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

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: Revascularization and osteogenesis are reinforced by M2 macrophage activation via the CCL2/CCR2 pathway. A and B) HUVECs and BMSCs proliferation assay under M2 macrophage activation. Created with BioRender.com . C) Migration assay and quantification of HUVECs. D) Tube formation assay and quantification of HUVECs. E and F) Early and later osteogenic differentiation of BMSC influenced by macrophage-induced microenvironment. Data are represented as means ± SD, ∗ p < 0.05 (vs Control), ∗∗ p < 0.01 (vs Control), ∗∗∗ p < 0.001 (vs Control), ∗∗∗∗ p < 0.0001 (vs Control); $ 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: Activation Assay, Proliferation Assay, Migration, Tube Formation Assay, Control

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

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: Revascularization and osteogenesis are reinforced by M2 macrophage activation via the CCL2/CCR2 pathway. A and B) HUVECs and BMSCs proliferation assay under M2 macrophage activation. Created with BioRender.com . C) Migration assay and quantification of HUVECs. D) Tube formation assay and quantification of HUVECs. E and F) Early and later osteogenic differentiation of BMSC influenced by macrophage-induced microenvironment. Data are represented as means ± SD, ∗ p < 0.05 (vs Control), ∗∗ p < 0.01 (vs Control), ∗∗∗ p < 0.001 (vs Control), ∗∗∗∗ p < 0.0001 (vs Control); $ 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: Activation Assay, Proliferation Assay, Migration, Tube Formation Assay, Control

Journal: Advanced Science

Article Title: Redirecting Monocyte Differentiation With Engineered Extracellular Vesicles for Glioma Immunotherapy

doi: 10.1002/advs.76910

Figure Lengend Snippet: C‐EVs‐mediated enhanced BBB permeability, excellent targeting ability, and brain tumor accumulation. (A) Schematic illustration of in vitro BBB model. (B) Transport ratio of liposome and EVs traverses the BBB model after different time periods ( n = 3). (C) Confocal images and relative fluorescence intensity of GL261‐IL13Rα2 cells in the lower chamber after treated with DiD‐labeled liposome or EVs for 24 h ( n = 5). Scale bar, 10 µm. (D) Flow cytometry analysis of fluorescence intensity of GL261‐IL13Rα2 in the lower chamber after incubated with EVs or EVs with anti‐CCR2 ( n = 3). (E) Schematic illustration of the 3D tumor spheroids and penetration of DiD‐labeled liposome or EVs into GL261‐IL13Rα2 tumor spheroids after 4 h incubation. Scale bar, 50 µm. (F) In vivo and ex vivo bioluminescence and fluorescence imaging of GL261‐IL13Rα2 tumor‐bearing mice and brain at 24 h after tail vein injection of DiR‐labeled EVs. Immunofluorescence staining of tumor‐bearing brain, dotted lines demarcate the tumor boundary (T), with adjacent normal brain tissue (N) shown for anatomical reference. Scale bars: 50 µm. (G and H) Representative fluorescence images (G) and their quantitative analysis (H) of GL261‐IL13Rα2‐bearing mice after i.v . injection with free DiR, DiR‐labeled liposome, or EVs at different time points. (I) Ex vivo images of the GL261‐IL13Rα2 bearing brain and their quantification of the fluorescence signal in the brain ( n = 3). (J) Immunofluorescence staining and the corresponding line profiles of the tumor‐bearing brain after tail vein injection of free DiR, DiR‐labeled liposome, or EVs. DAPI (blue) stained nuclei, and CD31 (green) labeled blood vessels. Scale bar, 50 µm. (K) C‐EVs bound to the membrane of GL261‐IL13Rα2 cells. Scale bar, 10 µm. (L) Degree of cellular uptake of EVs and C‐EVs in GL261 and GL261‐IL13Rα2 quantified by flow cytometry. (M) CLSM images of GL261‐IL13Rα2 cells incubated with EVs and C‐EVs at 4 h. Scale bar, 10 µm. (N) In vivo and ex vivo fluorescence imaging, and their quantification of the fluorescence signal of GL261‐IL13Rα2 tumor‐bearing mice and brain at 24 h after tail vein injection of DiR‐labeled EVs or C‐EVs. (O) Immunofluorescence staining of tumor‐bearing brain after i.v . injection with DiR‐labeled EVs or C‐EVs. Scale bars: 100 µm. Statistical analysis was performed by unpaired two‐tailed t ‐test (C,D and N) or one‐way ANOVA with Tukey's multiple comparisons tests (I). The experimental data were presented as mean ± S.E.M. ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Neutralizing antibodies against CCR2 (MedChemExpress, USA) were used in antibody‐blocking experiments.

Techniques: Permeability, In Vitro, Fluorescence, Labeling, Flow Cytometry, Incubation, In Vivo, Ex Vivo, Imaging, Injection, Immunofluorescence, Staining, Membrane, Two Tailed Test

Journal: Advanced Science

Article Title: RBM10 Deficiency Promotes Anti‐PD‐1 Resistance in LUAD via STING Alternative Splicing‐Driven CCL7 Signaling and Macrophage Polarization

doi: 10.1002/advs.202522159

Figure Lengend Snippet: CCL7‐CCR2 axis inhibition potentiates anti‐PD‐1 therapy efficacy in RBM10‐low LUAD. (A) mIHC analysis of CCR2 expression on macrophages in tumor tissues from LUAD patients with high and low RBM10 expression. (B) FC analysis of CCR2 expression on macrophages (live + CD45 + CD11b + F4/80 + CD206 + ) in tumor tissues from mice with high and low RBM10 expression (n = 3). (C) Treatment schema for LLC shRBM10 tumor‐bearing mice treated with anti‐PD‐1 antibody, RS102895 or not (n = 5). [Created in BioRender. Gao, W. (2026) https://BioRender.com/awmu085 ] (D) Subcutaneous tumor formation in C57BL/6 injected with shRBM10 LLC cells treated with anti‐PD‐1 antibody, RS102895 or not (n = 5). (E) Tumor growth curves of indicated groups. (F) Survival curves of indicated groups. (G,H) IHC staining of CCL7, CD8, CD68, CD86, and CD206 in tumor tissues from each group. All data are presented as the mean ± SEM (n ≥ 3). The P values in panels (B) were calculated using two‐tailed unpaired Student's t ‐test. The P values in panels (E) were calculated using two‐way ANOVA. Survival curves (F) were calculated using log‐rank test. ns (not significant), * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: In a separate cohort for CCR2 antagonist combination (Figure S11K), shRBM10 tumor‐bearing mice received daily intraperitoneal injections of RS102895 (5 mg/kg, MCE, HY‐18611) together with anti‐PD‐1 (twice weekly) or vehicle controls for 10 consecutive days.

Techniques: Inhibition, Expressing, Injection, Immunohistochemistry, Two Tailed Test