Background: Recently, green nanotechnology has emerged as a sustainable approach for producing metallic nanoparticles using phytochemicals. Objective: This study investigates the biosynthesis of silver nanoparticles (AgNPs) using extracts from roots of Angelica glauca and Paeonia suffruticosa plants, and evaluates their structural properties, antimicrobial activity, and anticancer potential. Methodology: The silver nanoparticles were synthesized via plant-based reduction of silver nitrate and characterized by using several technologies. Antimicrobial activity was assessed using the agar-well diffusion assay, anticancer activity against human breast cancer cells (MCF-7) was also evaluated. Results: GC–MS analysis of Paeonia suffruticosa extract revealed several bioactive phytochemicals. The results showed a color change from light yellow to dark brown, indicating the formation of silver nanoparticles. The absorption peak at approximately 420-440 nm, obtained from spectrophotometric analysis, confirmed the synthesis of these nanoparticles. The presence of strong, broad (FTIR) peaks also indicated the presence of hydroxyl (OH) groups. SEM & TEM results showed that most of the synthesized nanoparticles had an angular spherical structure and ranged in size from 30 to 98 nm. Bioactivity assays have shown that plant-derived silver nanoparticles exhibit significant antibacterial activity against both Gram-positive and Gram-negative bacteria, outperforming crude extracts alone. Conclusion: Silver nanoparticles synthesized using Angelica glauca and Paeonia suffruticosa root extracts represent versatile nanomaterials with potent antimicrobial and anticancer capabilities. Their properties point to promising prospects for future biomedical and pharmaceutical applications.