Reduced graphene oxide improves endothelial differentiation efficiency and angiogenesis ability of adipose-derived stem cells

Yao Jinfeng; Deng Mengzhao; Xie Tian; Chen Kan; Wang Haixia

doi:10.12307/2026.134

Abstract：BACKGROUND:Adipose-derived stem cells can differentiate into vascular endothelial cells in a specific induction environment and are ideal seed cells in the process of tissue engineering vascularization. OBJECTIVE:To explore the effect of reduced graphene oxide on the endothelial differentiation efficiency and angiogenesis ability of adipose-derived stem cells. METHODS:Firstly,reduced graphene oxide was prepared by green synthesis process and characterized by transmission electron microscopy,energy spectrum analysis and Fourier transform infrared spectroscopy.Different mass concentrations of reduced graphene oxide were co-cultured with adipose-derived stem cells,and the safe concentration range of reduced graphene oxide was determined by CCK8 assay.Then,the endothelial differentiation of adipose-derived stem cells was induced by reduced graphene oxide in a safe concentration range for different times.The expression of endothelial differentiation-related indicators of adipose-derived stem cells was detected by RT-qPCR and western blot assay.Finally,the tube formation and migration abilities of cells induced by reduced graphene oxide were evaluated by scratch test and tubule formation test. RESULTS AND CONCLUSION:(1)Reduced graphene oxide has a typical two-dimensional sheet structure,mainly containing carbon and oxygen elements,and characteristic absorption peaks related to oxygen-containing functional groups can be detected.(2)The results of CCK-8 assay showed that reduced graphene oxide at a concentration of 10 μg/mL or less had no effect on the cell viability of adipose-derived stem cells after 72 hours of culture.(3)The results of RT-qPCR and western blot assay show that reduced graphene oxide at a concentration of 10 μg/mL or less could significantly increase the expression of markers related to endothelial differentiation of adipose-derived stem cells after 6 and 9 days of induction,among which 5 μg/mL reduced graphene oxide induced 9 days,and the gene expression difference of endothelial differentiation markers was the greatest.(4)The results of scratch test and tubule formation test showed that compared with the blank group and the control group,the migration and tube formation abilities of cells in the reduced graphene oxide group were significantly enhanced.(5)The results show that green synthesized reduced graphene oxide can significantly improve the endothelial differentiation efficiency of adipose-derived stem cells,among which 5 μg/mL reduced graphene oxide induction for 9 days is the best time-concentration combination,and the induced differentiated cells have better angiogenesis function.