Fig. 6: In vitro redox homeostasis modulation and stem cell protection against oxidative stress. | Nature Communications

Fig. 6: In vitro redox homeostasis modulation and stem cell protection against oxidative stress.

From: Sono-activable and biocatalytic 3D-printed scaffolds for intelligently sequential therapies in osteosarcoma eradication and defect regeneration

Fig. 6

a Schematic illustration of the H2O2 redox homeostasis restore and O2-induced hypoxia relief of HS-ICTO under bone defect condition. b Time-dependent H2O2 elimination of HS and HS-ICTO-x (0.5, 1.0, and 2.0). c The produced O2 concentration was measured in the presence of HS and HS-ICTO-x (0.5, 1.0, and 2.0) and H2O2. d Representative CLSM images of live/dead staining of BMSCs seeded on 3D scaffolds with different treatments on day 5 and the corresponding CCK-8 assay result (n = 3 biologically independent replicates). e Representative CLSM images of phalloidin-Rhodamine stained BMSCs with different treatments and the corresponding cell size evaluation (n = 20, the size of twenty random cells from each group was calculated by ImageJ software). f TEM observation of the BMSCs after different treatments. The red arrow indicates damaged mitochondria, and the red star indicates swollen endoplasmic reticulum. g ALP staining of the BMSCs stimulated by osteogenic medium for 14 days with different treatments. h ARS staining of the BMSCs stimulated by osteogenic medium for 21 days with different treatments. i Quantitative analysis of the ALP and j ARS positive staining area (n = 3 biologically independent replicates). k Immunofluorescence staining of Runx2, BMP-2, and COLI expression of BMSCs stimulated by osteogenic medium for 14 days with different treatments. The images in (d–h, k) were representative of three independently repeated experiments from each group. l Quantitative analysis of the MFI of Runx2, BMP-2, and COLI immunofluorescence staining (n = 3 biologically independent replicates). m The Runx2, BMP-2, COLI, and ALP gene expression of BMSCs after different stimulation for 14 days measured by RT-qPCR (n = 3 biologically independent replicates). n Schematic illustration of HS-ICTO regulating H2O2 redox homeostasis and protecting BMSCs. Data are presented as mean ± SD, and ns represents no significant difference; statistical significance was calculated using one-way ANOVA followed by Tukey’s post hoc test for multiple comparisons; all tests were two-sided. In (l), a.u. indicates the arbitrary units. Source data are provided as a Source Data file.

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