Development of PEG-PLA/PLGA microparticles for pulmonary drug delivery prepared by a novel emulsification technique assisted with amphiphilic block copolymers

We developed a novel "spray dry-based" method for preparing surface-modified particle via "block copolymer-assisted" emulsification/evaporation for pulmonary drug delivery. The method included three steps: (1) o/w emulsion containing both hydrophobic polymers and amphiphilic block copolymers was obtained by emulsification of water and a polymer-containing organic solvent, (2) the o/w emulsion was misted with a nebulizer, and (3) the emulsion mists were dried by a heater. In this way, the hydrophobic polymers and the hydrophobic part of the amphiphilic block copolymers gradually tangled during the evaporation of organic solvents from the o/w emulsion. Consequently, the hydrophilic polymer chain was introduced on the particle surface. The particle surface can be easily modified although there are no reactive groups in the hydrophobic polymer molecules. We successfully obtained dry PEG-PLA/PLGA microparticles by controlling the weight ratio of the block copolymer and the hydrophobic polymer. The introduction of PEG to the particle surface involves an increase in the Zeta potential of the particles. Interestingly, the "dimpled" microparticles having a diameter of approximately 2 μm were obtained. The "dimpled" microparticles can serve as drug carriers for pulmonary drug delivery, because the particles have a large surface area. We expect that this novel surface-modification technique will enable efficient fabrication of particles in drug delivery systems.     

Structure and dynamics of a sponge phase in the methyl delta-aminolevulinate/monoolein/water/propylene glycol system

The structural effect caused by the addition of up to 16% (w/w) of the hydrochloride salt of delta-aminolevulinic acid (ALA, HOOC-CH2-CH2-CO-CH2-NH2HCl) or its methyl ester (m-ALA) to the sponge phase formed of monoolein/water/propylene glycol was investigated by means of crossed polarizers, small angle X-ray diffraction (SAXD) and nuclear magnetic resonance diffusometry (NMRD). Inspection with crossed polarizers revealed that additions of 4-16% (w/w) m-ALA transformed the isotropic bicontinuous sponge phase partly (4-10%) or completely (13 and 16%) into an anisotropic lamellar phase, indicating that m-ALA has a flattening effect on the bilayer curvature. The addition of 16% (w/w) ALA did not show any effect on the sponge phase. By addition of water to the anisotropic m-ALA samples, isotropic liquids were re-formed. The SAXD data for the isotropic liquids showed a diffuse Bragg peak and the NMRD self-diffusion coefficients for the drug (m-ALA) and the components of the original sponge phase (monoolein, water and propylene glycol) were shown to be essentially constant for 0-16% (w/w) added m-ALA. These results confirmed the hypothesis that the re-formed isotropic phases were indeed sponge phases. Water, for example, showed a diffusion coefficient of 3.1-3.9x10(-10)m(2)s(-1) in the sponge phase, compared to 5.3-5.7 x 10(-10)m2s(-1) in relevant water/propylene glycol solutions or 2.3 x 10(-9)m2s(-1) in pure water. The reduction can be explained as a consequence of the microstructure (congruent monoolein bilayer) of the sponge phase and of the viscosity effect caused by propylene glycol and m-ALA.     

Engineering proteinaceous colloidosomes as enzyme carriers for efficient and recyclable Pickering interfacial biocatalysis

Despite Pickering interfacial biocatalysis being a popular topic in biphasic biocatalysis, the development of water-in-oil (w/o) emulsion systems stabilized by single particles remains a challenge. For the first time, hydrophobized proteinaceous colloidosomes with magnetic-responsiveness are developed to function as both an enzyme carrier and emulsifier, achieving a breakthrough in protein-based w/o Pickering bioconversion. Enzyme-loaded protein colloidosomes are synthesized by a facile and mild method via emulsion templating. This system exhibits superior catalytic activity to other systems at the oil–water interface. Besides, feasible enzyme recovery and reusability ensure that this novel system can be employed as an efficient and eco-friendly recyclable platform.

Engineering proteinaceous colloidosomes with magnetic-responsiveness are designed as both enzyme carrier and emulsifier, achieving a breakthrough in protein-based w/o Pickering interfacial biocatalysis.     

