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Modulation of acute immune complex-mediated tissue injury by the presence of polyionic substances.

Considerable attention has been focused on the role of electrostatic charge in the pathogenesis of immune complex-mediated tissue injury. The authors have examined the ability of cationic (histone, polyhistidine, polyarginine) and anionic (polyanetholsulfonate) polyelectrolytes to modulate acute immune complex-mediated tissue injury. Tissue injury elicited in rats by the reversed dermal Arthus reaction was increased 26-43% by addition of polyelectrolytes to antibody prior to its intradermal injection. Kinetic studies using 111In-labeled neutrophils indicated that the enhanced tissue injury was not the result of increased influx of neutrophils. Infusion of polyethylene glycol-conjugated superoxide dismutase prior to induction of the Arthus reaction resulted in 40-68% suppression of tissue injury. Concomitant in vitro functional studies (enzyme secretion, O-2 and H2O2 generation, and chemiluminescence) of rat neutrophils demonstrated that addition of polyelectrolytes to preformed immune complexes (IgG-bovine serum albumin) resulted in marked increases in O-2, H2O2, and chemiluminescence, but no increases in enzyme secretion, compared with neutrophils stimulated with immune complexes alone. The cationic polyelectrolytes did not alter the capacity of preformed immune complexes to activate complement in vitro. These studies suggest that both cationic and anionic polyelectrolytes can increase the pathogenic potential of immune complexes and that this modulation is, at least in part, mediated by enhanced generation of toxic oxygen-derived metabolites by neutrophils.

Animals↗

The effect of support material and conditioning on wettability of PAH/PSS multilayer films.

An efficient method for characterizing wetting properties of heterogeneous surfaces produced by sequential adsorption of polyelectrolytes was developed. Three types of polyelectrolytes were used: polyallylamine hydrochloride (PAH), polyethyleneimine (PEI), both of a cationic type, and polysodium 4-styrenesulfonate (PSS), of an anionic type. Multilayer films were prepared by 'layer-by-layer' (LbL) deposition technique. Natural ruby mica, glass, titanium foil and silicon wafers were used as the support material for PE films. Wetting of polyelectrolyte films was determined experimentally by contact angle measurements, using technique of direct image analysis of shape of sessile drops. Periodic oscillations in contact angle values were observed for multilayers terminated by polycation and polyanion, respectively, and the variations in contact angle values strongly depended on the conditions of adsorption and multilayer treatment after deposition. Therefore, the influence of ionic strength of polyelectrolyte solution used for deposition on wetting of multilayer films was considered and also the effect of conditioning in different environments was investigated. It is usually assumed that film properties and stability strongly depend on the first layer which is used to anchor a multilayer at the surface of support material. To investigate influence of the first layer, PAH/PSS films were compared with more complex ones having PEI as the first layer with a sequence of PSS/PAH deposited on top of it.

Adsorption↗

Theory of delocalized ionic binding to polynucleotides: structural and excluded-volume effects.

A previously developed theory for the delocalized binding of ions to polyelectrolytes was restricted to point ions and a structurally rigid polyelectrolyte. For the binding of substances like oligolysines and polyamines to DNA, the restriction to point ions would appear not to be realistic. For the binding of ions to flexible chains like single-stranded polynucleotides, the restriction to a rigid polyelectrolyte may not be realistic. In this article, we assess the effect of relaxation of these two restrictions. Excluded volume among bound ions is modeled by a hard-rod potential in the context of the theory of a one-dimensional fluid. The possibility that a flexible chain folds in some manner in the immediate vicinity of a bound ion is modeled by allowing the mean spacing between charged groups on the polymer to become smaller as the number of bound ions increases. We compare our results with recent data on the binding of a series of oligolysines to single-stranded polynucleotides, which conflict with the predictions of the original theory of delocalized binding of point ions to rigid polyelectrolytes. Inclusion of excluded volume among bound ions does not significantly improve agreement with the data. Substantial improvement in the level of agreement is obtained when the polyion chain is assumed to be flexible. One of our conclusions is that the excluded-site description of anticooperativity, which was designed for the binding of ligands to discrete sites on a polymer chain, and which does not include the effect of ionic forces, should not be used in cases of delocalized binding of ions.

Binding Sites↗

Phosphophoryn, an "acidic" biomineralization regulatory protein: conformational folding in the presence of Cd(II).

