The current research explores the sustainable synthesis of AgNPs using 3,6-dihydroxyflavone, a bioactive flavonoid, and evaluates their multifunctional biological applications. The green-synthesized AgNPs were confirmed using UV-Vis spectroscopy, FTIR, XRD, SEM, TEM, EDS, DLS, and zeta potential analysis, confirming their crystalline nature, spherical morphology (22–28 nm), and moderate colloidal stability. The larvicidal potential of the AgNPs was assessed against An. stephensi, Cx. quinquefasciatus, and Ae. aegypti, with significant mortality observed at low concentrations, and LC50 values ranging from 9.10 to 12.62 µg/mL. Histological examination revealed severe midgut damage in treated larvae, indicating disruption of digestive and structural integrity. The AgNPs also exhibited strong antioxidant activity in DPPH and ABTS assays. Significant antibacterial activity was observed against both Gram-positive and Gram-negative test pathogens, and the compounds potentially inhibit biofilm formation. Furthermore, cytotoxicity studies demonstrated dose-dependent anticancer activity against MCF-7 cells while screening for minimal toxicity in normal Vero cells. The ecotoxicological assessment using Artemia salina confirmed that the AgNPs were safe to non-target aquatic organisms at effective larvicidal doses. In parallel, molecular docking showed strong affinity for 1MV8 (-7.35 kcal/mol, Ki 4.12 µM), supporting its role in the antibacterial, anti-biofilm, and anticancer effects of the synthesized AgNPs. Overall, this study highlights the potential of 3,6-dihydroxyflavone-mediated AgNPs as a promising green nanoplatform for mosquito control and biomedical applications.
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