il 1β  (Servicebio Inc)

Journal: International Journal of Molecular Medicine

Article Title: Triptolide exerts antiviral effects and alleviates influenza A-induced pneumonia by inhibiting the overactivation of absent in melanoma 2 signaling in immune cells

doi: 10.3892/ijmm.2026.5829

Figure Lengend Snippet: H1N1 infection affects cell viability, inflammatory cytokine secretion and interactions between HBEpiCs and THP-1 cells. (A) CCK-8 assay revealed that HBEpiC viability decreased in a concentration-dependent manner following H1N1 infection. (B) ELISA revealed that the levels of IL-1β, IL-6, TNF-α, and IL-8 in HBEpiCs decreased with increasing H1N1 infection. (C) CCK-8 assay indicated that supernatants from H1N1-infected HBEpiC cultures reduced the viability of THP-1 cells in a dose-dependent manner. (D) ELISA results suggested that the levels of inflammatory cytokines (IL-1β, IL-6, TNF-α and IL-8) in THP-1 cells were decreased following exposure to supernatants from H1N1-infected HBEpiC cultures. (E) Cell adhesion assay revealed that the number of THP-1 cells adhering to HBEpiCs increased with increasing H1N1 concentration (scale bar, 10 μ m). Arrow indicates THP-1 cells that remain attached to the surface of HBEpiCs, highlighting the adhesion interaction between the two cell types. (F) Transwell assay suggested that H1N1 infection enhanced the migration capacity of THP-1 cells, with increased migration observed at higher virus concentrations (scale bar, 50 μ m). The data are presented as the mean ± standard deviation; ** P<0.01, *** P<0.001 vs. control. H1N1, influenza A; HBEpiCs, human bronchial epithelial cells; ELISA, enzyme-linked immunosorbent assay; IL, interleukin; TNF-α, tumor necrosis factor-α; Con, control; MOI, multiplicity of infection.

Article Snippet: Cell supernatants were collected and analyzed using Human TNF-α High Sensitivity ELISA Kit [cat. no. EK182HS; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], Human IL-8 ELISA Kit [cat. no. EK108; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], a human IL-1β ELISA kit [EH0185; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] and IL-6 [cat. no. EK1217; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] according to the manufacturer's instructions.

Techniques: Infection, CCK-8 Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Cell Adhesion Assay, Transwell Assay, Migration, Virus, Standard Deviation, Control

Journal: International Journal of Molecular Medicine

Article Title: Triptolide exerts antiviral effects and alleviates influenza A-induced pneumonia by inhibiting the overactivation of absent in melanoma 2 signaling in immune cells

doi: 10.3892/ijmm.2026.5829

Figure Lengend Snippet: TP modulates the inflammatory response and immune cell activity in H1N1-infected HBEpiCs and THP-1 cells. (A) No significant changes were observed in HBEpiCs treated with various concentrations of TP (5, 10 and 20 nM) following H1N1 infection compared with the control. (B) After TP treatment, the levels of the inflammatory cytokines IL-1β, IL-6, TNF-α and IL-8 in HBEpiCs were markedly lower than those in the untreated group. (C) The viability of THP-1 cells pretreated with H1N1-infected HBEpiC culture supernatant decreased after TP treatment. (D) The levels of IL-1β, IL-6, TNF-α, and IL-8 in THP-1 cells were markedly lower after TP treatment. (E) The adhesion of THP-1 cells to HBEpiCs induced by H1N1 infection decreased in a dose-dependent manner with increasing TP concentration (scale bar, 10 μ m). Arrow indicates THP-1 cells that remain attached to the surface of HBEpiCs, highlighting the adhesion interaction between the two cell types. (F) The migration capacity of THP-1 cells was markedly reduced when the supernatant from H1N1-infected HBEpiC cultures was treated with TP (scale bar, 50 μ m). The data are presented as the mean ± standard deviation; * P<0.05, ** P<0.01, *** P<0.001 vs. control. TP, triptolide; H1N1, influenza A; HBEpiCs, human bronchial epithelial cells; IL, interleukin; TNF-α, tumor necrosis factor-α; Con, control.