The divalent cation-induced protein folding properties of the template macromolecule, bovine dentine phosphophoryn (BDPP), have been examined by 1H/31P/13C/113Cd-nmr spectroscopy. Cd(II) was employed, exploiting the sensitivity of 113Cd-nmr to ligand-binding interactions and kinetics. Cation binding was studied over the stoichiometric range of 0-50: 1 Cd(II): protein (mole ratio), well below the range of Cd(II) concentration required to induce protein precipitation. The stepwise titration of divalent cation-depleted phosphophoryn at pH 7.2 in H2O/D2O with 113CdCl2 revealed that (PSer)n, (PSerAsp)n, and (Asp)n polyelectrolyte cation-binding domains undergo two major transitions in their secondary and tertiary structures: the first transition, occurring between 1:10 and 1:1 Cd(II): protein stoichiometry, and the second, between 10:1 and 50:1. By monitoring the amide NH intensities, 31P-nmr chemical shift, and 13C Asp-C, resonances, it was concluded that Cd(II) ions exhibit a binding-site preference for polyelectrolyte cation-binding domains, in the order (PSer)n > (PSerAsp)n > (Asp)n This preference correlates with the degree of negative charge density for each sequence motif. Accompanying the backbone conformational transitions at the polyelectrolyte regions were conformational transitions in the flanking hinge domains, indicating that the hinge domains participate in the folding of the phosphophoryn molecule as divalent cation binding occurs at the polyelectrolyte domains. We were unsuccessful in detecting phosphophoryn-bound Cd(II) species by 113Cd-nmr because of chemical exchange modulation. However, using a smaller 21-residue peptide mimetic of phosphophoryn, we have observed three stoichiometric-dependent 113Cd resonances that differ in terms of the oxoanion coordination number. Our observation of multiple Cd(II) species in the presence of the peptide supports our contention that Cd(II) has many chemically distinct coordination sites on phosphophoryn, each in multiple equilibria with H2O, Cl-, and side-chain oxoatoms.

Amino Acid Sequence↗

Complexation between low-molecular-weight cationic ligands and negatively charged polymers as studied by capillary electrophoresis frontal analysis.

The potential of using capillary electrophoresis frontal analysis for the study of low-molecular-weight ligand-polyelectrolyte interactions was assessed. The interaction of the ligands 1-propylpyridinium bromide, 2-propylisochinolinium bromide, and paraquat with the polymer dextran sulfate was investigated as a function of polymer concentration and ionic strength of the buffer solution. Linear binding isotherms were obtained and association constants were determined. The complex formation was independent of the dextran sulfate concentration at low ionic strength. Ligand-polyelectrolyte interactions were strongly dependent on the ionic strength. The interaction of the divalent cation paraquat with the dextran sulfate was much stronger than the interactions of the monovalent cationic ligands with the polyelectrolyte. The binding data obtained were in accord with results obtained by equilibrium dialysis. Capillary electrophoresis frontal analysis has the potential to become a valuable tool for characterization of ligand-polyelectrolyte interactions in drug design as well as in other areas.

Cations↗

The role of the ionic environment in the orientation of nucleic acids in electric fields.

The relationship between polyelectrolyte theories based on linear charge density models and the electric-field induced orientation of the polyelectrolytes, poly(A), poly(C) and DNA is examined by varying their ionic environment with respect to ionic strength and acidity. The degree of counterion condensation on the polyelectrolytes predicted by the theories of Manning and Record is shown to be related linearly to the orientation as measured by their dichroism in the field. Micro-structural differences between poly(A) and poly(C) account for the differences in their dependence on the pH of the medium which affects the counterion condensation and thus the polarization in the orienting electric fields. The results consequently support recent treatments of linear polyelectrolytes having a high charge density which model them as smoothly charged linear polyions, but indicate that these models are insufficient to account for some of the effects of microstructural variations.

Journal Article↗

Protein binding properties of surface-modified porous polyethylene membranes.

