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Tris(hydroxymethyl)aminomethane (Tris) has been used as the buffer inbioactivity testing for over two decades and has become a standard choice forthe scientific community. While it is believed to be non-interacting, theextent of its interactions with titanium oxide surfaces has not beensystematically studied. Here, we use experimental (zeta potential measurements)and computational (molecular dynamics) approaches to evaluate the interactionof Tris with a rutile surface and how it affects the adsorption of othermolecules relevant in biomedical in vitro testing. We show that the interactionof Tris with the rutile surface is strong and significantly affects theinteraction of other organic residues with the surface. These stronginteractions are compounded by the Tris concentration in the in vitro testingprotocol which is much higher compared to other components. Our findingsindicate that the kinetics observed in in vitro tests will be stronglyinfluenced by the presence of Tris as a buffering agent when compared to thenatural CO2 buffer in blood. These results reveal that considering the so-farneglected active role of Tris in in vitro testing is critically needed and thatin vitro protocols using CO2 partial pressure as the buffering agent shouldyield more reliable results.

Interactions of Tris with rutile surfaces and consequences for in vitro bioactivity testing