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PubMed2003

Endothelial cells grown on thin polyelectrolyte mutlilayered films: an evaluation of a new versatile surface modification.

Endothelial cell seeding constitutes an appreciated method to improve blood compatibility of small-diameter vascular grafts. In this study, we report the development of a simple innovative technique based on multilayered polyelectrolyte films as cell adhesive substrates. Polyelectrolyte multilayered films ending by poly(sodium-4-styrenesulfonate)/poly(allylamine hydrochloride) (PSS/PAH) or poly(L-glutamic acid)/poly(D-lysine) (PGA/PDL) could enhance cell adhesion by modification of the physico-chemical properties of the surface. The biological responses of human umbilical vein endothelial cells seeded on the polyelectrolyte multilayer films, on PDL or PAH monolayers, and on control surfaces, were evaluated in terms of initial attachment, growth, cellular metabolic activity, endothelial phenotype, and adhesion. The results showed that polyelectrolyte multilayers neither induce cytotoxic effects nor alter the phenotype of the endothelial cells. The polyelectrolyte multilayered films enhanced initial cell attachment as compared to the polyelectrolyte monolayer. Cell growth observed on the films was similar to that on TCPS. Among the different coating tested, the film ending by PSS/PAH exhibited an excellent cellular biocompatibility and appeared to be the most interesting surface in terms of cellular adhesion and growth. Such films could be used to cover hydrophobic (cell resistant) substrates in order to promote cell colonization, thereby constituting an excellent material for endothelial cell seeding.

Biocompatible Materials↗
PubMed2004

Deposition and wetting characteristics of polyelectrolyte multilayers on plasma-modified porous polyethylene.

Hydrophilic and chemically reactive porous media were prepared by adsorbing functional polymers at the surface of sintered polyethylene membranes. Modification of the membrane was accomplished by first exposing the membrane to an oxygen glow discharge gas plasma to introduce an electrostatic charge at the membrane surfaces. Cationic polyelectrolyte polyethylenimine (PEI) was adsorbed from solution to the anionic-charged surface to form an adsorbed monolayer. The adsorption of a second anionic polyelectrolyte onto the PEI layer allows further modification of the membrane surface to form a polyelectrolyte-bilayer complex. The conformation and stability of the adsorbed monolayers and bilayers comprising the modified surface are probed as a function of the polymer structure, charge density, and solubility. Using X-ray photoelectron spectroscopy analysis, we demonstrate that the presence of the polyelectrolyte multilayers drastically increases the density and specificity of the functional groups at the surface, more than what can be achieved through the plasma modification alone. Also, using the wicking rate of deionized, distilled water through the porous membrane to gauge the interfacial energy of the modified surface, we show that the membrane wicking rate can be controlled by varying the chemistry of the adsorbing polyelectrolytes and, to a lesser extent, by adjusting the polarity or ionic strength of the polyelectrolyte solution.

Journal Article↗
PubMed2004

Characterization of polyelectrolyte multilayers by the streaming potential method.

Polyelectrolyte multilayer adsorption on mica was studied by the streaming potential method in the parallel-plate channel setup. The technique was calibrated by performing model measurements of streaming potential by using monodisperse latex particles. Two types of polyelectrolytes were used in our studies: poly(allylamine) hydrochloride (PAH), of a cationic type, and poly(sodium 4-styrenesulfonate) (PSS) of an anionic type, both having molecular weight of 70,000. The bulk characteristics of polymers were determined by measuring the specific density, diffusion coefficient for various ionic strengths, and zeta potential. These measurements as well as molecular dynamic simulations of chain shape and configurations suggested that the molecules assume an extended, wormlike shape in the bulk. Accordingly, the diffusion coefficient was interpreted in terms of a simple hydrodynamic model pertinent to flexible rods. These data allowed a proper interpretation of polyelectrolyte multilayer adsorption from NaCl solutions of various concentrations or from 10(-3) M Tris buffer. After completing a bilayer, periodic variations in the apparent zeta potential between positive and negative values were observed for multilayers terminated by PAH and PSS, respectively. These limiting zeta potential values correlated quite well with the zeta potential of the polymers in the bulk. The stability of polyelectrolyte films against prolonged washing (reaching 26 h) also was determined using the streaming potential method. It was demonstrated that the PSS layer was considerably more resistant to washing, compared to the PAH layer. It was concluded that the experimental data were consistent with the model postulating particle-like adsorption of polyelectrolytes with little chain interpenetration. It also was concluded that due to high sensitivity, the electrokinetic method applied can be effectively used for quantitative studies of polyelectrolyte adsorption, desorption, and reconformation.

