Synthesis, Characterization, and Anti-Glioblastoma Activity of Andrographolide–Iron Oxide Nanoparticles (AG-IONPs)

<b>Background</b>: Glioblastoma multiforme (GBM) is an aggressive primary brain malignancy associated with poor prognosis and limited therapeutic options. Nanoparticle-based drug delivery systems provide a promising strategy to enhance treatment efficacy by circumventing barriers such as the blood-brain barrier. This study was conducted to synthesize, characterize, and evaluate the in vitro anticancer potential of andrographolide-iron oxide nanoparticles (AG-IONPs) against GBM cells. <b>Methods:</b> Iron oxide nanoparticles (IONPs) were synthesized through co-precipitation and subsequently functionalized with andrographolide. Morphology, size, and surface charge were assessed by transmission electron microscopy (TEM), dynamic light scattering (DLS), and zeta potential analysis. Functionalization was confirmed by Fourier-transform infrared spectroscopy (FTIR) and UV-Vis spectroscopy. Nanoparticle stability was monitored over three months. Cytotoxicity toward DBTRG-05MG cells was evaluated using MTT assays at 24, 48, and 72 h, while anti-migratory effects were determined using scratch-wound assays. Results: TEM analysis revealed nearly spherical IONPs (7.0 ± 0.15 nm) and AG-IONPs (13.5 ± 1.25 nm). DLS indicated an increased hydrodynamic diameter following functionalization, while zeta potential values decreased from +21.22 ± 1.58 mV to +8.68 ± 0.87 mV. The successful incorporation of andrographolide was confirmed by FTIR and UV-Vis spectra. AG-IONPs demonstrated excellent colloidal stability for up to three months. Cytotoxicity assays revealed a dose- and time-dependent decrease in cell viability, with LC₅₀ values declining from 44.01 ± 3.23 μM (24 h) to 15.82 ± 2.30 μM (72 h). Scratch-wound assays further showed significant inhibition of cell migration relative to untreated controls. <b>Conclusions:</b> AG-IONPs exhibit favorable physicochemical properties, long-term stability, and potent anti-proliferative and anti-migratory effects against GBM cells in vitro. These findings support their potential as a multifunctional therapeutic platform, warranting further preclinical investigation.