Journal: Stem Cell Research & Therapy

Article Title: Tensile stress promotes the chondrogenic ability of condylar cartilage stem/progenitor cells in the temporomandibular joint via the Piezo1-Ca 2+ -Prkca pathway

doi: 10.1186/s13287-025-04439-7

Figure Lengend Snippet: The chondrogenic differentiation and proliferation of cathepsin K (Ctsk) + CSPCs are activated in the MA mouse model. A Representative type II collagen (Col2) immunofluorescence staining images of condyles from tdTomato; Ctsk-Cre mice at 3 and 14 days old. Arrows indicate Ctsk and Col2 double-positive cells in the condylar cartilage. Scale bar: 50 μm. B The schematic diagram displays the workflow of the MA model establishment in tdTomato; Ctsk-Cre mice. C The lower incisors of tdTomato; Ctsk-Cre mice were trimmed by 1 mm every 3 days for 2 weeks to achieve MA. D Representative Col2 immunofluorescence staining images of condyles from tdTomato; Ctsk-Cre mice after 1 and 2 weeks of Sham or MA operation. Scale bar: 100 μm. E Representative EdU immunofluorescence staining images of condyles from tdTomato; Ctsk-Cre mice after 1 and 2 weeks of Sham or MA operation. Scale bar: 100 μm. F Quantitative analysis of the percentage of Ctsk, Col2, and Ctsk, EdU double-positive cells in the condylar cartilage from tdTomato; Ctsk-Cre mice in the Sham and MA groups, n = 5. Data are presented as mean ± SD. *** p < 0.001; ns, not significant

Article Snippet: For immunohistochemistry staining, sections were incubated with type II collagen (Col2) (1:200; Boster, Wuhan, China) and sex-determining region Y-box transcription factor 9 (Sox9) (1:200, Proteintech, Wuhan, China) primary antibodies according to the protocol of the immunohistochemistry staining kit (MXB, China).

Techniques: Immunofluorescence, Staining

Journal: Stem Cell Research & Therapy

Article Title: Tensile stress promotes the chondrogenic ability of condylar cartilage stem/progenitor cells in the temporomandibular joint via the Piezo1-Ca 2+ -Prkca pathway

doi: 10.1186/s13287-025-04439-7

Figure Lengend Snippet: Prkca knockdown diminishes the chondrogenic effect of tensile stress on rat condylar CSPCs. A Western blot detection and the corresponding quantitative analysis of chondrogenic markers and Prkca in rat condylar CSPCs after 10% tensile loading for 48 h with siNC or siPrkca pretreatment in vitro, n = 3 biological replicates. Full-length blots are presented in Figure S3. B The schematic diagram shows the workflow of in vivo Prkca silencing in condylar CSPCs by locally injecting in vivo siPrkca into the joint capsules of rats. C qPCR detection and representative western blots of Prkca in the superficial layer of rat condylar cartilage with local in vivo siPrkca injection for 1 and 2 weeks, n = 6. Full-length blots are presented in Figure S4. D – F Representative safranin O/fast green and immunohistochemistry staining images of rat condyles after 1 and 2 weeks of MA with siNC or siPrkca injection. Dashed lines mark the superficial layer (SL) and the whole condylar cartilage (CC). Scale bar: 100 μm. G Quantitative analysis of the average SL and CC thickness, ratio of Col2-positive areas, and percentage of Sox9-positive cells in rat condylar cartilage after 1 and 2 weeks of MA with siNC or siPrkca injection, n = 5. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant

Article Snippet: For immunohistochemistry staining, sections were incubated with type II collagen (Col2) (1:200; Boster, Wuhan, China) and sex-determining region Y-box transcription factor 9 (Sox9) (1:200, Proteintech, Wuhan, China) primary antibodies according to the protocol of the immunohistochemistry staining kit (MXB, China).

Techniques: Knockdown, Western Blot, In Vitro, In Vivo, Capsules, Injection, Immunohistochemistry, Staining

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Towards Clinical Translation of In Situ Cartilage Engineering Strategies: Optimizing the Critical Facets of a Cell-Laden Hydrogel Therapy

doi: 10.1007/s13770-022-00487-9

Figure Lengend Snippet: Minimum expansion time frame to obtain chondrogenic hADSCs from the isolation phase. A Representative brightfield images of cryosections from pellet culture stained with SafraninO (in pink) and Haematoxylin (in purple) to identify cell’s nuclei. B Representative confocal images of cryosections from pellet immunostained with Phalloidin-RFP (Actin, in red), Collagen type 2 (Col 2, in cyan) and counterstained to detect cells nuclei (DAPI, in white). Superimposed channels are shown in the last row of panels (MERGE). For all the stainings, the cryosections were obtained from cells pelleted after 5 and 7 days of non-passaged proliferation, pushed into 3 weeks of chondrogenic differentiation. The calculated Collagen type 2 intensity from day 0 to day 21 in reported as Fold Increase (F.I.) in the day 21 panels of their corresponding groups

Article Snippet: After washing, samples were dropped in blocking solution (10% goat serum diluted in PBT) for 60 min and then incubated overnight at 4 °C with mouse anti-human Collagen type 2 (Col2) (#II6B3, DSHB) and goat anti-human Collagen type 1 (Col1) (# sc8784, Santa Cruz Biotechnology).