Adsorption↗
PubMed2005

Biocidal activity of a light-absorbing fluorescent conjugated polyelectrolyte.

Herein we describe studies that indicate a cationic conjugated polyelectrolyte shows biocidal activity against gram-negative bacteria (Escherichia coli, E. coli, BL21, with plasmids for Azurin and ampicillin resistance) and gram-positive bacterial spores (Bacillus anthracis, Sterne, B. anthracis, Sterne). These studies were carried out with aqueous suspensions of the conjugated polyelectrolyte, with the polyelectrolyte in supported formats and with samples in which the conjugated polyelectrolyte was coated on the bacteria. The results are interesting in that the biocidal activity is light-induced and appears effective due to the ability of the conjugated polyelectrolyte to form a surface coating on both types of bacteria. The effects observed here should be general and suggest that a range of conjugated polyelectrolytes in different formulations may provide a useful new class of biocides for both dark and light-activated applications.

Absorption↗
PubMed2005

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.

Journal Article↗
PubMed2006

On the indirect polyelectrolyte titration of cellulosic fibers. Conditions for charge stoichiometry and comparison with ESCA.

The effect of electrolyte (NaHCO3) concentration on the adsorption of poly-DADMAC (poly-diallyldimethylammonium chloride) onto cellulosic fibers with different charge profiles was investigated. Surface carboxymethylated fibers were obtained by grafting carboxymethyl cellulose (CMC) onto the fiber surface and bulk carboxymethylated fibers were obtained by reacting the fibers with monochloroacetic acid. It was shown that nonionic interactions do not exist between cellulose and poly-DADMAC, rather electrostatic interactions govern the adsorption. Charge stoichiometry prevails under electrolyte-free conditions, whereas surface charge overcompensation occurs at higher electrolyte concentrations. It was shown that charge stoichiometry prevails if the thickness of the electric double layer kappa(-1) was larger than the mean distance between the charges on the fiber surface, as predicted by polyelectrolyte adsorption theories, taking lateral correlation effects into account. In a second set of experiments the ESCA technique served to independently calibrate the polyelectrolyte titrations for determining the surface charge of cellulosic fibers. Various molecular masses of poly-DADMAC were adsorbed to carboxymethylated fibers having different charge profiles. The adsorption of low M(w) poly-DADMAC (7.0 x 10(3)), analyzed by polyelectrolyte titration, was about 10 times higher than that of the high M(w) poly-DADMAC (9.2 x 10(5)). Despite the difference in accessibility of these two polyelectrolytes to the fiber cell wall, ESCA surface analysis showed, as expected, only slight differences between the two polyelectrolytes. This gives strong credibility to the idea that surface charge content of cellulosic fibers can be analyzed by means of adsorption of a high-molecular-mass cationic polymer, i.e., by polyelectrolyte titration.

Adsorption↗
PubMed2004

Fabrication and characterization of polyelectrolyte microparticles with protein.

