Fig. 7
TSC-EVs induced proliferation and regenerative effects on MSCs by NGF/Akt activation. A NGFR mRNA level was analyzed by RT-PCR in MSCs treated with SFM or TSC-EVs for 24 h. B NGF in TSC-EVs was detected by western blot analysis. C Protein levels of Akt and p-Akt in MSCs with SFM or TSC-EVs treatment were determined by western blot and quantitated the expression level of each group. D Expression levels of NGFR gene and (E) protein levels of Akt and p-Akt were analyzed in MSCs with SFM, TSC-EV, and TSC-EV + anti-NGF. F Light microscopy images and (G) cell proliferation rate in MSCs with SFM, TSC-EV, and TSC-EV + anti-NGF. Scale bars, 200 μm. H Representative images of BrdU expression (green) in TSC-EV-treated MSCs with or without anti-NGF. Scale bars, 125 μm. I The number of BrdU-positive cells were represented by graph. J Representative photographs of wound scar area in mice treated with SFM, MSC-EV, TSC-EV, and TSC-EV + anti-NGF exposed MSCs. Scale bars, 0.5 mm. K The percentage of open area on skin wound was measured in each group. Data represented the mean and SD from four to eight mice per group. (*p < 0.05 and **p < 0.01) ALP activity (L) and calcium deposition (M) of MSCs treated with SFM, TSC-EV, and TSC-EV + anti-NGF were determined. N Representative images of SA-b-gal stained MSCs cultured with GM, SFM, TSC-EV, and TSC-EV + anti-NGF. Scale bars, 100 μm. O Stained cells were calculated in each group and there was no statistical significance in SFM, TSC-EV, and TSC-EV + anti-NGF. Error bars indicated the mean and SD. (n = 3; *p < 0.05, **p < 0.01, and ***p < 0.001)