Fig. 6: Prevention of ROS-related DNA damage and apoptosis in MSCs by Ru-Cu/EDHJ.
a Fluorescence images and (b) linear distribution of fluorescence intensity from DNA/RNA damage staining. Scale bar: 50 μm. c Fluorescence images, (d) spectrum images, and (e) quantitative analysis of fluorescence intensity from γ-H2A.X (n = 20 independent replicates), p(H2O2) < 0.0001, compared with Ctrl group; p(Ru-Cu/EDHJ-H2O2) < 0.0001, compared with H2O2 group. Scale bar: 50 μm. f Confocal scanning images from TUNEL. Scale bar: 50 μm. g Apoptosis analysis was performed using flow cytometry of Annexin V-FITC/PI stained hMSCs. h Quantitative fluorescence analysis from TUNEL (n = 30 independent replicates), p(H2O2) <0.0001, compared with Ctrl group; p(CeO2-H2O2) = 0.0052, p(Ru-Cu/EDHJ-H2O2) < 0.0001, compared with H2O2 group. i The cell percentages in stages of normal, early apoptosis, late apoptosis, and necrosis. j ALP staining after 3-day in vitro osteo-induction and AR staining after 21-day in vitro osteo-induction using the hMSCs. Quantitative analysis of (k) ALP (n = 3 independent replicates), p(H2O2) < 0.0001, compared with Ctrl group; p(Ru-Cu/EDHJ-H2O2) < 0.0001, compared with H2O2 group. Quantitative analysis of (l) AR (n = 3 independent replicates) in H2O2 treated hMSCs, p(H2O2) < 0.0001, compared with Ctrl group; p(Ru-Cu/EDHJ-H2O2) < 0.0001, compared with H2O2 group. m Schematic illustration of apoptosis and osteogenesis effects with Ru-Cu/EDHJ in high ROS-level microenvironment. Data are presented as means ± SD., *p < 0.05, **p < 0.01, ***p < 0.001, #p < 0.05, ##p < 0.01, ###p < 0.001, ns, no significance; statistical significance was calculated using one-way ANOVA followed by Tukey’s post-hoc test for multiple comparisons, all tests were two-sided. Experiments were repeated independently (a, c, d, f, g, j, l) three times with similar results. Source data are provided as a Source Data file.
