Layer-by-layer-assembled nanofilms prepared by different driving forces were directly fabricated onto a cell surface to control cellular inflammation. Polyelectrolyte multilayers (electrostatic interaction) induced an over 2.5-fold higher production of the inflammatory cytokine interleukin-6 (IL-6) than fibronectin (FN)-based multilayers (biological recognition), which did not show any significant production of IL-6. Moreover, thick FN-based nanofilms showed lower expression of IL-6 than thin ones. This is the first report of controlling cellular inflammation by directly prepared nanofilms on a cell surface.