Interaction between stabilized droplets of different phases in the same continuous phase of an aqueous three-phase system.

Water-in-water (W/W) emulsions, also called aqueous two-phase systems, are formed by mixing two incompatible polymers in water that phase separate into two distinct phases. They can be stabilized by addition of colloidal particles. Droplets of the dispersed phase can be used to compartmentalize ingredients and induce localized reactions. By mixing more types of incompatible polymers, emulsions containing droplets of different phases can be formed that can potentially capture different ingredients. Here the interaction between dispersed droplets of different types was studied by gently mixing a W/W emulsion containing droplets rich in dextran (DEX) dispersed in a continuous phase rich in polyethylene oxide with an emulsion containing droplets rich in fish gelatin (GEL) dispersed in the same continuous medium. Bis-hydrophilic microgels (MG) consisting of DEX grafted with poly(N-isopropylacrylamide) were added and their effect on the stability of each binary emulsion was investigated. Interestingly, when two very stable emulsions were gently mixed, droplets of different types were observed with confocal scanning laser microscopy to coalesce immediately upon contact. In this manner, Janus-type droplets were formed containing a DEX and a GEL compartment with no MG at the GEL/DEX interface that further associated into strings of alternating droplets. Contact angles between the different phases in emulsions with and without MG were compared and used to determine the effect of the microgels on the interfacial tension between the phases.

Modulation of circularly polarized luminescence by swelling of microgels functionalized with enantiopure [Ru(bpy)3]2+ luminophores.

Chemoresponsive microgels functionalized with enantiomeric Δ- or Λ-[Ru(bpy)3]2+ showed tunable chiroptical properties upon swelling and shrinking. The tuning is triggered by a modulation of the local mobility of [Ru(bpy)3]2+ upon addition of fructose, controlling interactions and distances between [Ru(bpy)3]2+ and phenylboronic acid.

Effect of charge on the stabilization of water-in-water emulsions by thermosensitive bis-hydrophilic microgels.

HYPOTHESIS: Molecular surfactants are not able to stabilize water-in-water (W/W) emulsions, unlike nano or micro-particles, which can achieve this in some cases. However, the effect of electrostatic interactions between particles on the emulsion stability has rarely been investigated. We hypothesize that introducing charges modifies the stabilization capacity of particles and renders it both pH- and ionic strength-dependent. EXPERIMENTS: Charge was introduced into bis-hydrophilic and thermoresponsive dextran/polyN-isopropylacrylamide microgels by replacing a small fraction of polyN-isopropylacrylamide with acrylic acid groups. The size of the microgels was obtained by dynamic light scattering. The stability and microstructure of dextran/poly(ethyleneoxide)-based W/W emulsions, was studied as a function of pH, NaCl concentration and temperature using confocal microscopy and by analytical centrifugation. FINDINGS: The swelling degree of charged microgels depends on the pH, ionic strength and the temperature. In the absence of salt, charged microgels do not adsorb at the interface and have little stabilizing effect even after neutralization. However, the interfacial coverage and the stability increase with rising concentration of NaCl. Saltinduced stabilization of these emulsions was also observed at 50 °C. Increasing the temperature strongly influences the emulsion stability at low pH.

Enzymatic cascade reaction in simple-coacervates.

The design of enzymatic droplet-sized reactors constitutes an important challenge with many potential applications such as medical diagnostics, water purification, bioengineering, or food industry. Coacervates, which are all-aqueous droplets, afford a simple model for the investigation of enzymatic cascade reaction since the reactions occur in all-aqueous media, which preserve the enzymes integrity. However, the question relative to how the sequestration and the proximity of enzymes within the coacervates might affect their activity remains open. Herein, we report the construction of enzymatic reactors exploiting the simple coacervation of ampholyte polymer chains, stabilized with agar. We demonstrate that these coacervates have the ability to sequester enzymes such as glucose oxidase and catalase and preserve their catalytic activity. The study is carried out by analyzing the color variation induced by the reduction of resazurin. Usually, phenoxazine molecules acting as electron acceptors are used to characterize glucose oxidase activity. Resazurin (pink) undergoes a first reduction to resorufin (salmon) and then to dihydroresorufin (transparent) in presence of glucose oxidase and glucose. We have observed that resorufin is partially regenerated in the presence of catalase, which demonstrates the enzymatic cascade reaction. Studying this enzymatic cascade reaction within coacervates as reactors provide new insights into the role of the proximity, confinement towards enzymatic activity.

