Effect of hydrogen peroxide on bovine serum albumin adsorption on Ti6Al4V alloy: A scanning Kelvin probe force microscopy study

Protein adsorption on the surface of implant materials greatly affects the performance of the implants, such as their stability as well as the release of metal ions from and the adhesion of cells to their surface. In addition, the production of extracellular H₂O₂ from the activation of inflammatory cells could interfere with protein–metal interactions and/or modify the conformation of adsorbed proteins. In this study, we utilised scanning Kelvin probe force microscopy (SKPFM) to visualise the impact of H₂O₂ on bovine serum albumin (BSA) adsorption on the positively polarised Ti6Al4V alloy in a phosphate-buffered saline (PBS) solution. We show that the negatively charged BSA adsorbs onto the surface of polished and anodically polarised Ti6Al4V in a dense layer with a continuous network-like morphology or cluster shape and reduces the variation in the total surface potential compared to that of blank Ti6Al4V. However, addition of H₂O₂ to the PBS solution interferes with the formation of the dense protein network, and only a thin and discontinuous protein layer adsorbs onto the surface of the Ti6Al4V alloy, lowering the total surface potential difference. The information presented in this work provides new insights into the adsorption distribution of proteins on metallic substrates in biomaterials field.