Coating of Sub-Micrometric Keratin Fibers on Titanium Substrates: A Successful Strategy for Stimulating Adhesion and Alignment of Fibroblasts and Reducing Bacterial Contamination

Abstract

Coatings are a versatile tool for modulation of the biological response of biomaterials; in particular, the use of biopolymers as coating material may improve cell interactions and tissue adhesion. Among others, keratin is a natural protein able to stimulate fibroblast cells effectively and has the ability to bind metal ions. Coatings of keratin fibers onto titanium substrates can improve soft tissue adhesion, eventually coupling topographical (contact guidance) and chemical stimulus through the alignment of the fibers along sub-micrometric grooves of the substrate. Sub-micrometric keratin fibers were obtained by electrospinning both in random and oriented arrangements (though a rotating collector); in addition, antibacterial properties were added by enrichment of the coating with silver ions. This type of coating can be of interest in transmucosal dental implants, where perimplantitis is often due to infection (biofilm formation) and disease worsening is due to inflammation of the surrounding soft tissue, which is guided by fibroblasts. Keratin fibres coatings were prepared and characterized by means of Field Emission scanning electron microscopy (FESEM), X-ray photoelectron spectroscopy (XPS), cell (gingival fibroblasts) and bacteria (S. aureus) culture tests. FESEM observations demonstrated the possibility to deposit keratin fibres onto titanium substrates in random or oriented arrangements effectively. Keratin fibres were able to increase fibroblast adhesion and proliferation. On randomly deposited keratin fibres, fibroblast cells were significantly biologically stimulated and showed high adhesion and proliferation, but not orientation ability; on the other hand, aligned keratin fibres on a grooved substrate were able to stimulate cells both from the topographical (orientation) and biological standpoint. Finally, Ag-doped keratin fibres coatings were able to reduce S. aureus adhesion significantly, maintaining high biocompatibility. Considering these results, keratin sub-micrometric fibres coatings are a promising strategy for stimulating fibroblasts and reducing bacterial contamination.