Nanostructured polyelectrolyte multilayer thin films electrostatically assembled alternately from such polymers as poly(allylamine hydrochloride) (PAH) and poly(acrylic acid) (PAA) were investigated for their in vitro cell interactions. Not surprisingly, NR6WT cells, a highly adhesive murine fibroblast cell line, attached to many different multilayer combinations tested. However, PAH/PAA multilayers constructed at pH deposition conditions of 2.0/2.0 were completely bioinert. Analogous cell interactions were observed with PAH/poly(methacrylic acid) (PAH/PMA), PAH/sulfonated poly(styrene) (PAH/SPS), and poly(diallyldimethylammonium chloride)/SPS (PDAC/SPS) systems, thereby suggesting a general trend in the fibroblasts' response to multilayers. Specifically, highly ionically stitched films attracted cells, whereas weakly ionically cross-linked multilayers, which swell substantially in physiological conditions to present richly hydrated surfaces, resisted fibroblast attachment. Thus, by manipulating the multilayer pH or ionic strength assembly conditions or both, which in turn dictate the molecular architecture of the thin films, one may powerfully direct a single multilayer combination to be either cell adhesive or cell resistant.
Polyelectrolyte multilayers of poly(acrylic acid) (PAA) and polyacrylamide (PAAm) were prepared via hydrogen-bonding interactions. These multilayers as assembled were stable at low pH but dissolved quickly in neutral pH water. We developed methods for stabilizing these multilayers to high pH through cross-linking by heating or UV-irradiation. Thermal treatment of the multilayers, which resulted in a partial imidization between carboxylic acid and amide groups, gave the multilayer good stability at high pH. In addition, we introduced photoreactive groups in the multilayer, which rendered the film insoluble after UV irradiation. Using these selective stabilization approaches, we have succeeded in micropatterning these films by ink-jet printing and photolithography to create subtractive patterns.
A newly discovered class of cell resistant surfaces, specifically engineered polyelectrolyte multilayers, was patterned with varying densities of adhesion ligands to control attachment of mammalian cells and to study the effects of ligand density on cell activity. Cell adhesive patterns were created on cell resistant multilayer films composed of poly(acrylic acid) and polyacrylamide through polymer-on-polymer stamping of poly(allylamine hydrochloride) PAH and subsequent reaction of the amine functional groups with an adhesion ligand containing RGD (Arg-Gly-Asp). These cell patterns demonstrated great promise for long-term applications since they remained stable for over 1 month, unlike ethylene glycol functional surfaces. By changing the stamping conditions of PAH, it was possible to alter the number of available functional groups in the patterned regions, and as a result, control the ligand density. Cell spreading, morphology, and cytoskeletal organization were compared at four different RGD densities. The highest RGD density, approximately 152 000 molecules/microm2, was created by stamping PAH at a pH of 11.0. Lowering the stamping ink pH led to patterns with lower ligand surface densities (83 000 molecules/microm2 for pH 9.0, 53,000 molecules/ microm2 for pH 7.0, and 25 000 molecules/microm2 for pH 3.5). An increasing number of cells attached and spread as the RGD density of the patterns increased. In addition, more cells showed well-defined actin stress fibers and focal adhesions at higher levels of RGD density. Finally, we found that pattern geometry affected cytoskeletal protein organization. Well-formed focal adhesions and cell-spanning stress fibers were only found in cells on wider line patterns (at least 25 microm in width).
Hydrogen-bonded multilayers comprised of polyacrylamide (PAAm) and a weak polyelectrolyte, such as poly(acrylic acid) (PAA) or poly(methacrylic acid) (PMA), were investigated for their surface-cell interactions. The assembled films were lightly cross-linked thermally or photochemically in order to render them stable in a physiological environment. Both PAA/PAAm and PMA/PAAm multilayers were found to exhibit a high resistance to the adhesion (cytophobicity) of mammalian fibroblasts, even with only a single bilayer coating. Protein adsorption to the multilayers, as revealed by surface plasmon resonance measurements, was greatly reduced for fibronectin and serum-containing medium. In situ swelling experiments indicate that the H-bonded multilayers are hydrogellike coatings capable of a high level of swelling in buffered solution. Utilizing the H-bonding nature of these multilayers, we were able to micropattern the films to create more complex cell-resistant/-adhesive surfaces. The long-term stability of the cell-resistant multilayers was found to be exceptionally good even under conditions (pH 7.4, buffered solution) where a high degree of swelling takes place. No degradation of the micropatterned films was observed over a period of a month, during which time the multilayer coatings remained highly resistant to cell-adhesion.
Bioinert polyelectrolyte multilayers comprised of poly(acrylic acid) and polyacrylamide were deposited on colloidal particles (1.7 µm in diameter) at low pH conditions by layer-by-layer assembly using hydrogenbonding interactions. The multilayer films were coated uniformly on the colloidal particles without causing any flocculation of the colloids, and the deposited films were subsequently cross-linked by a single treatment of a carbodiimide aqueous solution. The lightly cross-linked multilayer films show excellent stability at physiological conditions (pH 7.4, phosphate-buffered saline), whereas untreated multilayer films dissolved. The multilayer-coated surfaces, both on flat substrates and on colloidal particles, exhibit excellent resistance toward mammalian cell adhesion. With this new solution-based cross-linking method, bioinert H-bonded multilayer coatings offer potential for biomedical applications.
To develop high-energy density dielectric materials, hybrid BaTiO 3 particles with diblock copolymer shielding layers were fabricated and the dielectric properties of barium titanate-copolymer/polystyrene composites were examined. A significantly large breakdown strength of up to 222 V/μm at high permittivity was obtained. In our approach to formation of a robust polymer shell, polystyrene-blockpoly(styrene-co-vinylbenzylchloride) (PS-b-PSVBC) was introduced for the complete wrapping of the BT nanoparticles. The two-step processes to form polymer shielding and the resulting self-assembled layers result in excellent dielectric properties, where a charged layer is buried to minimize the development of the breakdown path and leakage current. Consequently, the maintained high breakdown strength, in spite of the large incorporation of BT particles to give high permittivity, afforded a noticeable energy density value of 9.7 J/cm 3 .
We propose and demonstrate a computational method for complex-field imaging from many noisy intensity images with varying defocus, using an extended complex Kalman filter. The technique offers dynamic smoothing of noisy measurements and is recursive rather than iterative, so is suitable for adaptive measurements. The Kalman filter provides near-optimal results in very low-light situations and may be adapted to propagation through turbulent, scattering, or nonlinear media.
We demonstrate a method for single-shot quantitative phase imaging based on the transport of intensity equation (TIE) in a volume holographic microscope (VHM). The VHM system uses a multiplexed volume hologram to laterally separate images from different focal planes. This axial intensity information is then used to solve the TIE and recover object phase quantitatively. Further, we show improved phase recovery by using five multiplexed gratings in one hologram.
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