In silico and in vitro Investigation of Osteogenesis Potentiality of Smilax china Hydro Alcoholic Root Extract on Human Osteoblast-like Cell Line (Saos2)-A Novel Approach

Background: Osteoporosis is a skeletal illness characterised by the breakdown of bone tissue's micro-architecture, which increases the risk of fractures by making bones more brittle. Nowadays, the herbal based therapies gaining more attraction towards osteoporosis for its low risk of side effects. Aim: The goal of this study is to prove Smilax china root extract’s osteogenesis ability towards osteoblast like cell line Saos2. Materials and Methods: Cytotoxicity parameters such as tetrazolium-based assay (MTT), Trypan Blue Exclusion (TBE) and morphometric analysis were used to assess its anti-apoptotic nature and dose fixation. The cells were differentiated using Osteogenic Differentiation Media (ODM), then alkaline phosphatase and Alizarin red staining were performed. Molecular docking was performed for the interaction of rutin and quercetin with Runx1 and runx2 by Auto dock vina. Results: Cytotoxicity assessments revealed that the Smilax china root extract showed 100% viability up to 200 µg/mL. Same results were obtained in trypan blue based viability test and morphometric analysis. Alkaline phosphatase and mineralisation were increased gradually at 50, 100 and 200 µg/mL of Smilax china root extract treated cells as dose dependent manner. In silico analysis revealed strong binding affinities of quercetin and rutin with Runx1 and Runx2 (Quercetin - Runx1 - (-7.9), Quercetin - Runx2 - (-7.4), Rutin - Runx1 - (-8.5) and Rutin - Runx2 - (-7.9)) suggesting their potential involvement in osteogenic regulation, although drug-likeness screening indicated the need for further optimization. Conclusion: He findings suggest that Smilax china root extract promotes osteogenic differentiation in Saos-2 cells and that its bioactive compounds may interact favorably with osteogenic transcription factors. Further studies are warranted to validate its therapeutic potential as a safe herbal candidate for osteoporosis management.