Covalently grafted polyethylene oxide) coatings have been widely studied for use in biomedical applications, particularly for the reduction of protein and other biomolecule adsorption. However, many of these studies have not characterized the hydrated structure of the coatings. This new study uses a combination of silica colloid probe interaction force measurements using atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) in order to determine the grafting density and hydrated layer structure of monomethoxy polyethylene oxide) aldehyde layers, covalently grafted onto amine plasma polymer surfaces, and their interactions with silica surfaces. For high grafting densities, purely repulsive interactions were measured as expected for densely grafted polymer brushes. These interactions could be described by theoretical expectations for compression of one polymer brush layer. However, at lower grafting densities, attractive interactions were observed at larger separation distances, originating from bridging interactions due to adsorption of the PEO chains on the surface of the silica colloid probe. This is a new finding indicating that the coupled PEO molecules have sufficient conformational freedom to interact strongly with an adjacent surface or, for example, protein molecules for which there is an affinity. The attractive interactions could be removed by grafting an additional PEO layer onto the silica colloid probe. Protein adsorption measurements confirmed that at high grafting densities, the amount of adsorbed protein on the PEO grafted surfaces was greatly reduced, to the order of the detection limit for the XPS technique.