Interfacial rheology of model water-air microgels laden interfaces: Effect of cross-linking.

HYPOTHESIS: The mechanical properties of model air/water interfaces covered by poly(N-isopropylacrylamide) microgels depend on the microgels deformability or in other words on the amount of cross-linker added during synthesis. EXPERIMENTS: The study is carried out by measuring the apparent dilational, the compression and the shear moduli using three complementary methods: (1) the pendant drop method with perturbative areas, (2) the Langmuir trough compression, and (3) shear rheology using a double wall ring cell mounted onto a Langmuir through. FINDINGS: In the range of surface coverages studied, the interfaces exhibit a solid-like behavior and elasticity goes through a maximum as a function of the surface pressure. This is observable whatever the investigation method. This maximum elasticity depends on the microgel deformability: the softer the microgels the higher the value of the moduli. The mechanical behavior of model interfaces is discussed, taking into account the core-shell structure of the particles and their packing at the interface.

Responsive microgels-based colloidosomes constructed from all-aqueous pH-switchable coacervate droplets.

HYPOTHESIS: Colloidosomes made of stimuli-responsive microgels offer the opportunity to design polymeric capsules with a hierarchical and tunable pore distribution. Coacervates stabilized by a microgel monolayer represent a unique strategy to build colloidosomes from all-aqueous emulsion drops, while exploiting the sequestration and dissolution properties of the coacervates. EXPERIMENTS: Methacrylated poly(N-isopropylacrylamide) (pNIPAM) microgels are used to stabilize coacervates made of an ampholyte polymer at a pH close to its isoelectric point. They are further cross-linked under UV-irradiation. The resulting assemblies are studied by means of confocal microscopy. Their permeability towards dextrans and nanoparticles is studied before and after dissolution of the coacervate. FINDINGS: PNIPAM microgels are found to stabilize the coacervates by adsorbing at their surface. Inter cross-linking the microgels results in the formation of an elastic colloidosome that persists after the coacervate dissolution and withstands surface deformations up to about 200%. The coacervate is exploited as a sequestrating core to entrap a water-soluble payload, which can be further released upon coacervate dissolution, while the membrane exhibits a size-selecting permeability. The membrane properties can also be switched by the volume phase transition of the microgels. Coacervate-embedded colloidosomes open new perspectives in the area of encapsulation/extraction and controlled transport of water-soluble/dispersed species.

Enhanced electrochemiluminescence at microgel-functionalized beads.

Bead-based assays are successfully combined with electrochemiluminescence (ECL) technology for detection of a wide range of biomarkers. Herein, we demonstrate a novel approach to enhance the ECL signal by decorating micrometric beads with [Ru(bpy)3]2+-grafted microgels (diameter ∼100 nm). Rapid and stable light emission was spatially resolved at the level of single functionalized beads. An enhancement of the ECL signal of microgel-labeled beads by 9-fold was observed in comparison to molecularly linked [Ru(bpy)3]2+ beads prepared by a sandwich immunoassay or an amide bond. Imaging the ECL signal at the single bead level shows that the size of the ECL-emitting layer is extended using the microgels. The reported method offers a great promise for the optimization of bead-based ECL detection and subsequent development of ECL microscopy.

Thermo-induced inversion of water-in-water emulsion stability by bis-hydrophilic microgels.

HYPOTHESIS: Stabilization of water-in-water (W/W) emulsions resulting from the separation of polymeric phases such as dextran (DEX) and poly(ethyleneoxide) (PEO) is highly challenging, because of the very low interfacial tensions between the two phases and because of the interface thickness extending over several nanometers. In the present work, we present a new type of stabilizers, based on bis-hydrophilic, thermoresponsive microgels, incorporating in the same structure poly(N-isopropylacrylamide) (pNIPAM) chains having an affinity for the PEO phase and dextran moieties. We hypothesize that these particles allow better control of the stability of the W/W emulsions. EXPERIMENTS: The microgels were synthesized by copolymerizing the NIPAM monomer with a multifunctional methacrylated dextran. They were characterized by dynamic light scattering, zeta potential measurements and nuclear magnetic resonance as a function of temperature. Microgels with different compositions were tested as stabilizers of droplets of the PEO phase dispersed in the DEX phase (P/D) or vice-versa (D/P), at different concentrations and temperatures. FINDINGS: Only microgels with the highest DEX content revealed excellent stabilizing properties for the emulsions by adsorbing at the droplet surface, thus demonstrating the fundamental role of bis-hydrophilicity. At room temperature, both pNIPAM and DEX chains were swollen by water and stabilized better D/P emulsions. However, above the volume phase transition temperature (VPTT ≈ 32 °C) of pNIPAM the microgels shrunk and stabilized better P/D emulsions. At all temperatures, excess microgels partitioned more to the PEO phase. The change in structure and interparticle interaction induced by heating can be exploited to control the W/W emulsion stability.

