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The charge structure function in electrolytes and polyelectrolytes.

The relation between charge and dielectric structure functions is examined in the case of electrolytes and polyelectrolytes. When the coupling constant is larger than unity, characteristic features should be observed on the dielectric structure function, in addition to the classical screening effect. We point our the existence of experimental conditions which provide direct information of the dielectric structure at shorter wavelength (lambda 1 nm). They correspond to zero average contrast in scattering experiments.

Journal Article↗

Dip patch clamp currents suggest electrodiffusive transport of the polyelectrolyte DNA through lipid bilayers.

Planar lipid bilayers formed from monolayers of diphytanoyl lecithin (DPhL) were found to interact with plasmid DNA (5.6 kbp; M(r) = 3.7 x 10(6)) leading to an increase in the conductance of the membrane. The association of DNA with a lipid bilayer greatly facilitates the transport of the small ions of the main salt KCl. The appearance of long-lived current levels, for instance, of 27.6 pA at Vm = +60 mV membrane voltage, where the actual contact (adsorption) is electrophoretically enhanced, suggests a locally conductive DNA/lipid interaction zone where parts of the DNA strand may be transiently inserted in the bilayer, leaving other parts of the DNA probably protruding out from the outer surface of the bilayer. At Vm = -60 mV, where DNA can be electrophoretically moved away from the membrane, the membrane current is practically zero. This current asymmetry is initially also observed at higher voltages, for instance at 200 mV. However, if the voltage sign (Vm = +200 mV) is changed after a transient positive current (approximately 15 pA) was observed, there is also now (at Vm = -200 mV) a finite negative current at the negative membrane voltage. Thus, it appears that at Vm = +200 mV the adsorbed parts of the polyelectrolyte DNA are not only transiently inserted in, but actually also electrophoretically pulled through, the porous zones onto the other membrane side leaving the bilayer structure basically intact. These data provide direct electric evidence for the electrophoretic transport of a highly charged and hydrated macromolecule, probably together with the associated gegen-ions, through the thin hydrophobic film of the lipid bilayer.

Diffusion↗

Acetylcholinesterase as polyelectrolyte: interaction with multivalent cationic inhibitors.

Influence of inorganic salts on the interaction of cobra venom acetylcholinesterase (EC 3.1.1.7) with hexamethonium and gallamine has been studied. The observed negative electrostatic salt effect in the dissociation constant of the enzyme-ligand complex, KD, has been described by equation pKD = pKD degrees-ZL psi +Z log[Me+Z] following from Manning's polyelectrolyte theory, where psi +Z is the fraction of condensed counterions Me+Z per one negative charge of the polyanionic enzyme. The ZL psi+Z values for the complex formation between native acetylcholinesterase and hexamethonium (ZL = +2) or gallamine (ZL = +3) were in quantitative agreement with those predicted by the theory making use of psi+1 = 0.50 found earlier from the influence of salts upon the hydrolysis of acetylcholine by the enzyme. Increase in the number of negative charges in acetylcholinesterase by its modification with pyromellitic dianhydride resulted in an increase of psi+1 to 0.6. The data show that the influence of salts on the electrostatic contribution to the energy of binding of cationic substrates and inhibitors by acetylcholinesterase can be quantitatively described proceeding from the counterion condensation model of Manning by using only one empirical parameter psi+1 for a given subtype or modified form of the enzyme.

Acetylcholine↗

Effect of polyelectrolytes on entry of Escherichia coli HB101 (pRI203) into HeLa cells.

The role of charged molecules in the entry mechanism of enteroinvasive bacteria was studied using Escherichia coli HB101 harboring a plasmid (pRI203) containing the Yersinia pseudotuberculosis invasion region as an experimental model. We investigated the effect of several anionic and cationic polyelectrolytes on the initial steps of infection of HeLa S3 cells by E. coli HB101 (pRI203). Experiments in which the polyions were added to cell monolayers together with bacteria showed that invasion was only slightly influenced by anions whereas cations strongly enhanced bacterial entry. DEAE-dextran, histone and poly-L-lysine were the most effective enhancers producing an up to five-fold increase in the number of both infected cells and internalized bacteria. Moreover, addition of the active polycations at different stages of infection demonstrated that their action took place during the attachment step, whereas internalization was not affected.

Cell Division↗

Polyplex-embedding in polyelectrolyte multilayers for gene delivery.

