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Recent advances in the use of magnetite nanoparticles for biomedical applications have led to special attention to these nanoparticles. The unique properties of magnetite nanoparticles such as superparamagnetism, low toxicity, and the ability to bond with biological molecules, are suitable for drug delivery, diagnostic methods and therapeutic approaches. The aim of this study was to synthesize magnetite nanoparticles with different biocompatible coatings and investigate their cytotoxicity. Magnetite nanoparticles were synthesized by co-precipitation method and the cytotoxicity of these nanoparticles was investigated with Hepatoma G2 cell using the MTT assay. Treated cells, did not showed any evident cell cycle arrest. The Fourier Transmission Infrared (FTIR) spectroscopy, X- ray powder Diffraction (XRD), Transmission Electron Microscopy (TEM) were evaluated. The results of XRD showed the coated magnetite nanoparticles were 10−12 nm and this size also achieved with TEM images. Synthesized magnetite nanoparticles with SiO2 and oleic acid coatings had lower cytotoxicity than other coatings.