Pickering emulsions stabilized by thermoresponsive oligo(ethylene glycol)-based microgels: Effect of temperature-sensitivity on emulsion stability.

HYPOTHESIS: The stability of emulsions stabilized by soft and responsive microgels and their macroscopic properties are governed by the microstructure of microgels, in particular their deformability. However, little is known about the role of the microgel chemistry, though it is expected that polymeric backbone with an amphiphilic structure is a requirement for their adsorption at the oil-water interface. EXPERIMENTS: A series of biocompatible, thermoresponsive and amphiphilic poly(oligoethylene glycol)methacrylate (pOEMA) microgels is synthesized, with varying hydrophobic-hydrophilic balance, or equivalent varying volume phase transition temperature (VPTT). Their behavior in the bulk phase and at solid interfaces is compared to their behavior at liquid interfaces, studied on flat and model interfaces by the pendant drop method, and on drops, in microgel-stabilized emulsions. FINDINGS: Controlling the composition of microgels by simply changing the number of ethylene oxide groups in the hydrophilic side chain allows a precise tuning of their VPTT in the range of 20-60 °C. Simultaneously, the swelling ratio and the deformability of the microgels increase by increasing the hydrophilicity, as a result of the polymerization process. Regardless of their hydrophilicity, all the swollen pOEMA microgels adsorb at the liquid interface and stabilize emulsions, whose flocculation state and mechanical stability depends on the microgel deformability. Unexpectedly, most emulsions remain stable upon heating above the VPTT of the microgels. Such feature highlights their extreme robustness, whose origin is discussed. This study opens new opportunities for the use of biocompatible Pickering emulsifiers.

Dynamic Covalent Chemistry Enables Reconfigurable All-Polysaccharide Nanogels.

Dynamic covalent bonds are established upon molecular recognition of sugar derivatives by boronic acid molecules. These reversible links can be used in a cross-linking method to fabricate polymer-based responsive nanosystems. Herein, the design of the first dynamic nanogels made entirely of polysaccharides (PS) is reported. Based on PS chains alternately modified with phenyl boronic acid groups and sugar moieties, these colloids self-assemble in physiological conditions and combine the biocompatible nature of their PS backbone with the reconfiguration capacities of their cross-linking chemistry. These dynamic nanogels are easily prepared, stable for a long time, pH responsive, and efficiently internalized by cancer cells.

Asymmetric Modification of Carbon Nanotube Arrays with Thermoresponsive Hydrogel for Controlled Delivery.

In this work, bipolar electrochemistry is used to perform wireless indirect electrodeposition of two different polymer coatings on both sides of carbon nanotube arrays. Using a thermoresponsive hydrogel on one side and an inert insoluble polymer on the other side, it is possible to generate, in a single step, a nanoporous reservoir with Janus character closed on one side by a thermoresponsive membrane. The thermoresponsive polymer, poly(N-isopropylacrylamide) (pNIPAM), is generated by the local reduction of persulfate ions, which initiates radical polymerization of NIPAM. Electrophoretic paint (EP) is chosen as an inert polymer. It is deposited by precipitation because of a local decrease in pH during water oxidation. Both polymers can be deposited simultaneously on opposite sides of the bipolar electrode during the application of the electric field, yielding a double-modified Janus object. Moreover, the length and thickness of the polymer layers can be controlled by varying the electric field and the deposition time. This concept is applied to vertically aligned carbon nanotube arrays (VACNTs), trapped inside an anodic aluminum oxide membrane, which can further be used as a smart reservoir for chemical storage and release. A fluorescent dye is loaded in the VACNTs and its release is studied as a function of temperature. Low temperature, when the hydrogel layer is in the swollen state, allows diffusion of the molecule. Dye release occurs on the hydrogel-modified side of the VACNTs. At high temperatures, when the hydrogel layer is in the collapsed state, dye release is blocked because of the impermeability of the pNIPAM layer. This concept paves the way toward the design of advanced devices in the fields of drug storage and directed delivery.

