A backpack-based myeloid cell therapy for multiple sclerosis.

Multiple sclerosis (MS) is an incurable autoimmune disease and is currently treated by systemic immunosuppressants with off-target side effects. Although aberrant myeloid function is often observed in MS plaques in the central nervous system (CNS), the role of myeloid cells in therapeutic intervention is currently overlooked. Here, we developed a myeloid cell-based strategy to reduce the disease burden in experimental autoimmune encephalomyelitis (EAE), a mouse model of progressive MS. We developed monocyte-adhered microparticles ("backpacks") for activating myeloid cell phenotype to an anti-inflammatory state through localized interleukin-4 and dexamethasone signals. We demonstrate that backpack-laden monocytes infiltrated into the inflamed CNS and modulated both the local and systemic immune responses. Within the CNS, backpack-carrying monocytes regulated both the infiltrating and tissue-resident myeloid cell compartments in the spinal cord for functions related to antigen presentation and reactive species production. Treatment with backpack-monocytes also decreased the level of systemic pro-inflammatory cytokines. Additionally, backpack-laden monocytes induced modulatory effects on TH1 and TH17 populations in the spinal cord and blood, demonstrating cross talk between the myeloid and lymphoid arms of disease. Backpack-carrying monocytes conferred therapeutic benefit in EAE mice, as quantified by improved motor function. The use of backpack-laden monocytes offers an antigen-free, biomaterial-based approach to precisely tune cell phenotype in vivo, demonstrating the utility of myeloid cells as a therapeutic modality and target.

Flowering in bursting bubbles with viscoelastic interfaces.

The lifetime of bubbles, from formation to rupture, attracts attention because bubbles are often present in natural and industrial processes, and their geometry, drainage, coarsening, and rupture strongly affect those operations. Bubble rupture happens rapidly, and it may generate a cascade of small droplets or bubbles. Once a hole is nucleated within a bubble, it opens up with a variety of shapes and velocities depending on the liquid properties. A range of bubble rupture modes are reported in literature in which the reduction of a surface energy drives the rupture against inertial and viscous forces. The role of surface viscoelasticity of the liquid film in this colorful scenario is, however, still unknown. We found that the presence of interfacial viscoelasticity has a profound effect in the bubble bursting dynamics. Indeed, we observed different bubble bursting mechanisms upon the transition from viscous-controlled to surface viscoelasticity-controlled rupture. When this transition occurs, a bursting bubble resembling the blooming of a flower is observed. A simple modeling argument is proposed, leading to the prediction of the characteristic length scales and the number and shape of the bubble flower petals, thus paving the way for the control of liquid formulations with surface viscoelasticity as a key ingredient. These findings can have important implications in the study of bubble dynamics, with consequences for the numerous processes involving bubble rupture. Bubble flowering can indeed impact phenomena such as the spreading of nutrients in nature or the life of cells in bioreactors.

Adsorption and Aggregation of Monoclonal Antibodies at Silicone Oil-Water Interfaces.

Monoclonal antibody (mAb) therapies are rapidly growing for the treatment of various diseases like cancer and autoimmune disorders. Many mAb drug products are sold as prefilled syringes and vials with liquid formulations. Typically, the walls of prefilled syringes are coated with silicone oil to lubricate the surfaces during use. MAbs are surface-active and adsorb to these silicone oil-solution interfaces, which is a potential source of aggregation. We studied formulations containing two different antibodies, mAb1 and mAb2, where mAb1 aggregated more when agitated in the presence of an oil-water interface. This directly correlated with differences in surface activity of the mAbs, studied with interfacial tension, surface mass adsorption, and interfacial rheology. The difference in interfacial properties between the mAbs was further reinforced in the coalescence behavior of oil droplets laden with mAbs. We also looked at the efficacy of surfactants, typically added to stabilize mAb formulations, in lowering adsorption and aggregation of mAbs at oil-water interfaces. We showed the differences between poloxamer-188 and polysorbate-20 in competing with mAbs for adsorption to interfaces and in lowering particulate and overall aggregation. Our results establish a direct correspondence between the adsorption of mAbs at oil-water interfaces and aggregation and the effect of surfactants in lowering aggregation by competitively adsorbing to these interfaces.

