In a marine environment, specific proteins are secreted by mussels and used as a bioglue to stick to a surface. These mussel proteins present an unusual amino acid 3,4-dihydroxyphenylalanine (known as DOPA). The outstanding adhesive properties of these materials in the sea harsh conditions have been attributed to the presence of the catechol groups present in DOPA. Inspired by the structure and composition of these adhesive proteins, we used dopamine-modified hyaluronic acid (HA-DN) prepared by carbodiimide chemistry to form thin and surface-adherent dopamine films. This conjugate was characterized by distinct techniques, such as nuclear magnetic resonance and ultraviolet spectrophotometry. Multilayer films were developed based on chitosan and HA-DN to form polymeric coatings using the layer-by-layer methodology. The nanostructured films formation was monitored by quartz crystal microbalance. The film surface was characterized by atomic force microscopy and scanning electron microscopy. Water contact angle measurements were also conducted. The adhesion properties were analyzed showing that the nanostructured films with dopamine promote an improved adhesion. In vitro tests showed an enhanced cell adhesion, proliferation and viability for the biomimetic films with catechol groups, demonstrating their potential to be used in distinct biomedical applications.