Fig 2: Regulation of neutrophil migration and apoptosis and macrophage polarization by the hybrid biomaterial in vitro. A) Quantitative flow cytometric analysis of neutrophil migration after the different treatments. B) Representative fluorescence images and quantitative analysis of neutrophil (red) migration to the lower chamber of transwell. Scale bars, 50 µm. C) Representative flow cytometry plots and quantitative analysis of neutrophil apoptosis. D) Western blot analysis of apoptosis-related proteins in neutrophils. E) Western blot analysis of the phenotypic markers in LPS-stimulated macrophages. Proinflammatory macrophage marker, iNOS; anti-inflammatory macrophage markers, CD206 and Arg-1. F) The levels of cytokines released by macrophages measured by enzyme-linked immunosorbent assay (ELISA). G) Representative fluorescence images of macrophage phenotypes after incubation with different pretreated neutrophils; iNOS (red), CD206 (green), and nuclei (blue). Scale bars, 50 µm. All data were generated from at least three independent experiments and are presented as the means ± standard deviation (SD). Statistical analysis was performed by one-way ANOVA. ns, not significant; *P < 0.05 and **P < 0.01.

Fig 3: Inflammatory response manipulation by Gel@fMLP/SiO2-FasL in critical-sized calvarial bone defects. A) Representative fluorescence images and the percentage of neutrophils (Ly-6G+ cells, green) that infiltrated the bone defects. Scale bars, 50 µm. B) Representative fluorescence images of apoptotic neutrophils (Ly-6G+ TUNEL+ cells) in bone defects; Ly-6G (red), TUNEL (green), and nuclei (blue). Scale bars, 50 µm. C) Representative fluorescence images of macrophage phenotypes in the bone defects. Scale bars, 50 µm. Quantitative analysis of iNOS+/CD206+ cells in each group. D) Representative fluorescence images of cytokine expression in bone defects. Scale bars, 50 µm. Quantitative analysis of TNF-a/TGF-ß expression levels in each group. n = 6 per group. The data are presented as the means ± SD. In (A), (C), and (D), statistical analysis was performed using one-way ANOVA. In (B), the CD206+ population of 4 d (in (C)) was analyzed using the Kruskal–Wallis H-test. ns, not significant; *P < 0.05 and **P < 0.01.

Fig 4: Inflammatory response realignment by Gel@fMLP/SiO2-FasL in diabetic cutaneous wounds. A) Quantitative analysis of neutrophil infiltration in diabetic cutaneous defects was performed by flow cytometry. B) Representative fluorescence images and the percentage of neutrophils (Ly-6G+ cells, green) in cutaneous defects. Scale bars, 50 µm. C) Representative flow cytometry plots and quantitative analysis of neutrophil apoptosis in cutaneous defects. D) Quantitative analysis of proinflammatory macrophages in cutaneous defects was performed by flow cytometry. E) Quantitative analysis of anti-inflammatory macrophages in cutaneous defects was performed by flow cytometry. F) Representative fluorescence images of macrophage phenotypes in cutaneous defects. Scale bars, 50 µm. Quantitative analysis of iNOS+/CD206+ cells in each group. G) Cytokines expression in cutaneous defects was analyzed by ELISA. n = 6 per group. The data are presented as the means ± SD. In (A)–(G), statistical analysis was performed by one-way ANOVA, except for TGF-ß expression (in (G)) which was analyzed by the Kruskal–Wallis H-test. ns, not significant; *P < 0.05 and **P < 0.01.

Fig 5: T17-MnO2 enhanced the immune response of BMDMs. (A) A schematic diagram of the synthesis of T17-MnO2. (B, C) Particle size distribution and zeta potential of T17-MnO2 were detected by the nanoparticle size analyzer. (D) LDH release of BMDMs was detected after T17-MnO2 treatment for 24 h. After treatment of MnCl2 (50 µmol/L), MnO2 (50 µmol/L), T17 (200 nmol/L), T17+MnCl2, and T17-MnO2 for 24 h, (E) the mRNA levels of Ifnb in BMDMs were detected by Real-time PCR, (F) the protein level of TBK1 in BMDMs was detected by Western blot, (G–I) and the protein levels of INOS, CD86 and CD206 were detected by flow cytometry. Data are presented as mean ± SD (n = 3). **P < 0.01, ***P < 0.001, ****P < 0.0001, ns, not significant.