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In vitro evaluation of immunomodulatory effects in composite hydrogel systems involving Nnat . (A) Top: Schematic representation of in vitro inflammatory model; Bottom: Morphological changes in macrophages observed via light microscopy. Scale bars: 100 μm. (B) Flow cytometric analysis of reactive oxygen species. (C) Phenotypic characterization of macrophage subtypes (M0/M1/M2) through surface marker detection. (D) Enhanced activation of NF-κB/P38 signaling pathways with elevated PI3K phosphorylation observed in Hydrogel + si- Nnat group versus hydrogel control. (E) <t>Quantitative</t> <t>PCR</t> analysis of gene expression. (F) Metabolic activity assessment through viability assays. (G) Apoptotic cell quantification. (H) TUNEL staining patterns across experimental groups. Scale bars: 50 μm. (I) Cytokine profile analysis using ELISA. (n = 5, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.)
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In vitro evaluation of immunomodulatory effects in composite hydrogel systems involving Nnat . (A) Top: Schematic representation of in vitro inflammatory model; Bottom: Morphological changes in macrophages observed via light microscopy. Scale bars: 100 μm. (B) Flow cytometric analysis of reactive oxygen species. (C) Phenotypic characterization of macrophage subtypes (M0/M1/M2) through surface marker detection. (D) Enhanced activation of NF-κB/P38 signaling pathways with elevated PI3K phosphorylation observed in Hydrogel + si- Nnat group versus hydrogel control. (E) <t>Quantitative</t> <t>PCR</t> analysis of gene expression. (F) Metabolic activity assessment through viability assays. (G) Apoptotic cell quantification. (H) TUNEL staining patterns across experimental groups. Scale bars: 50 μm. (I) Cytokine profile analysis using ELISA. (n = 5, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.)
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In vitro evaluation of immunomodulatory effects in composite hydrogel systems involving Nnat . (A) Top: Schematic representation of in vitro inflammatory model; Bottom: Morphological changes in macrophages observed via light microscopy. Scale bars: 100 μm. (B) Flow cytometric analysis of reactive oxygen species. (C) Phenotypic characterization of macrophage subtypes (M0/M1/M2) through surface marker detection. (D) Enhanced activation of NF-κB/P38 signaling pathways with elevated PI3K phosphorylation observed in Hydrogel + si- Nnat group versus hydrogel control. (E) <t>Quantitative</t> <t>PCR</t> analysis of gene expression. (F) Metabolic activity assessment through viability assays. (G) Apoptotic cell quantification. (H) TUNEL staining patterns across experimental groups. Scale bars: 50 μm. (I) Cytokine profile analysis using ELISA. (n = 5, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.)
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In vitro evaluation of immunomodulatory effects in composite hydrogel systems involving Nnat . (A) Top: Schematic representation of in vitro inflammatory model; Bottom: Morphological changes in macrophages observed via light microscopy. Scale bars: 100 μm. (B) Flow cytometric analysis of reactive oxygen species. (C) Phenotypic characterization of macrophage subtypes (M0/M1/M2) through surface marker detection. (D) Enhanced activation of NF-κB/P38 signaling pathways with elevated PI3K phosphorylation observed in Hydrogel + si- Nnat group versus hydrogel control. (E) <t>Quantitative</t> <t>PCR</t> analysis of gene expression. (F) Metabolic activity assessment through viability assays. (G) Apoptotic cell quantification. (H) TUNEL staining patterns across experimental groups. Scale bars: 50 μm. (I) Cytokine profile analysis using ELISA. (n = 5, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.)
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In vitro evaluation of immunomodulatory effects in composite hydrogel systems involving Nnat . (A) Top: Schematic representation of in vitro inflammatory model; Bottom: Morphological changes in macrophages observed via light microscopy. Scale bars: 100 μm. (B) Flow cytometric analysis of reactive oxygen species. (C) Phenotypic characterization of macrophage subtypes (M0/M1/M2) through surface marker detection. (D) Enhanced activation of NF-κB/P38 signaling pathways with elevated PI3K phosphorylation observed in Hydrogel + si- Nnat group versus hydrogel control. (E) <t>Quantitative</t> <t>PCR</t> analysis of gene expression. (F) Metabolic activity assessment through viability assays. (G) Apoptotic cell quantification. (H) TUNEL staining patterns across experimental groups. Scale bars: 50 μm. (I) Cytokine profile analysis using ELISA. (n = 5, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.)
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In vitro evaluation of immunomodulatory effects in composite hydrogel systems involving Nnat . (A) Top: Schematic representation of in vitro inflammatory model; Bottom: Morphological changes in macrophages observed via light microscopy. Scale bars: 100 μm. (B) Flow cytometric analysis of reactive oxygen species. (C) Phenotypic characterization of macrophage subtypes (M0/M1/M2) through surface marker detection. (D) Enhanced activation of NF-κB/P38 signaling pathways with elevated PI3K phosphorylation observed in Hydrogel + si- Nnat group versus hydrogel control. (E) <t>Quantitative</t> <t>PCR</t> analysis of gene expression. (F) Metabolic activity assessment through viability assays. (G) Apoptotic cell quantification. (H) TUNEL staining patterns across experimental groups. Scale bars: 50 μm. (I) Cytokine profile analysis using ELISA. (n = 5, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.)
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In vitro evaluation of immunomodulatory effects in composite hydrogel systems involving Nnat . (A) Top: Schematic representation of in vitro inflammatory model; Bottom: Morphological changes in macrophages observed via light microscopy. Scale bars: 100 μm. (B) Flow cytometric analysis of reactive oxygen species. (C) Phenotypic characterization of macrophage subtypes (M0/M1/M2) through surface marker detection. (D) Enhanced activation of NF-κB/P38 signaling pathways with elevated PI3K phosphorylation observed in Hydrogel + si- Nnat group versus hydrogel control. (E) Quantitative PCR analysis of gene expression. (F) Metabolic activity assessment through viability assays. (G) Apoptotic cell quantification. (H) TUNEL staining patterns across experimental groups. Scale bars: 50 μm. (I) Cytokine profile analysis using ELISA. (n = 5, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.)

