antibodies against collagen type ii  (Santa Cruz Biotechnology)

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: 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