Fig. 3

Evaluation of angiogenic and immunomodulatory properties of ASC-derived Exos in a mouse model of ischemic muscle injury (A-H). Ischemic muscle injury was induced by the ligation of the femoral artery. Mice were allocated into PBS; Normal Exos (Nor/Exo); Hypoxic Exos (Hyp/Exo); and Hyp/Exo + BLZ945 groups (each in 8). Laser speckle imaging indicated the changes in hind paw blood perfusion after 21 days (A). Analyses confirmed superior effects of Nor/Exo, especially Hyp/Exo, on the promotion of plantar perfusion (B). Immunofluorescence imaging of injured adductor muscles 3 weeks after injection of Exos (C and D). Data indicate the promotion of α-SMA⁺ and CD31⁺ vessels in mice that received ASC hypoxic and normoxic Exos compared to the PBS group (Scale bar: 150 µm). Simultaneous injection of BLZ945 blunted these effects (Counterstaining: Hoechst 33,342). Immunofluorescence imaging of CD31 in injured gastrocnemius muscles after injection of Exos (E and F; Scale bar: 100 µm). An average number of CD31⁺ vessels increased following the injection of hypoxic and normoxic Exos compared to the PBS mice. Again, BLZ945 blunted these effects. Flow cytometry analysis of F4/80 + macrophages (G) and M2 type CD206⁺ macrophages (H). One-Way ANOVA analysis with Tukey method. *p < 0.05, **p < 0.01, and ***p < 0.001. Adapted from [287].