 Toothbrushing, though aimed at biofilm removal, also affects the lubricative function of adsorbed salivary conditioning films (SCFs). Different modes of brushing (manual, powered, rotary–oscillatory or sonically driven) influence the SCF in different ways. Our objectives were to compare boundary lubrication of SCFs after different modes of brushing and to explain their lubrication on the basis of their roughness, dehydrated layer thickness, and degree of glycosylation. A pilot study was performed to relate in vitro lubrication with mouthfeel in human volunteers.