Journal: Journal of Translational Autoimmunity

Article Title: T cell proliferative response to a homocitrullinated peptide correlates with joint pathology in collagen induced arthritis

doi: 10.1016/j.jtauto.2025.100345

Figure Lengend Snippet: RA-Specific Serum IgG Antibodies in DBA/1J Mice. Sera were collected biweekly from DBA/1J mice immunized with bovine type II collagen (bCII) (arthritic: N = 9–18; non-arthritic: N = 17–37), or PBS (N = 14) and tested via indirect ELISA for A) anti-CitP, and B) anti-HomoCitP IgG antibodies. Graphs show the median [IQR] for each group, with a cut-off indicating the limit of detection (dashed line). Statistical analysis was performed using a mixed effects model with the Geisser-Greenhouse correction and Tukey's multiple comparisons test. The resulting p-values are indicated on the graphs.

Article Snippet: The anti-mCII ELISA kits were used according to manufacturer's protocol (2036T; Chondrex).

Techniques: Indirect ELISA

Journal: Bioactive Materials

Article Title: Microenvironment-educated MSC-EVs loaded injectable smart hydrogel for targeting senescent nucleus pulposus cells and inhibiting ferroptosis against intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.030

Figure Lengend Snippet: D-EVs Alleviate Cellular Senescence and Restore ECM anabolic/catabolic metabolism in Senescent NPCs. (A) The CCK8 assay was used to determine D-EVs concentrations on cell viability. (B) Flow cytometry analysis of proliferative capacity in the above group, and (C) quantitative analysis. (D) Representative ROS images of senescent NPCs treated with N-EVs, D-EVs, or D-EVs + GW4869. (E) Representative SA-β-Gal images of senescent NPCs treated with N-EVs, D-EVs, or D-EVs + GW4869, and (F) quantitative analysis. (G) Confocal analysis of γ-H2A with IF staining depicting DNA damage in the control, TBHP, N-Evs, or D-EVs group. (H) WB analysis of ECM metabolism–related and aging-related proteins in NPCs following treatment with Control, TBHP, N-Evs, or D-EVs. (I) Western blot analysis of p53, p21, and p16 in senescent NPCs treated with D-EVs, D-CM, or D-CM EV-dep . (J) Confocal analysis of COL2 with IF staining in the control, TBHP, N-EVs, or D-EVs group. The data were presented as mean ± SD. n = 3, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: For immunohistochemistry, the tissue sections were incubated overnight at 4 °C with primary antibodies against COL2 (1:1000, 28459-1-AP, Proteintech) and GPX4 (1:500, 381958, Zen-bio).

Techniques: CCK-8 Assay, Flow Cytometry, Staining, Control, Western Blot

Journal: Bioactive Materials

Article Title: Microenvironment-educated MSC-EVs loaded injectable smart hydrogel for targeting senescent nucleus pulposus cells and inhibiting ferroptosis against intervertebral disc degeneration

doi: 10.1016/j.bioactmat.2026.02.030

Figure Lengend Snippet: Histological evaluation of the D-EVs@Gel ROS in a rat IDD model. (A) Representative histological staining of intervertebral disc tissues (H&E, Safranin O, and Masson) at 4 and 8 weeks post-treatment. Scale bars: 1 mm. (B, E) Immunohistochemical staining and quantification for GPX4, a key inhibitor of ferroptosis, at 4 and 8 weeks post-treatment. (C, F) Immunohistochemical staining and quantification for COL2 at 4 and 8 weeks post-treatment. (D) Quantitative analyses of histological score at 4 and 8 weeks post-treatment. Scale bars: 500 μm. The data were presented as mean ± SD. n = 6, ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: For immunohistochemistry, the tissue sections were incubated overnight at 4 °C with primary antibodies against COL2 (1:1000, 28459-1-AP, Proteintech) and GPX4 (1:500, 381958, Zen-bio).

Techniques: Staining, Immunohistochemical staining

Journal: Bioactive Materials

Article Title: A dual-functional hydrogel integrating adhesive and lubricating interfaces for mitochondrial protection–Driven cartilage regeneration

doi: 10.1016/j.bioactmat.2026.02.051

Figure Lengend Snippet: Regulation of extracellular matrix metabolism by AdHy@Pae in LPS-induced chondrocytes. (A) Representative immunofluorescence images showing the expression of COL2A1, SOX9, MMP9, and ADAMTS5 in different treatment groups and corresponding (B) quantitative analysis. Data are shown as mean ± SD (n = 3, ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001).

