Fabrication and Characterization of Xylomannan/Fe(3)O(4) Nanocomposite to Improve Post-Vitrification Viability of Mouse Immature Oocytes

制备和表征木甘露聚糖/Fe(3)O(4)纳米复合材料以提高小鼠未成熟卵母细胞玻璃化冷冻后的存活率

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Abstract

BACKGROUND: Nano-biotechnology is a rapidly evolving field merging biology, engineering, and chemistry. Nanoparticles (NPs) with low toxicity and diverse properties, hold great promise for cryopreservation. Their ability to modify membrane permeability, optimize cryoprotectant entry and removal, tolerate freezing temperatures, enhance warming efficiency, and improve the heat transfer and thermodynamic properties of cryo-solution, makes them a game-changer for low-temperature preservation. This study investigated the potential of Fe(3)O(4)/Xylomannan nanocomposites (FXN) for improving mouse germinal vesicle vitrification. MATERIALS AND METHODS: This study is experimental research. Xylomannan, a polysaccharide derived from Enoki mushroom, exhibits cryoprotective properties. The nanocomposite was synthesized by coating Fe(3)O(4) NPs with Xylomannan. Characterization techniques, including X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FT-IR), and scanning electron microscopy (SEM) confirmed the successful synthesis and characterization of the Fe(3)O(4)/Xylomannan nanocomposite. Immature oocytes, GV DO (denuded oocytes) were obtained from 40 adult Naval Medical Research Institute (NMRI) mice and divided into 5 research groups (n-Vit, Vit, Vit+Fe(3)O(4), Vit+Xylo, and Vit+FXN), therefore, each experiment was repeated 5 times. RESULTS: The cytotoxicity of different concentrations (0.002, 0.004, 0.008 and 0.016%) of Xylomannan, Fe(3)O(4), and FXN was assessed on mouse embryo fibroblasts (MEF) at three time points (1, 2 and 7 days), using the MTT assay. While, the concentration 0.004% was the optimal dose of NC, and selected for the subsequent experiments, 0.008 and 0.016% concentrations showed significant reduction in cell viability (P=0.011). Overall, the present study demonstrated that the Fe(3)O(4)/Xylomannan nanocomposite significantly improves post-vitrification survival rate in immature oocytes compared to the other vitrification groups, with no significant difference compared to the control group. CONCLUSION: The findings of this study reveal the potential of Fe(3)O(4)/Xylomannan nanocomposite as a promising cryoprotective agent for improving oocyte vitrification.

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