Journal: Bioactive Materials
Article Title: Sustained oxygen-releasing hydrogel implants enhance flap regeneration by promoting mitochondrial biogenesis under mild hypoxia
doi: 10.1016/j.bioactmat.2025.04.010
Figure Lengend Snippet: Effects of cOMP-GelMA treatment on cell apoptosis and enhanced mitochondrial activity. (A) Representative immunofluorescence (IF) images from TUNEL assay of apoptotic cells. Scale bar = 2000 μm. (B) Quantification of TUNEL-positive apoptotic cells based on optical intensity measurements from the TUNEL assay images. n = 4–5 per group. (C) Schematic representation of the interaction between PGC-1α and NRF-1, highlighting their roles in mitochondrial biogenesis. (D) Representative IF images display co-staining of NRF-1 and PGC-1α mitochondrial activity markers. (E–H) Quantitative analysis of the expression levels of (E) NRF-1, (F) PGC-1α, and (G) quantification of cells positive for both markers; scale bar = 200 μm. n = 4–5 per group. (H) Ppargc1a mRNA expression in the subdermal plexus was assessed using qPCR, n = 5 per group. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 determined using one-way ANOVA.
Article Snippet: Following deparaffinization, antigen retrieval, and blocking the tissue sections were incubated with primary antibodies, including vascular endothelial growth factor (VEGF; 1:250, sc-57496, Santa Cruz Biotechnology, Inc., Texas, USA), myeloperoxidase (MPO; 1:250, ab188211, Abcam, Cambridge, UK), CD31 (1:100, ab28364, Abcam), alpha-smooth muscle actin (α-SMA; 1:100, 14395-1-AP, Proteintech Group Inc., Rosemont, USA), Nuclear Respiratory Factor 1 (NRF-1, 1:100, ab175932, Abcam), and PGC-1α (1:100, 66369-1-lg, Proteintech Group Inc.), Hypoxia Inducible Factor 1 Alpha (HIF-1a, 1:400, 48085S, Cell signaling, Massachusetts, USA), Cluster of Differentiation 68(CD68, 1:250, MCA341R, Bio-rad, California, USA), Phospho-nuclear factor kappa-light-chain-enhancer of activated B cells p65 (p-NFκB p65, 1:100, sc-136548, Santa Cruz Biotechnology), 8-hydroxy-2′-deoxyguanosine (8-OhdG, 1:200, sc-393871, Santa Cruz Biotechnology) and Superoxide dismutase 2, mitochondrial (SOD2, 1:50, sc-133134, Santa Cruz Biotechnology).
Techniques: Activity Assay, Immunofluorescence, TUNEL Assay, Staining, Expressing