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Commensal microflora coatings support adhesion and viability of human cells. (A) CLSM images showing the morphology and distribution of gingival fibroblasts, oral keratinocytes, and macrophages after 48 h on glass, Ti, and CHM. Scale bar: 100 μm. (B) Metabolic activity of (a) fibroblasts, (b) keratinocytes, and (c) macrophages after 48 h on the indicated surfaces. Cells were stained with green fluorescent Calcein AM to label viable cells and with red fluorescent <t>propidium</t> iodide (PI) to label nonviable (dead) cells. All experiments were conducted using three biological replicates (N = 3) and three technical replicates per group (n = 3). For CLSM, the biocompatibility assay was imaged at 10× magnification with a voxel size of 1.52 μm (x) × 1.52 μm (y) × 3 μm (z).
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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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Image Search Results


Commensal microflora coatings support adhesion and viability of human cells. (A) CLSM images showing the morphology and distribution of gingival fibroblasts, oral keratinocytes, and macrophages after 48 h on glass, Ti, and CHM. Scale bar: 100 μm. (B) Metabolic activity of (a) fibroblasts, (b) keratinocytes, and (c) macrophages after 48 h on the indicated surfaces. Cells were stained with green fluorescent Calcein AM to label viable cells and with red fluorescent propidium iodide (PI) to label nonviable (dead) cells. All experiments were conducted using three biological replicates (N = 3) and three technical replicates per group (n = 3). For CLSM, the biocompatibility assay was imaged at 10× magnification with a voxel size of 1.52 μm (x) × 1.52 μm (y) × 3 μm (z).

Journal: Bioactive Materials

Article Title: Commensal microbiota–coated biohybrid implants induce antibiofilm, osteogenic, and immunomodulatory responses in a human 3D immunocompetent model

doi: 10.1016/j.bioactmat.2026.06.006

Figure Lengend Snippet: Commensal microflora coatings support adhesion and viability of human cells. (A) CLSM images showing the morphology and distribution of gingival fibroblasts, oral keratinocytes, and macrophages after 48 h on glass, Ti, and CHM. Scale bar: 100 μm. (B) Metabolic activity of (a) fibroblasts, (b) keratinocytes, and (c) macrophages after 48 h on the indicated surfaces. Cells were stained with green fluorescent Calcein AM to label viable cells and with red fluorescent propidium iodide (PI) to label nonviable (dead) cells. All experiments were conducted using three biological replicates (N = 3) and three technical replicates per group (n = 3). For CLSM, the biocompatibility assay was imaged at 10× magnification with a voxel size of 1.52 μm (x) × 1.52 μm (y) × 3 μm (z).

Article Snippet: Ti- and CHM-integrated tissues, after coculture under sterile or biofilm-infected conditions, were longitudinally sectioned at the tissue–implant interface and stained with Calcein AM and propidium iodide (Invitrogen, Germany) for 30 min.

Techniques: Activity Assay, Staining

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