Bee Podmore Extract Modulates Osteogenic Differentiation of Metabolically Impaired Mesenchymal Stromal Cells

Equine metabolic syndrome (EMS) is a naturally occurring endocrine-metabolic disorder that compromises the survival, redox balance, and regenerative competence of adipose-derived mesenchymal stromal cells (EqASCs), limiting their value for autologous cell-based therapy. This study examined whether a hydroethanolic extract of Bee Podmore (BP), the chemically heterogeneous biomass of deceased Apis mellifera workers, could restore the osteogenic potential of EMS-derived EqASCs in vitro. The extract was characterized at the compound-class level (total phenolic content 45.4 µg GAE/mg, total flavonoid content 5.7 µg QE/mg, BCA-reactive protein equivalent 460 µg/mg, total lipid equivalent 470.3 µg cholesterol-equivalent/mg). Cytocompatibility screening in healthy control and EMS-derived EqASCs identified 60 µg/mL as a conservative, non-cytotoxic working concentration, which was applied to EMS-derived cells during osteogenic induction. Relative to untreated EMS cells, BP treatment restored alkaline phosphatase activity and increased Alizarin Red S-detectable mineralization. This functional recovery was accompanied by reactivation of a coordinated osteogenic gene-expression and protein-distribution program, with upregulation of RUNX2, BMP4, COL1A1, BGLAP, and SPP1 relative to untreated EMS cells. Together, these findings indicate that Bee Podmore, a multi-component apiary by-product, can partially reverse the osteogenic impairment characteristic of EMS-derived EqASCs, supporting its potential as a bioactive supplement to improve the regenerative competence of metabolically compromised stromal cells for future autologous cell-based therapeutic applications in horses.