Article Snippet: Cell supernatants were collected and analyzed using Human TNF-α High Sensitivity ELISA Kit [cat. no. EK182HS; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], Human IL-8 ELISA Kit [cat. no. EK108; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.], a human IL-1β ELISA kit [EH0185; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] and IL-6 [cat. no. EK1217; Hangzhou Multi Sciences (Lianke) Biotech Co., Ltd.] according to the manufacturer's instructions.

Techniques: Activity Assay, Infection, Control, Concentration Assay, Migration, Standard Deviation

Journal: Bioactive Materials

Article Title: Precisely regulated physically-crosslinked carriers enable synergetic release of bioactive factors for MSC-mediated cartilage regeneration

doi: 10.1016/j.bioactmat.2026.01.009

Figure Lengend Snippet: In vivo chondrogenesis of hydrogels incorporated with different combinations of PSF and KSF. (a) Alcian blue and Safranin O staining of different combinations after 21-day implantation. G1: 5 % β-sheet PSF +5 % β-sheet KSF; G2: 15 % β-sheet PSF +30 % β-sheet KSF; G3: 30 % β-sheet PSF +40 % β-sheet KSF; G4: 40 % β-sheet PSF +50 % β-sheet KSF. Scale bar = 200 μm. (b–d) Quantitative analysis of IL-1β, IL-6 and TNF-α surrounding defect cartilage 1 week and 3 weeks after operation. (e, f) Macroscopic and MRI observations of rat femoral condyles at week 6 and 12. Red circles and red arrows show the original defect zone under macroscope and MRI respectively. Scale bar = 1 mm. (g) Relative ratio of average optical density (versus G1) that shows the staining intensity of Alcian blue and Safranin O staining. (h) ICRS scoring of macroscopic evaluations. (i) The repaired cartilage using a nanoindentation instrument. Scale bar = 1 cm. (j, k) Reduced modulus and hardness of the regenerated cartilage. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; ∗∗∗∗p < 0.0001.

Article Snippet: The suspension was centrifuged at 10,000 rpm for 10 min at 4 °C, and the supernatant was collected, and Elisa assay was performed following the manufacturer's instructions for rat IL-1β, IL-6, and TNF-α ELISA kits (Elabscience, China).

Techniques: In Vivo, Staining

Journal: Bioactive Materials

Article Title: 3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis

doi: 10.1016/j.bioactmat.2026.01.043

Figure Lengend Snippet: Composite hydrogel promotes the polarization of BV2 cells to M2 types and alleviates PC12 cell pyroptosis in vitro inflammatory environment. (A) Representative western blots showing protein expression of iNOS and Arg-1 in each group, β-actin was utilized as a loading control. (B) Quantitative analysis of relative expression of iNOS and Arg-1. (C) Representative immunofluorescence images of CD68 positive and iNOS positive BV2 cells (scale bar: 20 μm). (D) Representative immunofluorescence images of CD68 positive and Arg-1 positive BV2 cells (scale bar: 20 μm). (E, F) Quantitative analysis of relative fluorescence intensity of iNOS and Arg-1. (G) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein associated with pyroptosis, β-actin was utilized as a loading control. (H) PI staining of PC12 cells in each group (scale bar, 100 μm). (I) Quantitative analysis of PI staining of PC12 cells (n = 3). (J) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA): The protein levels of IL-18 (Servicebio, Wuhan, China) and IL-1β (Servicebio, Wuhan, China) in the spinal cord tissue were detected using ELISA kits according to the manufacturer's protocol.

