Attachment of Fusobacterium nucleatum to fibronectin immobilized on gingival epithelial cells or glass coverslips

Abstract

To learn more about colonization of the oral epithelium by Fusobacterium nucleatum and the role of fibronectin in mediating adhesion of this microorganism, we studied attachment of this bacterium to cultured gingival epithelial cells that were coated with exogenous, purified plasma fibronectin. The three strains of F. nucleatum studied adhered in large numbers to epithelial cells that had been coated with fibronectin, compared with buffer-coated control cells. Bacterial adherence was also enhanced when epithelial cells were coated with whole human saliva. However, cells coated with saliva depleted of fibronectin did not facilitate adhesion of bacteria. Bacterial adhesion was restored when purified fibronectin was added back. We also tested adherence of bacteria to coverslips coated with fibronectin, saliva, and saliva depleted of fibronectin. The bacteria adhered to coverslips coated with fibronectin or whole human saliva, but did not adhere to coverslips coated with fibronectin-depleted saliva. Bacterial adhesion to coverslips was restored upon addition of purified fibronectin to the fibronectin-depleted saliva. Bacterial attachment to fibronectin-coated coverslips was found to be temperature-dependent, with maximal adhesion observed at 37 degrees C. Pre-treatment of F. nucleatum with soluble fibronectin inhibited attachment of the bacteria by 92%, whereas pre-treatment with bovine serum albumin had no effect. Pre-treatment of bacteria with laminin or type IV collagen caused moderate inhibition of attachment by 60% and 50%, respectively. Treatment of fibronectin-coated coverslips with Fab fragments of anti-fibronectin IgG blocked the attachment of F. nucleatum by 93%. Fab fragments of the other antisera tested had no inhibitory effect.(ABSTRACT TRUNCATED AT 250 WORDS)