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Bioactive glasses (BGs) have been used for bone formation and bone repair processes in recent years. This study investigated the titanium substitution effect on 58S BGs (Ti‐BGs) 60SiO2‐(36 − X)CaO‐4P2O5‐XTiO2 (X = 0, 3, and 5 mol.%) prepared by the sol‐gel technique, and the main goal was to find the optimum amount of titanium in Ti‐BGs. Synthesized BGs, which were investigated after immersion in simulated body fluid (SBF), were tested by X‐ray diffraction (XRD) analysis, Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy. Moreover alkaline phosphate (ALP) activity, 3‐(4,5dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay, and antibacterial studies were employed to investigate the biological properties of Ti‐BGs. According to the FTIR and XRD test results, hydroxyapatite (HA) formation on Ti‐BGs surfaces was confirmed. Meanwhile, the presence of 5 mol.% compared to 3 mol.% increased the HA grain distribution and their size on the Ti‐BGs surface. Additionally, MTT and ALP results confirmed that the optimal amount of titanium substitution in BG was 5 mol.%. Since 5 mol.% Ti incorporated BG (BG‐5) had the highest biocompatibility level, antibacterial properties, maximum cell proliferation, and ALP activity among the synthesized Ti‐BGs, it is presented as the best candidate for further in vivo investigations.

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