Techniques: In Vitro, Western Blot, Expressing, Control, Immunofluorescence, Fluorescence, Staining, Comparison

Journal: Bioactive Materials

Article Title: 3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis

doi: 10.1016/j.bioactmat.2026.01.043

Figure Lengend Snippet: The composite hydrogel inhibits post-SCI pyroptosis in neurons. (A) Immunofluorescence images of residual neurons existing in the anterior horn of the spinal cord (scale bar, 500 μm and 200 μm). (B) Immunofluorescence image of Caspase-1 expression of neurons 3 days after SCI (scale bar, 20 μm). (C) Quantitative analysis of relative fluorescence intensity of Caspase-1 protein expression in neurons within the specified groups (n = 3). (D) Immunofluorescence image of GSDMD-N expression of neurons 3 days after SCI (scale bar, 20 μm). (E) Quantitative analysis of relative fluorescence intensity of GSDMD-N protein expression in neurons within the specified groups (n = 3). (F) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein 3 days after SCI, GAPDH was utilized as a loading control. (G) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA): The protein levels of IL-18 (Servicebio, Wuhan, China) and IL-1β (Servicebio, Wuhan, China) in the spinal cord tissue were detected using ELISA kits according to the manufacturer's protocol.

Techniques: Immunofluorescence, Expressing, Fluorescence, Western Blot, Control, Comparison

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Hippocampal SIRT6 Expression Profile in a Lipopolysaccharide (LPS)-Induced Depression Model. (A) Western blot analysis of SIRT6 expression in the hippocampal brain tissues. D, day. n = 4 mice. (B) Immunostaining for SIRT6 in the hippocampal region three days post-LPS injection revealed an overall reduction in brain tissues, while its expression was concurrently increased specifically in microglia. This was quantified in two graphs showing total SIRT6 expression (left) and microglial SIRT6 expression (right), with arrowheads indicating SIRT6 localization in microglia. n = 4 mice. (C) Western blot analysis of SIRT6 expression in microglia sorted from the hippocampus by MCAS at 3 days post-LPS injection. n = 4 mice. Data are presented as mean ± SEM. Statistical analysis: One-way ANOVA with Tukey's post hoc test in A; Unpaired t -test for two-group comparisons in B-C.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Expressing, Western Blot, Immunostaining, Injection

