Journal: Arthritis Research & Therapy

Article Title: Roflumilast mitigates cartilage degeneration in osteoarthritis by activating CFTR-mediated anti-apoptotic signaling

doi: 10.1186/s13075-026-03872-0

Figure Lengend Snippet: Roflumilast attenuates ECM degradation and apoptosis in IL-1β-stimulated chondrocytes. a Cell viability of primary murine chondrocytes treated with increasing concentrations of roflumilast (0, 1, 5, 10, 20, 30, and 50 μM) for 48 h. Data are expressed as means ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for comparisons among multiple groups. b Intracellular cAMP levels in primary murine chondrocytes treated with roflumilast. Data are expressed as means ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for comparisons among multiple groups. c mRNA expression of Adamts5, Mmp13, Col2, and Acan in IL-1β-stimulated chondrocytes with or without roflumilast treatment, determined by qPCR. Data are expressed as means ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for comparisons among multiple groups. d - e Protein expression of Adamts5, Mmp13, Col2, and Acan in IL-1β-stimulated chondrocytes with or without roflumilast treatment, determined by Western blot. Data are expressed as means ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for comparisons among multiple groups. f - g Immunofluorescence staining for Col2, Acan, Mmp13, and Adamts5 in IL-1β-stimulated chondrocytes with or without roflumilast treatment. Data are expressed as means ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for comparisons among multiple groups. h - i TUNEL staining and flow cytometry for apoptosis in IL-1β-stimulated chondrocytes with or without roflumilast treatment. Data are expressed as means ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for comparisons among multiple groups. *P < 0.05 was considered statistically significant

Article Snippet: Membranes were blocked with 5% non-fat milk or BSA in TBST for 1 h at room temperature and then incubated overnight at 4 °C with primary antibodies: Col2 (#43306S, Cell Signaling Technology, 1:1000), Aggrecan (#A12045, Abclonal, 1:1000), MMP13 (#GB11247-1-100, Servicebio, 1: 1000), Adamts5 (#A2836, Abclonal, 1: 1000), Bax (#60267-1-Ig, Proteintech, 1: 1000), Bcl2 (#15071S, Cell Signaling Technology, 1: 1000), Akt (#9272S, Cell Signaling Technology, 1: 1000), p-AKT (#9271S, Cell Signaling Technology, 1: 1000), Caspase 3 (#9662S, Cell Signaling Technology, 1: 1000), Cftr (#20738-1-AP, Proteintech, 1: 1000), and Gapdh (#60004-1-Ig, Proteintech, 1: 1000).

Techniques: Analysis, Expressing, Western Blot, Immunofluorescence, Staining, TUNEL Assay, Flow Cytometry

Journal: Arthritis Research & Therapy

Article Title: Roflumilast mitigates cartilage degeneration in osteoarthritis by activating CFTR-mediated anti-apoptotic signaling

doi: 10.1186/s13075-026-03872-0

Figure Lengend Snippet: Roflumilast ameliorates cartilage erosion and alleviates OA progression. a - b Micro-CT analysis showing osteophyte formation, subchondral bone sclerosis, osteophyte numbers, subchondral bone volume, bone mineral density, trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp) in sham, DMM, and DMM with roflumilast treatment groups. Data are expressed as means ± SD (n=6 per group). Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for comparisons among multiple groups. c - e Toluidine blue staining, OARSI scores, chondrocyte number, cartilage thickness, and synovitis scores in sham, DMM, and DMM with roflumilast treatment groups. Data are expressed as means ± SD (n=6 per group). Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for comparisons among multiple groups. f Immunofluorescence staining for Col2/Acan and Mmp13/Adamts5 expression in DMM and DMM with roflumilast treatment groups. Data are expressed as means ± SD (n=6 per group). Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for comparisons among multiple groups. g TUNEL staining for chondrocyte apoptosis in DMM and DMM with roflumilast treatment groups. Data are expressed as means ± SD (n=6 per group). Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test for comparisons among multiple groups. *P < 0.05 was considered statistically significant

Article Snippet: Membranes were blocked with 5% non-fat milk or BSA in TBST for 1 h at room temperature and then incubated overnight at 4 °C with primary antibodies: Col2 (#43306S, Cell Signaling Technology, 1:1000), Aggrecan (#A12045, Abclonal, 1:1000), MMP13 (#GB11247-1-100, Servicebio, 1: 1000), Adamts5 (#A2836, Abclonal, 1: 1000), Bax (#60267-1-Ig, Proteintech, 1: 1000), Bcl2 (#15071S, Cell Signaling Technology, 1: 1000), Akt (#9272S, Cell Signaling Technology, 1: 1000), p-AKT (#9271S, Cell Signaling Technology, 1: 1000), Caspase 3 (#9662S, Cell Signaling Technology, 1: 1000), Cftr (#20738-1-AP, Proteintech, 1: 1000), and Gapdh (#60004-1-Ig, Proteintech, 1: 1000).

Techniques: Micro-CT, Analysis, Staining, Immunofluorescence, Expressing, TUNEL Assay

Journal: Arthritis Research & Therapy

Article Title: Roflumilast mitigates cartilage degeneration in osteoarthritis by activating CFTR-mediated anti-apoptotic signaling

doi: 10.1186/s13075-026-03872-0

Figure Lengend Snippet: Schematic illustration of the proposed mechanism by which roflumilast attenuates osteoarthritis progression. Roflumilast exerts chondroprotective effects by regulating CFTR-dependent signaling in chondrocytes. Activation of CFTR further engages the PI3K/Akt/Bcl-2 signaling axis, thereby reducing chondrocyte apoptosis and ECM degradation under inflammatory stimulation. In vivo, these effects may contribute to reduced cartilage degeneration, subchondral bone remodeling, and synovial inflammation in DMM-induced osteoarthritis

Article Snippet: Membranes were blocked with 5% non-fat milk or BSA in TBST for 1 h at room temperature and then incubated overnight at 4 °C with primary antibodies: Col2 (#43306S, Cell Signaling Technology, 1:1000), Aggrecan (#A12045, Abclonal, 1:1000), MMP13 (#GB11247-1-100, Servicebio, 1: 1000), Adamts5 (#A2836, Abclonal, 1: 1000), Bax (#60267-1-Ig, Proteintech, 1: 1000), Bcl2 (#15071S, Cell Signaling Technology, 1: 1000), Akt (#9272S, Cell Signaling Technology, 1: 1000), p-AKT (#9271S, Cell Signaling Technology, 1: 1000), Caspase 3 (#9662S, Cell Signaling Technology, 1: 1000), Cftr (#20738-1-AP, Proteintech, 1: 1000), and Gapdh (#60004-1-Ig, Proteintech, 1: 1000).

Techniques: Activation Assay, In Vivo

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