Interface of biomolecular condensates modulates redox reactions.

Biomolecular condensates mediate diverse cellular processes. The density transition process of condensate formation results in selective partitioning of molecules, which define a distinct chemical environment within the condensates. However, the fundamental features of the chemical environment and the mechanisms by which such environment can contribute to condensate functions have not been revealed. Here, we report that an electric potential gradient, thereby an electric field, is established at the liquid-liquid interface between the condensate and the bulk environment due to the density transition of ions and molecules brought about by phase separation. We find that the interface of condensates can drive spontaneous redox reactions in vitro and in living cells. Our results uncover a fundamental physicochemical property of the interface of condensates and the mechanism by which the interface can modulate biochemical activities.

Simple Estimate of the Potential Drop across an Amphiprotic Liquid-Liquid Interface.

Two immiscible liquids in contact with each other can have different internal electrostatic potentials. An associated electric double layer (EDL) therefore exists within each liquid. For amphiprotic liquids, the exchange of protons between the two liquids gives rise to two EDLs, a positively charged EDL in one of the liquids and negatively charged EDL in the other. Using the pKa and pKb of one liquid dissolved in the other and the pH equivalent within each amphiprotic liquid, we can estimate the potential drop, Δφ, between the interior of the two liquids, also known as the Galvani potential or liquid-liquid junction potential. This estimation is independent of surface charge and ionic strength. By using the ionic strength to find the thickness of the EDL, we also estimate the average electric field strength across the interface. For the special case of water (H2O) in contact with an immiscible alcohol (ROH), the potential drop across the interface from the water to the alcohol is Δφ = 2.303VT (pKb + pH - pKw - pH2OR), where VT is the thermal voltage at a given temperature T.

On-demand electrochemically controlled compound release from an ultrasonically powered implant.

On-demand drug delivery systems are promising for a wide range of therapeutic applications. When combined with wireless implants for controlled drug delivery, they can reduce overall dosage and side effects. Here, we demonstrate release of fluorescein from a novel on-demand release system for negatively charged compounds. The release system is based on a modified electroresponsive polypyrrole nanoparticulate film designed to minimize ion exchange with the stored compound - a major passive leakage mechanism. We further designed an ultrasonically powered mm-sized implant to electronically control the on-demand drug delivery system in vivo. Release kinetics are characterized both in vitro and in vivo in mice using fluorescein as a model drug, demonstrating the feasibility of wireless, controllable drug release using an ultrasonically powered implant.

Optimizing Coaxial Sonic Spray Geometry for Generating Water Microdroplets.

Sonic spray creates a stream of neutral and charged microdroplets without application of voltage, heating, laser irradiation, or corona discharge. The solvent of interest flows through an inner capillary (usually constructed of fused silica) that is surrounded by an outer stainless-steel tube through which a nebulizing gas flows under pressure. This technique has been widely used as the interface in mass spectrometric studies for chemical analysis and for understanding microdroplet chemistry. We have used light scattering to characterize the size distribution and density for water microdroplets as a function of several parameters, such as water quality, water flow rate, nebulizing gas pressure, and sonic sprayer geometry. We find that the size distribution of the microdroplets, which is critical to many applications, depends most sensitively on the distance between the inner and outer capillary outlets and the gas flow pressure. The best performance as measured by the smallness of the microdroplet diameters is obtained when the gas flow pressure is the highest and there is no separation distance, d, between the two capillary outlets. In addition, at d = 0 mm, the microdroplet diameter distribution is nearly independent of the water flow rate, indicating that studies under these conditions can be scaled up.

Microdroplets can act as electrochemical cells.

A water microdroplet in air or oil typically possesses an electric double layer (EDL) from the preferential adsorption of surface-bound ions at the periphery. We present the calculations of the ion gradients within a microdroplet at equilibrium, including systems containing buffers and water autoionization. These ion gradients are used to calculate the potential energy stored within the microdroplet. We consider how this stored potential energy can be utilized to drive chemical reactions, much like an electrochemical cell. Effective voltages as high as 111 mV are found for microdroplets having a low surface charge density (0.01 ions per nm2). Two sources of potential energy are investigated: (1) the electrostatic energy of the EDL of the microdroplet and (2) shifts in other chemical equilibria coupled to the main reaction through the EDL. A particularly important example of the latter is water autoionization, wherein the reaction of interest causes a flattening of the [H+] gradient within the EDL, resulting in a net recombination of H+ and OH- throughout the microdroplet. Numerical calculations are performed using a continuum model consisting of a balance between the electromigration and diffusion of ions throughout the microdroplet. Our treatment accounts for the autoionization of water and any chemical equilibrium of buffers present. The results are presented for uncharged water microdroplets with low amounts of salts and simple buffers in them. However, the calculational method presented here can be applied to microdroplets of any net charge, composed of any solvent, containing ions of any valence, and containing complex mixtures of chemical equilibria.

Effect of relative humidity on hydrogen peroxide production in water droplets.

Mist is generated by ultrasonic cavitation of water (Fisher Biograde, pH 5.5-6.5) at room temperature (20-25 °C) in open air with nearly constant temperature (22-25 °C) but varying relative humidity (RH; 24-52%) over the course of many months. Water droplets in the mist are initially about 7 µm in diameter at about 50% RH. They are collected, and the concentration of hydrogen peroxide (H2O2) is measured using commercial peroxide test strips and by bromothymol blue oxidation. The quantification method is based on the Fenton chemistry of dye degradation to determine the oxidation capacity of water samples that have been treated by ultrasonication. It is found that the hydrogen peroxide concentration varies nearly linearly with RH over the range studied, reaching a low of 2 parts per million (ppm) at 24% RH and a high of 6 ppm at 52% RH. Some possible public health implications concerning the transmission of respiratory viral infections are suggested for this threefold change in H2O2 concentration with RH.

