Fertility and Infertility Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Nano Drug Delivery Research Center, Faculty of Pharmacy, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Biopreserv Biobank. 2021 Jun;19(3):184-193. doi: 10.1089/bio.2020.0102. Epub 2020 Dec 22.
Oocyte vitrification, as a vital step in reproductive medicine, is strongly associated with lower development caused by cryodamaging factors, such as oxidative stress. In this study, we evaluated the antioxidative synergistic effects of Melatonin (Mel) and Resveratrol (RES) coencapsulated by solid lipid nanocarriers (SLNs) against the pure antioxidant combination (Mel+RES). In this research, the formation of Mel+RES-SLN was confirmed by Fourier-transformed infrared spectroscopy. The average mean diameter, size distribution, polydispersity index, and zeta potential of particles were measured by Zetasizer, and the morphology was evaluated by scanning electron microscopy. In addition, the encapsulation efficiency (EE%) or drug loading capacity (DL%) of the nanocapsule was determined by spectrophotometric methods. Germinal vesicle (GV)-stage oocytes harvested from 6- to 12-week-old female NMRI mice were randomly divided into seven groups for studies. In these groups, (0, 10 M + 0.5 μM, 10 M + 2 μM, or 10 M + 10 μM) of Mel+RES/Mel+RES-SLN were added into vitrification media. After thawing, oocytes were matured, fertilized, and cultured for 3 days. Extra/intracellular reactive oxygen species (ROS) levels were measured in maturation medium after 24 hours. Our results revealed a significant improvement in the normal morphology of warmed GV-stage oocytes, GV breakdown (GVBD) rate, Metaphase II (MII)-stage oocyte formation, fertilization rate, early embryo development, and a significant reduction in intra/extracellular ROS level when vitrification media was supplemented with the lowest Mel+RES-SLN concentration. studies also demonstrated that the highest concentration of Mel+RES-SLN was safe, without a detrimental effect on embryonic development upon treatment. In conclusion, the lowest concentration of Mel+RES-SLN supplementation in GV-stage oocyte vitrification media improved maturation, fertilization, and embryo development rate and decreased extra/intracellular ROS level through an enhanced/controlled intracellular penetration compared to the pure Mel+RES.
卵母细胞玻璃化是生殖医学中的一个重要步骤，它与冷冻损伤因素（如氧化应激）导致的发育率降低密切相关。在这项研究中，我们评估了褪黑素（Mel）和白藜芦醇（RES）包封在固体脂质纳米载体（SLNs）中的抗氧化协同作用，以对抗纯抗氧化剂组合（Mel+RES）。在这项研究中，通过傅里叶变换红外光谱证实了 Mel+RES-SLN 的形成。通过 Zetasizer 测量了颗粒的平均平均粒径、粒径分布、多分散指数和 Zeta 电位，并通过扫描电子显微镜评估了形态。此外，通过分光光度法测定纳米胶囊的包封效率（EE%）或载药量（DL%）。从 6-12 周龄 NMRI 雌性小鼠中收获生发泡（GV）期卵母细胞，并随机分为 7 组进行研究。在这些组中，将（0、10 M+0.5 μM、10 M+2 μM 或 10 M+10 μM）Mel+RES/Mel+RES-SLN 添加到玻璃化介质中。解冻后，使卵母细胞成熟、受精并培养 3 天。在 24 小时后测量成熟培养基中的细胞内/外活性氧（ROS）水平。我们的结果表明，当玻璃化培养基中添加最低浓度的 Mel+RES-SLN 时，变暖的 GV 期卵母细胞的正常形态、GV 破裂（GVBD）率、中期 II（MII）期卵母细胞形成、受精率、早期胚胎发育均得到显著改善，并且细胞内/外 ROS 水平显著降低。研究还表明，当以最高浓度处理时，Mel+RES-SLN 是安全的，对胚胎发育没有不利影响。总之，与纯 Mel+RES 相比，GV 期卵母细胞玻璃化培养基中添加最低浓度的 Mel+RES-SLN 可通过增强/控制细胞内渗透来提高成熟、受精和胚胎发育率，并降低细胞外/内 ROS 水平。