Sugar-responsive Pickering emulsions mediated by switching hydrophobicity in microgels.

HYPOTHESIS: Pickering emulsions stabilized by soft and responsive microgels can demulsify on demand upon microgel collapse. The concept has been explored with simple model microgels such as poly(N-isopropylacrylamide) (pNIPAM) and their derivatives, but the role of functionalization is largely unexplored. EXPERIMENTS: Saccharide-responsive phenylboronic-modified microgels are used as Pickering emulsion stabilizers. Emulsion stability and microgel organization at drop surface are studied as a function of saccharide concentration. Better insight into their behavior at interfaces is gained through adsorption kinetics and Langmuir film studies at air-water interface. FINDINGS: The functionalization of water-swollen microgels by phenylboronic functions imparts some hydrophobicity to the structure, at the origin of additional internal cross-links analogous which rigidify the structure compared to non-functionalized microgels, as proved by their slow adsorption kinetics and poor interfacial compressibility. Upon boronate ester formation with diol groups of the saccharide, the hydrophobic character of the phenylboronic acid decreases, increasing the adsorption kinetics and their interfacial compressibility. Emulsions are stable in the presence of saccharide, given the high deformability of the yet-hydrophilic microgels, and mechanically unstable with less deformable particles in low saccharide concentration. The hydrophobic-hydrophilic switch acts as a trigger to tune the microgel stabilizing properties.

Kinetics of spontaneous microgels adsorption and stabilization of emulsions produced using microfluidics.

The aim of the paper is to examine the adsorption kinetics of soft microgels and to understand the role of fundamental parameters such as electrostatics and deformability on the process. This knowledge is further exploited to produce microgel-stabilized emulsions using a co-flow microfluidic device. Uncharged microgels made of poly(N-isopropylacrylamide) are synthesized with variable cross-linker contents, and charged ones are produced by introducing pH sensitive co-monomers during the synthesis. The study is carried out by measuring the microgels adsorption kinetics by means of the pendant drop method. The surface pressure is derived from the previous results as a function of time and is measured as a function of the area compression using a Langmuir trough. Emulsions are produced using a microfluidic device varying the microgels concentration and their stability is visually assessed. The microgels deformability as well as higher particle concentrations favour their adsorption. The adsorption is not governed by diffusion, it is cooperative and irreversible. Conversely, the kinetics is slowed down for increasing cross-linking density. The presence of charges slows down the kinetics of adsorption. In the presence of electrolyte, the kinetics accelerates and becomes similar to the one of neutral microgels. The original features of microgel adsorption is highlighted and the differences with adsorption of polymers, star polymers, proteins, and polyelectrolytes are emphasized. Taking benefit from the adsorption kinetics, the required formulation conditions for producing microgel-stabilized emulsions using a co-flow microfluidic device are derived. There exists a critical concentration above which microgels spontaneously adsorb in a sufficient way to decrease the interfacial tension. This critical microgel concentration increases with the cross-linking density and is higher for charged microgels. Whatever the kinetics, the same surface pressure is finally reached. This peculiar behaviour is likely a consequence of the presence of dangling chains in the as-prepared microgels. Consequently, a microgel excess is required to produce emulsions using microfluidics where adsorption has to be spontaneous.

Self-coacervation of ampholyte polymer chains as an efficient encapsulation strategy.

