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Effects of microcurrent stimulation on chondrocyte proliferation in a 2-dimensional (2D) in vitro culture system. (A) Schematic illustration of chondrocytes cultured in 6-well plates under microcurrent stimulation. (B) Relative DNA content of chondrocytes in the 10-, 25-, 50-, 100-, and 200-μA groups compared with that of the control (Ctrl, 0 μA) group at day 9. (C) Relative optical density (OD) values of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 5, as determined by the CCK-8 assay. (D) <t>Representative</t> <t>Live/Dead</t> staining images of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups from days 1 to 9. (E) Quantitative analysis of cell viability in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (F) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining images and corresponding quantification of the EdU-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (G) Representative Ki67 staining images and corresponding quantification of the Ki67-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. Data are presented as mean ± SD ( n = 5). ns, not significant; *** P < 0.001; **** P < 0.0001.
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Effects of microcurrent stimulation on chondrocyte proliferation in a 2-dimensional (2D) in vitro culture system. (A) Schematic illustration of chondrocytes cultured in 6-well plates under microcurrent stimulation. (B) Relative DNA content of chondrocytes in the 10-, 25-, 50-, 100-, and 200-μA groups compared with that of the control (Ctrl, 0 μA) group at day 9. (C) Relative optical density (OD) values of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 5, as determined by the CCK-8 assay. (D) <t>Representative</t> <t>Live/Dead</t> staining images of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups from days 1 to 9. (E) Quantitative analysis of cell viability in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (F) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining images and corresponding quantification of the EdU-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (G) Representative Ki67 staining images and corresponding quantification of the Ki67-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. Data are presented as mean ± SD ( n = 5). ns, not significant; *** P < 0.001; **** P < 0.0001.
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Effects of microcurrent stimulation on chondrocyte proliferation in a 2-dimensional (2D) in vitro culture system. (A) Schematic illustration of chondrocytes cultured in 6-well plates under microcurrent stimulation. (B) Relative DNA content of chondrocytes in the 10-, 25-, 50-, 100-, and 200-μA groups compared with that of the control (Ctrl, 0 μA) group at day 9. (C) Relative optical density (OD) values of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 5, as determined by the CCK-8 assay. (D) <t>Representative</t> <t>Live/Dead</t> staining images of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups from days 1 to 9. (E) Quantitative analysis of cell viability in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (F) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining images and corresponding quantification of the EdU-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (G) Representative Ki67 staining images and corresponding quantification of the Ki67-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. Data are presented as mean ± SD ( n = 5). ns, not significant; *** P < 0.001; **** P < 0.0001.
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Effects of microcurrent stimulation on chondrocyte proliferation in a 2-dimensional (2D) in vitro culture system. (A) Schematic illustration of chondrocytes cultured in 6-well plates under microcurrent stimulation. (B) Relative DNA content of chondrocytes in the 10-, 25-, 50-, 100-, and 200-μA groups compared with that of the control (Ctrl, 0 μA) group at day 9. (C) Relative optical density (OD) values of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 5, as determined by the CCK-8 assay. (D) <t>Representative</t> <t>Live/Dead</t> staining images of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups from days 1 to 9. (E) Quantitative analysis of cell viability in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (F) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining images and corresponding quantification of the EdU-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (G) Representative Ki67 staining images and corresponding quantification of the Ki67-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. Data are presented as mean ± SD ( n = 5). ns, not significant; *** P < 0.001; **** P < 0.0001.
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Effects of microcurrent stimulation on chondrocyte proliferation in a 2-dimensional (2D) in vitro culture system. (A) Schematic illustration of chondrocytes cultured in 6-well plates under microcurrent stimulation. (B) Relative DNA content of chondrocytes in the 10-, 25-, 50-, 100-, and 200-μA groups compared with that of the control (Ctrl, 0 μA) group at day 9. (C) Relative optical density (OD) values of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 5, as determined by the CCK-8 assay. (D) <t>Representative</t> <t>Live/Dead</t> staining images of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups from days 1 to 9. (E) Quantitative analysis of cell viability in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (F) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining images and corresponding quantification of the EdU-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (G) Representative Ki67 staining images and corresponding quantification of the Ki67-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. Data are presented as mean ± SD ( n = 5). ns, not significant; *** P < 0.001; **** P < 0.0001.
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Effects of microcurrent stimulation on chondrocyte proliferation in a 2-dimensional (2D) in vitro culture system. (A) Schematic illustration of chondrocytes cultured in 6-well plates under microcurrent stimulation. (B) Relative DNA content of chondrocytes in the 10-, 25-, 50-, 100-, and 200-μA groups compared with that of the control (Ctrl, 0 μA) group at day 9. (C) Relative optical density (OD) values of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 5, as determined by the CCK-8 assay. (D) <t>Representative</t> <t>Live/Dead</t> staining images of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups from days 1 to 9. (E) Quantitative analysis of cell viability in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (F) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining images and corresponding quantification of the EdU-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (G) Representative Ki67 staining images and corresponding quantification of the Ki67-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. Data are presented as mean ± SD ( n = 5). ns, not significant; *** P < 0.001; **** P < 0.0001.
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Effects of microcurrent stimulation on chondrocyte proliferation in a 2-dimensional (2D) in vitro culture system. (A) Schematic illustration of chondrocytes cultured in 6-well plates under microcurrent stimulation. (B) Relative DNA content of chondrocytes in the 10-, 25-, 50-, 100-, and 200-μA groups compared with that of the control (Ctrl, 0 μA) group at day 9. (C) Relative optical density (OD) values of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 5, as determined by the CCK-8 assay. (D) <t>Representative</t> <t>Live/Dead</t> staining images of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups from days 1 to 9. (E) Quantitative analysis of cell viability in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (F) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining images and corresponding quantification of the EdU-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (G) Representative Ki67 staining images and corresponding quantification of the Ki67-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. Data are presented as mean ± SD ( n = 5). ns, not significant; *** P < 0.001; **** P < 0.0001.
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Effects of microcurrent stimulation on chondrocyte proliferation in a 2-dimensional (2D) in vitro culture system. (A) Schematic illustration of chondrocytes cultured in 6-well plates under microcurrent stimulation. (B) Relative DNA content of chondrocytes in the 10-, 25-, 50-, 100-, and 200-μA groups compared with that of the control (Ctrl, 0 μA) group at day 9. (C) Relative optical density (OD) values of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 5, as determined by the CCK-8 assay. (D) <t>Representative</t> <t>Live/Dead</t> staining images of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups from days 1 to 9. (E) Quantitative analysis of cell viability in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (F) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining images and corresponding quantification of the EdU-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (G) Representative Ki67 staining images and corresponding quantification of the Ki67-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. Data are presented as mean ± SD ( n = 5). ns, not significant; *** P < 0.001; **** P < 0.0001.
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Effects of microcurrent stimulation on chondrocyte proliferation in a 2-dimensional (2D) in vitro culture system. (A) Schematic illustration of chondrocytes cultured in 6-well plates under microcurrent stimulation. (B) Relative DNA content of chondrocytes in the 10-, 25-, 50-, 100-, and 200-μA groups compared with that of the control (Ctrl, 0 μA) group at day 9. (C) Relative optical density (OD) values of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 5, as determined by the CCK-8 assay. (D) Representative Live/Dead staining images of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups from days 1 to 9. (E) Quantitative analysis of cell viability in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (F) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining images and corresponding quantification of the EdU-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (G) Representative Ki67 staining images and corresponding quantification of the Ki67-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. Data are presented as mean ± SD ( n = 5). ns, not significant; *** P < 0.001; **** P < 0.0001.

