Deep Eutectic Solvent Eutectogels for Delivery of Broad-Spectrum Antimicrobials.

Gel-based wound dressings have gained popularity within the healthcare industry for the prevention and treatment of bacterial and fungal infections. Gels based on deep eutectic solvents (DESs), known as eutectogels, provide a promising alternative to hydrogels as they are non-volatile and highly tunable and can solubilize therapeutic agents, including those insoluble in hydrogels. A choline chloride:glycerol-cellulose eutectogel was loaded with numerous antimicrobial agents including silver nanoparticles, black phosphorus nanoflakes, and commercially available pharmaceuticals (octenidine dihydrochloride, tetracycline hydrochloride, and fluconazole). The eutectogels caused >97% growth reduction in Gram-positive methicillin-resistant Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa bacteria and the fungal species Candida albicans.

Margination of 2D Platelet Microparticles in Blood.

Margination describes the movement of particles toward the endothelial wall within blood vessels. While there have been several studies tracking the margination of spherical particles in blood, the behavior of anisotropic particle shapes is not well described. In this study 2D platelet particles which possess many attractive qualities for use as a drug delivery system, with their high surface area allowing for increased surface binding activity, were directly monitored and margination quantified. The margination propensity of 1 and 2 µm 2D platelet particles was contrasted to that of 2 µm spherical particles at apparent wall shear rates (WSRs) of 50, 100, and 200 s-1 by both directly tracking labeled particles using fluorescent microscopy as well as using small-angle X-ray scattering (SAXS). For fluorescence studies, margination was quantified using the margination parameter M, which describes the number of particles found closest to the walls of a microfluidic device, with an M-value of 0.2 indicating no margination. Increased margination was seen in 2D platelet particles when compared to spherical particles tested at all flow rates, with M-values of 0.39 and 0.31 seen for 1 and 2 µm 2D platelet particles, respectively, while 2 µm spherical particles had an M-value of 0.21. Similarly, margination was observed qualitatively using SAXS, with increased scattering seen for platelet particles near the microfluidic channel wall. For all particles, increased margination was seen at increasing shear rates.

Propulsion, deformation, and confinement response of hollow nanocellulose millimotors.

HYPOTHESIS: Micromotor and nanomotor particles are typically made using dense solid particles that can sediment or be trapped in confined flow environments. Creation of much larger motors should be possible if a very low-density system is used with sufficient strength to carry liquid and still experience propulsive motion. Light, dense millimotors should also be able to deform more than dense solid ones in constrictions. EXPERIMENTS: Millimotors are created from permeable capsules of bacterial cellulose that are coated with catalse-containing metal-organic frameworks, enabling reactive propulsion in aqueous hydrogen peroxide. The motion of the motors is quantified using particle tracking and the deformation is measured using microcapillary compression and flow through confined channels. FINDINGS: Two different propulsion mechanisms are dominant depending on the motor surface chemistry: oxygen bubbles are expelled from hydrophilic millimotors, driving motion via recoil force and buoyancy. Hydrophobic millimotors remain attached to growing bubbles and move by buoyancy alone. Despite their large size, the low-density capsules compress to pass through contractions that would impede and be blocked by solid motors. The sparse structure but relatively large size of the motors enables them to transport significant volumes of liquid using minimal solid mass as a motor support structure.

Development of excipients free inhalable co-spray-dried tobramycin and diclofenac formulations for cystic fibrosis using two and three fluid nozzles.

