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Fig. 5 | Inflammation and Regeneration

Fig. 5

From: Molecular hydrogen promotes wound healing by inducing early epidermal stem cell proliferation and extracellular matrix deposition

Fig. 5

H2 promoted in vivo and in vitro keratinocytes and fibroblasts cells viability by accelerating the re-epithelialization, migration and mitochondrial functions. A. Representative images of HaCat cell epithelization. Images were captured every two hours using a live cell imaging system, and the 0, 12 and 24 hour images were presented. B. The increased rate of the total colony area in 12 and 24 hours. C. Comparison of mitochondrial morphology 3 days post-wound for the D3H and D3C groups observed by electron microscope. The right two images in each panel are enlargements of the white frames in the left images. Red arrows indicate intact mitochondria, while yellow arrows indicate damaged mitochondria. D. Comparison of total mitochondria, intact mitochondria and damaged mitochondria numbers between D3H and D3C groups. E and G. Representative images of HUCP-MSCs and HPF fibroblasts migration at 0-h, 12-h, and 24-h time points in H2 medium and control groups. F, H Graphic analysis of HUCP-MSCs and HPF fibroblasts total moving distance in 24 h. I. Cell migration ability examined by wound-healing assay in H2 medium and control groups. J Wound-healing curve analysis under different conditions. K Heatmap and enrichment plot showing genes related to mitochondrial respiratory chain complex assembly, which is the top 1 GO enrichment function in D3H. L, M Top GO enrichment plots in D3H related to inner mitochondrial membrane organization and ATP synthesis-coupled proton transport, which indicates more intact and functional mitochondria in D3H. N GSEA GO enrichment plots enriched in Mt related functions for D3H (NES > 1.5). White and black dotted lines in E and G indicate the migration route of fibroblasts. Red dotted line in G. outlines the representative HUCP-MSCs. Data in B and D processed two-way ANOVA test, and data in F, H, and J processed unpaired t test. Data were plotted as mean ± SEM. *P value < 0.05; **P value <0.01; ***P value < 0.001; no stars for P value > 0.05; Scale bar in A and G = 200 μm, scale bar in C = 2 μm, scale bar in E = 100 μm, scale bar in I = 1000 μm

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