Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF inhibits NO, IL-6, and TNF-α production in RAW264.7 and MH-S cells. The cells were treated with JGF (50, 100, 150, 300, 600 μg/mL), 2-E (0.1 μM), DXT (10 μM), or LPS (0.1 μg/mL) for 24 h. ( A ) Cell viability was evaluated using crystal violet. ( B ) NO production was measured using the Griess assay. ( C-D ) IL-6 ( C ) and TNF-α ( D ) levels were determined by ELISA. EC 50 was calculated by CompuSyn software. Data was presented as mean ± standard deviation (SD) for groups (n = 3). Significant differences are denoted as ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Primary antibodies against IL-6 (Bioss, BS0379R, 1:500), TNF-α (Bioworld, BS1857, 1:300), and IL-1β (Bioss, BS6319R, 1:500) were applied overnight at room temperature.

Techniques: Griess Assay, Enzyme-linked Immunosorbent Assay, Software, Standard Deviation

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF downregulates 2-E-induced iNOS and COX-2 in RAW264.7 and MH-S cells. Cells were treated with JGF (0, 50, 200 μg/mL) or 2-E (0.1 μM) for 24 h. ( A ) Protein levels of iNOS and COX-2 in macrophages were measured by Western blot. ( B-C ) Quantification of iNOS and COX-2 in cells without ( B ) and with ( C ) 2-E stimulation, calculated using ImageJ. Actin was used as the internal control. The non-detected data showed as – or ND. Data are presented as mean ± SD (n = 3). Significant differences are denoted as ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against IL-6 (Bioss, BS0379R, 1:500), TNF-α (Bioworld, BS1857, 1:300), and IL-1β (Bioss, BS6319R, 1:500) were applied overnight at room temperature.

Techniques: Western Blot, Control

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF inhibits 2-E-induced phosphorylation of STAT3 in RAW264.7 and MH-S cells. Cells were treated with JGF (0, 50, 200 μg/mL) or 2-E (0.1 μM) for 3 h. ( A ) Protein levels of phosphorylated JAK2 and STAT3 were measured by Western blot. ( B-C ) Quantification of phosphorylated JAK2 and STAT3 in cells without ( B ) and with ( C ) 2-E stimulation, calculated using ImageJ. Actin was used as the internal control. Data are presented as mean ± SD (n = 3). Significant differences are denoted as ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against IL-6 (Bioss, BS0379R, 1:500), TNF-α (Bioworld, BS1857, 1:300), and IL-1β (Bioss, BS6319R, 1:500) were applied overnight at room temperature.

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF inhibits 2-E-induced phosphorylation of ERK1/2 in RAW264.7 and MH-S cells. Cells were treated with JGF (0, 50, 200 μg/mL) or 2-E (0.1 μM) for 3 h. ( A ) Protein levels of phosphorylated JNK1/2, ERK1/2, p38, and p65 were measured by Western blot. ( B-C ) Quantification of phosphorylated JNK1/2, ERK1/2, p38, and p65 in cells without ( B ) and with ( C ) 2-E stimulation, calculated using ImageJ. Actin was used as the internal control. Data are presented as mean ± SD (n = 3). Significant differences are denoted as ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: Primary antibodies against IL-6 (Bioss, BS0379R, 1:500), TNF-α (Bioworld, BS1857, 1:300), and IL-1β (Bioss, BS6319R, 1:500) were applied overnight at room temperature.

Techniques: Phospho-proteomics, Western Blot, Control

Journal: Journal of Traditional and Complementary Medicine

Article Title: Chemical characterization of Jing Guan Fang and its application in alleviating coronavirus envelope protein-induced proinflammatory responses in vitro and in vivo

doi: 10.1016/j.jtcme.2025.12.003

Figure Lengend Snippet: JGF reduces the 2-E-induced proinflammatory cytokines in vivo . ( A ) The experimental scheme for mouse exposure. ( B-F ) Levels of IL-6 ( B ), TNF-α ( C ), IFN-γ ( D ), IL-1β ( E ), and IL-12 ( F ) in lung tissue and serum were measured by ELISA. Data are presented as mean ± SD (n = 9 for serum, except DXT group n = 6; n = 6 for lung tissue, except DXT group n = 3) ( G ) Representative histological images of lung tissue stained with H&E and IHC images for IL-6, TNF-α, and IL-1β expression. ( H-J ) Quantification of IL-6 ( H ), TNF-α ( I ), and IL-1β ( J ) positive areas using ImageJ (n = 3). Significant differences between the control (CTL) group and other groups are denoted by ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Significant differences between the 2-E group and 2-E + JGF group are indicated by #p < 0.05, ##p < 0.01, ###p < 0.001.

Article Snippet: Primary antibodies against IL-6 (Bioss, BS0379R, 1:500), TNF-α (Bioworld, BS1857, 1:300), and IL-1β (Bioss, BS6319R, 1:500) were applied overnight at room temperature.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Staining, Expressing, Control

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