Separation of aminophenol isomers in polyelectrolyte multilayers modified PDMS microchip

The separation of three kinds of aminophenol isomers were achieved within 1 min in polyelectrolytes multilayers modified PDMS microchips by layer-by-layer assembly with electrochemical detection (EC). Two polyelectrolytes, poly(dially dimethyl ammonium chloride) (PDDA) and poly(sodium-4-styrene-sulfonate) (PSS) were used to form polyelectrolyte multilayers (PEMs). The surface characteristic of the modified microchip was studied by XPS. The electroosmotic flow (EOF) on PEMs modified PDMS microchips was more stable than that of the native PDMS microchips and the adsorption of samples was greatly reduced on PEMs modified PDMS microchips during the electrophoretic process. The column efficiencies on PEMs modified microchip were increased by 100 times and the signals enhanced by 2 times compared with those of native microchips. The separation conditions such as running buffer pH, running buffer concentration and separation voltage were also optimized.     

Phase separations in pH-responsive polyelectrolyte multilayers: charge extrusion versus charge expulsion

Polyelectrolyte multilayers with continuously variable amounts of ionizable weak acid functionality were prepared by blending ionizable and nonionizable polyelectrolytes in the deposition solutions. Diluting ionizable groups in this way yielded multilayers that were more structurally stable, shown by thickness and atomic force microscopy measurements, as their internal polymer charge was varied by the pH of the external solution. Multilayers prepared with opposite surface charge to that appearing within the bulk (as a result of ionization) were more stable, as were thinner films, both results suggesting the extrusion of bulk charge to the surface. These multilayers were able to control the direction and magnitude of electroosmotic flow in microfluidics systems. Multilayers bearing only one, diluted layer ofionizable material were surprisingly effective in this respect.     

Cationic poly(methacryl oxyethyl trimethylammonium) and its poly(ethylene glycol)‐grafted analogue as capillary coating materials in electrophoresis

Cationic polyelectrolytes were synthesized and used as semipermanent coating materials for capillaries in electrophoresis. The polyelectrolytes used were a homopolymer of poly(methacryl oxyethyl trimethylammonium chloride) (PMOTAC) and its poly(ethylene glycol) (PEG)‐grafted analogue. Two PMOTAC polyelectrolytes, with molar masses of 85,000 and 300,000 g/mol, and PEG‐grafted PMOTAC with a molar mass of 280,000 g/mol were synthesized and then characterized by size exclusion chromatography (SEC) and nuclear magnetic resonance (NMR) spectroscopy. Attachment of the polyelectrolytes to the wall of the fused silica capillary for electrophoresis caused the electroosmotic flow (EOF) to reverse. The polyelectrolyte coatings were tested over the pH range 2–11 at different buffer ionic strengths, and the most stable and strongest anodic EOFs were obtained at acidic pH values with low ionic strength buffers. Between runs the capillary is merely rinsed for 2 or 3 min with the background electrolyte solution. With the PMOTAC coatings at pH values ≤5, the RSDs of the EOFs were less than 2.9% after 60 injections. The effects of the molar mass of the polycation and of PEGylation of PMOTAC on the interactions between the polycations and basic proteins were studied at acidic pH values. The differences in the effective electrophoretic mobilities, resolution values, and plate numbers of the proteins with the different coatings were due to the EOF, as demonstrated through calculations of reduced mobilities, relative resolution values, and relative plate numbers. © 2007 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 45: 2655–2663, 2007     

Film stability during postassembly morphological changes in polyelectrolyte multilayers due to acid and base exposure

The mechanism of the transition from a continuous morphology to a porous morphology within polyelectrolyte multilayers (PEMs) of linear poly(ethylene imine) (LPEI) and poly(acrylic acid) (PAA) and poly(allylamine hydrochloride) (PAH) and PAA assembled by the layer-by-layer (LbL) technique is examined. These morphological changes were created by both acidic and basic postassembly treatments. Basic postassembly treatment is shown to create different types of porosity than acidic postassembly treatment. The morphological variation from the introduction of porosity to the collapse of these porous structures and the dissolution of films under postassembly treatments was observed by AFM, optical microscopy, quartz crystal microbalance (QCM), and SEM. These morphological transitions which are a result of structural rearrangement of weak polyelectrolytes due to pH changes are closely related to the neutralization of the polycations and the ionization of polyanions. Results obtained from FTIR spectroscopy and QCM confirm that polyelectrolytes are being selectively or partially released from the polyelectrolyte multilayers thin films (PEMs) in response to the pH treatment as a function of exposure time. In conclusion, here new information is presented about the structural reorganization found in a number of weak polyelectrolyte systems. This information will be useful in designing functional materials based on polyelectrolytes.     

