Physical topography modification is an approach to fabricate nanostructures surfaces with antimicrobial properties. Lithography-based technologies offer an effective technique to develop the desired sizes and geometry. The replica molding technique was employed to fabricate the PDMS nanostructures using the PMMA imaging layer and characterized using a FESEM and AFM. The cell viability of gram-positive bacteria on structural diminished by almost 80% and the cells were deformed and ruptured once attached to the structured surface. Thus, the PDMS structured surface enhanced the bactericidal properties of the film, which effectively inhibit bacterial attachment.