In this work, incorporation of plasmid DNA, pre-complexed with PEI, into polyelectrolyte multilayers has been studied to further develop platforms for local gene delivery. Polyplex embedding in synthetic and naturally degradable architectures was efficient for transfection of human hepato-cellular carcinoma cells.

Carcinoma, Hepatocellular↗

Photo-crosslinkable microcapsules formed by polyelectrolyte copolymer and modified collagen for rat hepatocyte encapsulation.

New anionic polyelectrolyte tetra-copolymers with photo-crosslinkable 4-(4-methoxycinnamoyl)phenyl methacrylate monomer in addition to a HEMA-MMA-MAA ter-copolymer system were synthesized. The tetra-copolymers were used to form photo-crosslinkable microcapsules with modified collagen by complex coacervation for rat hepatocytes encapsulation. The hepatocytes were encapsulated within a two-layered membrane comprising of modified collagen as the inner core and an outer photo-crosslinkable copolymer shell. Upon photo-crosslinking of the microcapsules with UV-Vis light irradiation, the mechanical strength and chemical stability of the microcapsules, and the cellular functions of the encapsulated hepatocytes were enhanced. Particularly, the mechanical stability of the microcapsules was dramatically strengthened. The new photo-crosslinkable tetra-copolymer formulation described in this article has opened a way to the development of hepatocyte microencapsulation technology for bioartifical liver assist device.

Animals↗

Tuning compliance of nanoscale polyelectrolyte multilayers to modulate cell adhesion.

It is well known that mechanical stimuli induce cellular responses ranging from morphological reorganization to mineral secretion, and that mechanical stimulation through modulation of the mechanical properties of cell substrata affects cell function in vitro and in vivo. However, there are few approaches by which the mechanical compliance of the substrata to which cells adhere and grow can be determined quantitatively and varied independent of substrata chemical composition. General methods by which mechanical state can be quantified and modulated at the cell population level are critical to understanding and engineering materials that promote and maintain cell phenotype for applications such as vascular tissue constructs. Here, we apply contact mechanics of nanoindentation to measure the mechanical compliance of weak polyelectrolyte multilayers (PEMs) of nanoscale thickness, and explore the effects of this tunable compliance for cell substrata applications. We show that the nominal elastic moduli E(s) of these substrata depend directly on the pH at which the PEMs are assembled, and can be varied over several orders of magnitude for given polycation/polyanion pairs. Further, we demonstrate that the attachment and proliferation of human microvascular endothelial cells (MVECs) can be regulated through independent changes in the compliance and terminal polyion layer of these PEM substrata. These data indicate that substrate mechanical compliance is a strong determinant of cell fate, and that PEMs of nanoscale thickness provide a valuable tool to vary the external mechanical environment of cells independently of chemical stimuli.

Acrylates↗

Wear reduction of orthopaedic bearing surfaces using polyelectrolyte multilayer nanocoatings.

This work explores the use of conformal polyelectrolyte multilayer (PEM) coatings for wear reduction of orthopedic bearing surfaces. These films, with easily tunable architectures, provide excellent adhesion to a wide variety of metallic, plastic, and ceramic substrates. For this study, PEM films, only a few hundred nanometers thick, were assembled by sequential adsorption of poly(acrylic acid) and poly(allylamine hydrochloride). It was observed that the pH of the polylectrolyte solutions used for film assembly needs careful consideration to avoid any adverse effects on film structure when exposed to physiological conditions of pH and ionic strength. The wear reducing capacity of these coatings in the presence of bovine calf serum-lubricant solution was established for metal/metal systems at the meso/microscale over 30 cycles of reciprocating motion, as well as for the commonly used metal/ultra-high molecular weight polyethylene (UHMWPE) system over 500,000 cycles of bi-directional motion in a macroscale pin-on-disk test. In the latter case, the use of the films reduced UHMWPE wear by up to 33% when compared with the uncoated control. This is the first clinically relevant laboratory demonstration of the wear-reducing ability of these films. Further optimization will be needed before this novel class of materials can be used by the orthopedic community.

Acrylic Resins↗

Proliferation and differentiation of human mesenchymal stem cell encapsulated in polyelectrolyte complexation fibrous scaffold.

