Aim: The aim of this study is to formulate miswak-synthesized silver nanoparticles and, its characterization and incorporate this into mouthwash along with xylitol, to test for its antimicrobial activity and cytotoxicity. Materials and methods: Silver nanoparticle was synthesized using miswak. The formed nanoparticle was characterized using transmission electron microscope. Mouthwash was formulated by incorporating it along with xylitol. Antimicrobial activity was assessed against Streptococcus mutans, Lactobacillus sp, Staphylococcus aureus, and Candida albicans. Cytotoxic testing was assessed on nauplli stage of developing shrimp. Results: The mouthwash showed efficient zone of diffusion against Staphylococcus aureus and Streptococcus mutans and a very strong antifungal activity. Silver nanoparticles were formed of size 5–25 nm and the particles were evenly scattered and was of spherical, angular, and rod shaped. The mouth rinse at higher concentrations showed mild toxicity. Conclusion: Silver nanoparticle formulated using miswak is strong antibacterial and antifungal agent. Xylitol is an efficient anticariogenic agent and it will reduce the bacterial load in the plaque samples. Thus, miswak-mediated silver nanoparticles incorporated into mouthwash along with xylitol can be an effective alternative to commercial mouthwash