In the course of studies of the interaction of proteins with TiO2 nanoparticles, we have investigated the role of the medium employed in cellular tests, by measuring the variation of ζ-potential vs pH in the range 2-9 and bovine serum albumin adsorption on TiO2 P25 in the presence of either HEPES or PBS as buffers, both mimicking the physiological pH, but with different chemical nature. The two buffers yield remarkably dissimilar surface charges and protein uptake, i.e., they impart different surface characteristics to the particles which could affect the contact with cells or tissues. This may account for dissimilar toxicological outcomes among in vitro tests and particularly between in vitro vs in vivo tests, considering the high amount of phosphate ions present in body fluids.