Water in oil (w/o) and double (w/o/w) emulsions prepared with spans: microstructure,stability, and rheology

The objective was to analyze the microstructure, stability, and rheology of model emulsions prepared with distilled water, refined sunflower oil, and different Spans (20, 40, 60, and 80) as emulsifiers. The effects of the water content and Span 60 concentration were studied. The lowest water contents led to w/o emulsions, whereas higher percentages gave w/o/w emulsions. Microscopy analysis showed that w/o/w emulsions of higher water contents had a lower number of internal water droplets. W/o emulsions were destabilized by coalescence and sedimentation, whereas creaming was observed in unstable w/o/w emulsions. In the last ones, the creaming stability decreased with increasing water content and enhanced with higher Span 60 concentration; the same effect was observed in their viscoelasticity: They were from unstable liquids to stable gels. Solid Spans (40 and 60) produced more consistent w/o/w emulsions at low water contents and more stable systems at high water percentages in comparison with liquid Spans (20 and 80).     

Phase Diagram,Electrical Conductivity,and Viscosity Study of Alpha-Sulfonated Methyl Esters Derived from Palm Stearin/1-Pentanol/Hydrocarbon/Water Systems

Microemulsion region of sodium salt alpha-sulfonated methyl ester derived from palm stearin (α-SMEPS)/1-pentanol/alkane/water systems was constructed and investigated at 30.0 ± 0.1°C. The water-in-oil (w/o) microemulsion, bicontinuous structure, and oil-in-water (o/w) microemulsion regions were determined by both conductivity and viscosity methods. Results showed that the microemulsion region decreases and moved towards (α-SMEPS f 1-pentanol) rich corner with the increase in hydrocarbon chain length of alkane from C8 to C16. The changes in alkane chain length from C16 to C8 were found to have shifted the o/w microemulsion f bicontinuous structure f w/o microemulsion transitions of α-SMEPS self-assembly to the rich water corner.     

Studies of cyclohexane/TX 100 + ethylpropionate/water system: effect of ethylpropionate as cosurfactant

 Pseudoternary phase diagram of cyclohexane/TX 100-ethylpropionate/water system was determined at 30 ^°C. One phase microemulsion was obtained over a small area. The surfactant/cosurfactant ratio was both kept at 1:1 and 1:2 w/w. Viscosity, conductance, adiabatic compressibility values at various temperatures show the expected trend. The addition of NaCl changes one phase microemulsion to Winsor II system and no Winsor III system was obtained. Contact angle data, conductance, etc., show oil continuous system at a constant surfactant weight fraction. The microstructure of Winsor IV microemulsion seems to be W/O. Received: 14 May 1996 Accepted: 23 October 1996     

The effect of changing the microstructure of a microemulsion on chemical reactivity

A kinetic study was carried out on various solvolytic reactions in water/ NH4OT /isooctane microemulsions. The NH4OT surfactant is a derivative of the sodium salt of bis(2-ethylhexyl) sulfosuccinate (NaOT or AOT), where the Na+ counterion has been replaced by NH4+. The counterion substitution effects the phase diagram of the system, and therefore, NH4OT-based microemulsions with high water content reaching values of W = 350 (W = [H2O]/[NH4OT]) can be obtained. The presence of high W values suggests a transition in the microemulsion microstructure from water-in-oil (w/o) to oil-in-water (o/w), as was confirmed by conductivity and 1H NMR self-diffusion measurements. The interpretation of the kinetic studies in terms of pseudophase formalism allows us to analyze the effect of the microemulsion on chemical reactivity, regardless of its microstructure. It has been confirmed that the values of the solvolytic rate constants at the interphase of oil-in-water microemulsions are similar to those obtained for aqueous SDS systems, showing that the hydration degree of the interphase of the oil-in-water microemulsions is independent of W. The influence of the surfactant counterion on the solvolytic rate constants was analyzed by comparing HOT-, NaOT-, and NH4OT-based microemulsions. An important influence on the rate constants caused by the changes in the structural properties of water has been observed as was confirmed by the water 1H NMR signals.     