This study aims to investigate the effect of physicochemical properties and aerosol performance of two (2FN) and three-fluid nozzles (3FN) on the inhalable co-formulation of tobramycin and diclofenac dry powders. Combination formulations of tobramycin and diclofenac at 2:1 and 4:1 w/w ratios were prepared at a laboratory scale using a spray dryer in conjunction with a 2FN or 3FN. Powder size, morphology, solid-state characteristics, and aerodynamic and dissolution properties were characterised. The nozzle types and the formulation composition influenced the yield, particle size, solid-state properties, aerosolization behaviour and dissolution of the co-spray dried formulations. In particular, using the 2FN the co-spray dried formulation of tobramycin and diclofenac at 2:1 w/w showed smaller particle size (D50, 3.01 ± 0.06 µm), high fine particle fractions (FPF) (61.1 ± 3.6% for tobramycin and 65.92 ± 3 for diclofenac) and faster dissolution with approx. 70% diclofenac released within 3 h and approx. 90% tobramycin was released within 45 min. However, the 3FN for the co-spray dried formulation of tobramycin and diclofenac at a 2:1 w/w ratio showed a larger particle size (D50, 3.42 ± 0.02 µm), lower FPF (40.6 ± 3.4% for tobramycin and 36.9 ± 0.84 for diclofenac) and comparative slower dissolution with approx. 60% diclofenac was released within 3 h and 80% tobramycin was released within 45 min. A similar trend was observed when the tobramycin to diclofenac ratio was increased to 4:1 w/w. Overall results suggest that spray drying with 2FN showed a superior and viable approach to producing excipients-free inhalable co-spray dried formulations of tobramycin and diclofenac. However, the formulation produced using the 3FN showed higher enrichment of hydrophobic diclofenac and an ability to control the tobramycin drug release in vitro.

Molecular and Colloidal Transport in Bacterial Cellulose Hydrogels.

Bacterial cellulose biofilms are complex networks of strong interwoven nanofibers that control transport and protect bacterial colonies in the film. The design of diverse applications of these bacterial cellulose films also relies on understanding and controlling transport through the fiber mesh, and transport simulations of the films are most accurate when guided by experimental characterization of the structures and the resultant diffusion inside. Diffusion through such films is a function of their key microstructural length scales, determining how molecules, as well as particles and microorganisms, permeate them. We use microscopy to study the unique bacterial cellulose film via its pore structure and quantify the mobility dynamics of various sizes of tracer particles and macromolecules. Mobility is hindered within the films, as confinement and local movement strongly depend on the void size relative to diffusing tracers. The biofilms have a naturally periodic structure of alternating dense and porous layers of nanofiber mesh, and we tune the magnitude of the spacing via fermentation conditions. Micron-sized particles can diffuse through the porous layers but cannot penetrate the dense layers. Tracer mobility in the porous layers is isotropic, indicating a largely random pore structure there. Molecular diffusion through the whole film is only slightly reduced by the structural tortuosity. Knowledge of transport variations within bacterial cellulose networks can be used to guide the design of symbiotic cultures in these structures and enhance their use in applications like biomedical implants, wound dressings, lab-grown meat, clothing textiles, and sensors.

Timothy Grass Pollen Induces Spatial Reorganisation of F-Actin and Loss of Junctional Integrity in Respiratory Cells.

Grass pollens have been identified as mediators of respiratory distress, capable of exacerbating respiratory diseases including epidemic thunderstorm asthma (ETSA). It is hypothesised that during thunderstorms, grass pollen grains swell to absorb atmospheric water, rupture, and release internal protein content to the atmosphere. The inhalation of atmospheric grass pollen proteins results in deadly ETSA events. We sought to identify the underlying cellular mechanisms that may contribute towards the severity of ETSA in temperate climates using Timothy grass (Phleum pratense). Respiratory cells exposed to Timothy grass pollen protein extract (PPE) caused cells to undergo hypoxia ultimately triggering the subcellular re-organisation of F-actin from the peri junctional belt to cytoplasmic fibre assembly traversing the cell body. This change in actin configuration coincided with the spatial reorganisation of microtubules and importantly, decreased cell compressibility specifically at the cell centre. Further to this, we find that the pollen-induced reorganisation of the actin cytoskeleton prompting secretion of the pro-inflammatory cytokine, interleukin-8. In addition, the loss of peri-junctional actin following exposure to pollen proteins was accompanied by the release of epithelial transmembrane protein, E-cadherin from cell-cell junctions resulting in a decrease in epithelial barrier integrity. We demonstrate that Timothy grass pollen regulates F-actin dynamics and E-cadherin localisation in respiratory cells to mediate cell-cell junctional integrity highlighting a possible molecular pathway underpinning ETSA events.

