An in vitro intestinal epithelial wound/repair model in which epithelium is stripped from villus tips and the wound is resealed during the following 60 minutes has previously been described. The process, termed epithelial restitution, results in part from the rapid migration of epithelial cells shouldering the wound over the denuded basement membrane. The present report examines the requirements for epithelial cell-basement membrane interactions during restitution in this model. Addition of heparin, soluble matrix components, or a variety of antibodies to matrix components (laminin; fibronectin; collagen I, III, IV) does not impair restitution. Although inhibition of protein synthesis alone also does not retard restitution, in the simultaneous presence of antibody to type III and IV collagen restitution is impeded as judged functionally and structurally. Preincubation of tissues with 20 mmol/L cis-OH-proline (a condition known to inhibit cellular secretion of newly synthesized collagen) similarly inhibited structurally and functionally defined restitution only if antibodies to type III and IV collagen were simultaneously present. These results suggest that collagen-epithelial cell interactions are important in restitution after injury, and if necessary, collagen can be produced locally and rapidly at the site of injury to allow restitution to normally proceed.