Coacervation is a phase separation process involving two aqueous phases, one solute-phase and one solute-poor phase. It is frequently observed among oppositely-charged polyelectrolyte systems. In this study, we focus on self-coacervation involving a single polymer chain and investigate its potential for encapsulation applications. Negatively charged polyacrylic acid polymer chains were partially cationized using diamine and carbodiimide chemistry affording ampholytes, named PAA-DA, with tunable charge ratio. When dispersed in water, at pH 7, PAA-DA was soluble but a phase separation occurs when decreasing pH close to the isoelectric point. Coacervation is found only for a given amine-to-acid ratio otherwise precipitation is observed. Increasing the pH above 4 yielded progressive destruction of the coacervates droplets via the formation of vacuoles within droplets and subsequent full homogeneous redispersion of PAA-DA in water. However, addition of calcium allowed increasing the coacervate droplet stability upon increasing the pH to 7 as the divalent ion induced gelation within droplets. Moreover, the coacervate droplets present the ability to spontaneously sequestrate a broad panel of entities, from small molecules to macromolecules or colloids, with different charges, size and hydrophobicity. Thanks to the reversible character of the coacervates, triggered-release could be easily achieved, either by varying the pH or by removing calcium ions in the case of calcium-stabilized coacervates. Self-coacervation presents the advantage of pathway-independent preparation, offering a real output interest in pharmacy, water treatment, food science or diagnostics.

Sealing hyaluronic acid microgels with oppositely-charged polypeptides: A simple strategy for packaging hydrophilic drugs with on-demand release.

A simple route to deliver on demand hydrosoluble molecules such as peptides, packaged in biocompatible and biodegradable microgels, is presented. Hyaluronic acid hydrogel particles with a controlled structure are prepared using a microfluidic approach. Their porosity and their rigidity can be tuned by changing the crosslinking density. These negatively-charged polyelectrolytes interact strongly with positively-charged linear peptides such as poly-l-lysine (PLL). Their interactions induce microgel deswelling and inhibit microgel enzymatic degradability by hyaluronidase. While small PLL penetrate the whole volume of the microgel, PLL larger than the mesh size of the network remain confined at its periphery. They make a complexed layer with reduced pore size, which insulates the microgel inner core from the outer medium. Consequently, enzymatic degradation of the matrix is fully inhibited and non-affinity hydrophilic species can be trapped in the core. Indeed, negatively-charged or small neutral peptides, without interactions with the network, usually diffuse freely across the network. By simple addition of large PLL, they are packaged in the core and can be released on demand, upon introduction of an enzyme that degrades selectively the capping agent. Single polyelectrolyte layer appears as a simple generic method to coat hydrogel-based materials of various scales for encapsulation and controlled delivery of hydrosoluble molecules.

Tuning Electrochemiluminescence in Multistimuli Responsive Hydrogel Films.

Luminescent and redox properties of stimuli-responsive hydrogel materials have been modulated by different external stimuli which trigger swelling or collapse of the polymer matrix. There is very rapid development in the field of such "smart" materials particularly combined with other sensing functionalities. Here, a poly(N-isopropylacrylamide) matrix incorporating covalently bound phenylboronic acids as a saccharide-sensing unit and redox-active [Ru(bpy)3]2+ luminophores was designed and exhibited multistimuli responsive electrochemical and luminescent switching behaviors. Redox activity of the films is reversibly changed by sequential stimuli (fructose and temperature) which control the swelling and the collapse of the films. Finally, electrogenerated chemiluminescence (ECL) is enhanced by a ∼16-fold factor during the film collapse induced by the temperature, whereas the swelling due to fructose provokes the decrease of the light emission. We demonstrate for the first time that ECL response correlates intrinsically with the swelling ratio and is finely modulated by both stimuli. The multistimuli responsive characteristics of such ECL-active hydrogels should find promising applications in biosensing, new luminescent materials, and logic gates in bioelectronic devices.

Organization of Microgels at the Air-Water Interface under Compression: Role of Electrostatics and Cross-Linking Density.

Poly(N-isopropylacrylamide) (pNIPAM) microgels are soft and deformable particles, which can adsorb at liquid interfaces. In the present paper, we study the two-dimensional organization of charged and quasi-neutral microgels with different cross-linking densities, under compression at the air-water interface and the transfer of the microgel monolayer onto a solid substrate at different surface pressures. At low cross-linking densities, the microgels form highly ordered hexagonal lattices on the solid substrate over large areas, with a unique lattice parameter that decreases continuously as the surface pressure increases. We thus prove that the microgel conformation evolves at the air-water interface. The microgels undergo a continuous transition from a highly flattened state at low surface coverage, where the maximal polymer segments are adsorbed at the interface, to entangled flattened microgels, and finally the thickening of the layer up to a dense hydrogel layer of compacted microgels. Moreover, two batches of microgels, with and without charges, are compared. The contribution of electrostatic interactions is assessed via changing the charge density of the microgels or modulating the Debye length. In both cases, electrostatics does not change the lattice parameter, meaning that, despite the microgel different swelling ratio, charges do not affect neither interactions between particles at the interface nor microgels adsorption. Conversely, the cross-linking density has a strong impact on microgel packing at the interface: increasing the cross-linking density strongly decreases the extent of microgel flattening and promotes the occurrence of coexisting hexagonally ordered domains with different lattice parameters.

Two-Dimensional Electrochemiluminescence: Light Emission Confined at the Oil-Water Interface in Emulsions Stabilized by Luminophore-Grafted Microgels.

We describe a method to confine electrochemiluminescence (ECL) at the oil-water interface of emulsion droplets that are stabilized by luminophore-grafted microgels. These hydrogel nanoparticles incorporating covalently bound Ru(bpy)32+ as the luminophore are irreversibly adsorbed at the interface of micrometric oil droplets dispersed in a continuous aqueous phase. We study the electrochemical and ECL properties of this multiscale system, composed of a collection of droplets in close contact in the presence of two types of model coreactants. ECL emission is observed upon oxidation of the coreactant and of the luminophore. ECL imaging confirms that light is emitted at the surface of oil droplets. Interestingly, light emission is observed more than 100 µm far from the electrode. It is possibly due to the interconnection between redox-active microgels, making an entangled two-dimensional network at the dodecane-water interface and/or to some optical effects related to the light propagation and refraction at different interfaces in this multiphasic system. Confining ECL in such an inhomogeneous medium should find promising applications in the study of compartmentalized systems, interfacial phenomena, sensors, and analysis of single oil droplets.

Modulation of Wetting Gradients by Tuning the Interplay between Surface Structuration and Anisotropic Molecular Layers with Bipolar Electrochemistry.

A new simple and versatile method for the preparation of surface-wetting gradients is proposed. It is based on the combination of electrode surface structuration introduced by a sacrificial template approach and the formation of a tunable molecular gradient by bipolar electrochemistry. The gradient involves the formation of a self-assembled monolayer on a gold surface by selecting an appropriate thiol molecule and subsequent reductive desorption by means of bipolar electrochemistry. Under these conditions, completion of the reductive desorption process evolves along the bipolar surface with a maximum strength localized at the cathodic edge and a decreasing driving force towards the middle of the surface. The remaining quantity of surface-immobilized thiol, therefore, varies as a function of the axial position, resulting in the formation of a molecular gradient. The surface of the bipolar electrode is characterized at each step of the modification by recording heterogeneous electron transfer. Also, the evolution of static contact angles measured with a water droplet deposited on the surface directly reveals the presence of the wetting gradient, which can be modulated by changing the properties of the thiol. This is exemplified with a long, hydrophobic alkane-thiol and a short, hydrophilic mercaptan.

Wireless Synthesis and Activation of Electrochemiluminescent Thermoresponsive Janus Objects Using Bipolar Electrochemistry.

In this work, bipolar electrochemistry (BPE) is used as a dual wireless tool to generate and to activate a thermoresponsive electrochemiluminescent (ECL) Janus object. For the first time, BPE allows regioselective growth of a poly(N-isopropylacrylamide) (pNIPAM) hydrogel film on one side of a carbon fiber. It is achieved thanks to the local reduction of persulfate ions, which initiate radical polymerization of NIPAM. By controlling the electric field and the time of the bipolar electrochemical reactions, we are able to control the length and the thickness of the deposit. The resulting pNIPAM film is found to be swollen in water at room temperature and collapsed when heated above 32 °C. We further incorporated a covalently attached ruthenium complex luminophore, Ru(bpy)32+, in the hydrogel film. In the second time, BPE is used to activate remotely the electrogenerated chemiluminescence (ECL) of the Ru(bpy)32+ moieties in the film. We take advantage of the film responsiveness to amplify the ECL signal. Upon collapse of the film, the ECL signal, which is sensitive to the distance between adjacent Ru(bpy)32+ centers, is strongly amplified. It is therefore shown that BPE is a versatile tool to generate highly sophisticated materials based on responsive polymers, which could lead to sensitive sensors.