Techniques: Isolation, Staining

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Towards Clinical Translation of In Situ Cartilage Engineering Strategies: Optimizing the Critical Facets of a Cell-Laden Hydrogel Therapy

doi: 10.1007/s13770-022-00487-9

Figure Lengend Snippet: Assessment of minimal hADSCs concentration required for chondrogenesis in cell-laden hydrogel bioscaffolds. A Representative confocal images of cryosections from bioscaffolds from the 3 different hADSCs/ml groups, assessed using immunostaining for DAPI, Actin and Collagen type 2 (Col 2). The cryosections has been obtained by cutting the samples along the z axis to provide spatial information from the top to the bottom of the bioscaffolds. B Superimposed high magnification of the selected white square areas (I and II) in ( A ). C The graphs show the quantification expressed as fold change relative to 1.25 hADCSs/ml group at day 21 post chondrogenesis. The Collagen type 2 (Col 2) intensity signal was calculated and averaged from 16 different ROI from the Collagen II stained cryosections (see Figure S7). D The graphs show the quantification of Glycosaminoglycan (GAG) content measured via the normalisation of GAG over total DNA present in the processed bioscaffolds and expressed as fold change relative to 1.25 hADSCs/ml group at day 21 post chondrogenesis. E Chondrogenic gene expression analysis: the bar graphs represent the fold changes calculated with 2 ΔΔCΤ method of Collagen type 2A1 (COL2A1), Aggrecan (ACAN), Sox9 (SOX9) and Collagen type 1A2 (COL1A2) markers in RT-qPCR assay, relative to 1.25 hADSCs/ml group at day 21 post chondrogenesis. GAPDH was used as the housekeeping gene. Graph bars represent standard error margin between three biological replicates. Statistical analysis was performed using an unpaired t-test

Article Snippet: After washing, samples were dropped in blocking solution (10% goat serum diluted in PBT) for 60 min and then incubated overnight at 4 °C with mouse anti-human Collagen type 2 (Col2) (#II6B3, DSHB) and goat anti-human Collagen type 1 (Col1) (# sc8784, Santa Cruz Biotechnology).

Techniques: Concentration Assay, Immunostaining, Staining, Expressing, Quantitative RT-PCR

Journal: Tissue Engineering and Regenerative Medicine

Article Title: Towards Clinical Translation of In Situ Cartilage Engineering Strategies: Optimizing the Critical Facets of a Cell-Laden Hydrogel Therapy

doi: 10.1007/s13770-022-00487-9

Figure Lengend Snippet: In situ stem cells-laden hydrogel therapy in a rabbit in vivo cartilage repair model. A Representative macroscopic pictures (Macro) and images from Haematoxylin and Eosin (H&E) stained paraffin sections from explanted samples of the indicated groups. Representative confocal images of paraffin sections from explanted samples of the indicated groups assessed using immunostaining for Collagen type 2 (Col 2, in cyan) and Collagen type 1 (Col 1, in red). Overimposed images of the two channels are shown in the Merge raw. B The graph shows the macroscopic score using the International Cartilage Repair Society (ICRS) system for the indicated groups, calculated at the end of the 8 weeks study on the explants. C The graph shows the microscopic score calculated at the end of the 8 weeks study on the HandE and Col1 and 2 stained paraffin sections. D The graph shows the percentage of the Collagen 1 (Col 1) and Collagen 2 (Col 2) positive areas. Graph bars represents the mean with standard deviation of 4 different regions along the entire diameter of the defect for each sample analysed calculated at the end of the 8 weeks study on the immunostained paraffin sections. E The graph shows the biomechanical evaluation using atomic force microscopy calculated at the end of the 8 weeks study on the explants and expressed as Young Modulus (kPa kilopascals). Statistical analysis was performed using unpaired t-test

Article Snippet: After washing, samples were dropped in blocking solution (10% goat serum diluted in PBT) for 60 min and then incubated overnight at 4 °C with mouse anti-human Collagen type 2 (Col2) (#II6B3, DSHB) and goat anti-human Collagen type 1 (Col1) (# sc8784, Santa Cruz Biotechnology).

Techniques: In Situ, In Vivo, Staining, Immunostaining, Standard Deviation, Microscopy

Journal: Materials

Article Title: Increased Adipogenic and Decreased Chondrogenic Differentiation of Adipose Derived Stem Cells on Nanowire Surfaces

doi: 10.3390/ma7042605

Figure Lengend Snippet: ( a ) Representative immunofluorescence images of ADSCs on PCL and NW for sox9 (green), actin (red) and nuclei (blue) after 1, 2, and 3 weeks of culture in chondrogenic conditions; ( b ) Representative immunofluorescence images of ADSCs on PCL and NW for col2 (green), actin (red) and nuclei (blue) after 1, 2, and 3 weeks of culture in chondrogenic conditions. Percentage of FITC-labeled sox9 ( a ) and col2 ( b ) were normalized by total number of cells within a particular image.

Article Snippet: The surfaces were then incubated in FITC-labeled secondary antibodies for Sox9 and Col2 (1:200 donkey antigoat IgG, Santa Cruz Biotechnology) for 45 min in the dark.

Techniques: Immunofluorescence, Labeling