The incorporation of proteins into microparticles fabricated by layer-by-layer adsorption of oppositely charged polyelectrolytes (dextran sulfate and protamine) on protein microaggregates was studied. Microaggregates with insulin were prepared by two different techniques: 1) formation of insoluble polyelectrolyte complex consisting of insulin and dextran sulfate (aggregate size of 7-20 micro m), or 2) salting out of insulin from solution by sodium chloride (aggregate size of 5-13 micro m). Microparticles varying in the number of cycles (from 1 to 8) of polyelectrolyte adsorption on protein aggregates were examined and compared. Morphology of the microparticles was studied by scanning electron and optical microscopy. It was shown that polyelectrolyte microparticles retained the shape and dimensions of the initial protein aggregates used as a template. Ultrasonication of microparticles obtained using salted out protein aggregates resulted in the formation of stable nanoparticles (100-200 nm). Regulation of protein release from the microparticles of both types by varying the number of polyelectrolyte adsorption cycles and pH of the medium was demonstrated. Insulin not bound to polyelectrolytes was released from the microparticles at pH values between 6 and 8, which corresponds to the pH of the human small intestine and ileum.

Adsorption↗
PubMed2006

Entropy and enthalpy of polyelectrolyte complexation: Langevin dynamics simulations.

We report a systematic study by Langevin dynamics simulation on the energetics of complexation between two oppositely charged polyelectrolytes of same charge density in dilute solutions of a good solvent with counterions and salt ions explicitly included. The enthalpy of polyelectrolyte complexation is quantified by comparisons of the Coulomb energy before and after complexation. The entropy of polyelectrolyte complexation is determined directly from simulations and compared with that from a mean-field lattice model explicitly accounting for counterion adsorption. At weak Coulomb interaction strengths, e.g., in solvents of high dielectric constant or with weakly charged polyelectrolytes, complexation is driven by a negative enthalpy due to electrostatic attraction between two oppositely charged chains, with counterion release entropy playing only a subsidiary role. In the strong interaction regime, complexation is driven by a large counterion release entropy and opposed by a positive enthalpy change. The addition of salt reduces the enthalpy of polyelectrolyte complexation by screening electrostatic interaction at all Coulomb interaction strengths. The counterion release entropy also decreases in the presence of salt, but the reduction only becomes significant at higher Coulomb interaction strengths. More significantly, in the range of Coulomb interaction strengths appropriate for highly charged polymers in aqueous solutions, complexation enthalpy depends weakly on salt concentration and counterion release entropy exhibits a large variation as a function of salt concentration. Our study quantitatively establishes that polyelectrolyte complexation in highly charged Coulomb systems is of entropic origin.

Computer Simulation↗
PubMed2006

Spatial distribution of polyelectrolyte and counterions in nanocapsules: a computer simulation study.

We study a spatial distribution of polyelectrolyte chains and counterions inside nanometer-size capsules by means of molecular dynamics simulation on the level of a colloidal model in which polyelectrolyte coils are modeled as soft charged spheres. The capsule shell is treated as a semipermeable membrane, impermeable for the polyelectrolyte chains, but allowing free diffusion of solvent molecules and counterions. As a result, counterions leak out from the capsule immersed into a fluid of low ionic strength. This counterion leakage leads to a formation of characteristic polyelectrolyte density profiles with the central plateau and large peaks at the wall. We show that a nonuniform distribution of the inner polyelectrolyte depends on the capsule radius, surface charge, concentration of encapsulated polyelectrolyte, and the volume fraction of capsules.

Adsorption↗
PubMed2006

Condensation of DNA-actin polyelectrolyte mixtures driven by ions of different valences.

Multivalent ions can induce condensation of like-charged polyelectrolytes into compact states, a process that requires different ion valences for different polyelectrolyte species. In this work we examine the condensation behavior in binary anionic polyelectrolyte mixtures consisting of DNA coils and F-actin rods in the presence of monovalent, divalent, and trivalent ions. As expected, monovalent ions do not condense either component and divalent ions selectively condense F-actin rods out of the polyelectrolyte mixture. For trivalent ions, however, we observe a microphase separation between the two polyelectrolytes into coexisting finite-sized F-actin bundles and DNA toroids. Further, by increasing the DNA volume fraction in the mixture, condensed F-actin bundles can be completely destabilized, leading to only DNA condensation within the mixture. We examine a number of possible causes and propose a model based on polyelectrolyte competition for ions.