A phase diagram of morphologies for anisotropic particles sculpted from emulsions.

HYPOTHESIS: A micron-scale oil-in-water emulsion droplet frozen in the presence of surfactants can be induced to eject the crystallizing solid from its liquid precursor. This dynamic process produces highly elongated solids whose shape depends critically on the rate of crystallization and the interfacial properties of the tri-phase system. EXPERIMENT: By systematically varying the surfactant concentration and cooling protocol, including quenching from different temperatures as well as directly controlling the cooling rate, we map out the space of possible particle morphologies as a function of experimental control parameters. These results are analyzed using a non-equilibrium Monte Carlo model where crystallization rate and interfacial energies can be specified explicitly. FINDINGS: Our model successfully predicts the geometry of the resulting particles as well as emergent phenomena including how the particle shape depends on nucleation site and deformation of the precursor droplet during crystallization.

Influence of Aqueous Phase Composition on Double Emulsion Stability and Colour Retention of Encapsulated Anthocyanins.

Water-in-oil-in-water (W1/O/W2) emulsions (double emulsions) have often been used for the encapsulation of bioactive compounds such as anthocyanins. Instability of both anthocyanins and double emulsions creates a need for a tailored composition of the aqueous phase. In this work, double emulsions with a gelled internal water phase were produced and monitored over a 20-day storage period. The effect of the electrolyte phase composition (varying electrolyte components, including adipic acid, citric acid, and varying concentration of potassium chloride (KCl)) on anthocyanin and double emulsion stability was analysed using colour analysis, droplet sizing, and emulsion rheology. The effect of electrolytes on colour retention was shown to differ between the primary W1/O emulsion and the secondary W1/O/W2 emulsion. Furthermore, droplet size analysis and emulsion rheology highlighted significant differences in the stability and structural behaviour of the emulsions as a function of electrolyte composition. In terms of colour retention and emulsion stability, a citrate-buffered system performed best. The results of this study highlight the importance of strict control of aqueous phase constituents to prevent anthocyanin degradation and maximise double emulsion stability. Additional experiments analysed the effect of pectin chemistry on the anthocyanin colour retention and leakage, finding no conclusive difference between the unmodified and amidated pectin.

Crystal Comets: A Geometric Model for Sculpting Anisotropic Particles from Emulsions.

Microscopic high aspect ratio particles have many applications including enhanced delivery of active ingredients and food stability. Here, we develop a simple, scalable process that produces particles with a continuously controllable aspect ratio. Oil-in-water emulsion droplets are quenched and crystallize in the presence of surfactants that facilitate the ejection of the solid oil phase from its liquid precursor. Tuning the ejection and crystallization rates to be comparable, by adjusting the surfactant concentration and quench depth, promotes anisotropic particle growth by continuously ejecting solidified oil from the precursor droplet as the crystallization proceeds. We predict the accessible morphologies using an analytical geometric model that indicates a nonconstant contact angle during the crystallization process. We see that the crystal aspect ratio is dependent on the surfactant concentration, which can be explained as a variation of the maximum growth angle achieved during crystallization.

Rectus femoris hyperreflexia contributes to Stiff-Knee gait after stroke.

