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PEG-pp@nMSC@MT hydrogel promoted macrophage polarization from M1 to M2 in the bone defect area. a) Schematic illustration for the immunomodulation function of the hydrogel. b) Immunofluorescent and c) Semi-quantitative analysis showing macrophage polarization in defect areas of 1 week (n = 3). Scale bar = 50 μm. d) mRNA expression of cytokines in tissue lysate (n = 3). e) Heatmap showing the cytokines release profile (IL-6, TNF- α , IL-1 β , IL-10, and IL-4) of tissue lysate (n = 3). f) Fluorescent confocal images, and g) Semi-quantitative analysis of immunofluorescent staining of iNOS and CD206 in macrophages (n = 3). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( a was created with bioRender. com).

Journal: Bioactive Materials

Article Title: MSC-mimicking nanovesicle embedded bio-adhesive hydrogel for dual immunomodulation and osteogenesis to promote maxillofacial bone regeneration

doi: 10.1016/j.bioactmat.2026.02.032

Figure Lengend Snippet: PEG-pp@nMSC@MT hydrogel promoted macrophage polarization from M1 to M2 in the bone defect area. a) Schematic illustration for the immunomodulation function of the hydrogel. b) Immunofluorescent and c) Semi-quantitative analysis showing macrophage polarization in defect areas of 1 week (n = 3). Scale bar = 50 μm. d) mRNA expression of cytokines in tissue lysate (n = 3). e) Heatmap showing the cytokines release profile (IL-6, TNF- α , IL-1 β , IL-10, and IL-4) of tissue lysate (n = 3). f) Fluorescent confocal images, and g) Semi-quantitative analysis of immunofluorescent staining of iNOS and CD206 in macrophages (n = 3). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( a was created with bioRender. com).

Article Snippet: Briefly, tissue sections underwent antigen retrieval (37 °C, 30 min), blocked by 5% bovine serum albumin (BSA, RT, 1 h), then sequentially incubated (4 °C) with lineage-specific probes: macrophage marker F4/80 (1:200, Cat. sc-52664, Santa Cruz Biotechnology) and CD68 (1:250, Cat. 14-0681-81, ThermoFisher, USA), CD206 (1:500, Cat. 24595T, Cell Signaling Technology, USA) and Arg-1 (1:250, Cat. 82975, Proteintech, China) for a M2 marker, iNOS (1:500, Cat. ab178945, Abcam, UK) and CD86 (1:300, Cat. DF6332, Affinity, China) for a M1 marker, RUNX2 (1:150, Cat. sc390351, Santa Cruz Biotechnology, USA) and OCN (1:150, Cat. sc390877, Santa Cruz Biotechnology, USA) for osteogenesis markers, VEGF (1:50, Cat. sc57496, Santa Cruz Biotechnology, USA) and CD31 (1:50, Cat. sc20071, Santa Cruz Biotechnology, USA) for angiogenesis markers, and CD146 (1:200, Cat. Ab75769, Abcam, UK) for stem cell surface marker, overnight at 4 °C.

Techniques: Expressing, Staining