Biocompatible 3D hierarchical flower-like iron-doped silver nanostructures as a platform for in vitro and in vivo drug delivery

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.