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This work presents the synthesis and characterization of a composite made of superparamagnetic iron oxide and hydroxyapatite nanoparticles (SPION/HAp) with a well-developed surface for loading anticancer drugs and for use in magnetic hyperthermia and local chemotherapy. The proposed material was obtained by an easy one-pot co-precipitation method with a controlled ratio of SPION to HAp. The morphology was studied by SEM and TEM, indicating rod-like structures for high HAp content in the composite and granule-like structures with increasing SPION. Its crystallinity, elemental composition, and functional groups were determined by X-ray diffraction, EDS, and FT-IR, respectively. The nanocomposite was then stabilized with citrates (CA), polyethylene glycol (PEG), and folic acid (FA) as agents to improve intracellular absorption, while turbidimetric studies confirmed that only citrates effectively stabilized the magnetic carriers to form a colloidal suspension. Subsequently, 5-fluorouracil (5-FU) was loaded into the magnetic carriers and tested in vitro using the L-929 cell line. The studies showed no cytotoxicity of the citrate-stabilized suspension against fibroblasts and some cytotoxicity after 5-FU release. In addition to in vitro studies, the composite was also tested on biomimetic membranes made of DOPC, DOPE, cholesterol, and DOPS lipids using Langmuir trough. The results show that the resulting suspension interacts with biomimetic membranes, while magnetic hyperthermia studies confirm effective heat generation to achieve therapeutic 42–46 °C and improve drug release from magnetic carriers. [ABSTRACT FROM AUTHOR]