Fig. 4
Promotion of angiogenesis by exosomes derived from apoptotic SHED. Graph represents the average proliferation of HUVECs measured via the CCK-8 assay at various time points. *indicates statistically significant differences between apoptotic and normal SHED exosomes at day 4 (p < 0.02), day 6 (p < 0.02), and day 8 (p < 0.01) (A). Migration of HUVECs assessed using an in vitro scratch assay (B). Scale bars = 500 μm. Quantification of gap closure (%) in B (C). * indicates statistically significant differences between apoptotic and normal SHED exosomes at 6 h (p < 0.02) and 9 h (p < 0.05). Detection of capillary-like network structures formed by HUVECs at 6 and 12 h using Matrigel tube formation (D). Scale bars = 500 μm. Quantification of tubular structures analyzed at 6 and 12 h using WimTube image analysis software (E). *indicates statistically significant differences in the total tube length (p = 0.002), total branching points (p < 0.001), and total loops (p = 0.018) between the apoptosis-induced SHED exosome group and the positive control or normal exosome groups. Immunofluorescence images of the tube network structures at 12 h with the upper panel indicating CD31 expression (red), the middle panel representing DAPI staining (blue), and the lower panel showing a merged image of CD31 and DAPI (F). Scale bars = 200 μm. Tube formation of HUVECs treated with anti-APN/CD13 and apoptotic SHED exosomes, as well as anti-MMP-2 and apoptotic SHED exosomes, showing reduced tube formation and branching. Scale bars = 100 μm (G).