Article Snippet: Rat chondrocytes (P1) were seeded in 24-well plates (5 × 10 4 cells per well) and cultured to ∼70–80% confluence, stimulated with LPS (10 μg mL −1 , 12 h) to induce an inflammatory phenotype, and then treated with hydrogel extracts (Hy, AdHy, AdHy@Pae) for 24 h. Cells were washed three times with PBS, fixed in 4% paraformaldehyde for 15 min, permeabilized with 0.1% Triton X-100 (Aladdin, T434386) for 10 min, and blocked with 5% BSA (Solarbio, SW3015) at room temperature for 1 h. Primary antibodies were applied overnight at 4 °C: COL2A1 (Proteintech, 28459-1-AP, 1:200), SOX9 (HUABIO, HA723548, 1:1000), MMP9 (HUABIO, ET1704-69, 1:200), and ADAMTS5 (HUABIO, HA722011, 1:100).

Techniques: Immunofluorescence, Expressing

Journal: Bioactive Materials

Article Title: A dual-functional hydrogel integrating adhesive and lubricating interfaces for mitochondrial protection–Driven cartilage regeneration

doi: 10.1016/j.bioactmat.2026.02.051

Figure Lengend Snippet: Expression analysis of cartilage-related proteins after treatment with AdHy@Pae. (A) Representative Western blot images showing ECM synthesis–related proteins (COL2A1, ACAN, COMP, and SOX9) and antioxidant markers (NRF2 and HO-1) in different groups. (B) Quantitative analysis of matrix-degrading enzymes (MMP13, ADAMTS5, ADAMTS1) and apoptosis-related proteins (BCL-2, Bax, and Caspase-3). Data are presented as mean ± SD (n = 3; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗∗∗P < 0.0001).

Article Snippet: Rat chondrocytes (P1) were seeded in 24-well plates (5 × 10 4 cells per well) and cultured to ∼70–80% confluence, stimulated with LPS (10 μg mL −1 , 12 h) to induce an inflammatory phenotype, and then treated with hydrogel extracts (Hy, AdHy, AdHy@Pae) for 24 h. Cells were washed three times with PBS, fixed in 4% paraformaldehyde for 15 min, permeabilized with 0.1% Triton X-100 (Aladdin, T434386) for 10 min, and blocked with 5% BSA (Solarbio, SW3015) at room temperature for 1 h. Primary antibodies were applied overnight at 4 °C: COL2A1 (Proteintech, 28459-1-AP, 1:200), SOX9 (HUABIO, HA723548, 1:1000), MMP9 (HUABIO, ET1704-69, 1:200), and ADAMTS5 (HUABIO, HA722011, 1:100).

Techniques: Expressing, Western Blot

Journal: Bioactive Materials

Article Title: A dual-functional hydrogel integrating adhesive and lubricating interfaces for mitochondrial protection–Driven cartilage regeneration

doi: 10.1016/j.bioactmat.2026.02.051

Figure Lengend Snippet: In vivo validation of the chondroprotective and mitochondrial regulatory effects of AdHy@Pae after therapy for 4 weeks. (A) Representative immunohistochemical staining of cartilage sections for COL2A1, SOX9, MMP3, and ADAMTS1 in different groups (Sham, PBS, Hy, AdHy, and AdHy@Pae). (B) Quantitative analysis of the immunohistochemical staining intensity showing relative expression levels of anabolic (COL2A1, SOX9) and catabolic (MMP3, ADAMTS1) markers. (C) TEM images of chondrocytes showing mitochondrial ultrastructure in each group, with high-magnification views (bottom panels) indicating cristae integrity and membrane morphology. Data are presented as mean ± SD (n = 3); ∗∗P < 0.01, ∗∗∗P < 0.001, ∗∗P < 0.0001.