Effect of alpha-lactalbumin on aerosol-OT phase structures in oil/water mixtures

The ability of water-soluble, globular proteins to tune surfactant/oil/water self-assemblies has potential for the formation of biocompatible microemulsions and also plays a role in protein function at biological interfaces. In this work, we examined the effect of the protein alpha-lactalbumin on Aerosol-OT (AOT) phase structures in equivolume mixtures of oil and 0.1 M brine. In this pseudo-ternary system, surfactants are free to move to either oil or water phase to adopt phase structures close to the spontaneous curvature of the surfactants. Using small-angle X-ray scattering, we observed that addition of this protein changed the spontaneous curvature of the surfactant monolayer substantially. In the absence of protein, AOT adopted a negative spontaneous curvature to form spherical w/o microemulsion droplets. When less than 1 wt % of alpha-lactalbumin was added into the system, the w/o droplets became nonspherical and larger in volume, corresponding to an increase in water uptake into the droplets. As the protein-to-surfactant ratio increased, protein, surfactant, and oil increasingly partitioned toward the aqueous phase. There the protein triggered the formation of o/w microemulsions with a positive spontaneous curvature. These protein-containing structures exhibited significant interparticle attraction. We also compared the influence of two oil types, isooctane and cyclohexane, on the protein/surfactant interactions. We propose that the more negative natural curvature of the AOT/cyclohexane monolayer in the absence of protein prevented protein incorporation within organic phase structures and consequently pushed the system self-assembly toward aqueous aggregate formation.     

Preparation of insulin-loaded PLA/PLGA microcapsules by a novel membrane emulsification method and its release in vitro

Uniform-sized biodegradable PLA/PLGA microcapsules loading recombinant human insulin (rhI) were successfully prepared by combining a Shirasu Porous Glass (SPG) membrane emulsification technique and a double emulsion-evaporation method. An aqueous phase containing rhI was used as the inner water phase (w1), and PLA/PLGA and Arlacel 83 were dissolved in a mixture solvent of dichloromethane (DCM) and toluene, which was used as the oil phase (o). These two solutions were emulsified by a homogenizer to form a w1/o primary emulsion. The primary emulsion was permeated through the uniform pores of a SPG membrane into an outer water phase by the pressure of nitrogen gas to form the uniform w1/o/w2 droplets. The solid polymer microcapsules were obtained by simply evaporating solvent from droplets. Various factors of the preparation process influencing the drug encapsulation efficiency and the drug cumulative release were investigated systemically. The results indicated that the drug encapsulation efficiency and the cumulative release were affected by the PLA/PLGA ratio, NaCl concentration in outer water phase, the inner water phase volume, rhI-loading amount, pH-value in outer water phase and the size of microcapsules. By optimizing the preparation process, the drug encapsulation efficiency was high up to 91.82%. The unique advantage of preparing drug-loaded microcapsules by membrane emulsification technique is that the size of microcapsules can be controlled accurately, and thus the drug cumulative release profile can be adjusted just by changing the size of microcapsules. Moreover, much higher encapsulation efficiency can be obtained when compared with the conventional mechanical stirring method.     

Etude de phases quaternaires Winsor IV. Diagrammes de phases et propriétés electriques

A comparative study of phase diagram features and electrical properties of Winsor IV phases (so-called microemulsions) led to define two types of quaternary systems involving water, a hydrocarbon, and an ionic surfactant/alcohol combination defined byk, the surfactant/alcohol mass ratio. Systems of the first type exhibit a Winsor IV domain consisting of two disjointed areas corresponding to water-in-oil (w/o) and oil-in-water (o/w) monophasic fluid transparent isotropic media. The w/o and o/w areas are separated by a composition zone over which exist viscous turbid long-range organized structures related to the o/w w/o phase inversion mechanism. In that case, over the w/o area, the low frequency electrical conductivity and permittivity undergo non-monotonous changes as the composition varies. From conductivity maxima and minima, it is possible to define in the general case two lines ₁ and ₂ separating three adjacent sub-areas to which can be assigned compositions representing pre-micellar entities, inverted swollen spherical micelles and micelles clusters. For systems of the second type, the w/o and o/w sub-areas merge so as to form a unique monophasic area, which implies that the w/o o/w phase inversion occurs through a progressive diffuse mechanism. In that case the conductivity exhibits much higher values than in the preceding situation, and its variations with composition allow to define two linesC _d andC _m partitioning the Winsor IV domain into three adjacent areas. AboveC _d , that is for low and medium water contents, the conductivity variations with water content follow equations derived from the Percolation and Effective Medium theories, which indicates that the w/o swollen spherical micelles are submitted to attractive interactions. Below C_m, i. e. in the water rich region, the conductivity decrease with water content results from the progressive dilution of the external aqueous phase of the o/w Winsor IV media. BetweenC _d andC _m , the Winsor IV media exhibit an anomalous conductive behaviour which suggests that they are neither w/o nor o/w systems. This region can be considered as the diffuse phase inversion zone over which the systems are in a hybrid state that could be depicted tentatively as resulting from the formation of equilibrium bicontinuous structures.

