Host-guest interactions have been increasingly explored for use in the dynamic physical cross-linking of polymeric precursors to form hydrogel networks. However, the orientation of guest motifs is restricted upon macromolecule conjugation. The implications of such restriction on both the kinetics and thermodynamics of the resulting host-guest supramolecular cross-links are poorly understood. Herein, guest cross-linking motifs from controlled regioisomers are demonstrated to yield distinct material properties. Moreover, the underlying phenomena point to a further unexpected impact of modular guest topology on the molecular scale in both the affinity and dynamics of supramolecular complex formation.