Rutile facet-dependent fibrinogen conformation: Why crystallographic orientation matters

Previous studies implied that single crystalline rutile surfaces have the ability to guide the functionality of adsorbed blood plasma proteins. However, a clear relation between the rutile crystallographic orientation and conformation of adsorbed proteins is still missing. Here, we examine the adsorption characteristics of human plasma fibrinogen (HPF) on atomically flat single rutile crystals with (110), (100), (101) and (001) facets. By direct visualization of individual protein molecules through atomic force microscopy (AFM) imaging, the distinct conformations of HPF were determined depending on rutile surface crystallographic orientation. In particular, dominant trinodular and globular conformation was found on (110) and (001) facets, respectively. The observed variations of HPF conformation were reasoned from the surface water contact angle and surface energy point of view. By analyzing AFM-based force measurements, statistically significant changes in surface energies of rutile surfaces covered with HPF were determined and linked to HPF conformation. Furthermore, the facet-dependent structural rearrangement of HPF was indirectly confirmed through deconvolution of high-resolution X-ray photoelectron spectroscopy (XPS) carbon and nitrogen spectra. The globular, and thus native-like HPF conformation observed on (001) facet, was reflected in the lowest level of amino group formation. We propose that the mechanism behind the crystallographic orientation-induced HPF conformation is driven by the facet-specific surface hydrophilicity and energy. From the biomedical material perspective, our results demonstrate that the conformation of HPF can be guided by controlling the crystallographic orientation of the underlying material surface. This might be beneficial to the field of titanium-based biomaterials design and development.