     

Interfacial composition and structural parameters of water/C12-s-C12 x 2Br/n-hexanol/n-heptane microemulsions studied by the dilution method

The interfacial composition of the stable water/C12-s-C12 x 2Br/n-hexanol/n-heptane microemulsions has been studied in detail by dilution method. The results showed a marked maximum amount of the n-hexanol populating on the surfaces of droplets (represented as a = n(a)i/n(s), where n(a)i and n(s) are respectively the moles of n-hexanol and gemini surfactant on the surface of droplets) with increasing water content. At a constant level of water addition (the molar ratio of water to surfactant W0 = 20), a decreased with increasing the spacer length in the C12-s-C12 x 2Br molecule. The structural parameters of a w/o microemulsion were also estimated by analyzing the data of dilution experiments, and we found that the radius of the water pool was very sensitive to the increment of water content. The radius of the water pool varied from 0.74 to 5.35 nm with increasing W0 from 10 to 50. The variation extent reached 4.61 nm. In the cases of water/CPC/n-butanol/isopropyl myristate and water/CTAB/n-butanol/isopropyl myristate, however, the corresponding variation extents were only 1.22 and 1.68 nm, respectively, when increasing comparable water content. The ratio of N(a)/N(2C), where N(a) and N(2C) are respectively the average numbers of n-hexanol and the total average numbers of alkyl chains of gemini surfactant populating on per droplet surface, decreased obviously with increasing water content at W0 > 15. This indicated that C12-2-C12 x 2Br favored to form large droplets that were suitable to solubilize more water.     

The influence of water on the photophysical and photochemical properties of Piroxicam in AOT/iso-octane/water reversed micelles

The photophysical properties of Piroxicam , a nonsteroidal anti-inflammatory drug (NSAID), were investigated at different pH_ext values in reversed micelles of Aerosol-OT (AOT) in iso -octane, using both steady-state and picosecond time-resolved fluorescence spectroscopy. In contrast with the very complex data obtained in aqueous media, where several prototropic species are in equilibrium, the reversed micellar system essentially favors two species. The absorption spectra shows only one isosbestic point at λ= 348 nm. Excited-state intramolecular proton transfer (ESIPT), also detected in water, is promoted at low water pool contents measured by ω₀= [H₂O]/[AOT]. A strongly shifted (λ_em= 470 nm) tautomeric emission is found. Upon the gradual increase of ω₀, striking differences with pH_ext are found. At pH_ext= 4, the drug preferentially locates itself in the interfacial region partitioning between a hydrophobic and a hydrophilic domain. Global analysis was applied to the decay data and the results were interpreted by the "two-state excited-state" formalism. At pH_ext= 7, the anionic species is prevalent and the probe locates itself deeper inside the water core of the reversed micelles. Thus, a strong dependence on water content is detected, approaching a behavior similar to that observed in free aqueous solutions.     