Actins↗
PubMed2006

Monte Carlo simulations of polyelectrolytes inside viral capsids.

Structural features of polyelectrolytes as single-stranded RNA or double-stranded DNA confined inside viral capsids and the thermodynamics of the encapsidation of the polyelectrolyte into the viral capsid have been examined for various polyelectrolyte lengths by using a coarse-grained model solved by Monte Carlo simulations. The capsid was modeled as a spherical shell with embedded charges and the genome as a linear jointed chain of oppositely charged beads, and their sizes corresponded to those of a scaled-down T=3 virus. Counterions were explicitly included, but no salt was added. The encapisdated chain was found to be predominantly located at the inner capsid surface, in a disordered manner for flexible chains and in a spool-like structure for stiff chains. The distribution of the small ions was strongly dependent on the polyelectrolyte-capsid charge ratio. The encapsidation enthalpy was negative and its magnitude decreased with increasing polyelectrolyte length, whereas the encapsidation entropy displayed a maximum when the capsid and polyelectrolyte had equal absolute charge. The encapsidation process remained thermodynamically favorable for genome charges ca. 3.5 times the capsid charge. The chain stiffness had only a relatively weak effect on the thermodynamics of the encapsidation.

Capsid↗
PubMed1998

Polyelectrolyte-induced domains in lipid bilayer membranes: the deuterium NMR perspective.

Domain formation in lipid bilayer membranes can occur through electrostatic interactions between charged lipids and oppositely charged polyelectrolytes, such as proteins or polynucleic acids. This review describes a novel method for examining such domains in lipid bilayers, based on 2H NMR spectroscopy. The 2H NMR spectrum of choline-deuterated phosphatidylcholine is sensitive to, and reports on, lipid bilayer surface charge. When a charged lipid bilayer is exposed to an oppositely charged polyelectrolyte, the latter binds electrostatically to the bilayer surface and attracts charged lipids into its vicinity. The resulting inhomogeneous charge distribution produces overlapping 2H NMR subspectra arising from phosphatidylcholine within charge-enriched versus charge-depleted regions. Such spectral details as the quadrupolar splittings and the relative intensities of the subspectra permit a complete analysis of the domain composition, size, and, within limits, lifetime. Using 2H NMR, domain formation in lipid bilayer membranes can be observed with both cationic and anionic polyelectrolytes, whether of natural or synthetic origin. Domain size and composition prove to be sensitive to the detailed chemical structure of both the polyelectrolyte and the charged lipids. Within the domains there is always a stoichiometric anion/cation binding ratio, indicating that the polyelectrolyte lies flat on the membrane surface. The amount of phosphatidylcholine within the domain varies as a function of its statistical availability, in accordance with the predictions of a recent thermodynamic model of domain formation. When the molecular weight of the polyelectrolyte is varied, the domain size alters in accordance with the predictions of classical polymer physics. As expected for a predominantly electrostatic phenomenon, the observed domains dissipate at high ionic strength.

Chemical Phenomena↗
PubMed2003

Annealing polyelectrolytes at charged interfaces.

The conformation of a weakly dissociating (annealing) polyelectrolyte chain end-tethered to a similarly or oppositely charged planar surface is analyzed in the framework of scaling arguments. For a similarly charged interface an analytical model is also utilized. We demonstrate that at low salt concentration in bulk solution there is a strong coupling between the polyelectrolyte conformation and its degree of ionization. In the case of an oppositely charged (adsorbing) surface, adsorption promotes ionization of the annealing polyelectrolyte. As a result, the adsorbed layer thickness decreases as a function of surface charge density more rapidly for an annealing polyelectrolyte than for a quenched one. In the case of a similarly charged (repulsive) surface the chain ionization is suppressed, and the annealing polyelectrolyte chain is less extended than the quenched one. Moreover, an increase in surface charge density leads to non-monotonous extension of the tethered polyelectrolyte.

Journal Article↗
PubMed2005

Effect of counterions on the swelling of spherical polyelectrolyte brushes.

