Scutellaria baicalensis root-derived silver nanoparticles for enhanced Baicalin’s biological activities to inhibit breast cancer cells

Abstract Breast cancer (BC) remains a major global health challenge, particularly aggressive subtypes such as triple-negative breast cancer (TNBC), which often exhibit poor therapeutic response and systemic toxicity. Baicalin (BA) possesses promising anticancer activity; however, its clinical application is limited by poor solubility, low bioavailability, and limited cellular uptake. The study aimed to synthesise baicalin-silver nanoparticles (BA-AgNPs) and to evaluate whether the nanoformulation could enhance the antioxidant, biocompatibility, and anticancer potential of BA against BC cells. BA-AgNPs were synthesised using a green reduction approach and characterised by UV–vis spectroscopy, FTIR, dynamic light scattering, zeta potential, SEM, and HR-TEM analysis. Antioxidant activity was evaluated using DPPH, FRAP, ABTS, H 2 O 2 , reducing power assay, and hydroxyl radical scavenging assays. Biocompatibility was assessed by hemolysis, and cytotoxicity was evaluated using L929, H9c2, NIH/3T3, and PBMCs. Anticancer activity was evaluated against MCF-7 and MDA-MB-231 cells using cell viability and neutral red uptake assay. Apoptosis was assessed using various fluorescence-based assays, DCHF-DA, annexin V-FITC/PI apoptosis assay, cell cycle analysis, a wound-healing assay, glucose consumption, nitric oxide, and NADH/NAD + assays. BA-AgNPs show an average hydrodynamic size of 169 nm, enhanced antioxidant activity, and selective cytotoxicity toward breast cancer cells with IC50 values of 90 µg/mL (MCF-7) and 25.9 µg/mL (MDA-MB-231), while showing low toxicity toward healthy cells. The study findings revealed increased ROS generation, mitochondrial membrane depolarization, lysosomal disruption, induction of apoptosis, cell cycle arrest, inhibition of migration, and disruption of redox homeostasis upon BA-AgNPs treatment, highlighting their potential as a promising nanoformulation for future breast cancer therapeutics.