In this study, we quantified the adsorption of immunoglobulin G (IgG) protein onto several polyelectrolyte-modified sintered porous polyethylene (PPE) membranes. The polymer surfaces had both cationic and anionic charges obtained via the adsorption of polyethylenimine (PEI) and polyacrylic acid (PAA), respectively, onto plasma-activated PPE. The amount of IgG adsorption was determined by measuring the gamma radiation emitted by [125I]-IgG radio labeled protein. By studying the impact of pH and ionic strength on IgG adsorption, we attempted to characterize the role and nature of the electrostatic interactions involved in the adsorption process to better understand how these interactions were influenced by the charge and structure of immobilized polyelectrolyte complexes at modified membrane surfaces. We were able to show that surface modification of PPE membranes with adsorbed PEI monolayers and PEI-PAA bilayers can greatly improve the IgG binding ability of the membrane under optimized conditions. We also showed that the observed improvement in the IgG binding is derived from electrostatic interactions between IgG and the polyelectrolyte surface. In addition, we found that the greatest IgG adsorption occurred when the IgG and the surface possessed predominantly opposite charges, rather than when the surface possessed the greatest electrostatic charge. Finally, we have found that the molecular weight of the terminating polyelectrolyte has a noticeable effect upon the electrostatic interactions between IgG and the PEI-PAA bilayer-modified PPE surfaces.

Acrylic Resins↗

Adsorption of bilayers and multilayers of cationic and anionic co-polymers of acrylamide on silicon oxide.

Silica surfaces were consecutively treated with copolymers of cationic and anionic polyacrylamides (C-PAM and A-PAM, respectively) and the layer-by-layer build-up was continuously monitored with the aid of stagnation point adsorption reflectometry (SPAR). Four different charge densities of the cationic polymer and one charge density of the anionic polymer were studied. The solid substrate used in the investigation was an oxidized Si wafer, the charge of which was varied by performing the measurements at different pH. Adsorption measurements were performed both in deionized water and with a background electrolyte concentration of 0.01 M NaCl. The results show that the adsorption of C-PAM at pH 6 was dominated by electrostatic interactions. However, a significant nonionic contribution to the adsorption of C-PAM on SiO(2) was detected-when the results of adsorption measurements conducted in deionized water and in 0.01 M NaCl were compared. At pH 9, the adsorption of C-PAM onto SiO(2) was found to be geometrically restricted since the adsorption stoichiometry between the polymer charges and the charges on the surface was less than 1 irrespective of the charge of the C-PAM. Adsorption of the A-PAM onto the C-PAM covered surface increases as a function of the adsorbed charges in the first layer. Experiments showed that it was possible to form multilayers of polyelectrolytes on the SiO(2) surface provided the charge of the C-PAM was high enough. The critical charge of the polyelectrolyte for the formation of multilayers was also dependent on the charge of the substrate; that is, the lower the surface charge the higher the critical charge of the C-PAM. The substrate affected the amount of polyelectrolyte adsorbed up to the fifth layer. For further layers there was almost a stoichiometric relationship between the charges of the polyelectrolytes in consecutive layers. Results from studies of the formed multilayers with a quartz crystal microbalance (QCM-D) indicated that there was a close correlation between energy dissipation into the multilayers and a decrease in the adsorption as detected with SPAR. This in turn indicates that a decrease in the reflectometer signal does not necessarily indicate a decrease in adsorption.

Journal Article↗

Interfacial properties of poly(maleic acid-alt-1-alkene) disodium salts at water/hydrocarbon interfaces.

The interfacial properties of poly(maleic acid-alt-1-alkene) disodium salts at hydrocarbon/water interfaces are determined. In all the studied systems, the interfacial tension decreases markedly with the polyelectrolyte concentration as the side-chain length increases. The results of the standard free energy of adsorption, DeltaG(ads)(0), are a linear function of the number of carbon atoms in the polyelectrolyte side chain. The contribution to DeltaG(ads)(0) per mol of methylene group varies from -0.64 to -0.52 kJ/mol for the n-octane/water to n-dodecane/water interfaces. DeltaG(ads)(0) data also reveal that the adsorption process is mainly determined by adsorption efficiency. Comparatively, the adsorption effectiveness seems to play a less important role. The theoretical interaction energies calculated for the insertion of one hydrocarbon molecule into the space formed by two neighboring polyelectrolyte side chains are in good agreement with the experimental results. The latter results are consistent with van der Waals-type interactions between the hydrocarbon molecules and the polyelectrolyte side chains.

Journal Article↗

Co-conditioning and dewatering of chemical sludge and waste activated sludge.

