The MDA-MB-231 cell line is derived from triple-negative breast cancer (TNBC), representing one of the most aggressive forms of breast cancer. Innovative therapeutic strategies, including s targeted therapies using nanocarriers, hold significant promise, particularly for difficult-to-treat cancers such as TNBC. Nanoparticles have transformed the medical field by serving as advanced drug delivery systems for cancer treatment. They play a critical role in overcoming the drug resistance often associated with cancer therapies. When utilized as drug delivery vehicles, nanoparticles can specifically target cancer cells and effectively reduce or eliminate multidrug resistance. Among them, chitosan-coated magnetic nanoparticles (MNPs) have been widely explored for the loading and controlled release of various anticancer agents. In this study, we evaluated the effects of dexamethasone-loaded chitosan-coated MNPs on MDA-MB-231 cell lines. Fourier transform infrared spectroscopy and scanning electron microscopy were employed to verify the successful loading of dexamethasone onto the nanoparticles. To assess cytotoxicity, empty nanoparticles, free drug, and drug-loaded nanoparticles were tested on the cells. The results indicated that empty nanoparticles exhibited no toxic effects. The IC50 value of the free drug was 123 µg/mL, while the IC50 value of the drug-loaded nanoparticles was significantly lower, at 63 µg/mL. These findings confirmed the successful conjugation of dexamethasone to the chitosan-coated MNPs, demonstrating substantial cytotoxic effects on breast cancer cells. Although dexamethasone has been reported to exhibit both tumor-suppressive and pro-metastatic effects, its specific impact on TNBC warrants further investigation in future studies.