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Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining <t>for</t> <t>Ly-6G</t> in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.
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Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining <t>for</t> <t>Ly-6G</t> in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.
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Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining <t>for</t> <t>Ly-6G</t> in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.
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Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining <t>for</t> <t>Ly-6G</t> in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.
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Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining <t>for</t> <t>Ly-6G</t> in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.
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Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining <t>for</t> <t>Ly-6G</t> in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.
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Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining <t>for</t> <t>Ly-6G</t> in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.
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Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining <t>for</t> <t>Ly-6G</t> in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.
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Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining for Ly-6G in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.

Journal: Bioactive Materials

Article Title: Skin-mimetic bilayer hydrogel normalizes diabetic wound healing by orchestrating inflammatory cell dynamics: An early intervention strategy

doi: 10.1016/j.bioactmat.2026.02.025

Figure Lengend Snippet: Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining for Ly-6G in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.

Article Snippet: Neutrophil recruitment was evaluated by immunohistochemical staining for Ly-6G (Servicebio, GB12229).

Techniques: Immunohistochemical staining, Staining, Saline, Expressing, Immunofluorescence, Generated, Standard Deviation