Preferred M2 Polarization by ASC-Based Hydrogel Accelerated Angiogenesis and Myogenesis in Volumetric Muscle Loss Rats

Figure 4

rASC-based hydrogel regulating inflammation, myogenesis, and vasculogenesis during muscle regeneration. (a) and (b) displayed the inflammation, angiogenic, and myogenic mRNA expression at 5 d and 2 w during muscle repair and regeneration. mRNA expressions were normalized to 18S for rat muscles. (c), (d) IF staining of Arg I (red) and CD68 (green) at 5 d and 7 d in wound areas. (e) Arg I+/CD68+ ratio manifested more prohealing M2 profile in the Hyd-ASC and Hyd-CM groups than in the hydrogel group. Scale bar = 20 μm. (f) The IF staining of α-SMA and CM-Dil-labeled ASCs at 2 w post injury. Scale bar = 100 μm. Nuclei in blue, ECs in green, and rASCs in red. (g) The pan macrophage marker CD68, M1 marker iNOS, M2 marker Arg I, endothelial cell marker α-SMA, myogenic markers Myogenin and MyoD, other prohealing-related protein like desmin and fibromodulin (FMOD), HGF, collagen I, inflammation protein TNF-α, signaling pathway protein AKT, and pAKT expressions were detected by Western blot. GAPDH was loaded as control. (h) Relative protein expression was analyzed by fold of GAPDH or ratio of phosphorylation to total protein. Data were mean ± SEM of three independent experiments. versus hydrogel group.

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