Softness matters: effects of compression on the behavior of adsorbed microgels at interfaces.

Deformable colloids and macromolecules adsorb at interfaces as they decrease the interfacial energy between the two media. The deformability, or softness, of these particles plays a pivotal role in the properties of the interface. In this study, we employ a comprehensive in situ approach, combining neutron reflectometry with molecular dynamics simulations, to thoroughly examine the profound influence of softness on the structure of microgel Langmuir monolayers under compression. Lateral compression of both hard and soft microgel particle monolayers induces substantial structural alterations, leading to an amplified protrusion of the microgels into the aqueous phase. However, a critical distinction emerges: hard microgels are pushed away from the interface, in stark contrast to the soft ones, which remain firmly anchored to it. Concurrently, on the air-exposed side of the monolayer, lateral compression induces a flattening of the surface of the hard monolayer. This phenomenon is not observed for the soft particles as the monolayer is already extremely flat even in the absence of compression. These findings significantly advance our understanding of the key role of softness on both the equilibrium phase behavior of the monolayer and its effect when soft colloids are used as stabilizers of responsive interfaces and emulsions.

Effects of Charge Density on Spread Hyperbranched Polyelectrolyte/Surfactant Films at the Air/Water Interface.

The interfacial structure and morphology of films spread from hyperbranched polyethylene imine/sodium dodecyl sulfate (PEI/SDS) aggregates at the air/water interface have been resolved for the first time with respect to polyelectrolyte charged density. A recently developed method to form efficient films from the dissociation of aggregates using a minimal quantity of materials is exploited as a step forward in enhancing understanding of the film properties with a view to their future use in technological applications. Interfacial techniques that resolve different time and length scales, namely, ellipsometry, Brewster angle microscopy, and neutron reflectometry, are used. Extended structures of both components are formed under a monolayer of the surfactant with bound polyelectrolytes upon film compression on subphases adjusted to pH 4 or 10, corresponding to high and low charge density of the polyelectrolyte, respectively. A rigid film is related to compact conformation of the PEI in the interfacial structure at pH 4, while it is observed that aggregates remain embedded in mobile films at pH 10. The ability to compact surfactants in the monolayer to the same extent as its maximum coverage in the absence of polyelectrolyte is distinct from the behavior observed for spread films involving linear polyelectrolytes, and intriguingly evidence points to the formation of extended structures over the full range of surface pressures. We conclude that the molecular architecture and charge density can be important parameters in controlling the structures and properties of spread polyelectrolyte/surfactant films, which holds relevance to a range of applications, such as those where PEI is used, including CO2 capture, electronic devices, and gene transfection.

Strikingly Different Roles of SARS-CoV-2 Fusion Peptides Uncovered by Neutron Scattering.

Coronavirus disease-2019 (COVID-19), a potentially lethal respiratory illness caused by the coronavirus SARS-CoV-2, emerged in the end of 2019 and has since spread aggressively across the globe. A thorough understanding of the molecular mechanisms of cellular infection by coronaviruses is therefore of utmost importance. A critical stage in infection is the fusion between viral and host membranes. Here, we present a detailed investigation of the role of selected SARS-CoV-2 Spike fusion peptides, and the influence of calcium and cholesterol, in this fusion process. Structural information from specular neutron reflectometry and small angle neutron scattering, complemented by dynamics information from quasi-elastic and spin-echo neutron spectroscopy, revealed strikingly different functions encoded in the Spike fusion domain. Calcium drives the N-terminal of the Spike fusion domain to fully cross the host plasma membrane. Removing calcium, however, reorients the peptide back to the lipid leaflet closest to the virus, leading to significant changes in lipid fluidity and rigidity. In conjunction with other regions of the fusion domain, which are also positioned to bridge and dehydrate viral and host membranes, the molecular events leading to cell entry by SARS-CoV-2 are proposed.

Reflectometry Reveals Accumulation of Surfactant Impurities at Bare Oil/Water Interfaces.

Bare interfaces between water and hydrophobic media like air or oil are of fundamental scientific interest and of great relevance for numerous applications. A number of observations involving water/hydrophobic interfaces have, however, eluded a consensus mechanistic interpretation so far. Recent theoretical studies ascribe these phenomena to an interfacial accumulation of charged surfactant impurities in water. In the present work, we show that identifying surfactant accumulation with X-ray reflectometry (XRR) or neutron reflectometry (NR) is challenging under conventional contrast configurations because interfacial surfactant layers are then hardly visible. On the other hand, both XRR and NR become more sensitive to surfactant accumulation when a suitable scattering length contrast is generated by using fluorinated oil. With this approach, significant interfacial accumulation of surfactant impurities at the bare oil/water interface is observed in experiments involving standard cleaning procedures. These results suggest that surfactant impurities may be a limiting factor for the investigation of fundamental phenomena involving water/hydrophobic interfaces.

Tuning Interfacial Properties and Processes by Controlling the Rheology and Structure of Poly( N-isopropylacrylamide) Particles at Air/Water Interfaces.