Journal: Research

Article Title: Effect of Microcurrent Stimulation on Cartilage Regeneration in Engineered Chondrocyte Spheroids

doi: 10.34133/research.1462

Figure Lengend Snippet: Effects of microcurrent stimulation on chondrocyte proliferation in a 2-dimensional (2D) in vitro culture system. (A) Schematic illustration of chondrocytes cultured in 6-well plates under microcurrent stimulation. (B) Relative DNA content of chondrocytes in the 10-, 25-, 50-, 100-, and 200-μA groups compared with that of the control (Ctrl, 0 μA) group at day 9. (C) Relative optical density (OD) values of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 5, as determined by the CCK-8 assay. (D) Representative Live/Dead staining images of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups from days 1 to 9. (E) Quantitative analysis of cell viability in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (F) Representative 5-ethynyl-2′-deoxyuridine (EdU) staining images and corresponding quantification of the EdU-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. (G) Representative Ki67 staining images and corresponding quantification of the Ki67-positive rate of chondrocytes in the Ctrl and 10-, 25-, 50-, 100-, and 200-μA groups at day 9. Data are presented as mean ± SD ( n = 5). ns, not significant; *** P < 0.001; **** P < 0.0001.

Article Snippet: Samples were washed with PBS and incubated in working solution containing 2 μM calcein-AM and 4 μM propidium iodide using a calcein-AM/propidium iodide Live/Dead staining kit (Beyotime Biotechnology, Shanghai, China) for 20 min at 37 °C in the dark.

Techniques: In Vitro, Cell Culture, Control, CCK-8 Assay, Staining, Analysis