Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Targeting EFEMP1 enhances chondrogenesis and inhibits hypertrophic differentiation in a spontaneous osteoarthritis mouse model

doi: 10.1007/s00109-026-02656-y

Figure Lengend Snippet: Effect of EFEMP1 knockdown by small interfering RNA (siRNA) on chondrocyte phenotype. A The human chondrocyte cell line C20A4 was transfected with either EFEMP1 siRNA (siEFEMP1) or scrambled siRNA (siCtrl). EFEMP1 and MMP-13 protein expression was analyzed by Western blotting. B Chondrocyte-related gene expression, including SOX9, aggrecan (ACAN), MMP13, type X collagen (COL10A1), and type II collagen (COL2A1), was assessed by quantitative PCR in siEFEMP1- and siCtrl-treated C20A4 cells. C Phosphokinase protein arrays were used to profile phosphorylation changes in intracellular proteins extracted from C20A4 chondrocytes. D Spot intensities exhibiting changes greater than ± 25% between groups were quantified using ImageJ software. Red lines indicate baseline levels from the siCtrl group. E Phosphorylation of p70S6K at threonine 421/serine 424 (T421/T424) was further examined by Western blot analysis. Data are presented as mean ± standard error of the mean (SEM). Statistical significance was determined using an independent-samples t-test. * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Samples were incubated overnight at 4 °C with primary antibodies against EFEMP1 (1:100, NBP1-77040; NOVUS, USA), SOX9 (1:50, sc-166505; Santa Cruz Biotechnology, USA), MMP-13 (1:100, GTX100665; GeneTex, USA), aggrecan fragments (1:100, AF1220; R&D Systems, USA), COL2A1 (1:100, ab34712; Abcam, UK), and COL10A1 (1:100, GTX37732; GeneTex, USA).

Techniques: Knockdown, Small Interfering RNA, Transfection, Expressing, Western Blot, Gene Expression, Real-time Polymerase Chain Reaction, Phospho-proteomics, Software

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Targeting EFEMP1 enhances chondrogenesis and inhibits hypertrophic differentiation in a spontaneous osteoarthritis mouse model

doi: 10.1007/s00109-026-02656-y

Figure Lengend Snippet: Immunofluorescence analysis of cartilage markers following EFEMP1 antibody treatment in STR/ort mice. A Representative immunofluorescence (IF) staining for EFEMP1, SOX9, MMP13, aggrecan fragments, COL2A1, and COL10A1 in the medial femoral condyle (MF) and medial tibial plateau (MT). Nuclei were counterstained with DAPI (blue), and articular cartilage regions are outlined by orange dashed lines. B Quantification of fluorescence intensity in articular cartilage regions of the femoral condyle and tibial plateau, with medial and lateral compartments combined. EFEMP1 and SOX9 signals were normalized to DAPI, whereas MMP13, aggrecan fragments, COL10A1, and COL2A1 signals were quantified as absolute FITC intensity. Data are presented as mean ± standard error of the mean (SEM). Statistical analysis was performed using an independent-samples t -test. P < 0.05, P < 0.01, and ** P < 0.001

Article Snippet: Samples were incubated overnight at 4 °C with primary antibodies against EFEMP1 (1:100, NBP1-77040; NOVUS, USA), SOX9 (1:50, sc-166505; Santa Cruz Biotechnology, USA), MMP-13 (1:100, GTX100665; GeneTex, USA), aggrecan fragments (1:100, AF1220; R&D Systems, USA), COL2A1 (1:100, ab34712; Abcam, UK), and COL10A1 (1:100, GTX37732; GeneTex, USA).

Techniques: Immunofluorescence, Staining, Fluorescence

Journal: PLoS ONE

Article Title: CCN Family Member 2/Connective Tissue Growth Factor (CCN2/CTGF) Has Anti-Aging Effects That Protect Articular Cartilage from Age-Related Degenerative Changes

doi: 10.1371/journal.pone.0071156

Figure Lengend Snippet: (A) Toluidine blue staining of frontal sections of medial portion of TG (a, c, e) and WT (b, d, f) littermate knee joints from 5- (a and b), 12- (c and d), and 24- (e and f) month-old male mice. TB staining indicated that WT articular cartilage was degraded with age, and finally the whole layer was lost after 24 months. (B) Immunohistochemical staining of aggrecan neoepitope in frontal sections of the medial portion of TG (a, c, e) and WT (b, d, f) littermate knee joints from 5- (a and b), 12- (c and d), and 24-month-old (e and f) mice. a’, b’, c’, d’, e’, and f’: higher magnification of the medial tibial plateau in the load-bearing region, indicated by the dotted box in a, b, c, d, e, and f, respectively. Bars: 200 µm. In the WT tibia (d and d’) even the remaining matrices were slightly degraded, as shown by the aggrecan neoepitope staining. TG cartilage showed a minor decrease in staining for the aggrecan neoepitope (a’, c’, and e’), but not severe as in the WT cartilage (b’, d’, and f’).

Article Snippet: Sections of knee joints were treated with bovine testicular hyaluronidase (25 mg/ml) for 30 minutes at room temperature for epitope retrieval and then immunostained with anti-CCN2 (Abcam, Cambridge, UK), anti-aggrecan neoepitope (Novus Biologicals, Littleton, CO), anti-type I (Millipore, Billerica, MA), type II (CII D3, ), and type X collagen (X53, both kindly provided by Dr. von der Mark, University of Erlangen) and anti-MMP13 antibodies (Millipore).

