Deposition of Antioxidant and Cytocompatible Caffeic Acid-Based Thin Films onto Ti6Al4V Alloys through Hexamethylenediamine-Mediated Crosslinking.

A promising approach for advanced bone implants is the deposition on titanium surfaces of organic thin films with improved therapeutic performances. Herein, we reported the efficient dip-coating deposition of caffeic acid (CA)-based films on both polished and chemically pre-treated Ti6Al4V alloys by exploiting hexamethylenediamine (HMDA) crosslinking ability. The formation of benzacridine systems, resulting from the interaction of CA with the amino groups of HMDA, as reported in previous studies, was suggested by the yellow/green color of the coatings. The coated surfaces were characterized by means of the Folin-Ciocalteu method, fluorescence microscopy, water contact angle measurements, X-ray photoelectron spectroscopy (XPS), zeta-potential measurements, and Fourier transform infrared spectroscopy, confirming the presence of a uniform coating on the titanium surfaces. The optimal mechanical adhesion of the coating, especially on the chemically pre-treated substrate, was also demonstrated by the tape adhesion test. Interestingly, both films exhibited marked antioxidant properties (2,2-diphenyl-1-picrylhydrazyl and ferric reducing antioxidant power assays) that persisted over time and were not lost even after prolonged storage of the material. The feature of the coatings in terms of the exposed groups (XPS and zeta potential titration evidence) was apparently dependent on the surface pre-treatment of the titanium substrate. Cytocompatibility, scavenger antioxidant activity, and antibacterial properties of the developed coatings were evaluated. The most promising results were obtained in the case of the chemically pre-treated CA/HMDA-based coated surface that showed good cytocompatibility and high reactive oxygen species' scavenging ability, preventing their intracellular accumulation under pro-inflammatory conditions; moreover, an anti-fouling effect preventing the formation of 3D biofilm-like bacterial aggregates was observed by scanning electron microscopy. These results open new perspectives for the development of innovative titanium surfaces with thin coatings from naturally occurring phenols for bone contact implants.