Journal: Bioactive Materials
Article Title: Spatiotemporally programming the immune-osteogenic cascade with a dual-immunomodulatory scaffold for functional bone regeneration
doi: 10.1016/j.bioactmat.2026.04.002
Figure Lengend Snippet: Immune regulation and endogenous bone regeneration mechanism investigation. A) Network diagram showing the number of interactions between six subclusters. B) KEGG enrichment analysis of the upregulated DEGs in DIBS group compared to the HA group. C) Circular visualization of related pathway–gene enrichment analysis. D) Heatmap of key gene regulation in specific pathways. E) qRT-PCR validation for key gene expression in specific pathways. F) The interaction networks showing the correlation of representative immunomodulatory genes (CCL2, CCL20, Sfrp1, and Stat3, etc.) with angiogenesis/osteogenesis and macrophage regulation gene sets. G) Flow cytometry analysis and quantification of CCR2 F4/80 macrophage in peripheral blood. H) Immunofluorescence staining analysis of macrophage polarization inside scaffolds (one week after intramuscular implantation). I) Macrophage proliferation assay in a CCR2-dependent manner. J and K) Macrophage polarization assay in a CCR2-dependent manner. Data are represented as means ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; $ p < 0.05 (vs group without inhibitor), $$ p < 0.01 (vs group without inhibitor), $$$ p < 0.001 (vs group without inhibitor), $$$$ p < 0.0001 (vs group without inhibitor). ns, not significant.
Article Snippet: Throughout the experimental period, sustained CCR2 inhibition was achieved via daily intraperitoneal injections (2 mg/kg) of the highly selective CCR2 inhibitor RS504393 (Cat. No. HY-15418, MCE).
Techniques: Quantitative RT-PCR, Biomarker Discovery, Gene Expression, Flow Cytometry, Immunofluorescence, Staining, Proliferation Assay