Journal: Bioactive Materials

Article Title: Exosome-functionalized photocrosslinked GelMA/HAMA hydrogel promotes facial nerve recovery via inflammatory microenvironment regulation

doi: 10.1016/j.bioactmat.2026.01.008

Figure Lengend Snippet: In vitro evaluation of immunomodulatory effects in composite hydrogel systems involving Nnat . (A) Top: Schematic representation of in vitro inflammatory model; Bottom: Morphological changes in macrophages observed via light microscopy. Scale bars: 100 μm. (B) Flow cytometric analysis of reactive oxygen species. (C) Phenotypic characterization of macrophage subtypes (M0/M1/M2) through surface marker detection. (D) Enhanced activation of NF-κB/P38 signaling pathways with elevated PI3K phosphorylation observed in Hydrogel + si- Nnat group versus hydrogel control. (E) Quantitative PCR analysis of gene expression. (F) Metabolic activity assessment through viability assays. (G) Apoptotic cell quantification. (H) TUNEL staining patterns across experimental groups. Scale bars: 50 μm. (I) Cytokine profile analysis using ELISA. (n = 5, ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.)

Article Snippet: Reaction systems (20 μL volume) were assembled in RNase-free tubes, incorporating cDNA templates, paired forward/reverse primers (10 μM concentration per primer), Vazyme's 2 × Taq Pro Universal SYBR qPCR Master Mix, and molecular-grade water.

Techniques: In Vitro, Light Microscopy, Marker, Activation Assay, Protein-Protein interactions, Phospho-proteomics, Control, Real-time Polymerase Chain Reaction, Gene Expression, Activity Assay, TUNEL Assay, Staining, Enzyme-linked Immunosorbent Assay