Abstract

Chirality influences essential biological processes such as molecular recognition and self- organization, impacting cell proliferation and differentiation mediated by interactions with the extracellular matrix (ECM). Despite extensive research on cell migration, the role of chirality on matrix-cell interactions has been largely overlooked. To explore this aspect, we engineered culture surfaces coated with natural collagen I or collagen-mimetic peptides (CMPs) with opposite chirality, i.e. with either L- or D-amino acids in their sequences. Here we show that D-surfaces impede epithelial keratocyte spreading, making cells more rounded, less adhesive, and slower. Further investigation through integrin inhibition assays and molecular dynamics simulations revealed that a chiral mismatch destabilizes the triple helix of heterochiral CMPs at the L/D junction. This study underscores the profound impact of ECM chirality on cellular behavior, providing new insights into the relationship between matrix (supra)molecular chirality, integrin-mediated molecular recognition, and cell migration dynamics.