Department of Physiology, University of Veterinary and Animal Sciences, Outfall Road, Lahore, 54000, Pakistan; Faculty of Veterinary and Animal Sciences, University of Poonch Rawalakot, Azad Jammu and Kashmir, Pakistan.
Animal Sciences Institute, National Agricultural Research Centre, Islamabad, 44000, Pakistan.
Theriogenology. 2024 Jan 15;214:166-172. doi: 10.1016/j.theriogenology.2023.10.022. Epub 2023 Oct 20.
The motility, health quality, and membrane disorders of spermatozoa are adversely affected during the process of semen cryopreservation due to the over-production of reactive oxygen species (ROS). Cerium oxide nanoparticles (CeONPs) possess properties to scavenge ROS either by mimicking specific antioxidants or by enhancing the activities of antioxidant enzymes. Therefore, we aimed at evaluating the effects of adding the CeONPs in the TRIS-citrate-yolk extender on in-vitro antioxidant enzyme activities, spermatozoa quality attributes, and in-vivo fertility of post-thaw Beetal buck spermatozoa. The CeONPs were prepared and characterized (UV-spectrophotometry, FTIR, and XRD). Semen samples, collected from bucks (n = 5), were distributed into five aliquots and diluted in an extender containing increasing concentrations of nanoparticles (0 μg/ml, called the control group, 25 μg/mL, 50 μg/mL, 75 μg/mL, and 100 μg/mL). At post-thaw, spermatozoa were evaluated for the above-mentioned attributes and the pregnancy rate by inseminating Beetal does (n = 252). Results demonstrated that CeONPs mitigated the detrimental effects of cryopreservation as ROS production and lipid peroxidation were lower (P < 0.001) in the 25, 50, and 75 μg/mL CeONPs-added groups compared to the control and 100 μg/ml CeONPs-added group. The addition of 25 μg/mL CeONPs improved (P < 0.001) the activities of superoxide dismutase, catalase, and peroxidase and the concentration of reduced glutathione (P < 0.001) compared to the other groups. In terms of sperm kinematics and velocity parameters, the groups added with the 25 and 50 μg/mL CeONPs exhibited higher total motility (P < 0.001), sperm progressive motility (P = 0.003), and rapid velocity (P < 0.001). The group added with the 50 μg/mL CeONPs had the highest (P = 0.04) average path velocity. The groups added with the 25 and 50 μg/mL CeONPs also exhibited higher plasma membrane integrity (P = 0.003), acrosomal integrity, and viability (P < 0.001) compared to the control group. The DNA integrity was also higher (P < 0.001) in all the CeONPs-added groups. The pregnancy rate was higher (P = 0.003) in the 25 (51.92 %) and 50 μg/mL CeONPs (58.33 %) groups compared to the other groups. Conclusively, our findings suggest that the inclusion of cerium oxide nanoparticles in the TRIS-citrate-yolk freezing extender can reduce the occurrence of cryopreservation-induced damages to Beetal's buck spermatozoa and ultimately enhance the pregnancy rate in does.
在精液冷冻保存过程中，由于活性氧（ROS）的过度产生，精子的运动性、健康质量和膜障碍都会受到不利影响。氧化铈纳米粒子（CeONPs）具有通过模拟特定抗氧化剂或增强抗氧化酶的活性来清除 ROS 的特性。因此，我们旨在评估在 TRIS-柠檬酸盐-卵黄稀释液中添加 CeONPs 对体外抗氧化酶活性、精子质量特性和解冻后贝塔尔雄鹿精子体内生育力的影响。CeONPs 通过紫外分光光度法、傅里叶变换红外光谱和 X 射线衍射进行了制备和表征。从雄鹿（n=5）采集精液样本，并分为五份，分别在含有不同浓度纳米粒子的稀释液中进行稀释（0μg/ml，称为对照组，25μg/ml、50μg/ml、75μg/ml 和 100μg/ml）。解冻后，通过向贝塔尔母鹿（n=252）授精来评估精子的上述属性和妊娠率。结果表明，CeONPs 减轻了冷冻保存的不利影响，因为与对照组和 100μg/ml CeONPs 添加组相比，ROS 产生和脂质过氧化在添加 25、50 和 75μg/ml CeONPs 的组中较低（P<0.001）。与其他组相比，添加 25μg/ml CeONPs 提高了超氧化物歧化酶、过氧化氢酶和过氧化物酶的活性以及还原型谷胱甘肽的浓度（P<0.001）。在精子运动学和速度参数方面，添加 25 和 50μg/ml CeONPs 的组表现出更高的总活力（P<0.001）、精子渐进性活力（P=0.003）和快速速度（P<0.001）。添加 50μg/ml CeONPs 的组的平均路径速度最高（P=0.04）。与对照组相比，添加 25 和 50μg/ml CeONPs 的组的质膜完整性（P=0.003）、顶体完整性和活力（P<0.001）也更高。所有添加 CeONPs 的组的 DNA 完整性也更高（P<0.001）。与其他组相比，添加 25μg/ml CeONPs （51.92%）和 50μg/ml CeONPs（58.33%）的组的妊娠率更高（P=0.003）。总之，我们的研究结果表明，在 TRIS-柠檬酸盐-卵黄冷冻液中添加氧化铈纳米粒子可以减少贝塔尔雄鹿精子冷冻保存诱导损伤的发生，并最终提高母鹿的妊娠率。
