Selectin-mediated adhesion in shear flow using micropatterned substrates: multiple-bond interactions govern the critical length for cell binding

Abstract

Receptor–ligand adhesive interactions play a pivotal role in diverse biological processes including inflammation and cancer metastasis. Cell adhesion is mediated by the molecular recognition of membrane-bound receptors by their cognate ligands on apposing cells. Cell–cell binding is regulated by distinct parameters such as the receptor–ligand binding kinetics, the tensile strength of individual bonds, the involvement of multiple bonds and their modulation by hydrodynamic shear. This work aims to investigate the interplay of these parameters on selectin-mediated cell adhesion in shear flow. We designed a microfluidic device that delivers cells in a single file over a receptor-functionalized substrate, thereby permitting accurate determination of the cell flux. The selectin(s) was presented on striped patches of fixed width and varying length. We identified the critical patch lengths of P- and L-selectin for the initiation of HL-60 cell binding in shear flow. This characteristic length is governed by the time required to form multiple-bond interactions, as revealed by a multiple-bond mathematical model. The number of bonds required to support cell binding increases with the applied shear stress (0.5–2 dyn cm⁻²) for L- but not P-selectin. This finding is explained by differences in the tensile strength of P- and L-selectin for PSGL-1. Our integrated experimental and mathematical approach advances our understanding of receptor-mediated cell adhesion in the vasculature. Detailed knowledge of how molecular interactions modulate macroscopic cell binding behavior pertinent to inflammation and metastasis would facilitate the development of promising diagnostic tools to combat these diseases.