Biogenic Silver Nanoparticles Mediated by Phytoconstituents of the Edible Spice Pimpinella anisum: Characterization and In Vitro, In Vivo, and In Silico Biological Evaluation.

This study evaluated the aqueous extract of Pimpinella anisum seeds (AE-PAS) and its green-synthesized silver nanoparticles (PA-AgNPs) using integrated in vitro, in vivo, and in silico approaches. LC-MS analysis identified 26 phytoconstituents, mainly sinapic acid (15.78%), hispidulin (15.35%), caffeic acid (11.09%), and rutin (8.86%). Total phenolic and flavonoid contents were 45.2 ± 2.1 mg GAE/g DE and 28.7 ± 1.5 mg QE/g DE, respectively. UV-Vis spectroscopy, scanning electron microscopy (SEM), Fourier-transform infrared (FTIR) spectroscopy, and X-ray diffraction (XRD) supported nanoparticle formation, indicating dry-state particle or agglomerate features of approximately 60-90 nm, a surface plasmon resonance (SPR) peak at 421 nm, 50.81% crystallinity, and a crystallite-domain size of 22.57 ± 1.2 nm. AE-PAS showed measurable antioxidant and albumin-denaturation-inhibitory responses (DPPH IC50 = 130.63 ± 1.3 µg/mL; albumin-denaturation IC50 = 721.79 ± 1.7 µg/mL). PA-AgNPs produced measurable inhibition zones in agar well diffusion screening, but antibacterial potency could not be established because MIC and MBC were not determined. The PA-AgNP DPPH result lacked a nanoparticle-only blank and should be regarded as an apparent assay-specific estimate. Hemolysis was concentration dependent, increasing from 5.20% at 50 µg/mL to 29.93% at 100 µg/mL. In behavioral models, PA-AgNPs (0.1 mg/kg) produced measurable antidepressant-like and analgesic responses; these findings represent preliminary behavioral evidence rather than confirmed efficacy. Molecular docking predicted favorable binding scores (-8.0 to -9.6 kcal/mol) for selected extract phytochemicals against MAO-A/B, COX-1/2, and TNF-α but did not establish target inhibition or a biological mechanism. Overall, the findings are preliminary and require comprehensive physicochemical characterization, appropriate silver-ion and material controls, quantitative antimicrobial testing, pharmacokinetic and mechanistic validation, and expanded safety assessment before biomedical translation.