Evaporation-induced foam stabilization in lubricating oils.

Foaming in liquids is ubiquitous in nature. Whereas the mechanism of foaming in aqueous systems has been thoroughly studied, nonaqueous systems have not enjoyed the same level of examination. Here we study the mechanism of foaming in a widely used class of nonaqueous liquids: lubricant base oils. Using a newly developed experimental technique, we show that the stability of lubricant foams can be evaluated at the level of single bubbles. The results obtained with this single-bubble technique indicate that solutocapillary flows are central to lubricant foam stabilization. These solutocapillary flows are shown to originate from the differential evaporation of multicomponent lubricants-an unexpected result given the low volatility of nonaqueous liquids. Further, we show that mixing of some combinations of different lubricant base oils, a common practice in the industry, exacerbates solutocapillary flows and hence leads to increased foaming.

Bubble Coalescence at Wormlike Micellar Solution-Air Interfaces.

Surfactants in aqueous solutions self-assemble in the presence of salt, to form long, flexible, wormlike micelles (WLM). WLM solutions exhibit viscoelastic properties and are used in many applications, such as for cosmetic products, drag reduction, and hydraulic fracturing. Understanding the coalescence stability of bubbles in WLM solutions is important for the development of WLM based products that require a stable dispersion of bubbles. In this paper, we investigate the thin film drainage dynamics leading up to the coalescence of bubbles at flat WLM solution-air interfaces. The salts and surfactant type and concentrations were chosen so as to have the viscoelastic properties of the tested WLM solutions span over 2 orders of magnitude in moduli and relaxation times. The various stages in drainage and coalescence, the formation of a thick region at the apex (a dimple), the thinning and washout of this dimple, and the final stages of drainage before rupture, are modified by the viscoelasticity of the wormlike micellar solutions. As a result of the unique viscoelastic properties of the WLM solutions, we also observe a number of interesting fluid dynamic phenomena during the drainage processes including elastic recoil, thin film ripping, and single-step terminal drainage.

Evaporation-induced Rayleigh-Taylor instabilities in polymer solutions.

Understanding the mechanics of detrimental convective instabilities in drying polymer solutions is crucial in many applications such as the production of film coatings. It is well known that solvent evaporation in polymer solutions can lead to Rayleigh-Bénard or Marangoni-type instabilities. Here, we reveal another mechanism, namely that evaporation can cause the interface to display Rayleigh-Taylor instabilities due to the build-up of a dense layer at the air-liquid interface. We study experimentally the onset time (tp) of the instability as a function of the macroscopic properties of aqueous polymer solutions, which we tune by varying the polymer concentration (c0), molecular weight and polymer type. In dilute solutions, tp shows two limiting behaviours depending on the polymer diffusivity. For high diffusivity polymers (low molecular weight), the pluming time scales as [Formula: see text]. This result agrees with previous studies on gravitational instabilities in miscible systems where diffusion stabilizes the system. On the other hand, in low diffusivity polymers the pluming time scales as [Formula: see text]. The stabilizing effect of an effective interfacial tension, similar to those in immiscible systems, explains this strong concentration dependence. Above a critical concentration, [Formula: see text], viscosity delays the growth of the instability, allowing time for diffusion to act as the dominant stabilizing mechanism. This results in tp scaling as (ν/c0)2/3. This article is part of the theme issue 'Stokes at 200 (Part 1)'.

The top 10 research priorities in cystic fibrosis developed by a partnership between people with CF and healthcare providers.

There remain many treatment uncertainties in cystic fibrosis (CF). With limited resources, research should focus on questions which are most important to the CF community. We conducted a James Lind Alliance Priority Setting Partnership in CF. Research questions were elicited and then prioritised in successive surveys. A workshop agreed the final top 10. Online methods avoided cross infection and widened participation. The elicitation survey had 482 respondents (1080 questions) and prioritisation survey 677 respondents. Participants were drawn equally from the patient and clinical communities globally. We have achieved a consensus on 10 research priorities which will be attractive to funders.