Article Snippet: Rat chondrocytes (P1) were seeded in 24-well plates (5 × 10 4 cells per well) and cultured to ∼70–80% confluence, stimulated with LPS (10 μg mL −1 , 12 h) to induce an inflammatory phenotype, and then treated with hydrogel extracts (Hy, AdHy, AdHy@Pae) for 24 h. Cells were washed three times with PBS, fixed in 4% paraformaldehyde for 15 min, permeabilized with 0.1% Triton X-100 (Aladdin, T434386) for 10 min, and blocked with 5% BSA (Solarbio, SW3015) at room temperature for 1 h. Primary antibodies were applied overnight at 4 °C: COL2A1 (Proteintech, 28459-1-AP, 1:200), SOX9 (HUABIO, HA723548, 1:1000), MMP9 (HUABIO, ET1704-69, 1:200), and ADAMTS5 (HUABIO, HA722011, 1:100).

Techniques: In Vivo, Biomarker Discovery, Immunohistochemical staining, Staining, Expressing, Membrane

Journal: Materials Today Bio

Article Title: Sequential delivery of sinigrin and dabigatran by an in situ self-stabilizing dynamic hydrogel attenuates intervertebral disc degeneration

doi: 10.1016/j.mtbio.2026.102827

Figure Lengend Snippet: DS@CAD ameliorates TNF-α-induced imbalance between anabolism and catabolism in HNPCs. (A) Heatmap visualization of RT‒qPCR data showing the expression of ECM synthesis markers (Col2a1, COL1 and Acan), degradation markers (Adamts4 and MMP13), and inflammation indicators (COX2 and iNOS) after different treatments. (B) Western blot analysis was used to evaluate ECM synthesis proteins (Col2a1, COL1 and Acan), degradation proteins (Adamts4 and MMP13), and inflammation markers (COX2 and iNOS) following different interventions. (C–H) Quantitative analysis of the Western blot data. (I, J) Alcian blue and Safranin O staining of NPCs after various treatments (scale bar: 200 μm). (K) IF of Col2a1, MMP13, and COX2 in NPCs after different treatments (scale bar: 20 μm). (L, M). Relative statistics of Alcian and Safranin O staining. (N–P) Statistical analysis of fluorescence intensity in the IF analysis. (n = 3; ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001).

Article Snippet: Cells in glass-bottomed dishes were sequentially treated: fixed with 4 % paraformaldehyde for 20 min, permeabilized with 0.5 % Triton X-100 for 15 min, and blocked with 5 % (w/v) bovine serum albumin for 30 min, before incubation with primary antibodies against Col2a1 (1:100, A19308, abclone, China), MMP13 (A1606, abclone, China), COX2 (27308-1-AP, Proteintech, China).

Techniques: Expressing, Western Blot, Staining, Fluorescence

Journal: Materials Today Bio

Article Title: Sequential delivery of sinigrin and dabigatran by an in situ self-stabilizing dynamic hydrogel attenuates intervertebral disc degeneration

doi: 10.1016/j.mtbio.2026.102827

Figure Lengend Snippet: DS@CAD directly promotes the anabolic activity of NPCs. (A): RT‒PCR results showing the expression of ECM synthesis markers (Acan, Col2a1, and SOX9). (B) ELISA analysis of COMP concentration. (C, D) Safranin O and Alcian blue staining of NPCs after various treatments. (E) Western blot results for ECM synthesis proteins. (F, G) IF of Acan and Col2a1 in NPCs after different treatments. (H, I) Statistical analysis of the Alcian and Safranin O staining results. (J) Quantitative analysis of the Western blot data. (K) Statistical analysis of the fluorescence intensity in the IF analysis. (n = 3; ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001).

Article Snippet: Cells in glass-bottomed dishes were sequentially treated: fixed with 4 % paraformaldehyde for 20 min, permeabilized with 0.5 % Triton X-100 for 15 min, and blocked with 5 % (w/v) bovine serum albumin for 30 min, before incubation with primary antibodies against Col2a1 (1:100, A19308, abclone, China), MMP13 (A1606, abclone, China), COX2 (27308-1-AP, Proteintech, China).

