Influence of the homogenisation procedure on the physicochemical properties of PLGA nanoparticles

Pilocarpine HCl-loaded PLGA nanoparticles were prepared by emulsification solvent evaporation. Three different stabilisers, polyvinylalcohol (PVA), Carbopol and Poloxamer were used, as well as mixtures thereof. The influence of the homogenisation pressure and number of cycles on the properties of nanoparticles were studied. Particle size was shown to depend on the stabiliser used. An increase of the homogenisation pressure or the number of cycles resulted in a decrease in particle size. The zeta potential value was influenced mainly by the nature of the stabiliser. Particles stabilised with poloxamer or PVA showed a slightly negative zeta potential value, while samples stabilised with carbopol possessed a more negative zeta potential, which became less negative after homogenisation. Drug encapsulation depended strongly on the stabiliser used. The higher drug entrapment of the carbopol-stabilised particles could be explained by an electrostatic interaction between the negatively charged carboxyl groups of carbopol and the positively charged, protonated pilocarpine. The drug release patterns of the particles prepared were quite similar. Differences between the release patterns of the homogenised particles could be attributed both to differences in size as well as drug encapsulation. Turbidimetric measurements suggested an interaction between mucin and PLGA nanoparticles exclusively stabilised with Carbopol.     

Interactions between drug delivery particles and mucin in solution and at interfaces

Cubosome particles were produced by fragmenting a cubic crystalline phase of glycerol monooleate and water in the presence of a stabilizing poly(ethylene oxide)-based polymer. The aim of our investigation was to study the interaction between these particles and mucin to gain information on how they would perform as a vehicle for mucosal drug delivery. Particle electrophoresis was used to investigate the interactions between particles and mucin in solution, and ellipsometry was utilized to study the interactions between particles and mucin-coated silica surfaces. The interaction studies were performed at relevant physiological conditions, and the pH and ionic strength were varied to gain more information about the driving forces for the interaction. The results from electrophoretic measurements showed that mucin in solution adsorbed to the particles at pH 4, whereas at pH 6 no clear interaction was detected. From ellipsometric measurements it was evident that the particles adsorb reversibly to a mucin-coated silica surface at pH 4, while no adsorption of particles could be detected at pH 6. The overall conclusion is that the interaction between these particles and mucin is weak and pH-dependent. These findings are in agreement with other investigations of the interactions between mucin and poly(ethylene oxide) chains.     

Entrapment of some compounds into biocompatible nano-sized particles and their releasing properties

Two types of biocompatible nanoparticles with an average diameter of around 200 nm were formed only by mixing hydrolysates of chitosan and carboxymethyl cellulose (CMC). Nanoparticle A was produced from chitosanase hydrolysate of chitosan and cellulase hydrolysate of carboxymethyl cellulose, and nanoparticle B was produced from lysozyme hydrolysate of chitosan and the carboxymethyl cellulose hydrolysate. Negatively charged or amphoteric compounds were first mixed with chitosan hydrolysate and then added to carboxymethyl cellulose hydrolysate to effectively entrap them in the particles. Positively charged compounds could also be effectively entrapped by mixing the hydrolysates and the compound in the reverse order. Negatively charged compounds with high molecular weights were maintained in the particles even at the higher pH levels than the pK(a) of the amino groups of chitosan. Entrapped compounds were gradually released from nanoparticle A by lysozyme treatment. In contrast, there was no release from nanoparticle B. These results indicate that nanoparticle A can be applied to controlled-release drug delivery systems, and that nanoparticle B is stably retained in the body without releasing the entrapped compounds.     

