Department of Embryology, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
Department of Cellular and Molecular Biology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran.
Reprod Sci. 2023 Jul;30(7):2122-2136. doi: 10.1007/s43032-022-01144-1. Epub 2022 Dec 23.
This study was conducted to evaluate the effects of static magnetic field (SMF) and nanoparticles (NPs) on the vitrification of cumulus-oocyte-complex (COC). To this end, the non-vitrified (nVit) and vitrified groups (Vit) that contain NPs, with or without SMF were labeled nVit_NPs, nVit_NPs_SMF, Vit_NPs, and Vit_NPs_SMF, respectively. The non-toxic dosages of NPs were first determined to be 0.008% w/v. The survival, apoptosis, and necrosis, mitochondrial activity, fertilization rate, subsequent-derived embryo development, and gene expressions were examined. The viability rates obtained by trypan blue and Anx-PI staining were meaningfully smaller in the Vit groups, compared to the nVit groups. The JC1 red/green signal ratios were reduced considerably in the Vit group, compared to the nVit. Transmission electron microscopy (TEM) was performed to assess the entry of the NPs into the oocytes. TEM images showed that NPs were present in nVit_NPs, and Vit_NPs. Thereafter, the effects of NPs and SMF on in vitro fertilization (IVF) were examined. The difference in blastocyst rates between nVit and Vit_NPs_SMF groups was significant. Finally, Nanog, Cdx2, Oct4, and Sox2 genes were evaluated. There were substantial differences in Cdx2 gene expressions between the Vit_NPs and nVit groups. The expression of Nanog in Vit was significantly higher than those of the Vit_NPs, Vit_NPs_SMF, and nVit groups. The data presented here provide deeper insight into the application of iron oxide nanoparticles in COC vitrification. It appears that using SMF and supplemented CPA by NPs inhibits cryoinjury and promote the embryo development capacity of vitrified-warmed COCs.
本研究旨在评估静磁场（SMF）和纳米颗粒（NPs）对卵丘-卵母细胞复合物（COC）玻璃化的影响。为此，分别标记了未玻璃化（nVit）和玻璃化组（Vit），其中含有 NPs，有无 SMF，标记为 nVit_NPs、nVit_NPs_SMF、Vit_NPs 和 Vit_NPs_SMF。首先确定 NPs 的无毒剂量为 0.008%w/v。检测了细胞存活率、凋亡、坏死、线粒体活性、受精率、随后衍生胚胎发育和基因表达。与 nVit 组相比，台盼蓝和 Anx-PI 染色的活力率在 Vit 组明显更小。与 nVit 组相比，JC1 红/绿信号比值在 Vit 组显著降低。进行透射电子显微镜（TEM）评估 NPs 进入卵母细胞的情况。TEM 图像显示，nVit_NPs 和 Vit_NPs 中存在 NPs。之后，检查了 NPs 和 SMF 对体外受精（IVF）的影响。nVit 和 Vit_NPs_SMF 组之间囊胚率的差异具有统计学意义。最后，评估了 Nanog、Cdx2、Oct4 和 Sox2 基因。Vit_NPs 和 nVit 组之间 Cdx2 基因表达存在显著差异。Vit 组中 Nanog 的表达明显高于 Vit_NPs、Vit_NPs_SMF 和 nVit 组。这里呈现的数据提供了对氧化铁纳米颗粒在 COC 玻璃化中应用的更深入了解。似乎使用 SMF 和补充有 NPs 的 CPA 可以抑制冷冻损伤并提高玻璃化-加热 COC 的胚胎发育能力。