The conditioning and dewatering behaviors of chemical and waste activated sludges from a tannery were studied. Capillary suction time (CST), specific resistance to filtration (SRF), and bound water content were used to evaluate the sludge dewatering behaviors. Zeta potentials were also measured. Experiments were conducted on each sludge conditioned and dewatered separately, and on the sludge mixed at various ratios. Results indicate that the chemical sludge was relatively difficult to be dewatered, even in the presence of polyelectrolyte. When the waste activated sludge was mixed with the chemical sludge at ratios of 1:1 and 2:1, respectively, the dewaterability of chemical sludge improved remarkably while the relatively better dewaterability of the waste activated sludge deteriorated only to a limited extent. As the mixing ratios became 4:1 and 8:1, the dewaterability of the mixed sludge was equal to that of the waste activated sludge. The optimal polyelectrolyte dosage for the mixed sludge was equal to or less than that of the waste activated sludge. It is proposed that the chemical sludges act as skeleton builders that reduce the compressibility of the mixed sludge whose dewaterability is enhanced. Bound water contents of sludge decreased at low polyelectrolyte dosage and were not significantly affected as polyelectrolyte dosage increased. Advantages and disadvantages of co-conditioning and dewatering chemical sludge and waste activated sludge were discussed.

Electrolytes↗

Cationic polymers for selectivity control in the capillary electrophoretic separation of inorganic anions.

The capillary electrophoretic (CE) separation of the inorganic anions bromide, chloride, nitrate, nitrite, fluoride, sulfate and phosphate is described in 0.005 mol/L sodium chromate electrolyte in the presence of soluble polydisperse ionic polymers (polyelectrolytes). The cationic polyelectrolytes used were as follows: poly(1,1-dimethyl-3,5-dimethylenepiperidinium) chromate, hexadimethrine chromate, poly(1,1-dimethyl-3,5-dimethylenepyrrolidinium) chromate and ((diethylamino)ethyl)-dextran chromate in the concentration range 0.004-0.6% (w/v). These polyelectrolytes were shown to be capable of reversing the direction of the electroosmotic flow as well as inducing changes in analyte electrophoretic mobility, separation selectivity, and resolution. Changes in electrophoretic mobility by as much as 25% were observed for the sulfate anion, and the resolution of fluoride and phosphate was enhanced by a factor of 7.8. In the presence of 0.05-0.17% w/v poly(1,1-dimethyl-3,5-dimethylenepyrrolidinium) chromate at pH 8, separation currents were found to increase only slightly as compared to an electrolyte containing equivalent amounts of sodium chromate. Electroosmotic flow was also found to be fairly constant (+/- 16%) in the pH range 6.55-10.02 for 0.01% (w/v) poly(1,1-dimethyl-3,5-dimethylenepiperidinium) chromate at an ionic strength of 0.04, compared to a 400% change in the absence of the polyelectrolyte. The reproducibility of the electroosmotic mobility was between 0.36 and 6% RSD, and analyte electrophoretic mobility was between 0.01 and 1.6% RSD. Peak height reproducibility was 0.2-8.0% RSD. Separation efficiencies were between 258,000 and 780,000 theoretical plates, and detection limits were between 4.4 x 10(-7) and 9.1 x 10(-6) mol/L.

Anions↗

Layer-by-layer engineering of biocompatible, decomposable core-shell structures.

The objective of the present investigation was to fabricate composite colloidal particles consisting of a sacrificial, decomposable template of biodegradable nature covered with biocompatible polyelectrolyte multilayers using the layer-by-layer sequential adsorption technique. Poly-dl-lactic acid and poly(dl-lactic-co-glycolic acid) were chosen to design the microparticulate template, and a preliminary feasibility study was carried out with poly(styrene sulfonate sodium)-poly(allylamine hydrochloride) as shell components. The properties of both core-shell and hollow structures obtained by core dissolution were characterized by confocal laser scanning microscopy, microelectrophoresis, scanning force microscopy, and scanning electron microscopy. The concept was then extended to biocompatible polyelectrolytes as shell wall building blocks to deduce stable hollow capsules with tailored properties. Uniform, complete coating with oppositely charged polyelectrolyte pairs was achieved for all the combinations investigated. The results demonstrate that polyester microparticles could serve as viable alternative components to conventionally employed templates to derive hollow capsules with defined size, shape, and shell thickness. With all the components used for fabrication being biocompatible, these polyelectrolyte capsules may find interesting applications in the fields of biology, biochemistry, biotechnology, and drug delivery.

