 groups.

Conclusions

Our results suggested that sperm ultrastructural morphology and motility were better preserved in the CoQ10 group during cryopreservation. In curcumin groups, DNA fragmentation and head defects were increased.

中文翻译：

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冷冻培养基中添加辅酶Q10和姜黄素对人精子冷冻保存的影响

背景

尽管人类精子的冷冻保存是常规且频繁的应用，但其效果远未达到预期的效果，因为冷冻和解冻会损害精子的活力、活力、顶体统一性和 DNA 完整性。

目标

在这项研究中，作者旨在研究在冷冻介质中添加抗氧化剂、辅酶 Q10 和姜黄素是否可以在人类精子的冷冻保存中提供更好的效果。

方法

40 名 18-45 岁健康男性的精液样本通过手淫收集在无菌容器中。研究中包括精子浓度（≥1500万/mL）和活力（渐进活力≥32％和总活力≥40％）正常参考值内的样本。精液样本平均分为五组并进行评估；i) 预冷冻精子悬浮液，ii) 冻融对照 (Ctrl)，在冷冻培养基中不添加任何补充剂，iii) 冻融后添加 0.25 mM 姜黄素 (Cur)，iv) 冻融后添加 25 µM 辅酶 Q10 (CoQ10) 和 v) 将冻融的姜黄素 (0.25 mM) 加辅酶 Q10 (25 µM) 补充剂 (CurCoQ10) 添加到冷冻培养基中。每组均进行液氮蒸气冷冻和快速解冻（ii-v）。精子活力、活力、顶体完整性、比较各组之间的 DNA 碎片率，并通过透射电子显微镜进行超微结构评估。另外，还测量了总抗氧化能力/总氧化能力值。

结果

根据 CASA 结果，与 Ctrl (7.1 ± 6.3)、Cur (6.4 ± 4.8) 和 CurCoQ10 (8.1 ± 7.7) 组相比，CoQ10 组 (9.4 ± 7.6) 的前向运动显着较高 (p < 0.05)。流式细胞术结果显示解冻后活力和顶体完整性值没有差异，但添加姜黄素的组中 DNA 碎片显着增加（p < 0.05）。解冻后在超微结构水平上，所有组均观察到顶体变化和顶体下缺陷。CoQ10 和 CurCoQ10 组保留了线粒体膜结构。

结论

我们的结果表明，在冷冻保存过程中，CoQ10 组的精子超微结构形态和活力得到了更好的保存。在姜黄素组中，DNA 断裂和头部缺陷增加。