The molecular mobility of cyclic molecules (e.g. α-cyclodextrins) threaded along a linear polymer chain (e.g. poly(ethylene glycol)) in polyrotaxanes is a unique feature for biomaterials with dynamic functionality. Surfaces with molecular mobility can be obtained by introducing polyrotaxanes. The molecular mobility of polyrotaxane-based surfaces can be modulated by changing the number of threaded cyclic molecules and modifying their functional groups. Biological ligands modified with α-cyclodextrins exhibit increased multivalent interactions with their receptors due to the molecular mobility of the latter. Furthermore, polyrotaxane-based surfaces not only improve the initial response of cells via multivalent interactions, but also affect cytoskeleton formation and the inherent quality of cells, including differentiation. Such polyrotaxane surfaces can emerge as new biointerfaces that can adapt to the dynamic biological nature.