Preferential adsorption of cell adhesive proteins from complex media on self-assembled monolayers and its effect on subsequent cell adhesion.

We examined the effect of surface chemistry on adsorption of fibronectin (Fn) and vitronectin (Vn) and subsequent cell adhesion, employing self-assembled monolayers (SAMs) of alkanethiols carrying terminal methyl (CH3), hydroxyl groups (OH), carboxylic acid (COOH), and amine (NH2). More Fn and Vn adsorbed to COOH- and NH2-SAMs than to CH3- and OH-SAMs from a mixture with bovine serum albumin (BSA) and from 2% fetal bovine serum. Adhesion of human umbilical vein endothelial cells (HUVECs) on CH3- and OH-SAMs preadsorbed with Fn and BSA decreased with decreasing adsorbed Fn; however, HUVECs adhered to COOH- and NH2-SAMs even in the presence of BSA at 1000-fold more than Fn in a mixture because of the preferential adsorption of Fn and/or displacement of preadsorbed BSA with Fn and Vn in a serum-containing medium. SAMs coated with a mixture of Vn and BSA exhibited adhesion of HUVECs regardless of surface functional groups. A well-organized focal adhesion complex and actin stress fibers were observed only for COOH- and NH2-SAMs when SAMs were preadsorbed with Vn and BSA. These results suggest that COOH- and NH2-SAMs allow for both cell adhesion and cell spreading because of the high density of cell-binding domains derived from adsorbed Vn. STATEMENT OF SIGNIFICANCE: Adsorption of cell adhesive proteins including fibronectin (Fn) and vitronectin (Vn) plays an important role in cell adhesion to artificial materials. However, for the development of biomaterials that contact with biological fluids, it is important to understand adsorption of Fn and Vn in complex media containing many kinds of proteins. Here, we focused on adsorption of Fn and Vn from complex media including mixed solution with albumin and fetal bovine serum, and its role on cell adhesion using self-assembled monolayers (SAMs). Our result demonstrates that SAMs carrying carboxylic acid or amine allow for both cell adhesion and cell spreading because of preferentially adsorbed Vn. The result provides insights into surface design of cell culture substrates and tissue engineering scaffolds.