Mucin-electrolyte interactions at the solid-liquid interface probed by QCM-D

The interaction between mucin and ions has been investigated by employing the quartz crystal microbalance technique with measurement of energy dissipation. The study was partially aimed at understanding the adsorption of mucin on surfaces with different chemistry, and for this purpose, surfaces exposing COOH, OH, and CH(3) groups were prepared. Mucin adsorbed to all three types of functionalized gold surfaces. Adsorption to the hydrophobic surface and to the charged hydrophilic surface (COOH) occured with high affinity despite the fact that in the latter case both mucin and the surface were negatively charged. On the uncharged hydrophilic surface exposing OH groups, the adsorption of mucin was very low. Another aim was to elucidate conformational changes induced by electrolytes on mucin layers adsorbed on hydrophobic surfaces from 30 mM NaNO(3). To this end, we investigated the effect of three electrolytes with increasing cation valance: NaCl, CaCl(2) and LaCl(3). At low NaCl concentrations, the preadsorbed layer expands, whereas at higher concentrations of NaCl the layer becomes more compact. This swelling/compacting of the mucin layer is fully reversible for NaCl. When the mucin layer instead is exposed to CaCl(2) or LaCl(3), compaction is observed at 1 mM. For CaCl(2), this process is only partially reversible, and for LaCl(3), the changes are irreversible within the time frame of the experiment. Finally, mucin interaction with the DTAB cationic surfactant in an aqueous solution of different electrolytes was evaluated with turbidimetry measurements. It is concluded that the electrolytes used in this work screen the association between mucin and DTAB and that the effect increases with increasing cation valency.     

Morphological and chemical features of nano and macroscale carbons affecting hydrogen peroxide decomposition in aqueous media

Stable film-forming nanocomposite particles with diameters ranging from 120 to 300 nm, based on polybutylmethacrylate (PBMA) and cellulose whiskers in water dispersions, were successfully synthesized in one step through mini-emulsion polymerization. The nanocomposite dispersion with a solid content of 25 wt.% and up to 5 wt.% of nanofiller loading was prepared by in situ polymerization, in the presence of the whiskers using dodecylpyridinium chloride (DPC), as a cationic surfactant, and 2,2-azobis(isobutyronitrile) (AIBN), as initiator. The electrostatic interaction between the positively charged droplets and negatively charged whiskers ensured the anchoring of the nanofiller around the polymer particles. The ensuing dispersions were characterized by Dynamic Light Scattering (DLS), ζ-Potential Measurements, and Field-Emission Scanning Electron Microscopy (FE-SEM). After the film formation process, the nanocomposite film exhibits a high transparency, denoting the good dispersion of the whiskers throughout the matrix.     

Mucin-chitosan complexes at the solid-liquid interface: multilayer formation and stability in surfactant solutions

The adsorption of a biologically important glycoprotein, mucin, and mucin-chitosan complex layer formation on negatively charged surfaces, silica and mica, have been investigated employing ellipsometry, the interferometric surface apparatus, and atomic force microscopy techniques. Particular attention has been paid to the effect of an anionic surfactant sodium, dodecyl sulfate (SDS), with respect to the stability of the adsorption layers. It has been shown that mucin adsorbs on negatively charged surfaces to form highly hydrated layers. Such mucin layers readily associate with surfactants and are easily removed from the surfaces by rinsing with solutions of SDS at concentrations > or =0.2 cmc (1 cmc SDS in 30 mM NaCl is equal to 3.3 mM). The mucin adsorption layer is negatively charged, and we show how a positively charged polyelectrolyte, chitosan, associates with the preadsorbed mucin to form mucin-chitosan complexes that resist desorption by SDS even at SDS concentrations as high as 1 cmc. Thus, a method of mucin layer protection against removal by surfactants is offered. Further, we show how mucin-chitosan multilayers can be formed.     

Effect of cationic polyacrylamides on the interactions between cellulose fibers

The interaction between cellulose fibers in the presence of cationic polyacrylamide (CPAM) was analyzed by rheology as a function of polyelectrolyte concentration, charge density, and molecular weight. CPAM was found to strongly influence the yield stress of cellulose suspensions; low doses of CPAM increased the yield stress, but at higher concentrations the yield stress declined. The charge density of the CPAM was the most significant factor in how yield stress responded to CPAM concentration; this effect was able to be normalized to a master curve by considering only the charged fraction of the polymer. The molecular weight of CPAM samples had some effect at high concentrations, but for lower CPAM doses the yield stress was independent of molecular weight over the range studied. The data suggest that CPAM modifies the interaction between cellulose surfaces via several mechanisms, with electrostatic interactions in the form of charge neutralization and charged patch formation dominating; polymer bridging and steric repulsion also influence the overall balance of forces between interacting cellulose fibers.     