Techniques: Activity Assay, Expressing, Enzyme-linked Immunosorbent Assay, Concentration Assay, Staining, Western Blot, Fluorescence

Journal: Materials Today Bio

Article Title: Sequential delivery of sinigrin and dabigatran by an in situ self-stabilizing dynamic hydrogel attenuates intervertebral disc degeneration

doi: 10.1016/j.mtbio.2026.102827

Figure Lengend Snippet: DS@CAD protects against IDD in a surgically induced model in vivo. (A) Procedural flow for in vivo experiments in rats (created using BioRender.com ). (B) Micro-CT and (C) MR images of the punctured disc segments at 4 weeks and 8 weeks posttreatment. (D) Representative images of H&E staining in each group at 4 and 8 weeks. (E) Representative images of SO&FG staining in each group at 4 and 8 weeks. (F) IHC staining image of Col2a1, MMP13 and COX2 at 8 weeks. (G, H) Differences in the DHI between each group at 4 and 8 weeks. (I) Quantitative analysis of the average grey value of IVDs. (J) Heatmap showing Pfirrmann grade differences within each group at 4 and 8 weeks. (K) Histological grades of the different groups at weeks 4 and 8. (n = 5; ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001).

Article Snippet: Cells in glass-bottomed dishes were sequentially treated: fixed with 4 % paraformaldehyde for 20 min, permeabilized with 0.5 % Triton X-100 for 15 min, and blocked with 5 % (w/v) bovine serum albumin for 30 min, before incubation with primary antibodies against Col2a1 (1:100, A19308, abclone, China), MMP13 (A1606, abclone, China), COX2 (27308-1-AP, Proteintech, China).

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

Journal: Materials Today Bio

Article Title: Sequential delivery of sinigrin and dabigatran by an in situ self-stabilizing dynamic hydrogel attenuates intervertebral disc degeneration

doi: 10.1016/j.mtbio.2026.102827

Figure Lengend Snippet: Molecular mechanism of DAB in ECM synthesis. (A) Volcano plot shows the DEG significance and fold change (CTR vs. DAB). (B) KEGG enrichment of the DEG pathways (CTR vs. DAB). (C) GSEA identified AMPK pathway enrichment. (D) Effects of DAB treatment duration (0, 10, 15, 30, 60, and 90 min) on protein levels in NPCs. (E) NPCs were treated with different concentrations of DAB (0, 0.5, 1.0, 5.0, and 10 μM). (F) Quantification of p-AMPK/AMPK protein levels. (G) P-AMPK and AMPK expression after 15 and 30 min of DAB ± Compound C treatment. (H) P-AMPK and AMPK expression following DAB treatment (1 and 10 μM) ± Compound C. (I) Quantification of p-AMPK/AMPK protein levels. (J–L) Col2a1 and Acan protein levels and IF results after DAB (5 and 10 μM) ± Compound C treatment. (M, N) Quantification and immunofluorescence analysis of Acan and Col2a1 protein expression. (n = 3; ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001).

Article Snippet: Cells in glass-bottomed dishes were sequentially treated: fixed with 4 % paraformaldehyde for 20 min, permeabilized with 0.5 % Triton X-100 for 15 min, and blocked with 5 % (w/v) bovine serum albumin for 30 min, before incubation with primary antibodies against Col2a1 (1:100, A19308, abclone, China), MMP13 (A1606, abclone, China), COX2 (27308-1-AP, Proteintech, China).

Techniques: Expressing, Immunofluorescence

Journal: Nature Communications

Article Title: Natural photosynthetic system for restoring homeostasis of animal organelle interaction network

