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Rational design and informed development of nontoxic antifouling coatings requires a thorough understanding of the interactions between surfaces and fouling species. With more complex antifouling materials, such as composites or zwitterionic polymers, there follows also a need for better characterization of the materials as such. To further the understanding of the antifouling properties of charge-balanced polymers, we explore the properties of layered polyelectrolytes and their interactions with charged surfaces. These polymers were prepared via self-initiated photografting and photopolymerization (SIPGP); on top of a uniform bottom layer of anionic poly(methacrylic acid) (PMAA), a cationic poly(2-dimethylaminoethyl methacrylate) (PDMAEMA) thickness gradient was formed. Infrared microscopy and imaging spectroscopic ellipsometry were used to characterize chemical composition and swelling of the combined layer. Direct force measurements by colloidal probe atomic force microscopy were performed to investigate the forces between the polymer gradients and charged probes. The swelling of PMAA and PDMAEMA are very different, with steric and electrostatic forces varying in a nontrivial manner along the gradient. The gradients can be tuned to form a protein-resistant charge-neutral region, and we demonstrate that this region, where both electrostatic and steric forces are small, is highly compressed and the origin of the protein resistance of this region is most likely an effect of strong hydration of charged residues at the surface, rather than swelling or bulk hydration of the polymer. In the highly swollen regions far from charge-neutrality, steric forces dominate the interactions between the probe and the polymer. In these regions, the SIPGP polymer has qualitative similarities with brushes, but we were unable to quantitatively describe the polymer as a brush, supporting previous data suggesting that these polymers are cross-linked.

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Herein, we present an easy-to-use protein and cell patterning method relying solely on pipetting, rinsing steps and illumination with a desktop lamp, which does not require any expensive laboratory equipment, custom-built hardware or delicate chemistry. This method is based on the adhesion promoter poly(allylamine)-grafted perfluorophenyl azide, which allows UV-induced cross-linking with proteins and the antifouling molecule poly(vinylpyrrolidone). Versatility is demonstrated by creating patterns with two different proteins and a polysaccharide directly on plastic well plates and on glass slides, and by subsequently seeding primary neurons and C2C12 myoblasts on the patterns to form islands and mini-networks. Patterning characterization is done via immunohistochemistry, Congo red staining, ellipsometry, and infrared spectroscopy. Using a pragmatic setup, patterning contrasts down to 5 µm and statistically significant long-term stability superior to the gold standard poly(l-lysine)-grafted poly(ethylene glycol) could be obtained. This simple method can be used in any laboratory or even in classrooms and its outstanding stability is especially interesting for long-term cell experiments, e.g., for bottom-up neuroscience, where well-defined microislands and microcircuits of primary neurons are studied over weeks.

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Biomaterials used in the ocular environment should exhibit specific tribological behavior to avoid discomfort and stress-induced epithelial damage during blinking. In this study, two macromolecules that are commonly employed as ocular biomaterials, namely, poly(vinylpyrrolidone) (PVP) and hyaluronan (HA), are compared with two known model glycoproteins, namely bovine submaxillary mucin (BSM) and α1-acid glycoprotein (AGP), with regard to their nonfouling efficiency, wettability, and tribological properties when freely present in the lubricant, enabling spontaneous adsorption, and when chemisorbed under low contact pressures. Chemisorbed coatings were prepared by means of photochemically triggered nitrene insertion reactions. BSM and AGP provided boundary lubrication when spontaneously adsorbed in a hydrophobic contact with a coefficient of friction (CoF) of ∼0.03-0.04. PVP and HA were found to be excellent boundary lubricants when chemisorbed (CoF ≤ 0.01). Notably, high-molecular-weight PVP generated thick adlayers, typically around 14 nm, and was able to reduce the CoF below 0.005 when slid against a BSM-coated poly(dimethylsiloxane) pin in a tearlike fluid.

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PURPOSE: To characterize the effect of lubricant composition and in vitro ageing on the coefficient of friction (CoF) of a wide range of commercially available soft contact lenses (SCLs). METHODS: The CoF of SCLs was characterized by means of microtribometry against a mucin-coated glass disk. One reusable (RU) silicone-hydrogel (SiHy) lens, senofilcon A, and two daily disposable (DD) lenses, etafilcon A (hydrogel) and nelfilcon A (hydrogel), were tested under different lubricant solutions, including a tear-like fluid (TLF) containing proteins and lipids. Five RU (balafilcon A [SiHy], comfilcon A [SiHy], etafilcon A [hydrogel], lotrafilcon B [SiHy], senofilcon A [SiHy]) and five DD (delefilcon A [SiHy], etafilcon A [hydrogel; two lens types], narafilcon A [SiHy], nelfilcon A [hydrogel]) lenses were tested before and after exposure to an in vitro ageing process, consisting of continuous immersion and withdrawal from TLF for 18 hours. The CoF in TLF was further compared to previously published data collected in a different lubricant. RESULTS: After in vitro ageing, three RU (balafilcon A, etafilcon A, comfilcon A) and three DD (delefilcon A, etafilcon A, nelfilcon A) lenses displayed a significant increase in CoF (P < 0.05). Lenses that contained poly (vinyl pyrrolidone; PVP) showed unaltered CoF after ageing. CONCLUSIONS: An in vitro methodology to simulate in vivo wearing of contact lenses has been proposed. The results suggest that certain lens materials show increased CoF after ageing, with potential clinical implications. The results indicate that the presence of a persistent wetting agent is of advantage to maintain a low CoF after prolonged wearing.