Biocompatible Materials↗

Specific interactions versus counterion condensation. 2. Theoretical treatment within the counterion condensation theory.

Polyuronates such as pectate and alginate are very well-known examples of biological polyelectrolytes undergoing, upon addition of divalent cations, an interchain association that acts as the junction of an eventually formed stable hydrogel. In the present paper, a thermodynamic model based on the counterion condensation theory has been developed to account for this cation-induced chain pairing of negatively charged polyelectrolytes. The strong interactions between cross-linking ions and uronate moieties in the specific binding site have been described in terms of chemical bonding, with complete charge annihilation between the two species. The chain-pairing process is depicted as progressively increasing with the concentration of cross-linking counterions and is thermodynamically defined by the fraction of each species. On these bases, the total Gibbs energy of the system has been expressed as the sum of the contributions of the Gibbs energy of the (single) chain stretches and of the (associated) dimers, weighted by their respective fractions 1 - theta and theta. In addition, the model assumes that the condensed divalent counterions exhibit an affinity free-energy for the chain, G(C)(aff,0), and the junction, G(D)(aff,0), respectively. Moreover, a specific Gibbs energy of chemical bonding, G(bond,0), has been introduced as the driving force for the formation of dimers. The model provides the mathematical formalism for calculating the fraction, theta, of chain dimers formed and the amount of ions condensed and bound onto the polyelectrolyte when two different types of counterions (of equal or different valence) are present. The effect of the parameter G(bond,0) has been investigated and, in particular, its difference from G(C,D)(aff,0) was found to be crucial in determining the distribution of the ions into territorial condensation and chemical bonding, respectively. Finally, the effect of the variation of the molar ratio between cross-linking ions and uronic groups in the specific binding sites, sigma0, was evaluated. In particular, a remarkable decrease in the amount of condensed counterions has been pointed out in the case of sigma0 = 1/3, with respect to the value of sigma0 = 1/4, characterizing the traditional "egg-box" structure, as a result of the drop of the charge density of the polyelectrolyte induced by complete charge annihilation.

Biopolymers↗

Protein-calcium carbonate coprecipitation: a tool for protein encapsulation.

A new approach of encapsulation of proteins in polyelectrolyte microcapsules has been developed using porous calcium carbonate microparticles as microsupports for layer-by-layer (LbL) polyelectrolyte assembling. Two different ways were used to prepare protein-loaded CaCO3 microparticles: (i) physical adsorption--adsorption of proteins from the solutions onto preformed CaCO3 microparticles, and (ii) coprecipitation--protein capture by CaCO3 microparticles in the process of growth from the mixture of aqueous solutions of CaCl2 and Na2CO3. The latter was found to be about five times more effective than the former (approximately 100 vs approximately 20 mug of captured protein per 1 mg of CaCO3). The procedure is rather mild; the revealed enzymatic activity of alpha-chymotrypsin captured initially by CaCO3 particles during their growth and then recovered after particle dissolution in EDTA was found to be about 85% compared to the native enzyme. Core decomposition and removal after assembly of the required number of polyelectrolyte layers resulted in release of protein into the interior of polyelectrolyte microcapsules (PAH/PSS)5 thus excluding the encapsulated material from direct contact with the surrounding. The advantage of the suggested approach is the possibility to control easily the concentration of protein inside the microcapsules and to minimize the protein immobilization within the capsule walls. Moreover, it is rather universal and may be used for encapsulation of a wide range of macromolecular compounds and bioactive species.

Adsorption↗

Electrostatic free energy of weakly charged macromolecules in solution and intermacromolecular complexes consisting of oppositely charged polymers.

When oppositely charged polyelectrolytes are mixed in water, attraction between oppositely charged groups may lead to the formation of polyelectrolyte complexes (associative phase separation, complex coacervation, interpolymer complexes). Theory is presented to describe the electrostatic free energy change when ionizable (annealed) (macro-)molecules form a macroscopic polyelectrolyte complex. The electrostatic free energy includes an electric term as well as a chemical term that is related to the dissociation of the ionic groups in the polymer. An example calculation for complexation of polyacid with polybase uses a cylindrical diffuse double layer model for free polymer in solution and electroneutrality within the complex and calculates the free energy of the system when the polymer is in solution or in a polyelectrolyte complex. Combined with a term for the nonelectrostatic free energy change upon complexation, a theoretical stability diagram is constructed that relates pH, salt concentration, and mixing ratio, which is in qualitative agreement with an experimental diagram obtained by Bungenberg de Jong (1949) for complex coacervation of arabic gum and gelatin. The theory furthermore explains the increased tendency toward phase separation when the polymer becomes more strongly charged and suggests that complexation of polyacid or polybase with zwitterionic polymer (e.g., protein) of the same charge sign (at the "wrong side" of the iso-electric point) may be due (in part) to an induced charge reversal of the protein.

Hydrogen-Ion Concentration↗

Diblock polyampholytes grafted onto spherical particles: Monte Carlo simulation and lattice mean-field theory.

Spherical brushes composed of diblock polyampholytes (diblock copolymers with oppositely charged blocks) grafted onto solid spherical particles in aqueous solution are investigated by using the primitive model solved with Monte Carlo simulations and by lattice mean-field theory. Polyampholyte chains of two compositions are considered: a copolymer with a long and a short block, A100B10, and a copolymer with two blocks of equal length, A50B50. The B block is end-grafted onto the surface, and its charge is varied, whereas the charge of the A block is fixed. Single-chain properties, radial and lateral spatial distributions of different types, and structure factors are analyzed. The brush structure strongly depends on the charge of the B block. In the limit of an uncharged B block, the chains are stretched and form an extended polyelectrolyte brush. In the other limit with the charges of the blocks compensating each other, the chains are collapsed and form a polyelectrolyte complex surrounding the particles. At intermediate charge conditions, a polyelectrolyte brush and a polyelectrolyte complex coexist and constitute two substructures of the spherical brush. The differences of the brush structures formed by the A100B10 and A50B50 polyampholytes are also analyzed. Finally, a comparison of the predictions of the two theoretical approaches is made.

Journal Article↗

Pore assembled multilayers of charged polypeptides in microporous membranes for ion separation.

In this study, highly permeable ion-selective membranes are prepared via immobilization of polyelectrolyte multilayer networks within the inner pore structure of a microporous (pore size = 0.2 microm) support. Electrostatic layer-by-layer assembly is achieved through alternate adsorption of cationic and anionic polyelectrolytes under convective flow conditions. To initiate pore assembly, the first layer consists of covalently bound charged polypeptides (poly(L-glutamic acid) (PLGA) or poly(L-lysine) (PLL)) establishing a charged support for subsequent adsorption. Nonstoichiometric immobilization of charged multilayers within a confined pore geometry leads to an enhanced volume density of ionizable groups in the membrane phase. This overall increase in the effective charge density allows for Donnan exclusion of ionic species (especially divalent co-ions) using microporous materials characterized by permeability values that exceed conventional membrane processes. Multilayer assemblies are fabricated using both PLGA/PLL and synthetic polyelectrolytes (poly(styrenesulfonate)/poly(allylamine)) in an attempt to compare the level of adsorption and separation properties of the resulting materials. The role of salt concentration in the carrier solvent on overall polyelectrolyte adsorption was examined to determine its effect on both solute (Cl-, SO4(2-), As(V)) and water transport. Constriction of the pore size induced by multilayer propagation was monitored through permeability measurements and dextran rejection studies at each stage of the deposition process.

Adsorption↗

Cell motility assays on tissue culture dishes via non-invasive confinement and release of cells.

In vitro cell migration assays are useful for screening bioactive agents that regulate angiogenesis, tumor metastasis, would healing, and immune responses by effecting changes in the rate of cell migration. Here we have developed a noninvasive in vitro migration assay that operates through release of confluent groups of cells initially confined within patterns of cell-resistant polyelectrolyte. Cell-resistant patterns of polyelectrolyte, separating groups of confluent cells, are rendered cell adhesive by adsorption of a second, cell adhesive polyelectrolyte of opposite charge; thereby, resulting in migration of cells into the separating regions. By dynamically controlling cell-surface interactions through self-assembly of cell-adhesive and cell resistant polyelectrolytes, this method eliminates the need to mechanically wound cells, as is done in current cell migration assays. The utility of this technique in identifying molecules and mechanisms that regulate cell migration is demonstrated by its application as an assay for the effects of platelet derived growth factors, cytoskeleton disrupting agents, and Merlin overexpression, on the migration of NIH 3T3 fibroblasts.

Animals↗