Journal: Arthritis Research & Therapy

Article Title: Meniscal and ligament modifications in spontaneous and post-traumatic mouse models of osteoarthritis

doi: 10.1186/s13075-020-02261-5

Figure Lengend Snippet: Representative images of meniscal pathology during spontaneous osteoarthritis development in Str/ort mice. a Toluidine blue, Collagen type II (Col2) and Sox9 immunolabelling in healthy CBA mouse meniscus, which can be divided into distinct regions: hyaline cartilage (H) surrounds an ossified region (O) and an outer fibrous (F) region. b – d Representative images from diseased menisci from Str/ort mouse knee joints with OA grades of 2 (mild), 4 (moderate) and 5 (severe). Toluidine blue staining showed a range of meniscal pathologies associated with OA development including an increase in the fibrous region (delineated by red lines), proteoglycan deposition and bone formation (red arrows). Col2- and sox9-positive cells (black arrows) were seen in the fibrous region of the meniscus with disease showed. Low mag = low magnification, scale bar = 100 μm; High mag = high magnification, scale bar = 50 μm. For orientation: femur at top of picture, tibia bottom

Article Snippet: Non-specific binding sites were blocked for 1 h (Col2: MoM Kit, Vector Labs, BMK-2202; Sox9: 10% v/v goat serum).

Techniques: Staining

Journal: Arthritis Research & Therapy

Article Title: Meniscal and ligament modifications in spontaneous and post-traumatic mouse models of osteoarthritis

doi: 10.1186/s13075-020-02261-5

Figure Lengend Snippet: Representative images of cruciate ligament changes with OA development in Str/ort mice. a Toluidine blue, Collagen type II (Col2) and Sox9 immunolabelling in healthy CBA mouse cruciate ligaments, with high magnification of insertion site into the tibia. b , c Cruciate ligaments from Str/ort mouse knee joints with OA grades of 2 (mild) and 5 (severe). Toluidine blue staining showed increased staining at the insertion site (high mag for panels b and c ) and within the ligament body (high mag for panel c ), with hypertrophy of local cells. Col2 deposition and sox-9 positive cells were also increased, especially in severe diseased joints. Low mag = low magnification, scale bar = 100 μm; High mag = high magnification, scale bar = 50 μm. For orientation: femur at top of picture, tibia bottom

Article Snippet: Non-specific binding sites were blocked for 1 h (Col2: MoM Kit, Vector Labs, BMK-2202; Sox9: 10% v/v goat serum).

Techniques: Staining

Journal: Arthritis Research & Therapy

Article Title: Meniscal and ligament modifications in spontaneous and post-traumatic mouse models of osteoarthritis

doi: 10.1186/s13075-020-02261-5

Figure Lengend Snippet: Representative images of collateral ligaments in OA Str/ort mouse knee joints. a Toluidine blue, collagen type II (Col2) and Sox9 immunolabelling in healthy CBA mouse collateral ligaments, with high magnification of the body of the ligament. b Collateral ligaments from Str/ort mouse knee joints with OA grades of 5 (severe). Toluidine blue staining showed increased staining with hypertrophy of local cells. Col2 deposition and sox9-positive cells were also increased. c Very severe OA joints showed severe changes including ossification within the body of the ligament (red arrows) and areas of col2 deposition and Sox9-expressing cells (black arrows). Low mag = low magnification, scale bar = 100 μm; High mag = high magnification, scale bar = 50 μm. For orientation: femur at top of picture, tibia bottom

Article Snippet: Non-specific binding sites were blocked for 1 h (Col2: MoM Kit, Vector Labs, BMK-2202; Sox9: 10% v/v goat serum).

