osteoclast differentiation  (TaKaRa)

Journal: Journal of functional foods

Article Title: A synbiotic medical food improves gut barrier function, reduces immune responses, and inhibits osteoclast activity in models of postmenopausal bone loss aligned with clinical outcomes

doi: 10.1016/j.jff.2025.107114

Figure Lengend Snippet: (A) A pictographic illustration of the workflow for this experiment, created in BioRender. Green, R. (2025) https://BioRender.com/seaeapz . Osteoclasts derived from Human PBMCs were grown on bone chips in osteoclastogenic media for 14 days with media collected at day 12 and cells were fixed and stained for TRAP at day 14. Media treatments included GALT model media (Vehicle), media control-treated GALT-conditioned basolateral media (GALT), or apical SBD111-treated GALT-conditioned basolateral media (MOI 10). Following fixation, cells were stained for TRAP and quantified using NOISe. ( B ) Representative images of TRAP+ osteoclasts grown on bone chips illustrating effects of treated media. ( C ) Example of NOISe quantification and segmentation abilities. ( D ) NOISe-based quantification of osteoclast number and ( E ) TRAP+ area. ( F ) CTX-1 levels (a soluble measure of resorbed bone) were measured by ELISA in media collected from the osteoclast culture 12 days post isolation. Bars indicate mean and SEM based on n = 3 for count and area measures ( D, E ). Boxplots represent median and interquartile range, with whiskers representing max and min values, of n = 4 independent experiments for bone resorption ( F ). Vehicle condition is shown to display the normal range of human PBMC-derived osteoclast resorption and differentiation measures. Significance between conditions was determined by one-way ANOVA with Tukey’s HSD and significant p values ( p < 0.05) are included presented.

Article Snippet: Cells were analyzed for tartrate-resistant acid phosphatase (TRAP) activity to quantify osteoclast differentiation as per manufacturer’s instructions (CAT# MK301, Takara; San Jose, CA).

Techniques: Derivative Assay, Staining, Control, Enzyme-linked Immunosorbent Assay, Isolation

Journal: Journal of functional foods

Article Title: A synbiotic medical food improves gut barrier function, reduces immune responses, and inhibits osteoclast activity in models of postmenopausal bone loss aligned with clinical outcomes

doi: 10.1016/j.jff.2025.107114

Figure Lengend Snippet: Impaired intestinal barrier integrity, as occurs during aging, obesity, and the menopause transition permits translocation of inflammatory microbial components into intestinal tissues. This activates the epithelium and local immune cells which further compromise barrier integrity, allowing translocation of additional inflammatory components. This cycle promotes systemic inflammation and promotes osteoclastogenesis and bone resorption through the migration of osteoclast-activating inflammatory immune cells and diffusion of osteoclast-modulating compounds to bone. SBD111 disrupts this cycle by enhancing intestinal barrier integrity and, in the case of disrupted barriers, by reducing inflammatory mediator secretion (T cell polarizing and neutrophil chemoattractant) by inflamed immune cells. Furthermore, SBD111 has shown the potential to produce an anti-osteoclastogenic environment through interactions with intestinal epithelial cells and underlying immune cells. Together these mechanisms may contribute to the bone-protective effects of SBD111 observed in groups of postmenopausal women. Image created in https://BioRender.com . Green, R. (2025) https://BioRender.com/sty1qzs .

Article Snippet: Cells were analyzed for tartrate-resistant acid phosphatase (TRAP) activity to quantify osteoclast differentiation as per manufacturer’s instructions (CAT# MK301, Takara; San Jose, CA).

Techniques: Translocation Assay, Migration, Diffusion-based Assay

Journal: eLife

Article Title: Psychological stress disturbs bone metabolism via miR-335-3p/Fos signaling in osteoclast

doi: 10.7554/eLife.95944

Figure Lengend Snippet: ( A ) Representative cytochemical tartrate-resistant acid phosphatase (TRAP) staining images, Scale bar, 200 um. ( B ) The amounts of osteoclasts (multinucleated TRAP-positive cells), n = 5. ( C ) The positive TRAP area of osteoclasts, n = 5. ( D ) Representative resorption pits, Scale bar, 200 um. ( E ) The area of resorbed surface, n = 6. ( F ) qRT-PCR quantification analysis of Acp5 , Nfatc1 , Mmp9 expression of RANKL-induced RAW264.7 transfected with mimic-NC, mimic-miR-335-3p, inhibitor-NC, or inhibitor-miR-335-3p, n = 3. ( G ) Representative IF images for FOS, NFATC1, and HOCHEST expression, Scale bar, 200 um. ( H ) Three-dimensional reconstruction images of femurs from agomir NC, agomir-mir-335-3p, antagomir NC, and antagomir-miR-335-3p mice. ( I ) Quantitative micro-CT analysis of femur trabecular bone, n = 3; scale bar, 1 mm. All data are presented as mean ± SD; *p<0.05, **p<0.01, ***p<0.001, by unpaired Student’s t -test. Figure 5—source data 1. Full dataset for .

Article Snippet: They were used for osteoclastic differentiation induction via RANKL (R&D Systems, USA; 462-TEC) (50 nM).

Techniques: Staining, Quantitative RT-PCR, Expressing, Transfection, Micro-CT

Journal: eLife

Article Title: Psychological stress disturbs bone metabolism via miR-335-3p/Fos signaling in osteoclast

doi: 10.7554/eLife.95944

Figure Lengend Snippet: ( A ) The ingenuity pathways analysis (IPA) of miR-335-3p and its predictive target gene. ( B ) Kyoto Encyclopedia of Genes and Genomes (KEGG) terms associated with targeted mRNA of differentially expressed miRNAs between groups obtained by database (miRDB and miRTarBase) prediction. ( D ) Firefly luciferase activity (n = 3). ( C ) Sequence alignment of miR-335-3p and its predictive target sites in 3′UTR of Fos . ( E ) Representative IF images for FOS, CTSK, and HOCHEST expression in the femur (scale bar, 50 um). ( F ) Representative cytochemical tartrate-resistant acid phosphatase (TRAP) staining images (scale bar, 200 um). ( G ) The area and ( H ) the number of osteoclasts (multinucleated TRAP-positive cells) (n = 4). ( I ) qRT-PCR quantification analysis of Acp5 , Nfatc1 , Mmp9 expression of RANKL-induced RAW264.7 transfected with inhibitor-NC, miR-335-3p-inhibitor, or miR-335-3p-inhibitor+T5224 (n = 3).All data are presented as mean ± SD; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, by one-way ANOVA with Tukey’s post hoc test. Figure 6—source data 1. Full dataset for .

Article Snippet: They were used for osteoclastic differentiation induction via RANKL (R&D Systems, USA; 462-TEC) (50 nM).

Techniques: Luciferase, Activity Assay, Sequencing, Expressing, Staining, Quantitative RT-PCR, Transfection

Journal: eLife

Article Title: Psychological stress disturbs bone metabolism via miR-335-3p/Fos signaling in osteoclast

doi: 10.7554/eLife.95944

Figure Lengend Snippet:

Article Snippet: They were used for osteoclastic differentiation induction via RANKL (R&D Systems, USA; 462-TEC) (50 nM).

Techniques: