Poly(acrylic acid) (PAA) brushes and poly(methacrylic acid) (PMAA) brushes on gold substrates were synthesized by surface-initiated atom-transfer radical polymerization of sodium acrylate and sodium methacrylate in water media at room temperature. Fourier transform infrared spectroscopy (FTIR) titration and contact angle titration methods were used in combination to investigate the dissociation behavior of these two brushes. Whereas FTIR titration gives effective bulk pKa values of the polyacid brushes (pKabulk of PAA brushes is 6.5-6.6 and pKabulk of PMAA brushes is 6.9-7.0), contact angle titration provides effective surface pKa of the brushes (pKasurf of PAA brushes is 4.4+/-0.01 and pKasurf of PMAA brushes is approximately 4.6+/-0.1). The difference between pKabulk and pKasurf suggests that acid groups further from the substrate surface are easier to ionize and have smaller pKa values. Although such behavior of weak polyelectrolyte brushes has been predicted by theoretical simulation, here we provide the first experimental evidence of this behavior.

Protein patterning was carried out using a simple procedure based on photolithography wherein the protein was not subjected to UV irradiation and high temperatures or contacted with denaturing solvents or strongly acidic or basic solutions. Self-assembled monolayers of poly(ethylene glycol) (PEG) on silicon surfaces were exposed to oxygen plasma through a patterned photoresist. The etched regions were back-filled with an initiator for surface-initiated atom transfer radical polymerization (ATRP). ATRP of sodium acrylate was readily achieved at room temperature in an aqueous medium. Protonation of the polymer resulted in patterned poly(acrylic acid) (PAA) brushes. A variety of biomolecules containing amino groups could be covalently tethered to the dense carboxyl groups of the brush, under relatively mild conditions. The PEG regions surrounding the PAA brush greatly reduced nonspecific adsorption. Avidin was covalently attached to PAA brushes, and biotin-tagged proteins could be immobilized through avidin-biotin interaction. Such an immobilization method, which is based on specific interactions, is expected to better retain protein functionality than direct covalent binding. Using biotin-tagged bovine serum albumin (BSA) as a model, a simple strategy was developed for immobilization of small biological molecules using BSA as linkages, while BSA can simultaneously block nonspecific interactions.

Various designs for coatings that resist the attachment of marine organisms are based on the concept of "ambiguous" surfaces that present both hydrophobic and hydrophilic functionalities as surface domains. In order to facilitate the optimal design of such surfaces, information is needed on the scale of the domains that the settling stages of marine organisms are able to distinguish. Previous experiments showed that Ulva zoospores settle (attach) in high numbers onto fluorinated monolayers compared to PEGylated monolayers. The main aim of the present study was to determine, when zoospores of the green alga Ulva are presented with a choice of fluorinated or PEGylated surfaces, what the minimum dimensions of the two types of surface are that zoospores can detect and consequently settle on. Silicon wafers were chemically modified to produce a pattern of squares containing alternating fluorinated and PEGylated stripes of different widths on either a uniform fluorinated or PEGylated background. Each 1 cm x 1 cm square contained stripes with widths of 500, 200, 100, 50, 20, 5, or 2 microm as well as an unpatterned square with a chemistry opposite that of the background. Spores were selective in choosing where to settle, settling at higher densities on fluorinated stripes compared to PEGylated stripes. However, the magnitude of response, and the consequences for settlement on patterned areas overall, was dependent on both the width of the stripes and the chemistry of the background. The data are discussed in relation to the ability of spores to "choose" favorable sites for settlement and the implications for the development of novel antifouling coatings.

Surface active block copolymers (SABCs) with amphiphilic side chains containing ethoxylated fluoroalkyl groups have previously demonstrated advantageous properties with regard to marine fouling resistance and release. While it was previously postulated that the ability of the block copolymer surface to undergo an environment-dependent transformation in surface structure aided this behaviour, protein adsorption characteristics of the surface were never explored. This study aims to expand our knowledge of protein interaction with the amphiphilic surface active block copolymer in an aqueous environment through experiments with bovine serum albumin (BSA), a widely utilized test protein. Fluorescence microscopy analysis using BSA labelled with fluorescein isothiocyanate (BSA-FITC) was performed on a SABC test surface to establish the polymer's protein adsorption resistance. Additionally, atomic force microscopy (AFM) based chemical force microscopy (CFM) was utilized to examine the force of adhesion of an AFM tip functionalized with strands of BSA protein with the SABC. No measurable force of adhesion was detected for 58% of the measurements of adhesion force taken for a BSA coated AFM tip interacting with the surface of the amphiphilic SABC in a PBS buffer. Furthermore, no measurements of force of adhesion were made in excess of 0.15 nN. This was in contrast to the non-zero mean adhesion force seen for several control surfaces in PBS buffer.

We use patterned poly(acrylic acid) (PAA) polymer brushes to explore the effects of surface chemistry and topography on cell-surface interactions. Most past studies of surface topography effects on cell adhesion have focused on patterned feature sizes that are larger than the dimensions of a cell, and PAA brushes have been characterized as cell repellent. Here we report cell adhesion studies for RBL mast cells incubated on PAA brush surfaces patterned with a variety of different feature sizes. We find that when patterned at sub-cellular dimensions on silicon surfaces, PAA brushes that are 30 or 15 nm thick facilitate cell adhesion. This appears to be mediated by fibronectin, which is secreted by the cells, adsorbing to the brushes and then engaging cell surface integrins. The result is detectable accumulation of plasma membrane within the brushes, and this involves cytoskeletal remodeling at the cell-surface interface. By decreasing brush thickness, we find that PAA can be ‘tuned’ to promote cell adhesion with down-modulated membrane accumulation. We exemplify the utility of patterned PAA brush arrays for spatially controlling the activation of cells by modifying brushes with ligands that specifically engage IgE bound to high affinity receptors on mast cells.