Novel cationic polyelectrolyte coatings for capillary electrophoresis

The use of bare fused silica capillary in CE can sometimes be inconvenient due to undesirable effects including adsorption of sample or instability of the EOF. This can often be avoided by coating the inner surface of the capillary. In this work, we present and characterize two novel polyelectrolyte coatings (PECs) poly(2‐(methacryloyloxy)ethyl trimethylammonium iodide) (PMOTAI) and poly(3‐methyl‐1‐(4‐vinylbenzyl)‐imidazolium chloride) (PIL‐1) for CE. The coated capillaries were studied using a series of aqueous buffers of varying pH, ionic strength, and composition. Our results show that the investigated polyelectrolytes are usable as semi‐permanent (physically adsorbed) coatings with at least five runs stability before a short coating regeneration is necessary. Both PECs showed a considerably decreased stability at pH 11.0. The EOF was higher using Good's buffers than with sodium phosphate buffer at the same pH and ionic strength. The thickness of the PEC layers studied by quartz crystal microbalance was 0.83 and 0.52 nm for PMOTAI and PIL‐1, respectively. The hydrophobicity of the PEC layers was determined by analysis of a homologous series of alkyl benzoates and expressed as the distribution constants. Our result demonstrates that both PECs had comparable hydrophobicity, which enabled separation of compounds with log P_o/w > 2. The ability to separate cationic drugs was shown with β‐blockers, compounds often misused in doping. Both coatings were also able to separate hydrolysis products of the ionic liquid 1,5‐diazabicyclo[4.3.0]non‐5‐ene acetate at highly acidic conditions, where bare fused silica capillaries failed to accomplish the separation.     

Polyelectrolyte multilayers and degradable polymer layers as multicompartment films

Polyelectrolyte multilayers are now a well established concept with numerous potential applications in particular as biomaterial coatings. To timely control the biological activity of cells in contact with a substrate, multicompartment films made of different polyelectrolyte multilayers deposited sequentially on the solid substrate constitute a promising new approach. In a first paper (Langmuir 2004, 20, 7298) we showed that such multicompartment films can be designed by alternating exponentially growing polyelectrolyte multilayers acting as reservoirs and linearly growing ones acting as barriers. In the present study, we first demonstrate however that these barriers composed of synthetic polyelectrolytes are not degraded despite the presence of phagocytic cells. We propose an alternative approach where exponentially growing poly(L-lysine)/hyaluronic acid (PLL/HA) multilayers, used as reservoirs, are alternated with biodegradable polymer layers consisting in poly(lactic-co-glycolic acid) (PLGA) and acting as barriers for PLL chains that diffuse within the PLL/HA reservoirs. We first show that these PLGA layers can be deposited alternatively with PLL/HA multilayers leading to polyelectrolyte multilayer/hydrolyzable polymeric layer films and acting as a reservoirs/barriers system. Bone marrow cells seeded on these films ending by a PLL/HA reservoir rapidly degrade it and internalize the PLL chains confined in this reservoir. Then the cells degraded locally the PLGA barrier and internalize the PLL localized in a lower (PLL/HA) compartment after 5 days of seeding. By changing the thickness of the PLGA layer, we hope to be able to tune the time delay of degradation. Such mixed architectures made of polyelectrolyte multilayers and hydrolyzable polymeric layers could act as coatings allowing us to induce a time scheduled cascade of biological activities. We are currently working on the use of comparable films with compartments filled by proteins or peptides and in which the degradation of the barriers results from a hydrolysis over tunable time scales.     

Biogenic Polyelectrolyte Multilayers on Poly(L-lactide) Films for Control of Osteoblast Adhesion

Cell adhesion and spreading are important events during cell-biomaterial interaction, which control survival, growth and differentiation of cells. Layer-by-layer technique was used to generate multilayer coatings for regulating adhesion of primary osteoblasts on biomaterials. Polyelectrolyte multilayers (PEM) were based on poly (ethylene imine) as primary polycation layer. PEM were then prepared from chitosan (CHI) as polycation and heparin (HEP), or sulfated HA (sHA) as polyanions. It was observed that attachment and spreading of primary osteoblasts (pOB) was highly dependent on the composition of multilayers, as well as pH values of polyelectrolyte solutions. Results presented in this paper may pave the way for application of PEM surface coatings for bone-contacting implant materials.     