Techniques: Staining, Immunohistochemical staining

Journal: Advanced Science

Article Title: A Compartmentalized Joint‐on‐chip (JoC) Model to Unravel the Contribution of Cartilage and Synovium to Osteoarthritis Pathogenesis

doi: 10.1002/advs.202500374

Figure Lengend Snippet: Generation of 3D cartilage micro‐tissues and Induction of an OA phenotype inside the JoC platform: a) Experimental plan, with brightfield images showing cartilage compartment. Scaler bar 300 µm. hACs embedded in a fibrin gel were seeded in the cartilage compartment on Day cart 0 and cultured up to Day cart 14. A cyclic HPC was applied for 7 days to induce OA traits. b) Gene expression analysis of cartilage micro‐constructs. Relative gene expression is shown, i.e., expression of gene of interest normalized against the expression of reference gene (GAPDH). Data are plotted as mean + SD (n ≥ 4); * p < 0.05, ** p <0.01, *** p <0.001. c) Confocal images showing the synthesis of collagen‐type II (magenta) and aggrecan (red) inside the cartilage micro‐tissues. Nuclei in blue (DAPI). Scale bar 20 µm. d) Gene expression analysis of cartilage micro‐constructs on Day cart 21, comparing control samples (“Static”) and mechanically stimulated samples (“HPC”). Relative gene expression is shown, i.e., expression of gene of interest normalized against the expression of reference gene (GAPDH). Data are plotted as mean + SD. n ≥ 6, * p < 0.05, ** p <0.01. e) Confocal images showing SOX9 (green), aggrecan (red) and MMP13 (magenta) inside the cartilage micro‐tissues, in static versus HPC samples. Scale bar 20 µm.

Article Snippet: Rabbit anti‐human collagen type‐II (dilution 1:200, abcam, ab34712) and mouse anti‐human aggrecan (dilution 1:100, Santa Cruz Biotech, sc‐33695) were used to evaluate the maturation of cartilage micro‐constructs at Day tot 14.

Techniques: Cell Culture, Gene Expression, Construct, Expressing, Control

Journal: Advanced Science

Article Title: A Compartmentalized Joint‐on‐chip (JoC) Model to Unravel the Contribution of Cartilage and Synovium to Osteoarthritis Pathogenesis

doi: 10.1002/advs.202500374

Figure Lengend Snippet: Effects of synovial inflammation on healthy cartilage micro‐tissues. a) Experimental plan. hACs were seeded in the cartilage compartment, while MΦs and SFBs in the synovium compartment as previously optimized. From Day tot 14 to Day tot 17, a pro‐inflammatory stimulation (100 ng mL −1 IFNγ, 100 ng mL −1 TNFα) was supplied to the synovium compartment. On Day tot 17, pro‐inflammatory stimulation was removed, and communication valves were open. Analyses were conducted on synovium and cartilage micro‐tissues on Day tot 21. b) Gene expression analysis of synovial micro‐constructs on Day tot 21, comparing non‐stimulated synovium micro‐tissues in communication with cartilage tissues (“Untreated”), and inflamed synovium micro‐tissues in communication with cartilage tissues (“Inf”). c) Gene expression analysis of cartilage micro‐constructs on Day tot 21, comparing cartilage micro‐tissues in communication with non‐stimulated synovium micro‐tissues (“Untreated”), and with inflamed synovium micro‐tissues (“Inf”). Relative gene expression is shown, i.e., expression of gene of interest normalized against the expression of reference gene (GAPDH). Data are plotted as mean + SD (n = 6, undetected values are not represented). * p <0.05, ** p <0.01. Dashed lines refer to cartilage monoculture under static conditions, with mix of media supplied from Day tot 17 (i.e., “Single culture” of Figure ). d) Confocal images of cartilage micro‐tissues, showing nuclei (DAPI, blue), aggrecan (red), and MMP13 (magenta), comparing micro‐tissues in communication with non‐stimulated synovium micro‐tissues (“Untreated”) and micro‐tissues in communication with inflamed synovium micro‐tissues (“Inf”). Scale bar 20 µm.

Article Snippet: Rabbit anti‐human collagen type‐II (dilution 1:200, abcam, ab34712) and mouse anti‐human aggrecan (dilution 1:100, Santa Cruz Biotech, sc‐33695) were used to evaluate the maturation of cartilage micro‐constructs at Day tot 14.

Techniques: Gene Expression, Construct, Expressing

Journal: Frontiers in Medicine

Article Title: Serum levels of EphA2 are elevated in knee osteoarthritis and associated with disease severity

doi: 10.3389/fmed.2025.1639458

Figure Lengend Snippet: Correlation between serum EphA2 levels and serum indicators of cartilage metabolism. Pearson correlation test was performed between EphA2 and (A) COMP, (B) Hyaluronan (HA), (C) MMP-13, (D) CTX-2, (E) PIICP, and (F) aggrecan in all osteoarthritis patients.

Article Snippet: Serum EphA2 (ml038095, Shanghai Enzyme-linked Biotechnology, Shanghai, China), COMP (DCMP0, R&D Systems), Hyaluronan (HA) (DHYAL0, R&D Systems), MMP-13 (SEKH-0259, Solarbio, China), CTX-2 (ml105642, Shanghai Enzyme-linked Biotechnology, Shanghai, China), PIICP (AB5126, EK-Bioscience), aggrecan (DY1220, R&D Systems), TNF- α (DTA00D, R&D Systems), IL-1β (DLB50, R&D Systems), IL-6 (D6050B, R&D Systems), IL-17A (D1700, R&D Systems), MDA (EU2577, FineTest), GSH (ml038257, Shanghai Enzyme-linked Biotechnology), GPX4 (ml060706, Shanghai Enzyme-linked Biotechnology), and ACSL4 (EH6088, FineTest) levels were measured using an ELISA kit, and absorbance at 450 nm was assessed using a microplate reader and each experiment run in duplicate ( ).

Techniques: