© 2020 Elsevier LtdHydroxyapatite (HA, Ca10(PO4)6(OH)2) is the main constituent mineral of bone and teeth in mammals. Due to its outstanding biocompatibility and osteoconductive capabilities, it is preferred for bone repair and replacement. Owing to high potential to have excellent biological properties, ternary ions-doped HAs have just begun to be investigated in the biomedical field and preparing multi-doped HAs is a fairly new approach. Boron (B, BO33-), strontium (Sr, Sr2+) and magnesium (Mg, Mg2+) provide a beneficial effect on bone growth, bone strength, biocompatibility and positively affect bone microstructure. The motivation of this study is taking advantages of the potential of the combine effects of these bivalent ions. In this study, 8 different compositions of BO33-, Sr2+, Mg2+ multi-doped HAs were synthesized by microwave irradiation method to investigate the structural, mechanical and biological features of bone substitutes. This is the first time we report the effect of boron, strontium and magnesium ions multi-doping on the structure of HA and its biological properties. Samples were sintered at 700, 900 and 1100 °C. The effect of varying ion contents and sintering temperature on structural and biological properties of the multi-doped samples was investigated. B, Sr and Mg ions were successfully doped into the HA structure according to X-Ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR) analyses. A biphasic structure was obtained with increasing amount of ion-doping. Increasing the sintering temperature affected the crystallinity and the density of the samples gradually. Vicker's microhardness and diametral strength of the samples increased at high sintering temperatures. B–Sr–Mg multi-doped HA promoted osteoblast-like Saos-2 cell proliferation, and as the sintering temperatures of the samples increased, the osteogenic differentiation level of the cultured cells also increased. Overall, results showed that the biological properties of HA were improved with the doping of Sr, Mg and B ions, and for bone implant applications samples sintered at 1100 °C were suggested to have potential as a biomaterial.