BACKGROUND: Stiff-Knee gait (SKG) after stroke is often accompanied by decreased knee flexion angle during the swing phase. The decreased knee flexion has been hypothesized to originate from excessive quadriceps activation. However, it is unclear whether hyperreflexia plays a role in this activation. The goal of this study was to establish the relationship between quadriceps hyperreflexia and knee flexion angle during walking in post-stroke SKG. METHODS: The rectus femoris (RF) H-reflex was recorded in 10 participants with post-stroke SKG and 10 healthy controls during standing and walking at the pre-swing phase. In order to attribute the pathological neuromodulation to quadriceps muscle hyperreflexia and activation, healthy individuals voluntarily increased quadriceps activity using electromyographic (EMG) feedback during standing and pre-swing upon RF H-reflex elicitation. RESULTS: We observed a negative correlation (R = - 0.92, p = 0.001) between knee flexion angle and RF H-reflex amplitude in post-stroke SKG. In contrast, H-reflex amplitude in healthy individuals in presence (R = 0.47, p = 0.23) or absence (R = - 0.17, p = 0.46) of increased RF muscle activity was not correlated with knee flexion angle. We observed a body position-dependent RF H-reflex modulation between standing and walking in healthy individuals with voluntarily increased RF activity (d = 2.86, p = 0.007), but such modulation was absent post-stroke (d = 0.73, p = 0.296). CONCLUSIONS: RF reflex modulation is impaired in post-stroke SKG. The strong correlation between RF hyperreflexia and knee flexion angle indicates a possible regulatory role of spinal reflex excitability in post-stroke SKG. Interventions targeting quadriceps hyperreflexia could help elucidate the causal role of hyperreflexia on knee joint function in post-stroke SKG.

Aggregation in viscoelastic emulsion droplet gels with capillarity-driven rearrangements.

Arrested, or partial, coalescence of viscoelastic emulsion droplets can occur when elastic resistance to deformation offsets droplet surface area minimization. Arrest is a critical element of food and consumer product microstructure and performance, but direct studies of structural arrest and rearrangement have been carried out using only two or three droplets at a time. The question remains whether the behavior of small numbers of droplets also occurs in larger, more realistic many-droplet systems. Here we study two-dimensional aggregation and arrested coalescence of emulsions containing ∼1000 droplets and find that the restructuring mechanisms observed for smaller systems have a large effect on local packing in multidroplet aggregates, but surprisingly do not significantly alter overall mass scaling in the aggregates. Specifically, increased regions of hexagonal packing are observed as the droplet solids level, and thus elasticity, is decreased because greater degrees of capillary force-driven restructuring are possible. Diffusion-limited droplet aggregation simulations that account for the restructuring mechanisms agree with the experimental results and suggest a basis for prediction of larger-scale network properties and bulk emulsion behavior.

Does kinematic gait quality improve with functional gait recovery? A longitudinal pilot study on early post-stroke individuals.

Typical clinical gait outcomes mostly focus on function; only sparse information exists on gait quality, i.e. symmetry or more natural gait patterns. It remains unclear whether functional gait recovery improves with gait quality, or whether these are two independent processes. The objective of this observational pilot study is to examine whether the gait quality improves with gait function (i.e. speed) over the course of early recovery. Full lower body gait kinematics were measured longitudinally in a clinical environment using wearable inertial measurement units. We recorded six individuals with subacute stroke (<1 month) for a total of 56 physical therapy sessions over the initial recovery stage of 12 weeks. We examined relations between gait symmetry in spatiotemporal, limb and joint kinematic parameters compared to gait function. We observed that overall gait symmetry improved with walking speed, but limb and joint kinematic parameters remained asymmetric at the maximum level of recovery (both p < 0.01). We also found that limb kinematic parameters (R2 = 41.9%) of the impaired side was preferentially associated with functional gait recovery over joint kinematics (R2 = 33.1%). These data suggest that our pilot cohort did not achieve "true" gait recovery despite achieving typical measures of recovery in gait speed and spatiotemporal symmetry. These initial results illustrate the multifaceted nature of recovery and justify further research on monitoring gait quality with a larger clinical study, providing insight for more effective training regimens.

Quantifying dosage of physical therapy using lower body kinematics: a longitudinal pilot study on early post-stroke individuals.

