Purpose: The presented study aimed to investigate the effects of Fe 3O 4 nanoparticles and static magnetic field on osteoblast and osteoclasts’ metabolic activity. Methods: Magnetic nanoparticles were prepared by a wet chemical co-precipitation process and analyzed using X-ray powder diffraction, high-resolution transmission electron microscope (HRTEM), dynamic light scattering (DLS), laser Doppler velocimetry, Raman and the Mössbauer spectroscopy. In vitro experiments were performed using MC3T3, 4B12 and RAW 264.7 cell lines. Cells were cultured in the presence of nanoparticles and with or without exposure to the magnetic field. Proteins were investigated with Western blotting and immunofluorescence and Western blot. Gene expression was analyzed with a quantitative real-time polymerase chain reaction. Results: Obtained particles were in the nano-range (average size around 50 nm) and had a spherical-like morphology. The typical hydrodynamic size was in the range 178– 202 nm and Zeta potential equaled – 9.51 mV. Mössbauer spectrum corresponds to the Fe+3 ions in tetrahedral (A) and Fe+3 and Fe+2 ions in octahedral (B) sites of Fe 3O 4. In vitro study revealed cytocompatibility and anti-inflammatory effects of fabricated nanoparticles. Furthermore, it was shown that nanoparticles combined with magnetic field exposure enhance osteogenic differentiation of MC3T3 cells by upregulation of RUNX-2 activity. Under the same experimental condition, nanoparticles and magnetic field decreased osteoclastogenesis of 4B12 by the induction of apoptosis through the mitochondrial-dependent pathway. Conclusion: Fe 3O 4 nanoparticles together with magnetic field can be applied for the fabrication of novel biomaterials for the treatment of bone disorders related to bone loss in which a balance between bone-forming and resorbing cells is disturbe