Impact of Compressibility on the Control of Bubble-Pressure Tensiometers.

An experimental and theoretical investigation is conducted to understand the role of compressibility on the quasi-static expansion and contraction of a bubble that is pinned at the opening of a small capillary. The results show that there are two regimes of expansion and contraction depending on the values of two dimensionless parameters which correspond to a dimensionless volume and maximum capillary pressure. In one regime, not all bubble sizes are accessible during expansion and contraction, and the bubbles exhibit a hysteretic behavior when cycling through expansion and contraction. We call this the bubble shape hysteresis. The magnitude of the bubble shape hysteresis is computed for a realistic range of the nondimensional parameters. In the other regime, the bubble size can be varied continuously, but compressibility can still make it difficult to smoothly control the size of the bubble. The theoretical analysis shows that compressibility affects the evolution of the bubbles, even when the bubble is smaller than a hemispherical cap. The analysis also provides the infusion and withdrawal rates that a syringe pump must supply to expand and contract the bubble at a desired rate, accounting for compressibility. The validity of the assumptions used in the model is verified by comparison against experimental data.

Antifoams in non-aqueous diesel fuels: Thin liquid film dynamics and antifoam mechanisms.

HypothesisFoaming in diesel fuels is not well understood and leads to operational challenges. To combat deleterious effects of foaming, diesel formulations can include additives called antifoams. Existing antifoams, unfortunately, are inherently ash-generating when combusted, with unknown environmental impacts. They are prohibited in certain countries, so identifying effective alternative ash-free antifoam chemistries is needed. ExperimentsWe conduct systematic characterization of foam stabilization and antifoaming mechanisms in diesel for two different antifoams (silicone-containing & ashless chemistries). Employing a custom technique combining single-bubble/single-antifoam-droplet manipulation with white light interferometry, we also obtain mechanistic insights into foam stability and antifoam dynamics. ResultsCoalescence times from both bulk foam and single bubble experiments confirm ashless antifoams are effective at reducing foaming, demonstrating the potential of ashless antifoams. Further, we perform single-antifoam-droplet experiments and obtain direct experimental evidence revealing the elusive antifoaming mechanisms. Interestingly, the silicone-containing and ashless antifoams seemingly function via two different mechanisms: spreading and dewetting respectively. This surprising finding refutes conventional wisdom that spreading is likely the only antifoam mechanism in diesels. These results and the reported experimental framework significantly enhance the scientific understanding of non-aqueous foams and will accelerate the engineering of alternative antifoam chemistries for non-aqueous systems.

Neutrophils bearing adhesive polymer micropatches as a drug-free cancer immunotherapy.

Tumour-associated neutrophils can exert antitumour effects but can also assume a pro-tumoural phenotype in the immunosuppressive tumour microenvironment. Here we show that neutrophils can be polarized towards the antitumour phenotype by discoidal polymer micrometric 'patches' that adhere to the neutrophils' surfaces without being internalized. Intravenously administered micropatch-loaded neutrophils accumulated in the spleen and in tumour-draining lymph nodes, and activated splenic natural killer cells and T cells, increasing the accumulation of dendritic cells and natural killer cells. In mice bearing subcutaneous B16F10 tumours or orthotopic 4T1 tumours, intravenous injection of the micropatch-loaded neutrophils led to robust systemic immune responses, a reduction in tumour burden and improvements in survival rates. Micropatch-activated neutrophils combined with the checkpoint inhibitor anti-cytotoxic T-lymphocyte-associated protein 4 resulted in strong inhibition of the growth of B16F10 tumours, and in complete tumour regression in one-third of the treated mice. Micropatch-loaded neutrophils could provide a potent, scalable and drug-free approach for neutrophil-based cancer immunotherapy.

