Materials used for medical applications may be modified and functionalized by using the direct laser interference patterning method. Thereby, periodic patterns such as lines, pillars, and lamellas can be fabricated on, eg, titanium and polystyrene (PS) with surface feature sizes down to the submicrometer range.On the one hand, line-like structures fabricated on titanium and polyimide directly influenced the growth behavior of tumor cells, human mesenchymal stem cells, and osteoblasts. Between 60% and 80% of the different cells followed the direction of the line-like structures. Thereby, no pronounced difference in cell numbers was observed compared to nonlaser-treated surfaces of the same materials.On the other hand, attachment of Staphylococcus epidermidis and Staphylococcus aureus bacteria was significantly reduced on isotropic micrometer and submicrometer structure-like pillars and lamella. Especially, lamella-like patterns on PS were least attractive to S. aureus bacteria even under in vivo conditions.