Silver-poly(methyl methacrylate) nanocomposite thin films were deposited on silicon substrates by means of matrix-assisted pulsed laser deposition. Atomic force microscopy revealed that the surface morphologies of the silver-poly(methyl methacrylate) nanocomposite thin films were dependent on the matrix-assisted pulsed laser evaporation processing parameters. Films produced using 4% (by weight of solvent) silver nanoparticles exhibited the highest antimicrobial activity. Antimicrobial activity of the thin films was dependent on the silver concentration, the microenvironment created by the polymer–silver interaction, and the surface potential.