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(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.

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

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 .

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