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: SIRT6 deficiency potentiates microglial activation in a LPS-induced depression model. (A) Immunostaining of GFAP and IBA1 in the hippocampus area in Sirt6 MCKO and Sirt6 fl/fl mice at day 5 post-LPS injection. n = 4 mice. (B-G) Quantification of (B) IBA1 + and (C) GFAP + cell counts, along with microglial morphology parameters: (D) number of branches, (E) average branch length, (F) total branch length, and (G) soma area. n = 4 mice. (H) qPCR analysis of TNF-α,IL-6, and IL-1β mRNA levels in in the microglial sorted from the hippocampus brain tissue. n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Activation Assay, Immunostaining, Injection, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Deletion of microglial Sirt6 inhibited the NRF2-HO1 signaling and worsened the peroxidation damage. (A) Gene Ontology (GO) analysis was performed on RNA-Seq data from microglia sorted from Sirt6 MCKO and Sirt6 fl/fl control mice. (B) TAC, MDA, SOD, and GSH/GSSG levels at 5 days after LPS injection. n = 4 mice. (C) Gene Set Enrichment Analysis (GSEA) of RNA-Seq data profiled from microglia sorted from Sirt6 MCKO and Sirt6 fl/fl control mice. (D-E) Analysis of NRF2-HO1 and associated signaling proteins in sorted microglia. Protein levels of NRF2, KEAP1, HO-1, NQO1, NLRP3, Cleaved Caspase-3, and Cleaved IL-1β were assessed by Western blot (D) and quantified (E). n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: RNA Sequencing, Control, Injection, Western Blot, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Knockout of the Nrf2 in microglial aggravated the depression behavior and facilitated the microglial activation. (A) Experimental timeline depicting the sequential administration of tamoxifen and LPS, followed by behavioral assessment. (B) Western blot analysis confirming the successful knockout of Nrf2 in microglial cells of Nrf2 MCKO mice. n = 3 mice. (C-D) Quantification of the duration of immobility in the (C) TST) and (D) FST. n = 8 mice. (E) Levels of TAC, MDA, SOD, and the GSH/GSSG ratio were measured in Nrf2 MCKO and Nrf2 fl/fl mice 3 days after LPS challenge. (F) Immunostaining of GFAP and IBA1 in the hippocampus area of Nrf2 MCKO and Nrf2 fl/fl mice 3 days after LPS injection. (G-L) Quantification of (G) IBA1 + and (H) GFAP + cell counts, along with microglial morphology parameters: (I) number of branches, (J) average branch length, (K) total branch length, and (L) soma area. n = 4 mice. (M) qPCR analysis of TNF-α, IL-6, and IL-1β mRNA levels in the microglial sorted from the hippocampus brain tissue. n = 4 mice. (N) Western blot analysis was conducted to determine the protein levels of SIRT6 and NRF2 downstream signaling components in primary microglia purified from the mouse hippocampus. n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Knock-Out, Activation Assay, Western Blot, Immunostaining, Injection, Purification, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Sirt6 regulated the microglial activation and the depression behavior through Nrf2-HO1 signaling in vivo (A) Experimental timeline of tamoxifen administration, AAV9-Cx3cr1-Nrf2 injection, LPS challenge, and behavioral tests in Sirt6 MCKO mice. (B) Western blot analysis of NRF2 levels after AAV9-Cx3cr1-Nrf2 injection. n = 3 mice. (C-D) Immobility time in TST (C) and FST (D). n = 8 mice. (E) TAC, MDA, SOD, and GSH/GSSG levels at 5 days after LPS injection. n = 4 mice. (F-L) Immunostaining of GFAP and IBA1 in the hippocampus area at 5 days after LPS injection (F); Quantification of (G) IBA1 + and (H) GFAP + cell counts, along with microglial morphology parameters: (I) number of branches, (J) average branch length, (K) total branch length, and (L) soma area. n = 4 mice. (M) qPCR analysis of TNF-α,IL-6, and IL-1β mRNA levels in the microglial sorted from the hippocampus brain tissue. n = 4 mice. (N) Western blot analysis of NRF2 downstream signaling component protein levels in microglia sorted from the hippocampus. n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Activation Assay, In Vivo, Injection, Western Blot, Immunostaining, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: Overexpression of Sirt6 in microglial improved the depression behavior and impeded the microglial activation (A) Experimental timeline depicting tamoxifen administration, followed by tail vein injection of AAV9-Cx3cr1-Sirt6, subsequent LPS challenge, and finally, a series of behavioral tests. (B) Western blot analysis of SIRT6 expression after AAV9-Cx3cr1-Sirt6 injection. n = 3 mice. (C-D) The immobility time was quantified in TST (C) and FST (D). n = 8 mice. (E) TAC, MDA, SOD, and GSH/GSSG levels at 5 days after LPS injection. (F-L) Immunostaining of GFAP and IBA1 in the hippocampus area at 5 days after LPS injection (F); Quantification of (G) IBA1 + and (H) GFAP + cell counts, along with microglial morphology parameters: (I) number of branches, (J) average branch length, (K) total branch length, and (L) soma area. n = 4 mice.(M) qPCR analysis of TNF-α,IL-6, and IL-1β mRNA levels in the microglial sorted from the hippocampus brain tissue. n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Over Expression, Activation Assay, Injection, Western Blot, Expressing, Immunostaining, Two Tailed Test

Journal: Neurobiology of Stress

Article Title: Microglial SIRT6 confers protection against neuroinflammation-associated depression through NRF2-HO1 signaling

doi: 10.1016/j.ynstr.2026.100804

Figure Lengend Snippet: UBCS039-mediated SIRT6 activation exerts antidepressant and anti-inflammatory effects (A) Schematic of the experimental timeline for UBCS039 administration, LPS injection, and behavioral testing. (B-C) The immobility time was measured in TST (B) and FST (C). n = 4 mice. (D-G) TAC (D), MDA (E), SOD (F), and GSH/GSSG (G) levels at 5 days after LPS injection. n = 4 mice. (H-I) Immunostaining of GFAP and IBA1 in the hippocampus area at 5 days after LPS injection (H); Quantification of IBA1 + and GFAP + cell counts, along with microglial morphology parameters: number of branches, average branch length, total branch length, and soma area (I). n = 4 mice. (J) qPCR analysis of TNF-α,IL-6, and IL-1β mRNA levels in the microglial sorted from the hippocampus brain tissue. n = 4 mice. Data are mean ± SEM. Statistical significance between two groups was determined by an unpaired two-tailed Student's t-test.

Article Snippet: Membranes were blocked with 5% non-fat milk in TBST for 1 h and incubated overnight at 4 °C with primary antibodies: SIRT6 (12486, Cell Signaling Technology), NRF2 (ab62352, Abcam), HO-1 (ab68477, Abcam), NQO1 (ab80588, Abcam), KEAP1 (8047, Cell Signaling Technology), NLRP3 (AG-20B-0014, AdipoGen), cleaved caspase-1 (4199, Cell Signaling Technology), cleaved IL-1β (83186, Cell Signaling Technology), and β-actin (A1978, Sigma-Aldrich).

Techniques: Activation Assay, Injection, Immunostaining, Two Tailed Test

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: Schematic diagram illustrates the mechanism of Mn-NC composite hydrogels for TMJ-OA treatment. Composite hydrogels can effectively protect chondrocytes from ROS damage, alleviate inflammatory reactions within the joint microenvironment, inhibit inflammation-induced chondrocyte apoptosis and ECM degradation, and simultaneously induce the regeneration and repair of subchondral bones via the enzyme-like activity of Mn-NC SAzymes and the release of Mn ions.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Activity Assay

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: Characterization of Mn-NC SAzymes. (a) Preparation of Mn-NC SAzymes. (b) TEM images of NC, Mn-NC-1, Mn-NC-5, and Mn-NC-10 SAzymes. (c) HAADF-STEM image of Mn-NC-10 SAzymes and the corresponding EDX elemental mappings of C, N, O, and Mn. Inset: the SAED pattern. (d) Aberration-corrected HAADF-STEM image of Mn-NC-10 SAzymes. (e) XRD patterns of NC, Mn-NC-1, Mn-NC-5, and Mn-NC-10 SAzymes. (f) Mn 2p and (g) Mn 3s XPS spectra of Mn-NC-10 SAzymes. (h) XANES and (i) FT EXAFS spectra of Mn-NC-10 SAzymes and reference samples at the Mn K-edge. (j) FT EXAFS spectra fitting curves of Mn-NC-10 SAzymes at the Mn K-edge (inset: structure model).

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques:

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: SOD/CAT-like activity and DFT calculations of the Mn-NC SAzymes. The SOD-like activity (a), H 2 O 2 scavenging ratio (b), and dissolved O 2 production curves (c) of NC, Mn-NC-1, Mn-NC-5, and Mn-NC-10 SAzymes. (d) •OH scavenging abilities of Mn-NC-10 evaluated by the electron spin resonance (ESR) spectroscopy. (e) Dissolved oxygen generation catalyzed by Mn-NC-10 at pH 6.6 under varying H 2 O 2 concentrations. (f) Michaelis-Menten kinetic curves of Mn-NC-10 derived from dissolved oxygen production by varying H 2 O 2 concentration. (g) Gibbs free-energy step diagram of the Mn-N 4 . (h) Gibbs free energy step diagram of N 4 C and Mn-N 4 -O in the CAT-like catalytic reaction. (i) Schematic of CAT-like catalytic reaction of N 4 C. (j) Schematic of CAT-like catalytic reaction of Mn-N 4 . (k) Differential charge profiles before and after introducing Mn-O to N 4 C. (l) Differential charge of H 2 O and O 2 before and after adsorption to N 4 C and Mn-N 4 -O (yellow represents gain of electrons, blue represents loss of electrons). TEM images (m), Mn ion releasing amount (n), changes of CAT-like activity (o), and changes of SOD-like activity (p) of Mn-NC-10 SAzymes incubated in PBS (pH = 7.0–7.6) for different time.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Activity Assay, Electron Paramagnetic Resonance, Spectroscopy, Derivative Assay, Concentration Assay, Adsorption, Incubation

