Magnetic graphene oxide improves the expression of some human cumulus cells biomarkers

Abstract

Purpose: Developing non-invasive, accurate methods for selecting high-implantation embryos is crucial for assisted reproductive technologies (ART). Cumulus cells (CCs) are reliable indicators of oocyte quality. This study investigated the effects of magnetic graphene oxide (MGO) on the viability, apoptosis, oxidative status, and proliferation-related proteins of human CCs in vitro. Methods: Human CCs were treated with MGO (1.5–400 µg/mL) for 24, 48, and 72 hours. Viability was assessed via MTT and Annexin V-FITC/PI flow cytometry. Expression of apoptosis-related (Bcl-2, Bax, cleaved/pro-caspase-3), antioxidant (Nrf2, SOD2), and cell-cycle (cyclin D1) proteins was analyzed using Western blotting. Additionally, Catalase (CAT) activity, Reduced glutathione (GSH), and total antioxidant capacity (TAC) were determined using colorimetric assays. Results: MGO showed dose- and time-dependent effects, with low concentrations showing high biocompatibility at 48 hours. Flow cytometry revealed MGO significantly increased cell viability from 88.06±1.7% (control) to 94.58±2.88%, 92.87±0.43%, 97.09±2.69%, and 97.61±1.72% at 3.125, 12.5, 50, and 100 µg/mL, respectively (p < 0.0001), while total apoptosis decreased from 11.05±0.55% to 4.96±0.27%, 6.74±0.46%, 2.50±0.21%, and 2.16±0.10% (p < 0.0001). MGO upregulated Bcl-2 (p < 0.001), Nrf2 (>2-fold, p < 0.0001), SOD2 (>2.5-fold, p < 0.0001), and cyclin D1 (1.5-fold, p < 0.01), but downregulated Bax (p < 0.05) and cleaved caspase-3 (p < 0.0001). CAT, GSH, and TAC were significantly elevated (p < 0.001). Conclusion: Low concentrations of MGO enhanced human CC viability, reduced apoptosis, and boosted antioxidant defenses and proliferation-related pathways. These findings suggest MGO’s biomedical potential in cell survival and anti-apoptotic effects on reproductive medicine research. However, more comprehensive studies are needed to determine its long-term safety and efficacy.