Interaction of DNA/polycation complexes with phospholipids: stabilizing strategy for gene delivery

The interaction between negatively charged lipid vesicles and positively charged DNA/polylysine complexes was studied. The interaction does not lead to release of DNA from the initial complexes. The particles formed are easy to prepare, they have slight negative charge, small dimensions and show good stability in physiological NaCl solution. Such properties might indicate that stabilization of the particles by lipid coating might be a potent strategy, alternative to PEGylation of DNA/polycation complexes. Interaction of DNA/polycation complex particles with lipid vesicles.     

Influence of the functionalization pattern of ethyl cellulose on the interactions with polystyrene latex particles in aqueous mixtures

The interactions between polystyrene latex particles and ethyl cellulose (EC) with different functionalization pattern have been investigated. 3-Mono-O-EC and EC with statistical functionalization pattern in the anhydroglucose units were studied in aqueous solutions and dispersions. EC belongs to a group of polymers that phase separate upon heating. The two types of EC showed large differences in phase separation temperature, which was explained as an effect of different interactions with water due to different functionalization pattern. Both types of EC did adsorb on polystyrene particles, which indicated a favorable interaction between EC and polystyrene latex particles, however, in a different manner depending on the structure of EC. The conventionally synthesized ethyl cellulose with statistical functionalization pattern formed much stronger networks with polystyrene latex particles than 3-mono-O-EC did. The lower phase separation temperature and the slightly higher molecular weight of the conventional ethyl cellulose gave it higher preference for interacting with polystyrene latex particles to form network. Throughout the study, comparison is made with other cellulose derivatives like ethyl(hydroxyethyl) cellulose (EHEC) and carboxymethyl cellulose (CMC).     

Interactions between chitosan-modified particles and mucin-coated surfaces

Lipid-based particles (Cubosome particles) were surface-modified by chitosan and the ratio between particles and chitosan was optimized to minimize the free chitosan concentration in the dispersion. The modified particles were characterized by electrophoretic measurements and the pH dependence of the zeta potential could be directly related to the protonation of chitosan. Interaction between the modified particles and mucin-coated silica surfaces were subsequently investigated in situ by ellipsometry to assess the mucoadhesive properties at physiologically relevant conditions. The result showed that a substantial amount of modified particles was adsorbed to mucin-coated silica surfaces at both pH 4 and pH 6, probably due to electrostatic interactions between amino groups in chitosan and negatively charged groups in mucin. Furthermore, the amount of bound particles decreased by less than 15% upon rinsing indicating relatively strong interactions. This investigation demonstrates that ellipsometry is a useful tool to study mucoadhesive properties of particles in the submicrometer range. Moreover, the novel chitosan-modified particles may be of interest for mucosal drug delivery applications.     

Mechanisms of Diffusion-Reduction Interaction of Microcrystalline Cellulose and Silver Ions

The preparation of microcrystalline cellulose samples containing intercalated silver(0) particles includes two main stages: diffusion of silver ions from aqueous AgNO₃ into the microcrystalline cellulose matrix, followed by reduction of the silver ions bound to cellulose macromolecules to Ag(0) and growth of Ag(0) particles inside the cellulose matrix and on its surface. Mechanisms of interaction of silver ions with cellulose as reducer and with other reducers and ligands are considered.     

Induced phase separation in low-ionic-strength cellulose nanocrystal suspensions containing high-molecular-weight blue dextrans

Spontaneous entropic phase separation phenomena occur in a wide range of systems containing highly anisotropic colloidal particles. Among these are aqueous suspensions of negatively charged cellulose I nanocrystals produced by sulfuric acid hydrolysis of native cellulose, which phase separate into isotropic and chiral nematic liquid-crystalline phases. Phase separation of an isotropic phase from a completely ordered nanocrystal suspension may be induced by the addition of salts or nonadsorbing macromolecules. In previous work (Edgar, C. D.; Gray, D. G. Macromolecules 2002, 35, 7400-7406), an isotropic phase was found to form over a period of several days when blue dextran (a sulfonated triazine dye, Cibacron blue 3G-A, covalently attached to high-molecular-weight dextran chains) was added to initially ordered suspensions. Here we report work showing that the observed phase separation was associated with the charged dye molecules attached to the dextran. The Cibacron blue 3G-A dye attached to blue dextran was found to induce greater phase separation than free (unbound) dye; at increasing ionic strength, depletion attractions due to the blue dextran increasingly contribute to the phase separation.     

