Figure 5. Antioxidant activity of nanoclay-organic bone sealant (NoBS).

A) Hydrogen peroxide and B) DPPH radical scavenging activity of methacrylated chitosan (MeGC) hydrogel, oBS, and NoBS (1.0% NC) with or without SAG (10 μM). C) Fluorescence microscopy images of H₂DCFDA-treated BMSCs grown with MeGC hydrogel, oBS, and NoBS (1.0% NC) with or without SAG (10 μM) in 0.2 mM H₂O₂-containing growth medium. Green fluorescence indicates DCF, which is the product resulting from the reaction between ROS and H₂DCFDA ROS indicator. Nuclei were stained with Hoechst 33342 (blue). Scale bar indicates 100 μm. Error bars indicate standard deviation (n = 3), ***p < 0.001 (ANOVA followed by Tukey’s post hoc test). D-G) Antioxidant activity of oBS, and NoBS (1.0% NC) on osteogenic activity of BMSCs was evaluated in the presence or absence of 0.2 mM, hydrogen peroxide. D) ALP staining and E) ALP activity were performed at day 4. F) Alizarin red S staining was carried out at day 14, and G) their quantification was also evaluated. Scale bars indicate 100 μm in D) and F). Error bars indicate standard deviations (three independent cultures, n = 3), *p < 0.05 (ANOVA followed by Tukey’s post hoc test)