Techniques: Staining, Expressing

Journal: Arthritis Research & Therapy

Article Title: Meniscal and ligament modifications in spontaneous and post-traumatic mouse models of osteoarthritis

doi: 10.1186/s13075-020-02261-5

Figure Lengend Snippet: Representative images of meniscus and ligament modifications during post-traumatic OA in DMM joints. a Staining of the contralateral control meniscus. b Toluidine blue staining showed meniscal changes in the DMM mice associated with OA development, including bone formation in the fibrous meniscal attachment site, proteoglycan and bone deposition (red arrow). Col2 deposition was also seen at the sites of high toluidine Blue staining with Sox9-expressing cells (black arrows). c Cruciate ligament insertion site in DMM joints showed areas of strong toluidine blue staining, concomitant with col2 deposition and Sox9 expression. Low mag = low magnification, scale bar = 100 μm; High mag = high magnification, scale bar = 50 μm

Article Snippet: Non-specific binding sites were blocked for 1 h (Col2: MoM Kit, Vector Labs, BMK-2202; Sox9: 10% v/v goat serum).

Techniques: Staining, Expressing

Journal: Science advances

Article Title: Bioenergetic-active exosomes for cartilage regeneration and homeostasis maintenance.

doi: 10.1126/sciadv.adp7872

Figure Lengend Snippet: Fig. 4. Bioenergetic-active exosomes promotes BMSC chondrogenic differentiation and chondrocyte’s homeostasis maintenance through the P2X7-mediated path- way. (A) Hematoxylin and eosin (H&E) (top)–, Toluidine blue (middle)–, Alcian blue (bottom)–stained sections of BMSC microspheres cocultured with N-EXO and Suc-EXO for 28 days. (B) Relative mRNA level of chondrogenic differentiation gene after stimulated by N-EXO and Suc-EXO for 7 days (bottom), 14 days (middle), and 21 days (top) in chondro- genic differentiation medium. (C) Relative mRNA level of Col2, ACAN, MMP9, and MMP13 of BMSCs in common medium. (D) The protein levels of MMP13, MMP9, and Col2 in BMSCs quantified by Western blot. (E) Relative mRNA level of PI3K (top), AKT (middle), and mTOR (down) after incubation with N-EXO, N-EXO, and KN62, Suc-EXO or Suc-EXO and KN62. (F) The protein levels of P2X7, P-mTOR, mTOR, P-S6K, S6K, P-AKT, and AKT in BMSCs quantified by Western blot. (G to I) Relative mRNA level of Col2 (G), ACAN (H), and MMP13 (I) in chondrocytes. (J) The protein levels of MMP13 and Col2 in chondrocytes quantified by Western blot. (K and L) Relative mRNA level of SIRT3 (K) and AMPK (L) in chondrocytes. (M) The protein levels of P2X7, SIRT3, p-AMPK, and AMPK in chondrocytes quantified by Western blot. Results in (B), (C), (E), (G) to (I), and (K) and (L) represent the mean ± SD (n = 3 biological repeats). Statistical analysis: *P < 0.05, **P < 0.01, and ***P < 0.001, analyzed using ANOVA with Tukey’s multiple comparisons test.

Article Snippet: 10, eadp7872 (2024) 18 October 2024 16 of 18 Proteintech, China), anti- MMP13 (1:1000; rabbit polyclonal, Proteintech, China), anti- Col2 (1:1000; rabbit polyclonal, Proteintech, China), anti–phospho- AMPK (1:1000; rabbit polyclonal, Proteintech, China), anti- SIRT3 (1:1000; rabbit polyclonal, ABclonal, China), anti- AMPK (1:1000; rabbit polyclonal, Proteintech, China), anti- P2RX7 (1:1000, rabbit polyclonal, ABclonal, China), anti- AKT(1:1000; rabbit monoclonal, Cell Signaling Technology (CST), USA), anti–phospho- AKT (Ser473) (1:2000; rabbit monoclonal, CST, USA), anti- calnexin (1:1000; rabbit polyclonal, Proteintech, China), anti- S6K (1:2000; rabbit monoclonal, Proteintech, China), anti–phospho- S6K (Thr389) (1:1000; rabbit monoclonal, CST, USA), anti–β- ACTIN (1:20,000; mouse monoclonal, Proteintech, China), and anti- GAPDH (1:50,000, mouse monoclonal, Proteintech, China).