Effect of Emulsification Method on the Properties of Lecithin‐ and PGPR‐Stabilized Water‐in‐Oil‐Emulsions

Water‐in‐oil (w/o) emulsions were prepared with phosphatidylcholine‐depleted lecithin or polyglycerol polyricinoleate (PGPR) as emulsifying agents. The effect of different laboratory emulsification devices and the effect of sodium chloride on particle size distribution, coalescence stability, and water droplet sedimentation were investigated. The properties of lecithin‐stabilized w/o emulsions were found to depend more strongly on the emulsifying method than those prepared with PGPR. The rotor‐stator system was not suitable for preparing stable w/o emulsions with lecithin. Whereas the addition of salt was essential to achieve coalescence‐stable emulsions prepared with PGPR, the presence of NaCl favored the coalescence of water droplets and phase separation in emulsions containing lecithin.     

Microheterogeneous solvation for aminolysis reactions in AOT-based water-in-oil microemulsions

A kinetic study was carried out on the aminolysis of p-nitrophenyl acetate (NPA) by n-decylamine (DEC), piperazine (PIP) and sarcosine (SAR) in AOT/isooctane/water (w/o) microemulsions. By using the pseudophase model both the rate constants at the interface, k2i, and the water microdroplet, k2w, can be obtained. The obtained results show that k2i increases together with the water content of the microemulsion, whereas k2w increases as the water content of the system decreases. In the aqueous microdroplet the predominant interaction Na+...OH2 causes a decrease in the strength of the hydrogen bonds and therefore facilitates the desolvation of the reagents as W decreases. This desolvation of the reagents causes the increase of k2w as W decreases. In the interface of the microemulsion the predominant interaction SO3-...HOH causes an increase in the electronic density on the water molecules and the consequent decrease in their efficiency in the solvation of the partial negative charge, which develops on the carbonyl oxygen atom in the transition state of the reaction. This decrease in the solvation causes k2i to decrease together with the water content of the system.     

Effect of Water Fraction on Rheological Properties of Waxy Crude Oil Emulsions

This article discusses the effect of water fraction on the rheological properties of waxy crude oil emulsions including gel point, yield stress, viscosity, and thixotropy. The experimental results reveal that the rheological behaviors of the w/o emulsion samples all intensify with the increase of water volume fraction within 60%. Of more significance is that a correlation for w/o emulsions between yield stress and water volume fraction is put forward with an average relative error of 6.75%. In addition, some mainstream viscosity prediction models of w/o emulsions are evaluated, and Elgibaly model is the best-fit for the emulsions in this study.     

Effect of Adding an Amphiphilic Solubilization Improver, Sucrose Distearate, on the Solubilization Capacity of Nonionic Microemulsions

We have studied the effect of adding sucrose distearate (2C(18)SE) on the solubilization capacity of microemulsions formed in the water/C(12)EO(6)/n-decane system. Upon addition of 2C(18)SE to the binary water/C(12)EO(6) system, a lamellar liquid crystal region developed. This suggests that the rigidity of the surfactant layer is strengthened. The solubilization of water and n-decane in the bicontinuous microemulsions increases about three times upon replacing 10% C(12)EO(6) with 2C(18)SE; besides, the HLB temperature is not greatly affected by 2C(18)SE. On the other hand, sucrose monostearate (C(18)SE) does not have such a function. The effect of added 2C(18)SE on the solubilization capacity of the discrete droplet-type o/w or w/o microemulsions was also studied. The efficiency of the solubilization-improving effect is reduced when the system is far from the HLB temperature. Copyright 2001 Academic Press.     

Phase behavior, microstructure transition, and antiradical activity of sucrose laurate/propylene glycol/the essential oil of Melaleuca alternifolia/water microemulsions

