Biofunctionalization of TiO₂ Surfaces with Self-Assembling Layers of Oligopeptides Covalently Grafted to Chitosan

In the field of tissue engineering, a promising approach to obtain a bioactive, biomimetic, and antibiotic implant is the functionalization of a "classical" biocompatible material, for example, titanium, with appropriate biomolecules. For this purpose, we propose preparing self-assembling films of multiple components, allowing the mixing of different biofunctionalities "on demand". Self-assembling peptides (SAPs) are synthetic materials characterized by the ability to self-organize in nanostructures both in aqueous solution and as thin or thick films. Moreover, ordered layers of SAPs adhere on titanium surface as a scaffold coating to mimic the extracellular matrix. Chitosan is a versatile hydrophilic polysaccharide derived from chitin, with a broad antimicrobial spectrum to which Gram-negative and Gram-positive bacteria and fungi are highly susceptible, and is already known in the literature for the ability of its derivatives to firmly graft titanium alloys and show protective effects against some bacterial species, either alone or in combination with other antimicrobial substances such as antibiotics or antimicrobial peptides. In this context, we functionalized titanium surfaces with chitosan grafted to EAK16-II (a SAP), obtaining layer-by-layer structures of different degrees of order, depending on the preparative stoichiometry and path. The chemical composition, molecular structure, and arrangement of the obtained biofunctionalized surfaces were investigated by surface-sensitive techniques such as reflection-absorption infrared spectroscopy (RAIRS) and state-of-the-art synchrotron radiation-induced spectroscopies as X-ray photoemission spectroscopy (SR-XPS), and near-edge X-ray absorption fine structure (NEXAFS). Furthermore, was demonstrated that surfaces coated with EAK and Chit-EAK can support hNPs cell attachment and growth.