doi: 10.1038/s41467-026-69825-y

Figure Lengend Snippet: a Schematic diagram of RNA-seq. Created in BioRender. Chen, P. (2026) https://BioRender.com/6rte857 . b Principal component analysis (PCA) of genes. c Heatmap showing differentially expressed genes in different groups; three biological replicates are shown. d Volcano plots of differentially expressed genes. P values were calculated using DESeq2 analysis with the Wald test. e Gene set enrichment analysis (GSEA) results of different groups. f Simultaneous imaging (left) and quantification (right) of cytosolic and mitochondrial Ca 2+ using Fluo-4/AM and Rhod-2/AM ( n = 3). g Immunofluorescence staining (left) and quantification (right) of CHOP and ATF4 protein levels ( n = 3). h Representative confocal image of human NPCs stained for calnexin (green, ER) and TOMM20 (red, mitochondria). Merged panel images were processed using Fiji/ImageJ (white dots indicate colocalized pixels). i Mander’s coefficients (M1) of ( h ). n = 3 biological replicates, 20 images were acquired and analyzed from each group. j Transmission electron microscopy (TEM) images of mitochondrial and ER morphology. The ER and mitochondria were graphically reconstructed: mitochondria (red); ER (green). k Quantification of the mitochondrial–ER contact length in ( j ). l Mitochondrial count per field from TEM in ( j ). m Quantification of ER lengths per field from ( j ). Measurements in ( k – m ) are from n = 3 biological replicates, with the normal group = 50 cells from 65 fields and the degeneration group = 50 cells from 63 fields. OMM: Outer mitochondrial membrane. n Calcein fluorescence (left) and quantification (right) of human NPCs ( n = 3). o Reactive oxygen species (ROS) staining (left) and quantification (right) of human NPCs ( n = 3). p JC-1 staining (left) and quantification (right) of human NPCs ( n = 3). All quantitative data are the mean ± s.d. n represents the number of biologically independent samples. P values were calculated using two-tailed Student’s t test ( f , g , i , and k – p ). Scale bars: 50 µm ( f – h , n – p ); 1 µm ( j ). Source data are provided as a Source Data file.

Article Snippet: For immunofluorescence analysis, the following antibodies were used: KRT19 (Proteintech, 10712-1-AP, 1:200), CD24 (Proteintech, 10600-1-AP, 1:200), CHOP (Proteintech, 15204-1-AP, 1:100); ATF4 (Proteintech, 60035-1-Ig, 1:200); Col II (Proteintech, 28459-1-AP, 1:800); aggrecan (Proteintech, 13880-1-AP, 1:200); MMP13 (Proteintech, 18165-1-AP, 1:200); ADAMTS-5 (Affinity Biosciences, DF13268, 1:200); TOMM20 (Abcam, ab56783, 1:200); Calnexin (Proteintech, 10427-2-AP, 1:200).

Techniques: RNA Sequencing, Imaging, Immunofluorescence, Staining, Transmission Assay, Electron Microscopy, Membrane, Fluorescence, Two Tailed Test

Journal: Nature Communications

Article Title: Natural photosynthetic system for restoring homeostasis of animal organelle interaction network

doi: 10.1038/s41467-026-69825-y

Figure Lengend Snippet: a , b Fluorescence staining images ( a ) and quantification ( b ) of aggrecan, Col II, CHOP, and ATF4 ( n = 4). c Representative fluorescence staining images of calnexin (green, ER) and TOMM20 (red, mitochondria). Merged panel images were processed using an ImageJ-based pipeline for quantifying MERCS (white dots indicate colocalized pixels). d Mander’s coefficients (M1) of ( c ). n = 4 biological replicates, 12 images were acquired and analyzed from each group. e TEM images of mitochondrial and ER morphology. The ER and mitochondria were graphically reconstructed: mitochondria (red); ER (green). f Quantification of the mitochondrial–ER contact length in ( e ). g Mitochondrial count per field from TEM in ( e ). h Quantification of ER lengths per field from ( e ). Measurements in ( f – h ) are from n = 4 biological replicates, with the sham group = 43 cells from 60 fields, the vehicle group = 48 cells from 62 fields, the NM-NTU with dark group = 55 cells from 59 fields, and the NM-NTU with light group = 49 cells from 61 fields. All quantitative data are the mean ± s.d. n represents the number of biologically independent samples. P values were calculated using one-way ANOVA ( b , d , f , g , h ). Scale bars: 400 µm ( a ); 50 µm ( c ); 1 µm ( e ). Source data are provided as a Source Data file.

Article Snippet: For immunofluorescence analysis, the following antibodies were used: KRT19 (Proteintech, 10712-1-AP, 1:200), CD24 (Proteintech, 10600-1-AP, 1:200), CHOP (Proteintech, 15204-1-AP, 1:100); ATF4 (Proteintech, 60035-1-Ig, 1:200); Col II (Proteintech, 28459-1-AP, 1:800); aggrecan (Proteintech, 13880-1-AP, 1:200); MMP13 (Proteintech, 18165-1-AP, 1:200); ADAMTS-5 (Affinity Biosciences, DF13268, 1:200); TOMM20 (Abcam, ab56783, 1:200); Calnexin (Proteintech, 10427-2-AP, 1:200).

Techniques: Fluorescence, Staining