Techniques: Staining, Western Blot, Incubation

Journal: Science advances

Article Title: Bioenergetic-active exosomes for cartilage regeneration and homeostasis maintenance.

doi: 10.1126/sciadv.adp7872

Figure Lengend Snippet: Fig. 7. Immunohistochemical and immunofluorescence assessment of cartilage regeneration. (A) Immunohistochemical staining for Col2, ACAN, MMP9, and MMP13. Relative staining intensity of ACAN (B), Col2 (C), MMP9 (D), and MMP13 (E) in cartilage samples at 6 and 12 weeks. (F) Immunofluorescence staining for SIRT3 and Col2 at 12 weeks. Results in (B) to (E) represent the mean ± SD (n = 3 technical repeats). Statistical analysis: **P < 0.01 and ***P < 0.001, analyzed using ANOVA with Tukey’s multiple comparisons test.

Article Snippet: 10, eadp7872 (2024) 18 October 2024 16 of 18 Proteintech, China), anti- MMP13 (1:1000; rabbit polyclonal, Proteintech, China), anti- Col2 (1:1000; rabbit polyclonal, Proteintech, China), anti–phospho- AMPK (1:1000; rabbit polyclonal, Proteintech, China), anti- SIRT3 (1:1000; rabbit polyclonal, ABclonal, China), anti- AMPK (1:1000; rabbit polyclonal, Proteintech, China), anti- P2RX7 (1:1000, rabbit polyclonal, ABclonal, China), anti- AKT(1:1000; rabbit monoclonal, Cell Signaling Technology (CST), USA), anti–phospho- AKT (Ser473) (1:2000; rabbit monoclonal, CST, USA), anti- calnexin (1:1000; rabbit polyclonal, Proteintech, China), anti- S6K (1:2000; rabbit monoclonal, Proteintech, China), anti–phospho- S6K (Thr389) (1:1000; rabbit monoclonal, CST, USA), anti–β- ACTIN (1:20,000; mouse monoclonal, Proteintech, China), and anti- GAPDH (1:50,000, mouse monoclonal, Proteintech, China).

Techniques: Immunohistochemical staining, Immunofluorescence, Staining

Journal: Molecular Medicine Reports

Article Title: Platelet-rich plasma protects rat chondrocytes from interleukin-1β-induced apoptosis

doi: 10.3892/mmr.2016.5767

Figure Lengend Snippet: List of antibodies used in western blotting.

Article Snippet: Permeabilization and blocking were performed by incubating the cells with 1% Triton-X 100 and 1% bovine serum albumin for 15 min. Cover slips were then incubated overnight at 4°C with rabbit polyclonal anti-COL2 primary antibodies (cat. no. Bs-11929R; dilution, 1:500; Bioss, Inc., Woburn, MA, USA).

Techniques: Western Blot

Journal: Molecular Medicine Reports

Article Title: Platelet-rich plasma protects rat chondrocytes from interleukin-1β-induced apoptosis

doi: 10.3892/mmr.2016.5767

Figure Lengend Snippet: Platelet-rich plasma (PRP) inhibits the expression of catabolic genes induced by interleukin (IL)-1β. (A) Quantitative polymerase chain reaction was performed 24 h following treatment of chondrocytes (N=3). (B) Western blotting detected downregulation of matrix metalloproteinases (MMPs), and upregulation of tissue inhibitor of metalloproteinases (TIMP), SRY-box 9 (SOX9) and collagen type II (COL2) in the IL-1β + PRP group compared with the IL-1β group. (C) Semi-quantification of blots confirmed the expression of anabolic proteins was increased by PRP (N=3). *P<0.05, vs. the IL-1β group. GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Permeabilization and blocking were performed by incubating the cells with 1% Triton-X 100 and 1% bovine serum albumin for 15 min. Cover slips were then incubated overnight at 4°C with rabbit polyclonal anti-COL2 primary antibodies (cat. no. Bs-11929R; dilution, 1:500; Bioss, Inc., Woburn, MA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot