Layer-by-layer polyelectrolyte films made of the naturally derived polysaccharides chitosan (CHI) and hyaluronic acid (HA) constitute a well-studied system for the development of cell-responsive biointerfaces. However, many cell lines exhibit decreased adhesion to CHI/HA multilayer films, particularly as the number of bilayers is increased. Here, our group demonstrates that films composed of glycol-modified chitosan exhibit significantly improved MC3T3 preosteoblast adhesion and viability compared to corresponding films consisting of unmodified CHI. These differences in cellular adhesion are likely due to differences in surface topography and roughness, as measured via atomic force microscopy (AFM), as well as in film chemistry and the water solubility of the cation, since both types of films exhibited similar: thickness, as measured via quartz crystal microbalance and AFM; wettability, as measured via contact angle; and serum protein adsorption, as measured via the bicinchoninic acid assay. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2012.