A biofunctional scaffold was constructed with human mesenchymal stem cells (hMSCs) encapsulated in polyelectrolyte complexation (PEC) fibers. Human MSCs were either encapsulated in PEC fibers and constructed into a fibrous scaffold or seeded on PEC fibrous scaffolds. The proliferation, chondrogenic and osteogenic differentiation of the encapsulated and seeded hMSCs were compared for a culture period of 5.5 weeks. Gene expression and extracellular matrix production showed evidences of chondrogenesis and osteogenesis in the cell-encapsulated scaffolds and cell-seeded scaffolds when the samples were cultured in the chondrogenic and osteogenic differentiation media, respectively. However, better cell proliferation and differentiation were observed on the hMSC-encapsulated scaffolds compared to the hMSC-seeded scaffolds. The study demonstrated that the cell-encapsulated PEC fibers could support proliferation and chondrogenic and osteogenic differentiation of the encapsulated-hMSCs. Together with our previous works, which demonstrated the feasibility of PEC fiber in controlled release of drug, protein and gene delivery, the reported PEC fibrous scaffold system will have the potential in composing a multi-component system for various tissue-engineering applications.

Biocompatible Materials↗

The preservation of phenotype and functionality of dendritic cells upon phagocytosis of polyelectrolyte-coated PLGA microparticles.

Biodegradable microparticles (MP) represent a promising and efficient delivery system for parenteral vaccination. Recently, MP have also been explored as tool for the ex vivo antigen loading of professional antigen-presenting cells such as dendritic cells (DC) to be used as cellular vaccines. The purpose of this study was to investigate various polycationic coatings on poly(lactide-co-glycolide) (PLGA) MP, with regard to their effect on phenotypic and functional maturation of monocyte-derived DC (MoDC) that had previously been loaded with the MP in vitro. The preparation and concomitant coating of the PLGA was performed by means of a solvent extraction/evaporation method using a recently developed microextrusion-based technique. The polyelectrolytes tested for MP coating encompassed aminodextran, chitosan, poly(ethylene imine) (PEI), poly(L-lysine) and protamine. Uncoated and differently coated PLGA MP were fed to immature MoDC, which ingested efficiently the different MP types irrespective of their surface coating. The MP-loaded immature MoDC were then matured with the help of a cytokine/PGE-2 maturation cocktail. Here, the presence of the ingested MP did not affect the MoDC maturation in terms of expression of the surface markers CD80, CD83, CD86, HLA-DR and MMR, irrespective of the MP surface coating. Importantly, none of the PLGA MP types alone induced significant maturation of MoDC in the absence of the maturation cocktail. MP-loaded and subsequently matured MoDC expressed high levels of the chemokine receptor CCR7, whose functional activity was evidenced by the migration of MoDC towards CCL21, irrespective of the presence of ingested MP. Further, MP-loaded and subsequently matured MoDC also secreted comparable amounts of IL-10 and IL-12p70, irrespective of the presence of ingested MP except for PEI-coated PLGA MP, which enhanced significantly the secretion of IL-12p70 in mature MoDC. In conclusion, phenotypic and functional maturation of MoDC by means of a maturation cocktail remained unchanged irrespective of the presence of previously ingested differently coated PLGA MP. This offers interesting perspectives for using these particulate systems together with entrapped antigens for ex vivo loading of MoDC in view of cellular immunotherapy.

Cell Proliferation↗

Encapsulation of glucose oxidase and an oxygen-quenched fluorophore in polyelectrolyte-coated calcium alginate microspheres as optical glucose sensor systems.

Microspheres coated with polyelectrolyte multilayers (PEM's) are being investigated for potential use as implantable biosensors-so-called "smart tattoos." In this work, the feasibility of this approach for glucose sensors was demonstrated by glucose oxidase encapsulated within calcium alginate microspheres, followed by entrapment of an oxygen-quenched ruthenium compound in the same microstructure. A novel feature of these microdevices is the formation of multilayer nanofilms on the surface of the microspheres, used to stabilize enzyme entrapment and control substrate diffusion. Confocal microscopy was used to confirm the stable encapsulation of sensor chemistry. The reversible response of sensors to step changes in glucose was observed, and preliminary experimental data were compared to theoretical predictions produced by a computational model. These findings demonstrate the promise of the described nanoengineering approach for production of functional implantable glucose sensor materials.

Alginates↗

Preparation of polyaniline-modified electrodes containing sulfonated polyelectrolytes using layer-by-layer techniques.