We investigate the swelling of colloidal spherical polyelectrolyte brushes in the presence of different counterions. The colloidal particles consist of a solid poly(styrene) core of ca. 100 nm diameter onto which linear polyelectrolyte chains are chemically grafted. Two types of polyelectrolyte chains have been used here: The cationic polyelectrolyte poly(2-(acryloyl)ethyltrimethylammonium chloride)) (PATAC) and the anionic poly(styrenesulfonate) (PSS). Both systems are dispersed in water and the degree of swelling of the surface layer is studied by dynamic light scattering. Adding more and more salt leads to a strong shrinking of the surface layer as expected for polyelectrolyte brushes. It is shown that data obtained at low ionic strength can be collapsed on suitable master curves for monovalent and divalent counterions, respectively. For some ions, however, high salt concentrations may lead to a re-swelling of the brush layer in case of the cationic systems. This points to specific interactions of the counterions with the PATAC chains. This strong specific interaction between the counterions and the attached polyelectrolyte may even lead to flocculation of the particles at intermediate salt concentration. Surprisingly, for iodide and magnesium counterions the solubility increases again if the salt concentration is raised to 1 mol/l. Hence, specific interaction leads to salting-out effects as well as to salting-in effects for these colloidal particles. All specific effects seen at high concentrations of added salt can be explained by the increase of the reduced excluded-volume parameter which is due to the adsorption of salt ions.

Colloids↗
PubMed1983

Charge determination of proteins with polyelectrolyte titration.

A recently developed photometric version of polyelectrolyte titration was applied for the determination of the number of charged residues on globular proteins. Based on the observation that oppositely charged polyelectrolytes form, in general, stoichiometric polyelectrolyte complexes, the protein solutions were incubated in excess with an oppositely charged polyelectrolyte, and the residual amount was back-titrated using o-toluidine blue for end point detection. It was found that within the range pH 2 to pH 9 the interaction of the polyelectrolytes, potassium polyvinylsulfate, polydiallylammonium chloride, and N-methylglycolchitosan iodide, with various proteins of known amino acid composition (ribonuclease A, trypsin, chymotrypsin A, pepsin, cytochrome c) occurs stoichiometrically through 1:1 ion pair interaction, irrespective of the spatial distribution of the interacting ionic sites. The close correspondence between the experimental data for the net charge and the calculated balance of ionized residues for the proteins at a given pH indicates that in the native structure of these proteins oppositely charged ionic functions are largely neutralized by the formation of intramolecular salt linkages. It is concluded that polyelectrolyte titration offers an easy access to the determination of the surface charge of proteins and other biopolymers. The data further support the notion of the importance of electrostatic cooperative interactions in biological systems.

Animals↗
PubMed1995

Salt effects on polyelectrolyte-ligand binding: comparison of Poisson-Boltzmann, and limiting law/counterion binding models.

The theory for salt dependence of the free energy, entropy, and enthalpy of a polyelectrolyte in the PB (PB) model is extended to treat the nonspecific salt dependence of polyelectrolyte-ligand binding reactions. The salt dependence of the binding constant (K) is given by the difference in osmotic pressure terms between the reactants and products. For simple 1-1 salts it is shown that this treatment is equivalent to the general preferential interaction model for the salt dependence of binding [C. Anderson and M. Record (1993) Journal of Physical Chemistry, Vol. 97, pp. 7116-7126]. The salt dependence, entropy, and enthalpy are compared for the PB model and one specific form of the preferential interaction coefficient model that uses counterion condensation/limiting law (LL) behavior. The PB and LL models are applied to three ligand-polyelectrolyte systems with the same net ligand charge: a model sphere-cylinder binding reaction, a drug-DNA binding reaction, and a protein-DNA binding reaction. For the small ligands both the PB and limiting law models give (In K vs. In[salt]) slopes close in magnitude to the net ligand charge. However, the enthalpy/entropy breakdown of the salt dependence is quite different. In the PB model there are considerable contributions from electrostatic enthalpy and dielectric (water reorientation) entropy, compared to the predominant ion cratic (release) entropy in the limiting law model. The relative contributions of these three terms in the PB model depends on the ligand: For the protein, ion release entropy is the smallest contribution to the salt dependence of binding. The effect of three approximations made in the LL model is examined: These approximations are (1) the ligand behaves ideally, (2) the preferential interaction coefficient of the polyelectrolyte is unchanged upon ligand binding, and (3) the polyelectrolyte preferential interaction coefficient is given by the limiting law/counterion-condensation value. Analysis of the PB model shows that assumptions 2 and 3 break down at finite salt concentrations. For the small ligands the effects on the slope cancel, however, giving net slopes that are similar in the PB and LL models, but with a different entropy/enthalpy breakdown. For the protein ligand the errors from assumptions 2 and 3 in the LL models do not cancel.