Comparison of polyelectrolyte multilayers built up with polydiallyldimethylammonium chloride and poly(ethyleneimine) from salt-free solutions by in-situ surface plasmon resonance measurements

Polyelectrolytes offer a widespread potential for the defined modification of planar inorganic or polymer surfaces. Essential parameters for the regular adsorption of subsequent polymer layers by electrostatic interactions are the charge of polyelectrolyte and of the outermost surface region, the surface of the substrate, and the molar mass of the polyelectrolyte. To study such effects in mono- and multilayers we used poly(diallyldimethylammonium chloride (PD) with a molar mass from 5000 to 400000 g/mol as a strong polycation and poly(ethyleneimine) (PEI) with 75000 g/mol as a weak polycation and poly(sodium styrenesulfonate) (PSS) from 70000 to 1Mio g/mol in the diluted and semi-diluted region. The characterization of the layers was performed by streaming potential, in-situ SPR and UV-Vis spectroscopy. Thereby the layer built up at the solid/liquid-interface could be followed and quantified at the molecular level. SPR revealed that the thicknesses of the multilayer depends strongly on pK values of the polyelectrolyte (strong or weak) and the molar masses. We observed a linear growth if both polyelectrolytes are strong and an exponential growth if one polyelectrolyte is weak. The thickness increased with higher molar masses of the polyelectrolytes. The process was followed in-situ in short time steps.     

Weak Polyelectrolytes as Nanoarchitectonic Design Tools for Functional Materials: A Review of Recent Achievements

The ionization degree, charge density, and conformation of weak polyelectrolytes can be adjusted through adjusting the pH and ionic strength stimuli. Such polymers thus offer a range of reversible interactions, including electrostatic complexation, H-bonding, and hydrophobic interactions, which position weak polyelectrolytes as key nano-units for the design of dynamic systems with precise structures, compositions, and responses to stimuli. The purpose of this review article is to discuss recent examples of nanoarchitectonic systems and applications that use weak polyelectrolytes as smart components. Surface platforms (electrodeposited films, brushes), multilayers (coatings and capsules), processed polyelectrolyte complexes (gels and membranes), and pharmaceutical vectors from both synthetic or natural-type weak polyelectrolytes are discussed. Finally, the increasing significance of block copolymers with weak polyion blocks is discussed with respect to the design of nanovectors by micellization and film/membrane nanopatterning via phase separation.     

Dynamic capillary coatings for electroosmotic flow control in capillary electrophoresis

Control of the electroosmotic flow (EOF) is critical for achieving optimal separations by capillary electrophoresis. For instance, manipulation of the EOF can yield either high resolution separations or rapid analyses. Dynamic capillary coatings are a simple and cost-effective approach to altering the EOF. The normal EOF can be slowed using buffer additives such as Mg²⁺ and hexamethonium which ion exchange onto the surface silanols to lower the effective wall charge. Alternatively, cationic polyelectrolytes or cationic surfactants can be used to establish a cationic coating on the capillary wall, which results in a reversed EOF. Practical considerations such as pH stability and reproducibility obtainable with an EOF modifier will be discussed.     

Layer by layer self-assembled polyelectrolyte multilayers with embedded phospholipid vesicles obtained by spraying: integrity of the vesicles

In a previous paper (Michel, M.; Vautier, D.; Voegel, J.-C.; Schaaf, P.; Ball, V. Langmuir 2004, 20, 4835), we showed that phospholipid vesicles can be incorporated into poly(glutamic-acid)/poly(allylamine) (PGA/PAH) multilayered polyelectrolyte films built by the alternated dipping of a surface in polyanion and polycation solutions. AFM imaging, quartz crystal microbalance, and ellipsometry suggested that the vesicles remain intact when adhering on the surface. In the present paper, we show that such films can also be realized by spraying both the polyelectrolyte solutions and the vesicles onto the surface. Using such vesicles filled with ferrocyanide ions, we prove by cyclic voltammetry that the sprayed vesicles remain intact when embedded in the multilayers. We show that multilayers containing two distinct layers of intact vesicles separated by several polyanion/polycation bilayers can also be constructed. Polyelectrolyte multilayers containing layers of phospholipid vesicles could act as reservoirs for drug or other biologically active molecules in controlled release bioactive coatings.     

pH-sensitive nanostructured architectures based on synthetic and/or natural weak polyelectrolytes

This study aims to evidence the influence of polyelectrolytes structure and the number of double layers on the properties of some new nanostructured architectures formed by layer-by-layer self-assembly of complementary weak polyelectrolytes on planar surfaces. For this purpose, we used chitosan and poly(allylamine hydrochloride) as polycations, and poly(acrylic acid) and poly(2-acrylamido-2-methylpropanesulfonic acid–co-acrylic acid) as polyanions. To get a direct image on the polyelectrolyte multilayers formation and properties, gravimetry, infrared spectroscopy, and atomic force microscopy have been used. The capacity of the polyions to overcompensate the complementary polyions charges, and thus to influence the swelling degree in water of thin films, was strongly influenced by the chain structure and flexibility. A special attention was paid to the responsiveness of the new composite materials to the pH of the swelling environment.     

