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The potential of external ions doped biomaterials has been extensively explored; however, the co-doped biomaterials remain typically unexplored for their biological properties for precise biomedical applications. The current study was aimed to explore the impact of structural features of Sr/Fe co-doped hydroxyapatite (HAp) bionanomaterial on osteoblastic proliferation and osteogenic differentiation for its application as a bone substitute. A 10 mol% isomorphous co-doping of strontium and iron with respect to calcium was carried into HAp in the solid solution. Raman spectroscopy verified the presence of major functional groups of apatite structure together with the carbonate peaks. The Sr/Fe co-doped HAp bionanomaterials showed slightly negative zeta potential (at neutral pH), versatile DLS particle size (140–205 nm), high BET surface area (186 m²/g), and narrow width pore size (13–19 nm). TG/DTA analysis …