Computer Graphics↗
PubMed2005

Interactions between self-assembled polyelectrolyte shells and tumor cells.

Layer-by-layer self-assembled polyelectrolyte shells are a new class of micro/nanocapsules with unique physicochemical properties for potential applications in drug/gene delivery. The objective of this study was to investigate the interactions of polyelectrolyte shells ( approximately 1 mum in diameter) with MCF-7 breast cancer cells and identify key parameters that affect such interactions. Tailoring of surface properties of polyelectrolyte shells was achieved by choosing different outermost layer materials, including cationic polymers, anionic polymers, and lipid bilayers. Different surface compositions led to a wide range of electrostatic potentials from -46 to +47 mV in phophate-buffered saline buffer. Confocal microscopy studies showed that the polyelectrolyte shells were internalized into the cell cytoplasm, but not into the nuclei. Correlation of cell uptake with shell surface compositions was complicated by the adsorption of serum proteins on the surface of polyelectrolyte shells, particularly polycation-coated shells. To prevent protein adsorption, poly(ethylene glycol) (PEG) grafted poly(ethyleneimine) (PEI) copolymers (1:1, 1:5, 1:10 graft ratios) were synthesized and introduced on the shell surface. Shells coated with PEI-PEG copolymers effectively reduced protein adsorption whereas PEI-PEG copolymers with lower graft ratios achieved higher cell uptake efficiency after 24 h of incubation with MCF-7 cells.

Biocompatible Materials↗
PubMed2001

Micropolarity and microviscosity of amitriptyline and dextran sulfate/carrageenan-amitriptyline systems: the nature of polyelectrolyte-drug complexes.

The polarity and viscosity of the microenvironment of aggregates of the cationic amphiphilic drug amitriptyline and dextran sulfate (DxS)/carrageenan-amitriptyline aggregates in aqueous solution were investigated by means of steady state and time-resolved fluorescence. For the latter systems, equilibrium dialysis and capillary viscometry were also used. The micropolarity as detected by pyrene indicated the formation of amitriptyline aggregates, both with and without polyelectrolyte, having properties similar to "traditional" cationic micelles. The pyrene lifetime in the amitriptyline- and amitriptyline-polyelectrolyte aggregates was long ( approximately 230 and approximately 300 ns, respectively), indicating that pyrene was well protected from oxygen quenching, especially so in the latter case. The microviscosity of the amitriptyline aggregates themselves, and in the presence of polyelectrolyte, was high, as indicated by intramolecular excimer formation of 1,3-di(1-pyrenyl)-propane (P3P), rotational diffusion fluorescence depolarization of 1,6-diphenyl-1,3,5-hexatriene (DPH), and intramolecular rotational relaxation about bonds of [p-(dimethylamino)benzylidene]-malonitrile (BMN). These results and the concurrent decrease of the bulk viscosity indicate that the polyelectrolyte, acting as polycounterion, is tightly wrapped around the amitriptyline aggregates. Amitriptyline hence behaves in accordance with accepted models of cationic surfactant-polyelectrolyte interaction.

Amitriptyline↗