Preclinical characterization of macrophage-adhering gadolinium micropatches for MRI contrast after traumatic brain injury in pigs.

The choroid plexus (ChP) of the brain plays a central role in orchestrating the recruitment of peripheral leukocytes into the central nervous system (CNS) through the blood-cerebrospinal fluid (BCSF) barrier in pathological conditions, thus offering a unique niche to diagnose CNS disorders. We explored whether magnetic resonance imaging of the ChP could be optimized for mild traumatic brain injury (mTBI). mTBI induces subtle, yet influential, changes in the brain and is currently severely underdiagnosed. We hypothesized that mTBI induces sufficient alterations in the ChP to cause infiltration of circulating leukocytes through the BCSF barrier and developed macrophage-adhering gadolinium [Gd(III)]-loaded anisotropic micropatches (GLAMs), specifically designed to image infiltrating immune cells. GLAMs are hydrogel-based discoidal microparticles that adhere to macrophages without phagocytosis. We present a fabrication process to prepare GLAMs at scale and demonstrate their loading with Gd(III) at high relaxivities, a key indicator of their effectiveness in enhancing image contrast and clarity in medical imaging. In vitro experiments with primary murine and porcine macrophages demonstrated that GLAMs adhere to macrophages also under shear stress and did not affect macrophage viability or functions. Studies in a porcine mTBI model confirmed that intravenously administered macrophage-adhering GLAMs provide a differential signal in the ChP and lateral ventricles at Gd(III) doses 500- to 1000-fold lower than those used in the current clinical standard Gadavist. Under the same mTBI conditions, Gadavist did not offer a differential signal at clinically used doses. Our results suggest that macrophage-adhering GLAMs could facilitate mTBI diagnosis.

Alzheimer's and Parkinson's disease therapies in the clinic.

Alzheimer's disease (AD) and Parkinson's disease (PD) are the most prevalent neurodegenerative diseases, affecting millions and costing billions each year in the United States alone. Despite tremendous progress in developing therapeutics that manage the symptoms of these two diseases, the scientific community has yet to develop a treatment that effectively slows down, inhibits, or cures neurodegeneration. To gain a better understanding of the current therapeutic frontier for the treatment of AD and PD, we provide a review on past and present therapeutic strategies for these two major neurodegenerative disorders in the clinical trial process. We briefly recap currently US Food and Drug Administration-approved therapies, and then explore trends in clinical trials across the variables of therapy mechanism of disease intervention, administration route, use of delivery vehicle, and outcome measures, across the clinical phases over time for "Drug" and "Biologic" therapeutics. We then present the success rate of past clinical trials and analyze the intersections in therapeutic approaches for AD and PD, revealing the shift in clinical trials away from therapies targeting neurotransmitter systems that provide symptomatic relief, and towards anti-aggregation, anti-inflammatory, anti-oxidant, and regeneration strategies that aim to inhibit the root causes of disease progression. We also highlight the evolving distribution of the types of "Biologic" therapies investigated, and the slowly increasing yet still severe under-utilization of delivery vehicles for AD and PD therapeutics. We then briefly discuss novel preclinical strategies for treating AD and PD. Overall, this review aims to provide a succinct overview of the clinical landscape of AD and PD therapies to better understand the field's therapeutic strategy in the past and the field's evolution in approach to the present, to better inform how to effectively treat AD and PD in the future.

A biomimetic chip to assess subcutaneous bioavailability of monoclonal antibodies in humans.