Binding of mucin to water-soluble and surface-grafted boronate-containing polymers

The binding of mucin to water-soluble copolymers of N,N-dimethylacrylamide and N-acryloyl-m-aminophenylboronic acid grafted on the surface of glass is studied. Atomic force microscopy studies show that many graft copolymer islands 20–200 nm in diameter and 50 nm in height occur on the modified surface of flat glass. Owing to the presence of phenyl boronate groups, the copolymer behaves as a weak polyelectrolyte (pK _a = 9.0) and, in the grafted state in an aqueous solution, experiences reversible transitions between states with higher and lower degrees of ionization. As evidenced by spectral correlation interferometry, this phenomenon brings about a change in the thickness of the grafted layer by approximately 0.5 nm. The ability of phenyl boronate groups to form cyclic esters with diol and polyol groups results in complexation of the soluble copolymer with mucin oligosaccharides and entails the appearance of slowly growing submicron particles formed by similarly charged polymers. The specificity of complexation is confirmed by dissolution of particles in the presence of fructose: a saccharide with a strong affinity for phenyl boronate groups. The binding of mucin to glass, which is chemically modified with the above copolymer, leads to formation of an adsorption layer with a thickness of 1.2–1.8 nm. Thus, boronate-containing copolymers are suitable for preparing carriers with controllable adsorption properties with respect to polyols, including mucinlike proteins of cellular glycocalyxes.     

Adsorption and viscoelastic properties of cationic xylan on cellulose film using QCM-D

The adsorption and viscoelastic properties of cationic xylan layers adsorbed from an aqueous electrolyte solution (NaCl 0, 1, 10, 100 mM) on a cellulose model surface were studied using quartz crystal microbalance with dissipation (QCM-D). Three cationic xylans with different charge densities were used (molecular weight, 9,600 g/mol with degrees of substitution, DS = 0.150, 0.191, and 0.259). The influences of the electrolyte concentration and charge density of cationic xylan on its adsorption onto a cellulose surface were investigated. Low charged cationic xylan was substantially more efficient in surface adsorption on cellulose compared to high charged cationic xylan at a low concentration of electrolytes. Adsorption of low charged cationic xylan decreased with increases in electrolyte concentration. However, adsorption of high cationic xylan increased with electrolyte concentration. The conformation and viscoelastic properties of the layers were interpreted by modeling the data under the assumption that the layers can be explained by the a Voigt model. Low charged cationic xylan adsorbed relatively weakly onto the cellulose surface, and formed a thicker, softer layer than high charged cationic xylan. On the other hand, high charged cationic xylan formed a thinner adsorption layer onto the cellulose surface.     

Interactions of β-Lactoglobulin with Bovine Submaxillary Mucin vs. Porcine Gastric Mucin: The Role of Hydrophobic and Hydrophilic Residues as Studied by Fluorescence Spectroscopy

The aim of this study was to investigate binding interactions between β-lactoglobulin (BLG) and two different mucins, bovine submaxillary mucins (BSM) and porcine gastric mucin (PGM), using intrinsic and extrinsic fluorescence spectroscopies. Intrinsic fluorescence spectra showed an enhanced decrease of fluorescence intensity of BLG at all pH conditions when BLG was mixed with PGM rather than with BSM. We propose that, unlike BSM, the tertiary structure of PGM changes and the hydrophobic regions are exposed at pH 3 due to protonation of negatively charged residues. Results suggest that PGM also facilitated the structural unfolding of BLG and its binding with PGM by a hydrophobic interaction, especially at acidic pH, which was further supported by extrinsic fluorescence spectroscopy. Hydrophobic interaction is suggested as the dominant interaction mechanism between BLG and PGM at pH 3, whereas electrostatic interaction is the dominant one between BLG and BSM.     