Nonionic sucrose ester microemulsions composed of sucrose laurate (SL), propylene glycol (PG) and water were prepared with the essential oil of Melaleuca alternifolia, commonly known as tea tree oil (TTO), as oil phase to investigate the phase behavior, microstructure, and antiradical activity. The pseudo-ternary phase diagrams were constructed to elucidate the phase behavior of the microemulsion formations at different weight ratios of surfactant and cosurfactant (S_m = SL/PG) of 1:1, 2:1, and 3:1. The extension of the microemulsion zone was found to be strongly dependent on the S_m ratios. The single phase microemulsion domain, especially o/w microemulsion region increased when S_m ratio is increased from 1:1 to 3:1 and no liquid crystalline structure was observed for all formulations studied. Microstructural aspects were studied by electrical conductivity and pulsed gradient spin echo (PGSE) NMR measurements along water titration line L28 (R_o = 2:8). The results from these combined techniques were in good agreement in regard to the microstructure transition points. The microstructural inversion of w/o to bicontinuous microemulsions occurred at 30 wt.% water while the transition from bicontinuous to o/w structure occurred at 55 wt.% water. The physical stability on storage temperature and time was examined by dynamic light scattering after the centrifuge test and freeze–thaw cycles. The droplet size was kept almost the same without any phase separation, providing less temperature-sensitivity up to 70 °C and good stability for 3 months at room temperature. The chemical profile and radical scavenging activity of TTO in o/w microemulsions was evaluated by means of gas chromatography–mass spectroscopy (GC–MS) and 2,2′-diphenyl-1-picrylhydrazyl free radical (DPPH) scavenging method, respectively. The major abundant constituents of crude TTO, monoterpene alcohols (terpinen-4-ol (41.65%), α-terpineol (3.18%)) and hydrocarbons (γ-terpinene (22.95%), α-terpinene (10.16%)) were identified and the composition percentage of each constituent was calculated form the GC peak areas by normalization method. The DPPH scavenging activity of TTO microemulsion was lower than pure TTO because the SL surfactant may obstruct the interaction between the TTO and DPPH, reducing the number of effective collisions.     

Influence of CTAB/alkanol/cyclohexane w/o microemulsions on the basic hydrolysis of crystal violet

The basic hydrolysis of crystal violet has been studied in w/o microemulsions of the CTAB/alkanols/cyclohexane system (alkanols: 1-butanol and 1-hexanol). The reaction can be considered to occur in the water phase of the droplets and from the rate constant the apparent dielectric constant of the water phase was determined. The cyclohexane incorporation in the system produces a decrease in the effective dielectric constant of the water phase and in the specific conductivity.     

In Situ Immobilization of Ultrafine Particles Synthesized in a Water/Oil Microemulsion

We investigated the in situ immobilization of ultrafine particles synthesized in a water/oil (w/o) microemulsion to silica for its possible application to supported metal catalysts. ZnS particles immobilized to silica by the ME method were consistent with those synthesized in a w/o microemulsion. Therefore, ZnS particles in a w/o microemulsion could be immobilized to silica without aggregation by this method. The relationship between the method of synthesizing Rh ultrafine particles in a w/o microemulsion and the diameter and diameter distribution of Rh particles immobilized to silica was studied. Rh-SiO(2) catalysts with a sharp diameter distribution could be prepared by immobilizing Rh-hydrazine complex particles because these complex particles would be very stable in a w/o microemulsion. The Rh particle diameters of Rh-SiO(2) catalysts prepared by changing the amount of silica produced were almost identical. Accordingly, the Rh particle diameter of Rh-SiO(2) catalysts could be controlled independently of Rh content by the ME method. Copyright 2001 Academic Press.     

Validation of a gas chromatography/thermal conductivity detection method for the determination of the water content of oxygenated solvents

A gas chromatographic (GC) method for the determination of the water content of acetone solutions containing methyl isobutyl ketone (MIBK) and other by-products of MIBK synthesis was developed and validated. Linearity is demonstrated (R2=0.9999) over a broad range of water concentrations in acetone ranging from 0.04 to 15.2% (w/w). The quantitation limit (QL) of this technique was found to be 0.346% (w/w) and the detection limit (DL) was estimated to be 13ppm. Validation of this analytical method's accuracy against a coulometric Karl Fischer titration (KFT) method showed excellent agreement between these two techniques. The GC/TCD method is accurate to within a relative error of 3.49% with respect to the KFT method for water concentration above the QL. However, the analysis of six samples below the QL showed an average relative error of 10.96%. Consequently, the KFT method is recommended for the moisture analysis of samples for water concentrations below 0.35% (w/w). Excellent precision is realized for water concentration above the QL (RSD<0.8%). Good precision (RSD=1.1%) was obtained for the sample with the lowest water concentration investigated (0.04%, w/w). Repeatability, robustness and excellent specificity of this technique are demonstrated.