Energy transfer between conjugated polyelectrolytes in layer-by-layer assembled films

We present a study of Fo?rster resonance energy transfer (FRET) between two emissive conjugated polyelectrolytes (CPEs) in layer-by-layer (LbL) self-assembled films as a means of examining their organization and architecture. The two CPEs are a carboxylic acid functionalized polyfluorene (PFl-CO(2)) and thienylene linked poly(phenylene ethynylene) (PPE-Th-CO(2)). The PFl-CO(2) presents a maximum emission at 418 nm, while the PPE-Th-CO(2) has an absorption λ(max) centered at 431 nm, in sufficient proximity for effective FRET. Several LbL films have been constructed using varied concentrations of the deposition solutions and identity of the buffer layers separating the two emissive layers, using a system of either weak polyelectrolytes, poly(allylamine hydrochloride) (PAH)/poly(sodium methacrylate) (PMA), or strong polyelectrolytes, poly(diallylammonium chloride) (PDDA)/poly(styrene sulfonate) sodium (PSS). The efficiency of FRET has been monitored using fluorescence spectroscopy. Initially, the fluorescence of the PFl-CO(2) (E(g) ～ 3.0 eV), which emits at 420 nm, is quenched by the lower band gap PPE-Th-CO(2) (E(g) ～ 2.5 eV). For films using the PAH/PMA system as buffer bilayers and deposited from 1 mM solutions, the PFl-CO(2) fluorescence is progressively recovered as the number of intervening buffer bilayers is increased. Ellipsometry measurements indicate that energy transfer between the two emissive layers is efficient to a distance of ca. 7 nm.     

Micropatterning of polymer thin films with pH-sensitive and cross-linkable hydrogen-bonded polyelectrolyte multilayers

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.     

毛细管电泳中影响径向电场控制电渗的主要因素

利用自制的二维电场毛细管电泳系统研究了不同因素对径向电场控制电渗能力的影响,发现缓冲液的ｐＨ值、浓度、种类以及管壁表面状态、管径等对电渗的电场调控有关键性的影响。有趣的是,添加剂不影响电场的调控能力,而杯芳烃涂层毛细管却能提高电渗对径向电场的响应能力。利用这种涂层效应有可能实现较高ｐＨ值下电渗的电场调控。     

Correlating the compliance and permeability of photo-cross-linked polyelectrolyte multilayers

Photo-cross-linkable polyelectrolyte multilayers were made from poly(allylamine) (PAH) and poly(acrylic acid) (PAA) modified with a photosensitive benzophenone. Nanoindentation, using atomic force microscopy (AFM) of these and unmodified PAH/PAA multilayers, was used to assess their mechanical properties in situ under an aqueous buffer. Under the conditions employed (and a 20 nm radius AFM tip), reliable nanoindentations that appeared to be decoupled from the properties of the silicon substrate were obtained for films greater than 150 nm in thickness. A strong difference in the apparent modulus was observed for films terminated with positive as compared to negative polyelectrolytes. Films terminated with PAA were more glassy, suggesting better charge matching of polyelectrolytes. Multilayers irradiated for up to 100 min showed a smooth, controlled increase in the modulus with little change in the water contact angle. The permeability to iodide ion, measured electrochemically, also decreased in a controlled fashion.     

Modulation of hemocompatibility of polysulfone by polyelectrolyte multilayer films

Polyelectrolyte multilayer (PEM) films have been recently applied to surface modification of biomaterials. Cellular interactions with PEM films consisted of weak polyelectrolytes are greatly affected by the conditions of polyelectrolyte deposition, such as pH of polyelectrolyte solution. Previous studies indicated that the adhesion of several types of mammalian cells to PAH/PAA multilayer films was hindered by low pH and high layer numbers. The objective of this study is to evaluate whether the hemocompatibility of polysulfone can be modulated by deposition of poly(allylamine hydrochloride) (PAH)/poly(acrylic acid) (PAA) multilayer films. PAH/PAA multilayer films with different layer numbers were assembled onto polysulfone at either pH 2.0 or pH 6.5. The number of platelet adhesion and the morphology of adherent platelets were determined to evaluate hemocompatibility of modified substrates. Compared to non-treat polysulfone, the PEM films developed at pH 2.0 decreased platelet adhesion, while those built at pH 6.5 enhanced platelet deposition. Platelet adhesion was found positively correlated to polyclonal antibodies binding to surface-bound fibrinogen. The extent of platelet spreading was increased with layer numbers of PEM films, suggesting that the adherent platelets on thick PEM films were prone to activation. In conclusion, PAH/PAA films with few layers developed at pH 2.0 possessed better hemocompatibility compared to other substrates.     

