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Calcium phosphates, due to their similarity to the inorganic fraction of mineralized tissues, are of great importance in treatment of bone defects. In order to improve the biological activity of hydroxyapatite (HAP), its fluoride-substituted modification (FAP) was synthesized using the sol-gel method and calcined at three different temperatures in the range of 800–1200 °C. Physicochemical and biological properties were evaluated to indicate which material would support bone regeneration the best. X-ray diffraction (XRD), scanning electron microscopy (SEM) with energy dispersive X-ray spectrometry (EDS), and Fourier transform infrared spectroscopy (FTIR) revealed that fluoride ions were incorporated into the apatite lattice structure. In studies it was found that fluorapatite sintered at the highest temperature had the lowest porosity, no internal pores and the highest density. In vitro ion reactivity assessments showed that during the 28-day immersion of the samples in the simulated body fluid, the uptake of calcium and phosphorus ions was inversely correlated to the calcination temperature. All tested materials were non-toxic since the cytotoxicity MTT assay demonstrated that the viability of preosteoblast cells incubated with sample extracts was high. Fluorapatite sintered at 800 °C was determined to be of optimal porosity and fluoride release capacity and then used in cell proliferation studies. The results showed that it significantly shortened the doubling time and thus enhanced the proliferation of osteogenic cells, as compared to the fluoride solutions and control group. Therefore, this material is proposed for the use in orthopedic applications and bone tissue engineering.