Influencing the adhesion properties and wettability of mucin protein films by variation of the environmental pH.

Mucins, the main component of the mucus secretions of goblet and epithelial cells, are known for exhibiting a different behaviour in accordance with their surrounding environment (i.e. among others the environmental pH), which induces a drastic change in their measured mechanical properties. In this work, we have first employed Atomic Force Microscopy (AFM) in Force Spectroscopy mode to evaluate the adhesion of porcine mucin films at the nanoscale, and the changes caused in this particular factor by a pH variation between 7.0 and 4.0, both quite common values in biological conditions. Measurements also involved additional varying factors such as the indenting tip chemistry (hydrophobic vs hydrophilic), its residence time on the measured film (0, 1 and/or 2 seconds), and increasing pulling rates (ranging from 0.1 up to 10 µm/s). A second approach regarded the macroscale behaviour of the films, due to their potential applicability in the development of a new set of stimuli-responsive biomaterials. This was possible by means of complementary Wilhelmy plate method (to test the wetting properties) and cell proliferation studies on films previously exposed to the corresponding pH solution. According to our results, treatment with lowest pH (4.0) provides porcine mucin with a more hydrophilic character, showing a much stronger adhesion for analogous chemistries, as well as enhanced capability for cell attachment and proliferation, which opens new pathways for their future use and consideration as scaffold-forming material.

Adhesion, unfolding forces, and molecular elasticity of fibronectin coatings: An atomic force microscopy study.

Fibronectin is an extracellular matrix protein that is involved in cell adhesion, growth, migration, differentiation, and wound healing. Fibronectin coatings are currently used in many laboratories for biomedical and biotechnology purposes. In this study we have investigated the adhesion and mechanical properties of fibronectin coatings. The coatings were also used to study the role of the residence time and the influence of the loading rate in nonspecific interactions. The results showed that the adhesion force between silica and fibronectin increased with loading rate delivering similar values for residence times of 1 and 2 s. Further analysis indicated that the distance to the transition state was about 0.5 nm. Moreover, the adhesion force did not vary with the loading rate for contact time of 0 s. The unfolding of fibronectin domains also depended of the Dwell time (no unfolding events were observed for zero residence time). Applied loads of 2 nN were able to stretch the fibronectin layer up to 200 nm and to unfold the three fibronectin domains, which were similar for a Dwell time of 1 and 2 s. However, the unfolding length increased with loading rate: below 2.5 µm s-1 the obtained lengths matched the value of FN I (13.5 nm), while for higher speeds the measured values corresponded to the lengths of FN II (18 nm) and FN III (27 nm). This investigation has answered and opened new questions about the mechanical stability and function of fibronectin coatings. The results have also raised theoretical questions about the difference between specific and nonspecific interactions to be addressed in future work.