Fig. 2: In vitro characterization of BMP2 and BMP2/7-EV impact on calcification and alkaline phosphatase activity.

A Alizarin red assay with MC3T3-E1 cells treated with MSC-EV, BMP2-EV, or BMP2/7-EV for 16 days versus untreated control. Zoomed in image of each well included below. Scale bar 100 µm. B Alkaline phosphatase activity assay with MC3T3-E1 cells treated with MSC-EV, BMP2-EV, or BMP2/7-EV for 10 days versus untreated control. C Quantification (mean +/- SD at optical density 405 nm) of alizarin red assay (A) at day 16. Statistical significance determined by one-way ANOVA with Tukey post hoc analysis. (n = 6). D Quantification (mean normalized to mg protein +/- SD) of alkaline phosphatase activity (B) at 3, 6, 10, or 14 days post-treatment. Statistical significance compared was determined by two-way ANOVA with Tukey post hoc analysis. Statistical significance for each group matches the color of the line. (n = 5).