Polyaniline (PAni) has been used frequently for the construction of biosensors. However, a prime limitation is its instability at basic or neutral pH because of the loss of its electrochemical activity and conductivity. In this study, three available sulfonated polyanions: Nafion, poly(vinyl sulfonate) (PVS), and poly(styrene sulfonate) (PSS) serving as the counterion and providing an acidic microenvironment to stabilize PAni, are used to fabricate a sensor for ammonium ion detection. Nafion used to be a common ion-sensitive membrane due to its high proton conductivity. However, its high cost and limited solubility has constrained its uses. PVS and PSS are water-soluble polymers, easily incorporating with PAni to form the composites. Surface analysis by electron spectroscopy for chemical analysis (ESCA) and scanning electron microscope (SEM), and the electrochromic property for the PAni composites provided the convenient tools to characterize the electrode fabrication. On the aspect of sensing the ammonium ions, the modified electrodes exhibited electroactivity of PAni in ammonium ion detection and also showed the linear dependence of reduction current on the ammonium ion concentration. The pH effect on the sensing response was also evaluated and found insignificant to the response (ranging from pH 6.9-7.6). For increasing the stability of the electrodes, the diazo-resin (DAR) was introduced to the coat on the outmost layer and then cured by UV irradiation, giving the covalent network between the layers of polyelectrolytes. The PSS-doped PAni electrode was found to perform detection sensitivity in the linear range of 0-100mM of ammonium ion concentration.

Adsorption↗

Ion hydration: Implications for cellular function, polyelectrolytes, and protein crystallization.

Only oppositely charged ions with matching absolute free energies of hydration spontaneously form inner sphere ion pairs in free solution [K.D.Collins, Ions from the Hofmeister series and osmolytes: effects on proteins in solution and in the crystallization process, Methods 34 (2004) 300-311.]. We approximate this with a Law of Matching Water Affinities which is used to examine the issues of (1) how ions are selected to be compatible with the high solubility requirements of cytosolic components; (2) how cytosolic components tend to interact weakly, so that association or dissociation can be driven by environmental signals; (3) how polyelectrolytes (nucleic acids) differ from isolated charges (in proteins); (4) how ions, osmolytes and polymers are used to crystallize proteins; and (5) how the "chelate effect" is used by macromolecules to bind ions at specific sites even when there is a mismatch in water affinity between the ion and the macromolecular ligands.

Crystallization↗

Lysozyme purification from tobacco extract by polyelectrolyte precipitation.

Tobacco is widely used as a model plant for feasibility studies of recombinant protein production from transgenic plants. However, dealing with large quantities of biomass to recover recombinant proteins is a challenge for down-stream processing. In this study, the effect of isoelectric precipitation on native tobacco protein was first studied. Among the three acids studied, hydrochloric acid is shown to be more effective than acetic or citric acid, and at pH 4, 60% of native tobacco protein was precipitated by HCl. Egg white lysozyme was used as the model protein to test the feasibility of polyelectrolyte precipitation in protein recovery from tobacco extract. Precipitation of lysozyme at pH 7 was shown ineffective probably because of the interference of polyphenolic acids. However, after isoelectric precipitation at pH 5 poly(acrylic) acid (PAA) was shown to precipitate 85% of the soluble lysozyme when the polymer dosage was increased to 1.5 mg polymer/mg lysozyme, while negligible amounts of native tobacco protein was co-precipitated. Lysozyme precipitation by PAA in tobacco extract obtained at pH 5 was also studied, and lysozyme yield was significant improved.

Chemical Precipitation↗

Polyelectrolyte multilayer capsules as vehicles with tunable permeability.

This review is devoted to a novel type of polymer micro- and nanocapsules. The shell of the capsule is fabricated by alternate adsorption of oppositely charged polyelectrolytes (PEs) onto the surface of colloidal particles. Cores of different nature (organic or inorganic) with size varied from 0.1 to 10 mum can be used for templating such PE capsules. The shell thickness can be tuned in nanometer range by assembling of defined number of PE layers. The permeability of capsules depends on the pH, ionic strength, solvent, polymer composition, and shell thickness; it can be controlled and varied over wide range of substances regarding their molecular weight and charge. Including functional polymers into capsule wall, such as weak PEs or thermosensitive polymers, makes the capsule permeability sensitive to correspondent external stimuli. Permeability of the capsules is of essential interest in diverse areas related to exploitation of systems with controlled and sustained release properties. The envisaged applications of such capsules/vesicles cover biotechnology, medicine, catalysis, food industry, etc.

Adsorption↗

Electrokinetic fingerprinting of grafted polyelectrolyte layers--a theoretical approach.

