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Inhibitory effect on osteoclast differentiation and bone <t>resorption.</t> (A) RAW264.7 cells were cultured in DMEM high glucose for 5 days in the presence of RANKL (50 ng/mL) with CBD, taurine, and both. (B) RAW264.7 cells were seeded in CaP-coated 48-well plates and treated with 50 ng/mL RANKL in the presence of CBD, taurine, and both. (C) TRAP-positive multinucleated cells (TRAP+ MNCs) were counted after fixation and staining for TRAP. (D) Pit area was measured. The results are presented as means ± SD (n=3). *** p <0.001 indicates significant differences from CBD+taurine treated groups with CBD and taurine alone.
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Inhibitory effect on osteoclast differentiation and bone <t>resorption.</t> (A) RAW264.7 cells were cultured in DMEM high glucose for 5 days in the presence of RANKL (50 ng/mL) with CBD, taurine, and both. (B) RAW264.7 cells were seeded in CaP-coated 48-well plates and treated with 50 ng/mL RANKL in the presence of CBD, taurine, and both. (C) TRAP-positive multinucleated cells (TRAP+ MNCs) were counted after fixation and staining for TRAP. (D) Pit area was measured. The results are presented as means ± SD (n=3). *** p <0.001 indicates significant differences from CBD+taurine treated groups with CBD and taurine alone.
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EN’s role in osteoclast differentiation and function. Raw264.7 cells were pre-treated with EN (10 and 20 μg/mL) for 1 h, followed by RANKL (40 ng/mL). ( A ) The area of <t>resorption</t> pits formed by mature osteoclast cells was evaluated using a bone resorption assay and visualized under a light microscope at 40× magnification (Scale bar. 100 μm). ( B ) The pit area measurements, quantified in pixels, are depicted in a bar graph. ( C ) The formation of actin rings in mature osteoclast cells was observed using a LSM 700 laser scanning confocal microscope, highlighting F-actin staining (upper line) and DAPI staining (bottom line) at 100× magnification (Scale bar = 200 μm). Data represent mean ± SEM ( n ≥ 3). One-way analysis of variance (ANOVA) was utilized, followed by the Bonferroni post hoc test for pairwise comparisons. *** p < 0.001; n.s. = not significant. DMSO: dimethyl sulfoxide.
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EN’s role in osteoclast differentiation and function. Raw264.7 cells were pre-treated with EN (10 and 20 μg/mL) for 1 h, followed by RANKL (40 ng/mL). ( A ) The area of <t>resorption</t> pits formed by mature osteoclast cells was evaluated using a bone resorption assay and visualized under a light microscope at 40× magnification (Scale bar. 100 μm). ( B ) The pit area measurements, quantified in pixels, are depicted in a bar graph. ( C ) The formation of actin rings in mature osteoclast cells was observed using a LSM 700 laser scanning confocal microscope, highlighting F-actin staining (upper line) and DAPI staining (bottom line) at 100× magnification (Scale bar = 200 μm). Data represent mean ± SEM ( n ≥ 3). One-way analysis of variance (ANOVA) was utilized, followed by the Bonferroni post hoc test for pairwise comparisons. *** p < 0.001; n.s. = not significant. DMSO: dimethyl sulfoxide.
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Inhibitory effect on osteoclast differentiation and bone resorption. (A) RAW264.7 cells were cultured in DMEM high glucose for 5 days in the presence of RANKL (50 ng/mL) with CBD, taurine, and both. (B) RAW264.7 cells were seeded in CaP-coated 48-well plates and treated with 50 ng/mL RANKL in the presence of CBD, taurine, and both. (C) TRAP-positive multinucleated cells (TRAP+ MNCs) were counted after fixation and staining for TRAP. (D) Pit area was measured. The results are presented as means ± SD (n=3). *** p <0.001 indicates significant differences from CBD+taurine treated groups with CBD and taurine alone.

Journal: Biomolecules & Therapeutics

Article Title: Combination of Cannabidiol with Taurine Synergistically Treated Periodontitis in Rats

doi: 10.4062/biomolther.2024.104

Figure Lengend Snippet: Inhibitory effect on osteoclast differentiation and bone resorption. (A) RAW264.7 cells were cultured in DMEM high glucose for 5 days in the presence of RANKL (50 ng/mL) with CBD, taurine, and both. (B) RAW264.7 cells were seeded in CaP-coated 48-well plates and treated with 50 ng/mL RANKL in the presence of CBD, taurine, and both. (C) TRAP-positive multinucleated cells (TRAP+ MNCs) were counted after fixation and staining for TRAP. (D) Pit area was measured. The results are presented as means ± SD (n=3). *** p <0.001 indicates significant differences from CBD+taurine treated groups with CBD and taurine alone.

Article Snippet: The bone resorption pit assay was carried out using a Bone Resorption Test Kit (CSF-BRA-48 KIT, Cosmo Bio) in plates coated with calcium phosphate.

Techniques: Cell Culture, Staining

EN’s role in osteoclast differentiation and function. Raw264.7 cells were pre-treated with EN (10 and 20 μg/mL) for 1 h, followed by RANKL (40 ng/mL). ( A ) The area of resorption pits formed by mature osteoclast cells was evaluated using a bone resorption assay and visualized under a light microscope at 40× magnification (Scale bar. 100 μm). ( B ) The pit area measurements, quantified in pixels, are depicted in a bar graph. ( C ) The formation of actin rings in mature osteoclast cells was observed using a LSM 700 laser scanning confocal microscope, highlighting F-actin staining (upper line) and DAPI staining (bottom line) at 100× magnification (Scale bar = 200 μm). Data represent mean ± SEM ( n ≥ 3). One-way analysis of variance (ANOVA) was utilized, followed by the Bonferroni post hoc test for pairwise comparisons. *** p < 0.001; n.s. = not significant. DMSO: dimethyl sulfoxide.

Journal: Molecules

Article Title: A 70% Ethanol Neorhodomela munita Extract Attenuates RANKL-Induced Osteoclast Activation and H 2 O 2 -Induced Osteoblast Apoptosis In Vitro

doi: 10.3390/molecules29081741

Figure Lengend Snippet: EN’s role in osteoclast differentiation and function. Raw264.7 cells were pre-treated with EN (10 and 20 μg/mL) for 1 h, followed by RANKL (40 ng/mL). ( A ) The area of resorption pits formed by mature osteoclast cells was evaluated using a bone resorption assay and visualized under a light microscope at 40× magnification (Scale bar. 100 μm). ( B ) The pit area measurements, quantified in pixels, are depicted in a bar graph. ( C ) The formation of actin rings in mature osteoclast cells was observed using a LSM 700 laser scanning confocal microscope, highlighting F-actin staining (upper line) and DAPI staining (bottom line) at 100× magnification (Scale bar = 200 μm). Data represent mean ± SEM ( n ≥ 3). One-way analysis of variance (ANOVA) was utilized, followed by the Bonferroni post hoc test for pairwise comparisons. *** p < 0.001; n.s. = not significant. DMSO: dimethyl sulfoxide.

Article Snippet: The bone resorption assay was conducted using a bone resorption assay kit (CSR-BRA-48KIT; COSMOBIO, Tokyo, Japan), following the manufacturer’s instructions with slight modifications.

Techniques: Light Microscopy, Microscopy, Staining