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Bioadhesive micropatterns, capable of laterally confining cells to a 2D lattice, have proven effective in simulating the in vivo tissue environment. They reveal fundamental aspects of the role of adhesion in cell mechanics, proliferation, and differentiation. Here we present an approach based on photochemistry for the fabrication of synthetic polymer micropatterns. Perfluorophenyl azide (PFPA), upon deep-UV exposure, forms a reactive nitrene capable of covalently linking to a molecule that is in close proximity. PFPA has been grafted onto a backbone of poly(allyl amine), which readily forms a self-assembled monolayer on silicon wafers or glass. A film of polystyrene was applied by spin-coating, and by laterally confining the UV exposure through a chromium-on-quartz photomask, monolayers of polymers could be immobilized in circular microdomains. Poly(vinylpyrrolidone) (PVP) was attached to the background to form a barrier to nonspecific protein adsorption and cell adhesion. Micropatterns were characterized with high-lateral-resolution time-of-flight secondary ion mass spectrometry (TOF-SIMS), which confirmed the formation of polystyrene domains within a PVP background. Fluorescence-microscopy adsorption assays with rhodamine-labeled bovine serum albumin demonstrated the nonfouling efficiency of PVP and, combined with TOF-SIMS, allowed for a comprehensive characterization of the pattern geometry. The applicability of the micropatterned platform in single-cell assays was tested by culturing two cell types, WM 239 melanoma cells and SaOs-2 osteoblasts, on micropatterned glass, either with or without backfilling of the patterns with fibronectin. It was demonstrated that the platform was efficient in confining cells to the fibronectin-backfilled micropatterns for at least 48 h. PVP is thus proposed as a viable, highly stable alternative to poly(ethylene glycol) for nonfouling applications. Due to the versatility of the nitrene-insertion reaction, the platform could be extended to other polymer pairs or proteins and the surface chemistry adapted to specific applications.

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We report on the preparation and characterization of thin polyampholytic hydrogel gradient films permitting pH-controlled protein resistance via the regulation of surface charges. The hydrogel gradients are composed of cationic poly(2-aminoethyl methacrylate hydrochloride) (PAEMA), and anionic poly(2-carboxyethyl acrylate) (PCEA) layers, which are fabricated by self-initiated photografting and photopolymerization (SIPGP). Using a two-step UV exposure procedure, a polymer thickness gradient of one component is formed on top of a uniform layer of the oppositely charged polymer. The swelling of the gradient films in water and buffers at different pH were characterized by imaging spectroscopic ellipsometry. The surface charge distribution and steric interactions with the hydrogel gradients were recorded by direct force measurement with colloidal-probe atomic force microscopy. We demonstrate that formation of a charged polymer thickness gradient on top of a uniform layer of opposite charge can result in a region of charge-neutrality. This charge-neutral region is highly resistant to non-specific adsorption of proteins, and its location along the gradient can be controlled via the pH of the surrounding buffer. The pH-controlled protein adsorption and desorption was monitored in real-time by imaging surface plasmon resonance, while the corresponding redistribution of surface charge was confirmed by direct force measurements.

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A versatile, photochemical surface-modification approach using nitrene-insertion reactions has been employed to develop an ultrathin, two-component, polymer-gradient coating. Perfluorophenyl azide (PFPA) acted as the photosensitive moiety, forming a nitrene radical upon 254 nm UV exposure. Cationic poly(allyl amine) was grafted with PFPA and surface-anchored onto silicon wafers by means of electrostatic self-assembly. After spin-coating of polystyrene (PS), the substrate was illuminated from behind a moving shutter, thereby controlling the azide-to-nitrene conversion degree across the substrate, and leading to a gradually varying PS density after rinsing. Backfilling with poly(vinyl pyrrolidone) (PVP) and re-exposing to UV light formed a two-component polymer-density gradient. The composition varied linearly following exposure to a linear UV exposure profile, as determined with spectroscopic ellipsometry (ELM) and X-ray photoelectron spectroscopy (XPS). High-spatial-resolution, time-of-flight secondary ion mass spectrometry (ToF-SIMS) revealed a high degree of mixing between the two incompatible polymers on the micrometer scale. The dynamic water-contact angle (dCA) was found to depend strongly on the sample history, suggesting adaptive properties of the coating, which was further confirmed by angle-resolved XPS (ARXPS). To confirm the applicability of the system for biological investigations, gradients were exposed to zoospores of the macrofouling alga Ulva linza , and a critical PS composition of 70% was identified, above which settlement started to increase. It has been shown that a two-component polymer-density gradient can provide a high-throughput platform for determining critical surface properties of polymer blend materials.

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