Electrokinetic fingerprinting (EF) was introduced by Marlow and Rowell [Marlow BJ, Rowel RL. Langmuir 1990;6:1088] for the comprehensive characterization of charged particle surfaces. Afterwards, EF was applied by many groups for the characterization of "hard" (i.e. non-swelling) surfaces. However, the advantages of EF could not yet utilized for the characterization of grafted polyelectrolyte layers (PL) since the theoretical background was not yet elaborated. A theory for the characterization of PL at complete dissociation of the functional groups was developed by Ohshima [Adv Colloid Interface Sci 1995;62:189] and later extended by Dukhin et al. [Dukhin S, Zimmermann R, Werner C. J Colloid Interface Sci 2005;286:761] for any degree of dissociation. Further progress in the characterization of soft surfaces may be achieved by combining EF and surface conductivity (SC) measurements. Both theory and experiment demonstrate that integrated measurements of SC and apparent zeta potential zeta(a) in broad ranges of pH and ionic strength provide information about Donnan potential Psi(D), surface charge, pK and surface potential Psi(0), while the interpretation is more uncertain, when only zeta(a) is measured. This advanced method of PL characterization is established for PL grafted on flat surfaces. When PL are formed on spherical particles, the SC may be measured by means of conductometry and/or dielectric spectroscopy. However, the current theories can only be applied within a rather narrow range of the practically relevant conditions. To overcome this limitation, an unified approach to the theory of electrophoresis for spherical particles with grafted PL was elaborated taking into account the existence of two different electrokinetic models for soft surfaces. While one model is focused on hydrodynamic permeability of soft surface and disregards surface current, another model considers the surface current and disregards electrokinetic water transport within the soft surface layer. Unification became possible through generalization of the capillary osmosis theory over soft surfaces.

Electric Conductivity↗

pH-sensitive polyelectrolyte complex gel microspheres composed of chitosan/sodium tripolyphosphate/dextran sulfate: swelling kinetics and drug delivery properties.

Porous chitosan (CS) polyelectrolyte complex (PEC) hydrogel microspheres were prepared via either wet phase-inversion or ionotropic crosslinking with sodium tripolyphosphate (Na+ - TPP) and dextran sulfate (DS). The resulting microspheres were characterized using scanning electron microscopy (SEM) and elemental analysis (EA). The controlled release behavior of ibuprofen (IBU) from these microspheres was investigated. The PEC microspheres were about 700-950 microm in diameter with large pores and open porous structure. The CS/TPP/DS microspheres resisted hydrolysis in strong acid and biodegradation in enzymatic surroundings. The swelling kinetics for CS microspheres was close to Fickian diffusion, whereas those for CS/TPP and CS/TPP/DS were non-Fickian. Furthermore, the equilibrium water content (EWC) and water diffusion coefficient (D) increased with the pH of the media. The release profiles of IBU from CS/TPP/DS microspheres were slow in simulated gastric fluid (SGF, pH 1.4) over 3 h, but nearly all of the initial drug content was released in simulated intestinal fluid (SIF, pH 6.8) within 6 h after changing media. Overall the results demonstrated that CS/TPP/DS microspheres could successfully deliver a hydrophobic drug to the intestine without losing the drug in the stomach, and hence could be potential candidates as an orally administered drug delivery system.

Administration, Oral↗

Layer-by-layer polyelectrolyte coating of low molecular weight poly(lactic acid) nanoparticles.

Low molecular weight (M(w)) poly(L-lactic acid) (PLA) nanoparticles were coated with polyelectrolytes (PEs) by layer-by-layer (LbL) technique using a filtration approach. Poly(allylamine hydrochloride) and poly(sodium 4-styrenesulfonate) were applied as PEs in coating. LbL coating is aimed to use in producing (nano)particulate drug delivery systems with improved biocompatibility and sustained or targeted release of drug substances. Nanoparticles of rapidly biodegradable polymers, like the low M(w) PLA, open up a possibility to control the release of the encapsulated substance by the coating, but set challenges to the coating process due to increased aggregation tendency and degradation rate of the polymer. When the core PLA nanoparticles were prepared by nanoprecipitation, surface properties of the nanoparticles were affected by solvent selection. Successful LbL coating of the PLA nanoparticles was obtained only with chloroform, but not with dichloromethane as the solvent during nanoprecipitation. Reason for this was found to be the more charged surface of the nanoparticles prepared with chloroform compared to the nanoparticles prepared with dichloromethane.

Electrolytes↗