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In this study, we report the synthesis and characterization of NiFe2O4 and CoFe2O4 nanoparticles (NPs) which are widely used in the biomedical area. There is still limited knowledge how the properties of these materials are influenced by different chemical routes. In this work, we investigated the effect of heat treatment over cytotoxicity of cobalt and niquel ferrites NPs synthesized by sol-gel method. Then the samples were studied using transmission electron microscopy (TEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), vibrating sample magnetometer (VSM), Fourier Transform Infrared Spectroscopy Analysis (FTIR), and X-ray fluorescence (XRF). The average crystallite sizes of the particles were found to be in the range of 20–35 nm. The hemocompatibility (erythrocytes and leukocytes) was checked. Cytotoxicity results were similar to those of the control test sample, therefore suggesting hemocompatibility of the tested materials.