In this study, we exploit a facile, one-pot method to prepare MCM-41 type mesoporous silica nanoparticles decorated with silver nanoparticles (Ag-MSNs). Silver nanoparticles with diameter of 2–10 nm are highly dispersed in the framework of mesoporous silica nanoparticles. These Ag-MSNs possess an enhanced antibacterial effect against both Gram-positive and Gram-negative bacteria by preventing the aggregation of silver nanoparticles and continuously releasing silver ions for one month. The cytotoxicity assay indicates that the effective antibacterial concentration of Ag-MSNs shows little effect on human cells. This report describes an efficient and economical route to synthesize mesoporous silica nanoparticles with uniform silver nanoparticles, and these nanoparticles show promising applications as antibiotics.