Interaction forces between waterborne bacteria and activated carbon particles

Activated carbons remove waterborne bacteria from potable water systems through attractive Lifshitz-van der Waals forces despite electrostatic repulsion between negatively charged cells and carbon surfaces. In this paper we quantify the interaction forces between bacteria with negatively and positively charged, mesoporous wood-based carbons, as well as with a microporous coconut carbon. To this end, we glued carbon particles to the cantilever of an atomic force microscope and measured the interaction forces upon approach and retraction of thus made tips. Waterborne Raoultella terrigena and Escherichia coli adhered weakly (1-2 nN) to different activated carbon particles, and the main difference between the activated carbons was the percentage of curves with attractive sites revealed upon traversing of a carbon particle through the bacterial EPS layer. The percentage of curves showing adhesion forces upon retraction varied between 21% and 69%, and was highest for R. terrigena with positively charged carbon (66%) and a coconut carbon (69%). Macroscopic bacterial removal by the mesoporous carbon particles increased with increasing percentages of attractive sites revealed upon traversing a carbon particle through the outer bacterial surface layer.     

Normal and friction forces between mucin and mucin-chitosan layers in absence and presence of SDS

Employing the colloidal probe AFM technique we have investigated normal and friction forces between flat mica surfaces and silica particles coated with mucin and combined mucin-chitosan layers in presence and absence of anionic surfactant, SDS, in 30 mM NaCl solution. We have shown that the normal interactions between mucin coated mica and silica surfaces are dominated by long-range steric repulsion on both compression and decompression. Friction forces between such mucin layers are characterized by a low effective friction coefficient, mu(eff)=0.03+/-0.02, which is lower than the value of 0.13+/-0.02 observed when chitosan layers were adsorbed. Forces between combined mucin-chitosan layers have also been measured. Adsorption of chitosan on mucin results in considerable compaction of the layer, and development of attractive forces detectable on separation. Friction between mucin-chitosan layers in 30 mM NaCl solution is high, with mu(eff) approximately 0.4. Adsorption of additional mucin to this layer results in no improvement with respect to lubrication as compared to the mucin-chitosan layer, and mu(eff) approximately 0.4 is observed. We argue that the layers containing both mucin and chitosan are not strictly layered but rather strongly entangled. As a result attractive interactions between oppositely charged moieties of sialic acid residues from mucin and amine groups from chitosan residing on the opposing surfaces contribute to the increased friction. The effects of SDS on normal and friction forces between combined mucin-chitosan layers were also investigated. The relation between surface interactions and friction properties is discussed.     

Electrokinetic phenomena of soft particles

A review is given on the recent developments of theories of electrokinetics of soft particles (i.e., hard particles covered with an ion-penetrable surface layer of polyelectrolytes) with particular emphasis on the electrophoretic relaxation effects for highly charged soft particles and the effects of inhomogeneous distribution of polymer segments in the surface layer. The electrostatic interaction between soft particles is also discussed. Not only the interaction before contact of the surface layers of the interacting particles but also the interaction after their contact is considered.     

Synthesis and characterization of amphoteric latex particles

Stable monodisperse amphoteric latex particles were prepared by the semibatch surfactant-free emulsion copolymerization of methyl methacrylate and methacrylic acid (MAA) initiated by 2,2'-azobis(2-amidinopropane) dihydrochloride (V-50). These submicron particles have a net positive charge, which is attributed to the ionized amino group at low pH. In contrast, they become negatively charged owing to the ionized carboxyl group at high pH. There exists a pH at which these particles exhibit a net charge of zero (pI). At a constant level of V-50, the pI value of these latices decreases with increasing amount of MAA used in the polymerization recipe. The effect of pH on the colloidal stability of these amphoteric latices toward the addition of the negatively charged latex was investigated. The resultant coagulation kinetics was used to study the electrostatic interaction between the amphoteric particles and negatively charged particles.