Journal: Molecular Therapy. Nucleic Acids

Article Title: miR-140 Attenuates the Progression of Early-Stage Osteoarthritis by Retarding Chondrocyte Senescence

doi: 10.1016/j.omtn.2019.10.032

Figure Lengend Snippet: Chondrocyte Senescence Correlates with Human OA Pathogenesis (A) Representative micrographs of H&E and Safranin O staining of normal and OA cartilage (scale bars, 200 μm). (B) The modified Mankin score of normal and OA cartilage. (C) Representative micrographs of immunohistochemical staining of p16 INK4a , p21, p53, and γH2AX in normal and OA cartilage (scale bars, 100 μm for p16 INK4a , p21, and p53; 50 μm for γH2AX). (D–G) The percentage of (D) p16 INK4a -, (E) p21-, (F) p53-, and (G) γH2AX-positive chondrocytes in normal and OA cartilage. (H) Percentage of normal and OA cartilage-derived chondrocytes in the G0/G1, S, and G2/M phases of the cell cycle, determined by flow cytometric analysis. (I) Representative micrographs of SA-βGal staining in normal and OA cartilage-derived chondrocytes (scale bars, 200 μm). (J) Percentage of normal and OA cartilage-derived chondrocytes positive for SA-βGal. E-OA, early-stage OA; ML-OA, middle- to late-stage OA. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: SA-βGal activity was measured using a staining kit (Beyotime, Nantong, Jiangsu, China) and a mammalian SA-βGal assay kit (Thermo Fisher Scientific, Waltham, MA, USA), according to the manufacturer’s protocols.

Techniques: Staining, Modification, Immunohistochemical staining, Derivative Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: miR-140 Attenuates the Progression of Early-Stage Osteoarthritis by Retarding Chondrocyte Senescence

doi: 10.1016/j.omtn.2019.10.032

Figure Lengend Snippet: Pretransfection with miR-140 Inhibits IL-1β-Induced Human Chondrocyte Senescence (A) PCR analysis of the relative expression level of miR-140, 24 h after IL-1β stimulation or transfection with miR-Scr or miR-140 in normal cartilage-derived chondrocytes. (B–F) Percentage of chondrocytes in the G0/G1, S, and G2/M phases of the cell cycle (B), representative micrographs of SA-βGal staining (scale bars, 200 μm) and percentage of SA-βGal-positive chondrocytes (C), gene (D) and protein (E) expression of p16 INK4a , p21, and p53, representative micrographs of γH2AX staining (scale bars, top: 100 μm; bottom: 20 μm), and the percentage of γH2AX-positive chondrocytes (F) after treatment with normal media (Cont), treatment with IL-1β, pretransfection with miR-Scr + IL-1β, or pretransfection with miR-140 + IL-1β. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: SA-βGal activity was measured using a staining kit (Beyotime, Nantong, Jiangsu, China) and a mammalian SA-βGal assay kit (Thermo Fisher Scientific, Waltham, MA, USA), according to the manufacturer’s protocols.

Techniques: Expressing, Transfection, Derivative Assay, Staining

Journal: Scientific Reports

Article Title: Reversal of senescence-associated beta-galactosidase expression during in vitro three-dimensional tissue-engineering of human chondrocytes in a polymer scaffold

doi: 10.1038/s41598-021-93607-9

Figure Lengend Snippet: Illustration of the study groups and time-points of evaluation of SA-βgal, p16INK4a and p21 in two-dimensional (2D) and three-dimensional (3D) thermo-reversible gelation polymer (3D-TGP) cultures.

Article Snippet: For SA-βgal measurement, cells from the 2D and 3D-TGP cultures were stained using a SPiDER-βgal cellular senescence plate assay kit (Dojindo Laboratories, Japan) to measure their SA-βgal.

Techniques: Polymer

Journal: Scientific Reports

Article Title: Reversal of senescence-associated beta-galactosidase expression during in vitro three-dimensional tissue-engineering of human chondrocytes in a polymer scaffold

doi: 10.1038/s41598-021-93607-9

Figure Lengend Snippet: Mean fluorescence intensity (ΔG MFI) of the expression of SA-βgal evaluated by flow cytometry in 2D compared to 3D-TGP at different durations of culture with 2D-cultured chondrocytes (evaluation I and II) showing higher levels of SA-βgal as culture, while the 3D-cultured cells (evaluation III and IV) showed very low levels of SA-βgal throughout the culture period. ( A ) Sample 1; ( B ) Sample 2 (FSC (High) ); ( C ) Sample 2 (FSC (Low) ); ( D ) Sample 3 (FSC (High) ); ( E ) Sample 3 (FSC (Low) ). I, II, III and IV denote the Senescence-associated beta-galactosidase (SA-βgal) evaluation time-points as described in Fig. .

Article Snippet: For SA-βgal measurement, cells from the 2D and 3D-TGP cultures were stained using a SPiDER-βgal cellular senescence plate assay kit (Dojindo Laboratories, Japan) to measure their SA-βgal.

Techniques: Fluorescence, Expressing, Flow Cytometry, Cell Culture

Journal: Scientific Reports

Article Title: Reversal of senescence-associated beta-galactosidase expression during in vitro three-dimensional tissue-engineering of human chondrocytes in a polymer scaffold

doi: 10.1038/s41598-021-93607-9

Figure Lengend Snippet: Gating in the flow cytometric analysis of osteoarthritic chondrocytes grown in 2D and 3D-TGP with the SA-βgal expression in two heterogeneous populations sorted by flow cytometry (FSC high versus FSC low ) with the 2D-grown cells showing higher SA-βgal expression than 3D-TGP cultured cells on both day 26 and day 42 of culture ( A ) Day 26; ( B ) Day 42. II, III and IV denote the Senescence-associated beta-galactosidase (SA-βgal) evaluation time-points as described in Fig. .

Article Snippet: For SA-βgal measurement, cells from the 2D and 3D-TGP cultures were stained using a SPiDER-βgal cellular senescence plate assay kit (Dojindo Laboratories, Japan) to measure their SA-βgal.

Techniques: Expressing, Flow Cytometry, Cell Culture