<p>The current study explores the potential of eugenol as a therapeutic agent for bone regeneration through an integrative approach combining in silico and in vitro analyses. Network pharmacology and molecular docking was employed to investigate the molecular targets and pathways associated with eugenol in the context of bone-related diseases, revealing its multi-targeted therapeutic potential. Based on these findings, composite biomaterials were fabricated using Sodium Alginate, Chitosan, and β-Tricalcium phosphate, infused with Eugenol, to develop a bioactive scaffold for bone tissue engineering. The fabricated scaffolds were characterized using Fourier Transform Infrared Spectroscopy, X-Ray Diffraction, Thermogravimetric Analysis, Scanning Electron Microscopy, and compression testing to evaluate their physicochemical and mechanical properties. Additionally, porosity, swelling behaviour, in vitro degradation and bio-mineralization were assessed to determine the scaffold’s suitability for bone regeneration. The porous structure, swelling ratio, and controlled degradation profile indicated favourable properties for cell infiltration and tissue integration. Cytocompatibility of the scaffolds was evaluated using the MTT assay on MG-63 osteoblastic cell lines, demonstrating high cell viability and biocompatibility. Overall, the Eugenol-infused Alginate/Chitosan/β-Tricalcium phosphate composite scaffold exhibits promising potential as a multifunctional biomaterial for bone tissue engineering applications.</p> Graphical Abstract <p></p>