Subcutaneous (subQ) injection is a common route for delivering biotherapeutics, wherein pharmacokinetics is largely influenced by drug transport in a complex subQ tissue microenvironment. The selection of good drug candidates with beneficial pharmacokinetics for subQ injections is currently limited by a lack of reliable testing models. To address this limitation, we report here a Subcutaneous Co-Culture Tissue-on-a-chip for Injection Simulation (SubCuTIS). SubCuTIS possesses a 3D coculture tissue architecture, and it allows facile quantitative determination of relevant scale independent drug transport rate constants. SubCuTIS captures key in vivo physiological characteristics of the subQ tissues, and it differentiates the transport behavior of various chemically distinct molecules. We supplemented the transport measurements with theoretical modeling, which identified subtle differences in the local absorption rate constants of seven clinically available mAbs. Accounting for first-order proteolytic catabolism, we established a mathematical framework to assess clinical bioavailability using the local absorption rate constants obtained from SubCuTIS. Taken together, the technology described here broadens the applicability of organs-on-chips as a standardized and easy-to-use device for quantitative analysis of subQ drug transport.

Physicochemical characteristics of droplet interface bilayers.

Droplet interface bilayer (DIB) is a lipid bilayer formed when two lipid monolayer-coated aqueous droplets are brought in contact within an oil phase. DIBs, especially post functionalization, are a facile model system to study the biophysics of the cell membrane. Continued advances in enhancing and functionalizing DIBs to be a faithful cell membrane mimetic requires a deep understanding of the physicochemical characteristics of droplet interface bilayers. In this review, we provide a comprehensive overview of the current scientific understanding of DIB characteristics starting with the key experimental frameworks for DIB generation, visualization and functionalization. Subsequently we report experimentally measured physical, electrical and transport characteristics of DIBs across physiologically relevant lipids. Advances in simulations and mathematical modelling of DIBs are also discussed, with an emphasis on revealing principles governing the key physicochemical characteristics. Finally, we conclude the review with important outstanding questions in the field.

Dewetting characteristics of contact lenses coated with wetting agents.

HYPOTHESIS: Although wetting agents have been developed to limit tear film dewetting over contact lenses, systematic analyses correlating wetting agent properties to mechanisms of the tear film destabilization are not readily available. Clarifying destabilization characteristics across key physio-chemical variables will provide a rational basis for identifying optimal wetting agents. EXPERIMENTS: We employ an in-house, in vitro platform to comprehensively evaluate drainage and dewetting dynamics of five wetting agents across seventeen different formulations and two model tear film solutions. We consider the film thickness evolution, film thickness at breakup, dewetted front propagation, and develop correlations to contact angle to compare the samples. FINDINGS: Zwitterionic wetting agents effectively stabilize the tear film by reducing the film thickness at the onset of dewetting, and delaying the propagation of dewetted regions across the lens. Furthermore, tuning wetting agent surface concentrations and utilizing binary mixtures of wetting agents can enhance wetting characteristics. Finally, despite disparities in wetting agent molecular properties, the time to dewet 50% of the lens scales linearly with the product of the receding contact angle and contact angle hysteresis. Hence, we fundamentally establish the importance of minimizing the absolute contact angle and contact angle hysteresis for effective wetting performance.

Surface energy and separation mechanics of droplet interface phospholipid bilayers.

Droplet interface bilayers are a convenient model system to study the physio-chemical properties of phospholipid bilayers, the major component of the cell membrane. The mechanical response of these bilayers to various external mechanical stimuli is an active area of research because of its implications for cellular viability and the development of artificial cells. In this article, we characterize the separation mechanics of droplet interface bilayers under step strain using a combination of experiments and numerical modelling. Initially, we show that the bilayer surface energy can be obtained using principles of energy conservation. Subsequently, we subject the system to a step strain by separating the drops in a step-wise manner, and track the evolution of the bilayer contact angle and radius. The relaxation time of the bilayer contact angle and radius along with the decay magnitude of the bilayer radius were observed to increase with each separation step. By analysing the forces acting on the bilayer and the rate of separation, we show that the bilayer separates primarily through the peeling process with the dominant resistance to separation coming from viscous dissipation associated with corner flows. Finally, we explain the intrinsic features of the observed bilayer separation by means of a mathematical model comprising the Young-Laplace equation and an evolution equation. We believe that the reported experimental and numerical results extend the scientific understanding of lipid bilayer mechanics, and that the developed experimental and numerical tools offer a convenient platform to study the mechanics of other types of bilayers.