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The Mn-NC composite hydrogels protect chondrocytes from H 2 O 2 -induced damage by attenuating ROS. (a) Live/dead staining of rat chondrocytes after incubation with different hydrogels for 24 h, scale bar = 200 μm. (b) Cell viability of rat chondrocytes after incubation with different hydrogels for 1, 2, and 3 days. (c) Cell viability, (d) Intracellular ROS content, and (e) Bright-field and ROS staining of rat chondrocytes after incubation with different hydrogels under H 2 O 2 -contaning condition for 24 h, scale bar = 50 μm. Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Control group, #p < 0.05, ##p < 0.01, and ###p < 0.001 suggests a significant difference in comparison to the Control + H 2 O 2 group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Staining, Incubation, Comparison, Control

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The Mn-NC composite hydrogels protect chondrocytes from IL-1β-induced damage and promote subchondral bone remodeling by stimulating osteogenic differentiation of human mesenchymal stem cells. Cell viability (a) and expressions of inflammatory factors TNF-α (b), IL-6 (c), and inflammation-related proteins (d) in chondrocytes after stimulation with IL-1β. The expressions of inflammatory factors including TNF-α (e), iNOS (f), IL-6 (g), IL-1β (h) and corresponding proteins (i) from rat macrophages (NR8383) co-cultured with Control, Control + H 2 O 2 , NAC + H 2 O 2 , CH + H 2 O 2 , CH@Mn2+H 2 O 2 groups. (j) ALP-positive area staining images from Control, CH, and CH@Mn2 groups at 7 days and corresponding quantitative ALP activity. Scale bar = 200 μm. (k) Expression of osteogenesis-related genes and proteins including ALP, BMP2, OPN from Control, CH, and CH@Mn2 groups at 7 days. Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Control group, # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the Control + IL-1β, Control + H 2 O 2 or CH group. ▽ p < 0.05, ▽▽ p < 0.01, and ▽▽▽ p < 0.001 suggests a significant difference in comparison to the NAC + IL-1β or NAC + H 2 O 2 group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Cell Culture, Control, Staining, Activity Assay, Expressing, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The Mn-NC composite hydrogels inhibit chondrocyte apoptosis and ECM degradation in rats after IL-1β stimulation. Nucleus, apoptosis, and cytoskeleton staining of chondrocytes (a) and flow cytometry analysis (b) after IL-1β stimulation, scale bar = 50 μm. Expressions of apoptosis-related genes and proteins including Bax and Bcl2 (c) and ECM degeneration related genes and proteins including Adamts5 and MMP13 (d) from Control, Control + IL-1β, NAC + IL-1β, CH + IL-1β, and CH@Mn2+IL-1β groups. Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Control group. # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the Control + IL-1β group. ▽ p < 0.05, ▽▽ p < 0.01, and ▽▽▽ p < 0.001 suggests a significant difference in comparison to the NAC + IL-1β group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Staining, Flow Cytometry, Control, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The in vivo therapeutic effect of Mn-NC composite hydrogels on TMJ-OA after 2 weeks. (a) Schematic diagram of in vivo experiments of hydrogel treatment of TMJ-OA. HE staining (b), SO/FG staining (c), immunohistochemical staining of COLII (d) and MMP13 (e), immunofluorescence staining of CD86 (f) and CD206 (g) at the synovial, and immunofluorescence staining of IL-1β (h) in the TMJ of rats after 2 weeks of treatment, scale bar = 100 μm. (i) Histological OARSI score of articular cartilage after 2 weeks of treatment and quantitative data of immunohistochemical positive area and proportion of immunofluorescence positive cells in (c–h). Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Sham group. # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the MIA group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: In Vivo, Staining, Immunohistochemical staining, Immunofluorescence, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The in vivo therapeutic effect of Mn-NC composite hydrogels on TMJ-OA after 8 weeks. HE staining (a), SO/FG staining (b), immunohistochemical staining of COLII (c) and MMP13 (d), immunofluorescence staining of CD86 (e) and CD206 (f) at the synovial, and immunofluorescence staining of IL-1β (g) in the TMJ of rats after 8 weeks of treatment, scale bar = 100 μm. (h) Histological OARSI score of articular cartilage after 8 weeks of treatment and quantitative data of immunohistochemical positive area and proportion of immunofluorescence positive cells in (b–g). Data represent means ± SD, n = 3. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 indicates a significant difference in comparison to the Sham group. # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the MIA group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: In Vivo, Staining, Immunohistochemical staining, Immunofluorescence, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: The in vivo subchondral bone repair and regeneration effect of Mn-NC composite hydrogels on TMJ-OA after 2 and 8 weeks. (a, b) The temporomandibular joints of rats were collected at 2 and 8 weeks, three views of the condyles reconstructed by micro-CT scan (coronal, horizontal, and sagittal view), scale bar = 1 mm. Quantitative statistics of trabecular thickness (Tb. Th) (c), trabecular separation (Tb. Sp) (d), and ratios of bone volume (BV/TV) (e) from Sham, MIA, MIA + CH, MIA + CH@Mn2 at 2 and 8 weeks. (f) Trap staining images and quantification analysis after 2 weeks of hydrogels treatment. (g) Trap staining images and quantification analysis after 8 weeks of hydrogels treatment. (h) OPN staining images and quantification analysis after 8 weeks of hydrogels treatment. Data represent means ± SD, n = 3. ∗p < 0.05 and ∗∗p < 0.01 indicates a significant difference in comparison to the Sham group. # p < 0.05, ## p < 0.01, and ### p < 0.001 suggests a significant difference in comparison to the MIA group.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: In Vivo, Micro-CT, Staining, Comparison