Erratum: Effect of Relative Humidity on Hydrogen Peroxide Production in Water Droplets - CORRIGENDUM.

[This corrects the article DOI: 10.1017/qrd.2021.6.].

Effects of Weak Electrolytes on Electric Double Layer Ion Distributions.

Many common experimental systems have electric double layers containing weak electrolytes, including systems with buffers. The pH at the boundary of the diffuse layer is an important parameter for determining the physicochemical state of the system, including surface charge density. We show that the Boltzmann equilibrium relation can be used as an exact solution for weak electrolyte electric double layers. Using these results, we provide a closed-form relation for the maximum pH change in a buffered electric double layer, in terms of the boundary potential. Importantly, our results suggest that equilibrium electric double layer concepts developed for strong electrolytes can be expanded to include weak electrolytes.

Simple model for the electric field and spatial distribution of ions in a microdroplet.

It is well established that the chemistry in microdroplets has been found to be radically different from reactions in bulk, particularly in the case of water. It has also been established that there is a threshold size for microdroplets to behave differently than droplets near the 10 µm diameter range. We present a three-dimensional electrostatic treatment in the spirit of the Gouy-Chapman model for double layers at interfaces. Our treatment predicts a strong concentration of charged molecules toward the surface of the droplet. As the droplet size deceases, the majority of the volume of the liquid experiences a large DC electric field. Such electric fields are highly unusual in a conducting fluid such as water. We believe that this unique environment helps to explain the reaction rate acceleration and new chemistry that have been observed in microdroplets compared to bulk phase.

CONCENTRATION GRADIENTS INSIDE MICRODROPLETS.

Small water microdroplets in microfluidic systems have a high surface charge density resulting from charged surfactants. As a result, an electric double layer forms inside the droplet. Depletion of ions from the center of the droplet to form the double layer can shift the concentration of ions dramatically from that of the microdroplet precursor solution. Here we show numerical solutions to the Gouy-Chapman model in spherical coordinates. Some notable effects include: 1) large percentages of the microdroplet volume experience very large DC electric fields; 2) many ions get forced into a Stern layer giving dramatically different conditions from the bulk.

Electrically controlled drug release using pH-sensitive polymer films.

Drug delivery systems (DDS) that allow spatially and temporally controlled release of drugs are of particular interest in the field of drug delivery. These systems create opportunities for individually tailored doses of drugs to be administered as well as reduce side effects by localizing the initial drug dose to the organ of interest. We present an electroresponsive DDS in the form of a bioresorbable nanocomposite film which operates at low voltages (<-2 V). The method is based on electrochemically generating local pH changes at an electrode surface to induce dissolution of a pH-sensitive polymer, which is used as the carrier material. We previously demonstrated this proof-of-concept using a poly(methyl methacrylate-co-methacrylic acid) (co-PMMA) copolymer commercially marketed as Eudragit S100 (EGT). However, as EGT is soluble at a pH above 7, experiments were performed in isotonic saline solutions (pH ∼ 6.4). In this work, we have synthesized co-PMMA with a variety of monomer ratios to shift the solubility of the copolymer to higher pH values, and developed a polymer that can be used under physiologically relevant conditions. The generalizability of this system was demonstrated by showing controlled release of different drug molecules with varying parameters like size, hydrophobicity, and pKa. Fluorescein, a hydrophilic model compound, meloxicam, a hydrophobic anti-arthritic medication, curcumin, a small molecule with anti-cancer therapeutic potential, and insulin, a polypeptide hormone used in the treatment of hypoglycemia, could all be released on demand with minimal leakage. The drug loading achieved was ∼32 wt% by weight of the co-polymer.

Visible light photoswitching of conjugated polymer nanoparticle fluorescence.

Conjugated polymer nanoparticles doped with a reverse photochromic dye exhibit highly quenched fluorescence that can be reversibly activated by controlling the form of the photochrome with visible light.

Functionalization of conjugated polymer nanoparticles for fluorescence photomodulation.

The emission of conjugated polymer nanoparticles (CPNs or Pdots) is often tailored for specific uses by functionalizing CPNs with dyes that act as fluorescence resonance energy transfer (FRET) acceptors. A number of dye functionalization methods for CPNs have been developed, ranging from simple noncovalent doping to covalent attachment. We seek to develop guidelines for when noncovalent doping is acceptable and when covalent attachment is necessary to achieve the desired result. We present results of CPNs functionalized with photochromic spirooxazines by four different methods: simple doping, doping with an amphiphilic coating polymer, covalent functionalization prior to CPN formation, and covalent functionalization after CPN formation. The different CPNs are evaluated in terms of their fluorescence photomodulation properties to determine how the preparation method affects the CPN-dye photophysical interactions. Doping preparations yield the most efficient quenching of CPN emission due to shorter donor-acceptor distances in these CPNs compared to those with covalently tethered dyes. Aging studies reveal that the photochromic dyes in doped samples degrade over time to a far greater extent than those in covalently functionalized samples. These results suggest that dye-doped CPNs are appropriate for short-term experiments where highly efficient FRET is desired while covalent dye functionalization is a better choice for experiments executed over an extended time frame.