Abstract
Nanostructured platforms have attracted significant attention as promising drug carriers due to their unique properties and therapeutic efficiency. This work aims to prepare three-dimensional flower-like iron-doped silver nanostructures (HFAg-Fe) and evaluate their potential as nanocarriers for Methotrexate (MTX), a first-generation anticancer and autoimmune drug. The HFAg-Fe were synthesized, modified, and characterized by FE-SEM, FT-IR, XRD, AFM, and elemental analyses. Drug loading and release were studied using UV–vis spectroscopy, showing an MTX loading capacity of 86% and pH-responsive behavior rapid release under acidic tumor conditions and slower release at physiological pH, potentially reducing side effects. Hemolysis and cytotoxicity assays indicated that HFAg-Fe/MTX possessed superior biocompatibility and anticancer efficacy compared to free MTX, while fluorescence microscopy confirmed efficient uptake by cells. Cell cycle analysis showed treatment-induced arrest, mainly at the S phase. X-ray micro-computed Tomography revealed significant tumor volume reduction and preferential accumulation of the nanocarrier at the tumor site. These findings highlight the potential of HFAg-Fe nanostructures as safe drug delivery systems, where 4-aminothiophenol serves as a linker to facilitate drug loading and release, potentially enhancing outcomes in cancer therapy.