Figure 6: Effects of combined treatment of Ac-PGP and hEPC on the cutaneous wound healing and neovascularization.

(a) Excisional skin wounds were introduced by 8-mm biopsy punch on the dermal skin of BALB/CA-nu/nu mice, followed by treatment with HBSS, Ac-PGP (0.1 μM), CM-DiI labeled-hEPC injection, and the combination of Ac-PGP (0.1 μM) and hEPCs injection. Representative images at indicated days during the recovery period are shown. (b) Quantitative analysis of the wound area at indicated days in comparison with the original wound during the recovery period is shown. The wound area was measured by using Image J software program (version 1.50i). Data indicate mean ± SD. *p < 0.05, **p < 0.01, *** < 0.001 versus HBSS, ^#p < 0.05 Ac-PGP + hEPC versus Ac-PGP or hEPCs (n = 8). (c) Representative images of mouse skin wound samples at day 6 after immunostaining with anti-CD31 (green) or α-SMA (red) antibodies are shown. Nuclei were counterstained with DAPI (blue). Bar = 100 μm. (d) Quantitative analysis of CD31-positive blood vessels in the dermal area of wound is shown. *p < 0.05 versus HBSS, ^#p < 0.05 Ac-PGP + hEPCs versus Ac-PGP or hEPCs (n = 8). (e) Quantitative analysis of α-SMA-positive vessels in the dermal area of wound is shown. *p < 0.05 versus HBSS, ^#p < 0.05 Ac-PGP + hEPCs versus Ac-PGP or hEPCs (n = 8). (f) Representative images of day 6 mouse skin wound samples injected with CM-DiI-labeled hEPCs with or without the combination of Ac-PGP treatment (0.1 μM) are shown. Nuclei were counterstained with DAPI (blue). Bar = 50 μm. (g) Quantitative analysis of CM-DiI-positive hEPCs in dermal area of day 3, 6, 9, and 12 skin wound samples is shown. Data indicate mean ± SD. ** < 0.01 Ac-PGP + hEPC versus hEPCs (n = 6).