Mechanistic Insights into Ipriflavone's Role in Postmenopausal Osteoporosis through Integrated Computational and in vitro Techniques

Background: Postmenopausal osteoporosis affects a large percentage of the female population worldwide. Understanding the molecular pathways is critical for effective treatment. This study aimed to assess ipriflavone's efficacy in treating postmenopausal osteoporosis using computational techniques and in vitro evidence. Materials and Methods: Ligands were prepared for docking while receptors were readied via protein preparation. Molecular docking was performed to generate ligand-receptor complexes which underwent molecular dynamics simulation using GROMACS. Simulation trajectories were analyzed to gauge binding stability. Finally, ligand activity against MG-63 osteosarcoma cells was evaluated through MTT and Alizarin Red assays to measure viability and calcium uptake respectively. Results: The results of the study indicated that ipriflavone can modulate several genes associated with postmenopausal osteoporosis, including ESR1, ESR2, CYP19A1, LGMN, CASP3, BMP4, TNFRSF1A, and CTSK. Among these genes, ipriflavone showed the highest binding affinity with estrogen receptors (ESR1 and ESR2) with a docking score of -8.7 kcal/mol. Molecular dynamics analysis confirmed the stability of the ipriflavone complex for up to 200 ns. Pathway analysis using KEGG revealed the specific pathways modulated by ipriflavone. Furthermore, in vitro experiments using the MG63 cell line demonstrated that ipriflavone is non-toxic and can increase calcium uptake, indicating its potential osteogenic effect. The MTT assay supported the safety of ipriflavone treatment in MG63 cells. Conclusion: This study illuminates ipriflavone's anti-osteoporotic mechanism. According to this study, ipriflavone may modify estrogen receptors and treat postmenopausal osteoporosis. More in vivo investigations are needed to prove ipriflavone's biological potency and efficacy in a more complex physiological environment.