fahimeh kabiri
1 
, tahereh foroutan
1* , roghiyeh Pashaei-Asl
2,3, maryam Pashaei-Asl
4,5* , Mohammad Ganjian
6, vahid vatanpour
71 Department of Animal Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran
2 Interdisciplinary Research Development Center, Iran University of Medical Sciences, Tehran, Iran
3 Department of Biochemistry, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
4 Department of Reproductive Biology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran
5 Women’s Reproductive Health Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
6 Department of Electronic Technology, TU Ilmenau, Ilmenau, Germany
7 Faculty of Chemistry, Kharazmi University, Tehran, Iran
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.