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Emdogain[R] interaction with extracellular matrix proteins and their effects on cell adhesion, spreading and migration Narani, Nazanin

Abstract

Studies over the past twenty years have demonstrated that the cells of Hertwig's epithelial root sheath secrete enamel matrix proteins and that these proteins are involved in the formation of acellular extrinsic fiber cementum during development and regeneration of periodontal tissues (Slavkin and Boyde, 1975; Hammarstrom et al., 1997b). However, the cellular and molecular mechanisms involved in the process of periodontal regeneration induced by enamel matrix proteins are poorly understood. This in vitro study was designed to investigate Emdogain's interaction with extracellular matrix (ECM) proteins and regulation of adhesion, spreading and migration of epithelial cells and periodontal ligament fibroblasts. We examined the binding interaction between Emdogain and the ECM proteins, fibronectin; collagen types I and IV; and laminin I using ELISA. Binding of Emdogain to fibronectin and collagen type I (10 µg/ml) was inversely related to Emdogain coating concentration and resulting precipitation. That is, fibronectin and collagen type I bound to non-precipitated Emdogain and precipiation of Emdogain at higher concentrations did not bind fibronectin or collagen type I. Type IV collagen and laminin I did not bind to any concentration of Emdogain-coated surfaces. Epithelial cells plated on Emdogain-coated surfaces (1-30,000 µg/ml) in the presence of cylcoheximide, showed minimal adhesion (5%) compared to type I collagen-coated surfaces (positive control) and no spreading was found at 2 hours. When fibronectin or collagen type I (10 µg/ml) was used along with Emdogain, epithelial cell adhesion was significantly enhanced (3-5 fold higher compared to Emdogain alone). PDL fibroblast adhesion and spreading on Emdogain (1-30,000 µg/ml) was concentration-dependent with the maximum adhesion and spreading on 10,000 µg/ml (40% and 10% of positive control at 2 hours, respectively). Fibronectin or collagen type I (10 µg/ml) used along with Emdogain for surface coating enhanced adhesion to a level that was comparable with positive control. Cell spreading was also improved and reached 30% compared to 70% on positive control. Collectively these data indicate that Emdogain supports adhesion and spreading of PDL fibroblasts but not epithelial cells. Addition of ECM proteins enhances cell binding to Emdogain that is more significant for PDL fibroblasts compared to epithelial cells.

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