Introduction Recently, metal nanoparticles, especially silver nanoparticles (AgNPs), are important in the medical and therapeutic applications. The purpose of this study was to the synthesis of silver nanoparticle using Asparagus khorasanesis extract and comparative evaluation of antimicrobial, cytotoxic and apoptotic effects of synthesized and commercial silver nanoparticles. Materials and Methods In this descriptive study, AgNPs were synthesized using Asparagus khorasanesis extract. Its physical-chemical properties were investigated by UV-vis spectroscopy, transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-Ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) and energy-dispersive X-ray spectroscopy (EDX) techniques. The antimicrobial activity of synthesized and commercial AgNPs were evaluated by microdilution method. In addition, their cytotoxic and apoptotic effects on breast cancer cell line (MCF-7) was performed using MTT and flow cytometric methods, respectively at concentrations of 3. 125 to 100 μ g/mL. Results The synthesized AgNPs showed an absorbance peak of 438 nm in UV-vis. The results of SEM, TEM, FTIR, XRD, and EDX showed that the synthesized AgNPs had a spherical structure with a mean size of 12. 96 nm and with 90. 5% purity. The microdilution results showed that synthesized AgNPs had more significant antimicrobial effects in comparison of commercial AgNPs. Moreover, the MTT and flow cytometric results showed that synthesized AgNPs in 50 and 100 μ g/ml concentration exhibited more cytotoxic and apoptotic properties than commercial silver nanoparticles. Conclusion The results of this study showed that the synthesized AgNPs have significantly biological effects more than commercial AgNPs, and therefore these nanoparticles can be considered as drug candidates.