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: Scientific Reports

Article Title: 3D Bioprinting of osteochondral tissue substitutes – in vitro -chondrogenesis in multi-layered mineralized constructs

doi: 10.1038/s41598-020-65050-9

Figure Lengend Snippet: Assays used for gene expression analysis via qRT-PCR (Thermo Fisher Scientific, USA).

Article Snippet: The PCR was performed with 1–2 µl cDNA using TaqMan™ Fast Advanced Master Mix for the TaqMan gene expression assay kits COL2, ACN, COMP, COL10, COL1, SOX9 (Table ); an Applied Biosystems 7500 cycler was used with the following protocol: Initial pre-heating at 50 °C (02:00) (mm:ss) and denaturation at 95 °C 00:20, denaturation at 95 °C for 00:03 and annealing at 60 °C for 00:30 (40 cycles).

Techniques: Expressing, Amplification

Journal: Scientific Reports

Article Title: 3D Bioprinting of osteochondral tissue substitutes – in vitro -chondrogenesis in multi-layered mineralized constructs

doi: 10.1038/s41598-020-65050-9

Figure Lengend Snippet: Cytoskeletal morphology, distribution and chondrogenic phenotype of hCh in biphasic interwoven algMC-CPC scaffolds. ( A ) Fluorescence staining of cell nuclei (blue: DAPI) and cytoskeletal F actin filaments (green: phalloidin) at day 28, scale bar = 500 µm. Cells tended to migrate out of the gel and colonize CPC surface (arrows) in ctrl condition (left, day 28) while redifferentiated cells remained inside the algMC matrix phase (right, day 28). ( B ) Fluorescence staining of cell nuclei (blue: DAPI), the chondrogenic markers ACN (top, red) or COL2 (bottom, turquoise) and cytoskeletal F actin filaments (green: phalloidin) in chondrocyte clusters of interwoven algMC-CPC scaffolds in close distance to CPC after 21 days, scale bar = 100 µm.

Article Snippet: The PCR was performed with 1–2 µl cDNA using TaqMan™ Fast Advanced Master Mix for the TaqMan gene expression assay kits COL2, ACN, COMP, COL10, COL1, SOX9 (Table ); an Applied Biosystems 7500 cycler was used with the following protocol: Initial pre-heating at 50 °C (02:00) (mm:ss) and denaturation at 95 °C 00:20, denaturation at 95 °C for 00:03 and annealing at 60 °C for 00:30 (40 cycles).

Techniques: Fluorescence, Staining