BACKGROUND: While therapy is an important part of the recovery process, there is a lack of quantitative data detailing the "dosage" of therapy received due to the limitations on in/outpatient accessibility and mobility. Advances in wearable sensor technology have allowed us to obtain an unprecedented glimpse into joint-level kinematics in an unobtrusive manner. The objective of this observational longitudinal pilot study was to evaluate the relations between lower body joint kinematics during therapy and functional gait recovery over the first three months after stroke. METHODS: Six individuals with subacute stroke (< 1 month) were monitored for a total of 59 one-hour physical therapy sessions including gait and non-gait activities. Participants donned a heart rate monitor and an inertial motion capture system to measure full lower body joint kinematics during each therapy session. Linear mixed regression models were used to examine relations between functional gait recovery (speed) and activity features including total joint displacements, defined as amount of motion (AoM), step number, change in heart rate (∆HR), and types of tasks performed. RESULTS: All activity features including AoM, step number, types of tasks performed (all p < 0.01), and ∆HR (p < 0.05) showed strong associations with gait speed. However, AoM (R2 = 32.1%) revealed the greatest explained variance followed by step number (R2 = 14.1%), types of tasks performed (R2 = 8.0%) and ∆HR (R2 = 5.8%). These relations included both gait and non-gait tasks. Contrary to our expectations, we did not observe a greater relation of functional recovery to motion in the impaired limb (R2 = 27.8%) compared to the unimpaired limb (R2 = 32.9%). CONCLUSIONS: This proof-of-concept study shows that recording joint kinematics during gait therapy longitudinally after stroke is feasible and yields important information for the recovery process. These initial results suggest that compared to step number, more holistic outcome measures such as joint motions may be more informative and help elucidate the dosage of therapy.

Correction: Comparison of bulk and microfluidic methods to monitor the phase behaviour of nanoparticles during digestion of lipid-based drug formulations using in situ X-ray scattering.

Correction for 'Comparison of bulk and microfluidic methods to monitor the phase behaviour of nanoparticles during digestion of lipid-based drug formulations using in situ X-ray scattering' by Ben J. Boyd et al., Soft Matter, 2019, 15, 9565-9578.

Comparison of bulk and microfluidic methods to monitor the phase behaviour of nanoparticles during digestion of lipid-based drug formulations using in situ X-ray scattering.

The performance of orally administered lipid-based drug formulations is crucially dependent on digestion, and understanding the colloidal structures formed during digestion is necessary for rational formulation design. Previous studies using the established bulk pH-stat approach (Hong et al. 2015), coupled to synchrotron small angle X-ray scattering (SAXS), have begun to shed light on this subject. Such studies of digestion using in situ SAXS measurements are complex and have limitations regarding the resolution of intermediate structures. Using a microfluidic device, the digestion of lipid systems may be monitored with far better control over the mixing of the components and the application of enzyme, thereby elucidating a finer understanding of the structural progression of these lipid systems. This work compares a simple T-junction microcapillary device and a custom-built microfluidic chip featuring hydrodynamic flow focusing, with an equivalent experiment with the full scale pH-stat approach. Both microfluidic devices were found to be suitable for in situ SAXS measurements in tracking the kinetics with improved time and signal sensitivity compared to other microfluidic devices studying similar lipid-based systems, and producing more consistent and controllable structural transformations. Particle sizing of the nanoparticles produced in the microfluidic devices were more consistent than the pH-stat approach.

Geometry and kinetics determine the microstructure in arrested coalescence of Pickering emulsion droplets.

