Neha Verma, Apurva Prajapati, Shilpa Kumari, Hitesh D Patel, Kanu Priya
Bone fragility and reduced bone mass are hallmarks of osteoporosis, which is characterized by microarchitectural deterioration. The anti-osteoporotic properties of fraction F1, which was separated from Holoptelea integrifolia, were evaluated in the present study using MG 63 osteoblastic cells. Analysis using GC-MS to find the bioactive components in fraction F1. Cell viability, alkaline phosphatase activity, mineralization assays, and in silico validation using molecular simulations to target the important osteoporotic proteins BMP2 and CCR2 were among the assays used to measure biological activity. It was discovered that cell viability increased with increasing concentrations, and the MG-63 cells had strong cytocompatibility and proliferation ability. Specifically, at 48 h, MTT-derived metabolic activity ranged from 72.48% of the untreated control at 1 µg/mL to 132.57% at 320 µg/mL, with the first concentration exceeding the untreated-control value observed at 20 µg/mL. Further, the ALP activity increased slightly from 0.611 to 0.615 U/mL at 7 days and from 0.616 to 0.620 U/mL at 14 days. The assay also showed slightly higher optical density at 14 days compared to the control, indicating increased calcium deposition. The main phytoconstituents in fraction F1 demonstrated strong binding affinity to BMP2 and CCR2 proteins in molecular docking tests, indicating positive outcomes in interacting with different osteogenic and inflammatory pathways. The molecular docking results were validated using 100 ns molecular simulations, which demonstrated that the complexes generated could bind. Therefore, the aforementioned findings clearly demonstrate that fraction F1 has potential for osteoblast differentiation, mineralization, and proliferation through protein interactions, indicating promising outcomes for its development as an anti-osteoporotic agent in future research.