Surface (bio)-functionalization of metallic materials: How to cope with real interfaces?

Metallic materials are an important class of biomaterials used in various medical devices, owing to a suitable combination of their mechanical properties. The (bio)-functionalization of their surfaces is frequently performed for biocompatibility requirements, as it offers a powerful way to control their interaction with biological systems. This is particularly important when physicochemical processes and biological events, mainly involving proteins and cells, are initiated at the host -material interface. This review addresses the state of "real interfaces" in the context of (bio)-functionalization of metallic materials, and the necessity to cope with it to avoid frequent improper evaluation of the procedure used. This issue is, indeed, well-recognized but often neglected and emerges from three main issues: (i) ubiquity of surface contamination with organic compounds, (ii) reactivity of metallic surfaces in biological medium, and (iii) discrepancy in (bio)-functionalization procedures between expectations and reality. These disturb the assessment of the strategies adopted for surface modifications and limit the possibilities to provide guidelines for their improvements. For this purpose, X-ray photoelectrons spectroscopy (XPS) comes to the rescue. Based on significant progresses made in methodological developments, and through a large amount of data compiled to generate statistically meaningful information, and to insure selectivity, precision and accuracy, the state of "real interfaces" is explored in depth, while looking after the two main constituents: (i) the bio-organic adlayer, in which the discrimination between the compounds of interest (anchoring molecules, coupling agents, proteins, etc) and organic contaminants can be made, and (ii) the metallic surface, which undergoes dynamic processes due to their reactivity. Moreover, through one of the widespread (bio)-functionalization strategy, given as a case study, a particular attention is devoted to describe the state of the interface at different stages (composition, depth distribution of contaminants and (bio)compounds of interest) and the mode of protein retention. It is highlighted, in particular, that the occurrence or improvement of bioactivity does not demonstrate that the chemical schemes worked in reality. These aspects are particularly essential to make progress on the way to choose the suitable (bio)-functionalization strategy and to provide guidelines to improve its efficiency.