Arrested coalescence occurs in Pickering emulsions where colloidal particles adsorbed on the surface of the droplets become crowded and inhibit both relaxation of the droplet shape and further coalescence. The resulting droplets have a nonuniform distribution of curvature and, depending on the initial coverage, may incorporate a region with negative Gaussian curvature around the neck that bridges the two droplets. Here, we resolve the relative influence of the curvature and the kinetic process of arrest on the microstructure of the final state. In the quasistatic case, defects are induced and distributed to screen the Gaussian curvature. Conversely, if the rate of area change per particle exceeds the diffusion constant of the particles, the evolving surface induces local solidification reminiscent of jamming fronts observed in other colloidal systems. In this regime, the final structure is shown to be strongly affected by the compressive history just prior to arrest, which can be predicted from the extrinsic geometry of the sequence of surfaces in contrast to the intrinsic geometry that governs the static regime.

Soft Bacterial Cellulose Microcapsules with Adaptable Shapes.

Microcapsules with controlled stability and permeability are in high demand for applications in separation and encapsulation. We have developed a biointerfacial process to fabricate strong, but flexible, porous microcapsules from bacterial cellulose at an oil-water emulsion interface. A broad range of microcapsule sizes has been successfully produced, from 100 µm to 5 cm in diameter. The three-dimensional capsule microstructure was imaged using confocal microscopy, showing a cellulose membrane thickness of around 30 µm that is highly porous, with some pores larger than 0.5 µm that are permeable to most macromolecules by free diffusion but can exclude larger structures like bacteria. The mechanical deformation of cellulose microcapsules reveals their flexibility, enabling them to pass through constrictions with a much smaller diameter than their initial size by bending and folding. Our work provides a new approach for producing soft, permeable, and biocompatible microcapsules for substance encapsulation and protection. The capsules may offer a replacement for suspended polymer beads in commercial applications and could potentially act as a framework for artificial cells.

Polymerization of cubosome and hexosome templates to produce complex microparticle shapes.

Cubic and hexagonal liquid crystalline particles, or cubosomes and hexosomes, are used as templates to polymerize various monomers to produce particles with unique micron-scale geometric shapes. Emulsion droplets containing water, ethanol, and the lipid glyceryl monooleate are suspended in a yield stress fluid and used to produce shapes based on cubic and hexagonal symmetry by slow crystallization. The underlying liquid crystalline ordering of the aqueous lipid system drives symmetric shape formation, while its amphiphilicity allows incorporation of various organic monomers. Photopolymerization of monomers in the lipid templates creates polymeric particles shaped like polyhedra based on cubic symmetry as well as biconical cylinders based on hexagonal symmetry, and their shape is preserved after template removal. Product particle shapes are controlled by varying the structures and hydrophobicity of the monomers, as they control formation of different phases and microstructures. Monomer polarity determines whether the template can exhibit hexagonal phase, as when divinylbenzene is used, or cubic phase, when di(ethylene glycol) dimethacrylate is used. The monomers also control the microstructure of the final particles produced, forming rigid shapes composed of linked polymer nanospheres when divinylbenzene or di(ethylene glycol) dimethacrylate are used, and soft hydrogel particles when N,N'-methylenebisacrylamide is used.

Large Hexosomes from Emulsion Droplets: Particle Shape and Mesostructure Control.

Soft, rotationally symmetric particles of dispersed hexagonal liquid crystalline phase are produced using a method previously developed for cubosome microparticle production. The technique forms hexosome particles via removal of ethanol from emulsion droplets containing monoolein, water, and one of the various hydrophobic molecules: vitamin E, hexadecane, oleic acid, cyclohexane, or divinylbenzene. The unique rotational symmetry of the particles is characterized by optical microscopy and small-angle X-ray scattering to link particle phase, shape, and structure to composition. Rheology of the soft particles can be varied independently of shape, enabling control of transport, deformation, and biological response by controlling composition and molecular structure of the additives. The direct observations of formation, and the resultant hexosome shapes, link the particle-scale and mesoscale properties of these novel self-assembled particles and broaden their applications. The micron-scale hexosomes provide a route to understanding the effects of particle size, crystallization rate, and rheology on the production of soft particles with liquid crystalline structure and unique shape and symmetry.