In vitro Evaluation of Osteogenic Potential of Foxtail Millet (Setaria italica linn.) Extract on MG-63 Cell Line

Background: Bone-related diseases are a significant global health concern. Foxtail millet (FM) ( Setaria italica Linn .), an ancient cereal grain, possesses established nutritional and therapeutic properties. Recent studies highlight its potential in bone health, yet a comprehensive evaluation of its osteogenic effects using in vitro models is limited. Methods: Ethanolic extract of FM was prepared and subjected to physicochemical analysis. Cell viability was assessed using the (4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay (MTT) assay. Osteogenic potential was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) analysis of alkaline phosphatase (ALP), osteocalcin (OCN), and runt-related transcription factor 2 (RUNX-2). β-actin served as the housekeeping gene for normalization. Results: MG-63 cells were treated with FM extract at concentrations of 150 µg/mL and 74 µg/mL, with Cipcal (calcium carbonate) at 100 µg/mL and 20 µg/mL as positive controls. Gene expression analysis revealed a complex regulatory pattern. The FM extract downregulated ALP messenger RNA (mRNA) expression at both concentrations (0.228-fold at 150 µg/mL and 0.538-fold at 74 µg/mL) and upregulated RUNX-2 mRNA (1.30-fold at 150 µg/mL and 1.65-fold at 74 µg/mL) and OCN mRNA (1.26-fold at 74 µg/mL). Cipcal also exhibited inconsistent effects, downregulating RUNX-2 (0.587-fold) and ALP (0.275-fold) at 100 µg/mL. Conclusion: FM extract shows significant osteogenic activity, showing its potential as a natural therapeutic for bone-related disorders such as osteoporosis. Comprehensive in vivo studies and clinical trials are needed to confirm its efficacy for bone regeneration.