Shear stress rosettes capture the complex flow physics in diseased arteries.

Wall shear stress (WSS) is an important parameter in arterial mechanobiology. Various flow metrics, such as time averaged WSS (TAWSS), oscillatory shear index (OSI), and transWSS, have been used to characterize and relate possible WSS variations in arterial diseases like aneurysms and atherosclerosis. We use a graphical representation of WSS using shear rosettes to map temporal changes in the flow dynamics during a cardiac cycle at any spatial location on the vessel surface. The presence of secondary flows and flow reversals can be interpreted directly from the shape of the shear rosette. The mean WSS is given by the rosette centroid, the OSI by the splay around the rosette origin, and the transWSS by its width. We define a new metric, anisotropy ratio (AR), based on the ratio of the length to width of the shear rosette, to capture flow bi-directionality. We characterized the flow physics in controls and patient specific geometries of the ascending aorta (AA) and internal carotid artery (ICA) that have fundamentally different flow dynamics due to differences in the Reynolds and Womersley numbers. The differences in the flow dynamics are well reflected in the shapes of the WSS rosettes and the corresponding flow metrics.

Foam stability in filtered lubricants containing antifoams.

Lubricant formulations are filtered to remove deleterious particulate matter. An unintended consequence of this important process is the detrimental effect of fine filtration on the foaming performance of lubricants with antifoam additives. Here we outline a method to study this phenomenon in detail by probing the coalescence stability of single bubbles in filtered antifoam laden lubricants. Initially, we establish the validity of Garrett's hypothesis for the tested antifoam laden lubricants. Subsequently, we show that the bubble stability in filtered lubricants are positively correlated to the number of filtration cycles - with the most dramatic changes in bubble stability accompanying the initial few cycles of filtration. Further, we show that post filtration, the stability of bubbles in lubricants is inversely correlated to the pore size of the filter and the volume fraction of antifoam in the lubricant prior to filtration. The results also reveal that in the presence of antifoam additives, the bubble coalescence times span multiple Rayleigh distributions. We also provide visual evidence that shows the tested antifoams employ a bridging-stretching mechanism to rupture non-aqueous foams. Finally, a simple probabilistic model is introduced that helps in analyzing the distribution of coalescence times of single bubbles to obtain insights into the volume fraction of antifoams in the lubricant. We believe these results are valuable in guiding the design of lubricants with robust and superior foaming performance.

Single bubble and drop techniques for characterizing foams and emulsions.

The physics of foams and emulsions has traditionally been studied using bulk foam/emulsion tests and single film platforms such as the Scheludko cell. Recently there has been a renewed interest in a third class of techniques that we term as single bubble/drop tests, which employ isolated whole bubbles and drops to probe the characteristics of foams and emulsions. Single bubble and drop techniques provide a convenient framework for investigating a number of important characteristics of foams and emulsions, including the rheology, stabilization mechanisms, and rupture dynamics. In this review we provide a comprehensive discussion of the various single bubble/drop platforms and the associated experimental measurement protocols including the construction of coalescence time distributions, visualization of the thin film profiles and characterization of the interfacial rheological properties. Subsequently, we summarize the recent developments in foam and emulsion science with a focus on the results obtained through single bubble/drop techniques. We conclude the review by presenting important venues for future research.

Hyperspectral imaging for dynamic thin film interferometry.

Dynamic thin film interferometry is a technique used to non-invasively characterize the thickness of thin liquid films that are evolving in both space and time. Recovering the underlying thickness from the captured interferograms, unconditionally and automatically is still an open problem. Here we report a compact setup employing a snapshot hyperspectral camera and the related algorithms for the automated determination of thickness profiles of dynamic thin liquid films. The proposed technique is shown to recover film thickness profiles to within 100 nm of accuracy as compared to those profiles reconstructed through the manual color matching process. Subsequently, we discuss the characteristics and advantages of hyperspectral interferometry including the increased robustness against imaging noise as well as the ability to perform thickness reconstruction without considering the absolute light intensity information.