By combining controlled experiments on single interfaces with measurements on solitary bubbles and liquid foams, we show that poly( N-isopropylacrylamide) (PNIPAM) microgels assembled at air/water interfaces exhibit a solid to liquid transition changing the temperature, and that this is associated with the change in the interfacial microstructure of the PNIPAM particles around their volume phase transition temperature. We show that the solid behaves as a soft 2D colloidal glass, and that the existence of this solid/liquid transition offers an ideal platform to tune the permeability of air bubbles covered by PNIPAM and to control macroscopic foam properties such as drainage, stability, and foamability. PNIPAM particles on fluid interfaces allow new tunable materials, for example foam structures with variable mechanical properties upon small temperature changes.

Physico-chemical foundations of particle-laden fluid interfaces.

Particle-laden interfaces are ubiquitous nowadays. The understanding of their properties and structure is essential for solving different problems of technological and industrial relevance; e.g. stabilization of foams, emulsions and thin films. These rely on the response of the interface to mechanical perturbations. The complex mechanical response appearing in particle-laden interfaces requires deepening on the understanding of physico-chemical mechanisms underlying the assembly of particles at interface which plays a central role in the distribution of particles at the interface, and in the complex interfacial dynamics appearing in these systems. Therefore, the study of particle-laden interfaces deserves attention to provide a comprehensive explanation on the complex relaxation mechanisms involved in the stabilization of fluid interfaces.

Microcapsule Buckling Triggered by Compression-Induced Interfacial Phase Change.

There is an emerging trend toward the fabrication of microcapsules at liquid interfaces. In order to control the parameters of such capsules, the interfacial processes governing their formation must be understood. Here, poly(vinyl alcohol) films are assembled at the interface of water-in-oil microfluidic droplets. The polymer is cross-linked using cucurbit[8]uril ternary supramolecular complexes. It is shown that compression-induced phase change causes the onset of buckling in the interfacial film. On evaporative compression, the interfacial film both increases in density and thickens, until it reaches a critical density and a phase change occurs. We show that this increase in density can be simply related to the film Poisson ratio and area compression. This description captures fundamentals of many compressive interfacial phase changes and can also explain the observation of a fixed thickness-to-radius ratio at buckling, [Formula: see text].

Interfacial Assembly of Surfactant-Decorated Nanoparticles: On the Rheological Description of a Colloidal 2D Glass.

We address the rheology of assemblies of surfactant-decorated silica nanoparticles irreversibly adsorbed at the gas/liquid interface. Positively charged surfactant molecules (such as CTAB) bind to silica nanoparticle surfaces, and the resulting particle-surfactant complexes adsorb at gas/liquid interfaces. The surfactant molecules control the wettability of such decorated nanoparticles and their adsorption. The interparticle forces can be tuned by changing the surfactant concentration Cs. Increasing Cs, in addition to a decrease of the particles wettability, leads to an increase of the area fraction of particles at the interface. Oscillatory shear measurements (strain- and frequency-sweep) have been performed. Here, we explore the effect of the surfactant concentration Cs. At high enough Cs, the interface is highly packed, and an overall solidlike response is observed, with 2D glass properties.

Foams stabilised by mixtures of nanoparticles and oppositely charged surfactants: relationship between bubble shrinkage and foam coarsening.

We have studied foams stabilised by surfactant-decorated nanoparticles adsorbed at the bubble surfaces. We show that the controlled compression of a single bubble allows one to understand the coarsening behavior of these foams. When bubbles are compressed, the particles become tightly packed in the surface layer. They lose their mobility, and the interface becomes solid-like when the jammed state is reached. Further compression leads to interfacial buckling characterised by crumpled surfaces. We find that the surface concentration of particles at which the jamming and the buckling transitions occur are independent of the surfactant concentration. This is a surprising feature. It suggests that the surfactants are mandatory to help the particles adsorb at the interface and that they change the equilibrium surface concentration of the decorated particles. But they do not affect the surface properties once the particles are adsorbed. We measured the compression elastic modulus of the surface in the jammed state and found it to be compatible with the Gibbs condition for which the spontaneous dissolution of bubbles is arrested. Due to this effect, the coarsening process of a foam composed of many close-packed bubbles occurs in two steps. In the first step, coarsening is slow and coalescence of the bigger bubbles is observed. In the second step, a number of very small bubbles remains, which exhibit crumpled surfaces and are stable over long times. This suggests that foam coarsening is arrested once the smallest bubbles become fully covered after the initial shrinking step.

Equation of state and adsorption dynamics of soft microgel particles at an air-water interface.

Understanding the adsorption dynamics of soft microgel particles is a key step in designing such particles for potential applications as stimuli-responsive Pickering stabilizers for foams or emulsions. In this study we experimentally determine an equation of state (EOS) for poly (N-isopropylacrylamide) (PNIPAM) microgel particles adsorbed onto an air-water interface using a Langmuir film balance. We detect a finite surface pressure at very low surface concentration of particles, for which standard theories based on hard disk models predict negligible pressures, implying that the particles must deform strongly upon adsorption to the interface. Furthermore, we study the evolution of the surface pressure due to the adsorption of PNIPAM particles as a function of time using pendant drop tensiometry. The equation of state determined in the equilibrium measurements allows us to extract the adsorbed amount as a function of time. We find a mixed-kinetic adsorption that is initially controlled by the diffusion of particles towards the interface. At later stages, a slow exponential relaxation indicates the presence of a coverage-dependent adsorption barrier related to crowding of particles at the interface.

