Evaluation of Moringa oleifera and Zingiber officinale Extracts in an Oxidative Stress-Induced MC3T3-E1/RAW264.7 Co-Culture Model of Osteoporosis

Background Osteoporosis is characterized by disrupted bone remodeling caused by impaired osteoblast–osteoclast communication under oxidative stress. This study aimed to evaluate the therapeutic potential of Moringa oleifera and Zingiber officinale extracts using an oxidative stress-induced osteoblast–osteoclast direct co-culture model. Methods An integrated approach combining LC–HRMS metabolomic profiling, network pharmacology, molecular docking, and molecular dynamics simulations was employed to identify representative phytochemicals and predict their therapeutic targets in osteoporosis. Moreover, an oxidative stress-induced direct co-culture model of MC3T3-E1 and RAW264.7 cells was used to evaluate the effects of ethanolic and aqueous extracts of Moringa oleifera and Zingiber officinale , administered individually and in combination. Results Computational analyses identified quercetin and 6-gingerol as representative bioactive compounds targeting key regulators associated with oxidative stress and bone remodeling. An optimized co-culture system using 500 μM H₂O₂ increased RANKL production by preosteoblasts while preserving cell viability. Under these conditions, both extracts suppressed the expression of the osteoclastogenic transcription factor NFATc1 and DC-STAMP gene expression, whereas their effects on collagen type I, osteopontin, and cathepsin K were variable. Conclusion The findings demonstrate the anti-osteoporotic potential of Moringa oleifera and Zingiber officinale and the utility of the co-culture model for studying early osteoblast–osteoclast crosstalk.