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Nanometer-sized structures have been found to interact with cell function and development. In tissue engineering, the fabrication of bioactive devices which mimic physiologic conditions has a key role in eliciting specific cellular responses and in guaranteeing long term success of implants. To this regard the layer by layer (LBL) self assembly technique is an efficient method to develop nanostructured thin films. This technique was used to assemble biomimetic coatings containing fibronectin, an adhesive glycoprotein of the extracellular matrix (ECM). The deposited films were then tested for the response of a line of human osteoblast-like cells in order to evaluate their potential for bone tissue repair purposes. The assembled films resulted to be effective in improving cell adhesion and proliferation. Therefore, this technique shows a high potential for the optimization of the surface properties of biomaterials.