Mechanism of formation and construction of self-assembled myoglobin/hyaluronic acid multilayer films: an electrochemical QCM, impedance, and AFM study.

Self-assembled multilayer films of hyaluronic acid (HA) and the protein myoglobin (Mb) were built up layer by layer on Au covered quartz crystal microbalance (AuQCM) electrode substrates. Film formation and growth were monitored by an electrochemical quartz crystal microbalance (EQCM), and the step-by-step construction was investigated through quantification of the mass variation corresponding to adsorption of each monolayer together with cyclic voltammetry. A decrease of friction at the liquid/electrode interface was observed, indicating that the electrode surface becomes less rough after deposition of several monolayers. The properties of the {HA/Mb}(n) films were evaluated by electrochemical impedance spectroscopy (EIS). Modeling of the impedance spectra shows smoothing of the modified electrode surface with reorganization of the film structure after few monolayers, and the contribution of each layer to the electron transfer process was analyzed. The smoothing of the surfaces and the structural differences between successive bilayers was confirmed by morphological observations by using atomic force microscopy.