Journal: Bioactive Materials

Article Title: Nanozyme hydrogels remodel pathological microenvironment for temporomandibular joint osteoarthritis therapy via inhibiting MAPK signal pathway

doi: 10.1016/j.bioactmat.2026.01.031

Figure Lengend Snippet: Underlying mechanism of Mn-NC SAzymes composite hydrogels in treating TMJ-OA. (a) The volcano plot showing the DEGs of IL-1β VS Control. (b) The volcano plot showing the DEGs of CH@Mn2 VS IL-1β. (c) Differential gene clustering thermogram of extracellular matrix organization (gray), regulation of inflammatory response (green) and cell adhesion molecule binding (purple) pathway. (d) Gene Ontology (GO) enrichment analysis of CH@Mn2 VS IL-1β. (e) The KEGG enrichment result of the DEGs of IL-1β VS Control. (f) The KEGG enrichment result of the DEGs of CH@Mn2 VS IL-1β. (g) Differential gene clustering thermogram of the MAPK pathway. (h) Activation of MAPK pathway after IL-1β stimulation. n = 3.

Article Snippet: Additionally, the sections were stained with antibodies specific to anti-CD86, CD206, and IL-1β (Boster Biological Technology Co., Ltd., Wuhan, China) for immunofluorescence analysis.

Techniques: Control, Binding Assay, Activation Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: M2-exo@HI promotes in vitro microglia polarization, BBB repair and neuroprotection. (a) Flow cytometry analysis of M1-type (CD86 + ) and M2-type microglia (CD163 + ) following treatment with different formulations. (b,c) Percentages of CD86 + and CD163 + microglia populations (n = 3). (d–g) The cytokine levels of IL-10, TGF-β, TNF-α, and IL-1β in treated microglia (n = 3). (h) Fluorescence microscopy images showing erythrophagocytosis by microglia across treatment groups. (i) Schematic of the in vitro BBB model assessing FITC-dextran permeability using a transwell assay. (j) Quantitative analysis of FITC-dextran penetration (n = 7). (k) Flow cytometry analysis of neuronal apoptosis across treatments (n = 3). (l) Quantitative analysis of neuronal apoptosis (n = 3). Data are presented as mean ± SD. Statistical significance was tested by one-way ANOVA with Tukey's multiple comparisons test.

Article Snippet: ELISA kits included mouse TNF- α ELISA kit (Solarbio), mouse IL-10 ELISA kit (Elabscience), mouse Hp ELISA kit (Elabscience), mouse IL-1β ELISA kit (Solarbio), and TGF-β ELISA kit (Solarbio).

Techniques: In Vitro, Flow Cytometry, Fluorescence, Microscopy, Permeability, Transwell Assay