Characterization of stimuli-sensitive polymers for biomedical applications

Stimuli-sensitive polymers were synthesized by copolymerizing varying ratios of N-isopropyl acrylamide(NIPAAm) and acrylic acid(AAc). The influence of polyelectrolytes on the lower critical solution temperatures(LCSTs) of these temperature/pH sensitive polymers was investigated in the pH range of 2-12. Polyelectrolyte complexes were prepared by mixing poly(NIPAAm-co-AAc) as anionic polyelectrolyte with poly(allyl amine)(PAA) or poly(L-lysine)(PLL) as cationic polyelectrolytes, respectively. Back titration was performed to determine the pK_a values of PAAc in poly(NIPAAm-co-AAc) and to study the effect of comonomer ionization on the cloud point temperature. The effect of polyelectrolyte complex formation on the conformation of PLL was studied as a function of temperature by means of circular dichroism(CD). The swelling ratio of poly(NIPAAm-co-AAc) hydrogels as a function of pH at various temperature was obtained by measuring the weight of the hydrogels in buffer solutions. The LCSTs of the poly(NIPAAm-co-AAc) were strongly affected by pH, polyelectrolyte solutes, AAc content, and charge density. The influence of more hydrophobic PLL as a polyelectrolyte on the cloud point of PNIPAAm/water in the copolymer was stronger than that of poly(allyl amine)(PAA). Indomethacin was loaded into these hydrogels, and controlled release of this molecule from the hydrogel was determined under various temperature and pH conditions using UV/Vis spectrophotometry.     

Surface plasmon resonance spectroscopy study of electrostatically adsorbed layers

The use of 4-(dimethylamino)pyridine to form an adhesion layer for the adsorption of anionic polyelectrolytes on gold surfaces is investigated. In situ surface plasmon resonance spectroscopy is used to monitor the changes in thickness of the adsorbed layers as a function of pH changes. Weak (poly(acrylate)) and strong (poly(styrenesulfonate)) polyelectrolytes have been studied. Although 4-(dimethylamino)pyridine is weakly bound to gold, it is not displaced by these polyelectrolytes and acts as an adhesion layer. The relationship of the interaction of anionic polyelectrolytes with 4-(dimethylamino)pyridine-modified planar gold and gold nanoparticles is discussed.     

In-situ investigation of the adsorption of globular model proteins on stimuli-responsive binary polyelectrolyte brushes

Binary brushes constituted from two incompatible polymers can be used in the form of ultrathin polymeric layers as a versatile tool for surface engineering to tune physicochemical surface characteristics such as wettability, surface charge, chemical composition, and morphology and furthermore to create responsive surface properties. Mixed brushes of oppositely charged weak polyelectrolytes represent a special case of responding surfaces that are sensitive to changes in the pH value of the aqueous environment and therefore represent interesting tools for biosurface engineering. The polyelectrolyte brushes used for this study were composed of two oppositely charged polyelelctrolytes poly(2-vinylpyridine) (P2VP) and poly(acrylic acid) (PAA). The in-situ properties and surface characteristics such as as surface charge, surface tension, and extent of swelling of these brush layers are functions of the pH value of the surrounding aqueous solution. To test the behavior of the mixed polylelctrolyte brushes in contact with biosystems, protein adsorption experiments with globular model proteins were performed at different pH values and salt concentrations (confinement of counterions) of the buffer solutions. The influence of the pH value, buffer salt concentration, and isoelectric points (IEP) of the brush and protein on the adsorbed amount and the interfacial tension during protein adsorption as well as the protein adsorption mechanism postulated in reference to recently developed theories of protein adsorption on polyelectrolyte brushes is discussed. In the salted regime, protein adsorption was found to be similar to the often-described adsorption at hydrophobic surfaces. However, in the osmotic regime the balance of electrostatic repulsion and a strong entropic driving force, "counterion release", was found to be the main influence on protein adsorption.