Association of oppositely charged polyelectrolyte and surfactant in solution: equilibrium and nonequilibrium features.

The association of polyelectrolytes and surfactants bearing charges of opposite signs has been for long time considered under an equilibrium framework. However, this is far to provide a true description of the physico-chemical rules of the association process when kinetically arrested nonequilibrium states are formed. This is the result of specific interaction pathways between the polyelectrolyte chains and the surfactant molecules determined by the mixing procedure and the mixture composition. In fact, the specific characteristics of the method used for the mixture can induce local inhomogeneities in the mixture composition which can govern the properties of the obtained supramolecular aggregates, driving to the systems to a situation far from the true equilibrium. This topical review tries to provide to the reader a general perspective of the role of the nonequilibrium aspects in the control of the polyelectrolyte-surfactant association process, and how these impact on the obtained supramolecular nanoassemblies, and their properties.

Unravelling the orientation of the inositol-biphosphate ring and its dependence on phosphatidylinositol 4,5-bisphosphate cluster formation in model membranes.

HYPOTHESIS: Inositol phospholipids are well known to form clusters in the cytoplasmic leaflet of the plasma membrane that are responsible for the interaction and recruitment of proteins involved in key biological processes like endocytosis, ion channel activation and secondary messenger production. Although their phosphorylated inositol ring headgroup plays an important role in protein binding, its orientation with respect to the plane of the membrane and its lateral packing density has not been previously described experimentally. EXPERIMENTS: Here, we study phosphatidylinositol 4,5-bisphosphate (PIP2) planar model membranes in the form of Langmuir monolayers by surface pressure-area isotherms, Brewster angle microscopy and neutron reflectometry to elucidate the relation between lateral (in-plane) and perpendicular (out-of-plane) molecular organization of PIP2. FINDINGS: Different surface areas were explored through monolayer compression, allowing us to correlate the formation of transient PIP2 clusters with the change in orientation of the inositol-biphosphate headgroup, which was experimentally determined by neutron reflectometry.

Investigation on the relationship between lipid composition and structure in model membranes composed of extracted natural phospholipids.

HYPOTHESIS: Unravelling the structural diversity of cellular membranes is a paramount challenge in life sciences. In particular, lipid composition affects the membrane collective behaviour, and its interactions with other biological molecules. EXPERIMENTS: Here, the relationship between membrane composition and resultant structural features was investigated by surface pressure-area isotherms, Brewster angle microscopy and neutron reflectometry on in vitro membrane models of the mammalian plasma and endoplasmic-reticulum-Golgi intermediate compartment membranes in the form of Langmuir monolayers. Natural extracted yeast lipids were used because, unlike synthetic lipids, the acyl chain saturation pattern of yeast and mammalian lipids are similar. FINDINGS: The structure of the model membranes, orthogonal to the plane of the membrane, as well as their lateral packing, were found to depend strongly on their specific composition, with cholesterol having a major influence on the in-plane morphology, yielding a coexistence of liquid-order and liquid-disorder phases.

Probing the effect of the capping polyelectrolyte on the internal structure of Layer-by-Layer decorated nanoliposomes.

HYPOTHESIS: The internal organization of polyelectrolyte layers deposited on colloidal templates plays a very important role for the potential applications of these systems as capsules for drug delivery purposes. EXPERIMENTS: The mutual arrangement of oppositely charged polyelectrolyte layers upon their deposition on positively charged liposomes has been studied by combining up three different scattering techniques and Electronic Spin Resonance, which has provided information about the inter-layer interactions and their effect on the final structure of the capsules. FINDINGS: The sequential deposition of oppositely charged polyelectrolytes on the external leaflet of positively charged liposomes allows modulating the organization of the obtained supramolecular structures, impacting the packing and rigidity of the obtained capsules due to the change of the ionic cross-linking of the multi-layered film as a result of the specific charge of the last deposited layer. The possibility to modulate the properties of the LbL capsules by tuning the characteristics of the last deposited layers offers a very interesting route for the design of materials for encapsulation purposes with their properties controlled almost at will by changing the number of deposited layers and their chemistry.

Control of the structure and morphology of polypeptide/surfactant spread films by exploiting specific interactions.

We demonstrate control of the structure and morphology of polypeptide/surfactant films at the air/water interface as a function of the maximum compression ratio of the surface area, exploiting a recently developed film formation mechanism that requires minimal quantities of materials involving the dissociation of aggregates. The systems studied are poly(L-lysine) (PLL) or poly(L-arginine) (PLA) with sodium dodecyl sulfate (SDS), chosen because the surfactant (i) interacts more strongly with the latter polypeptide due to the formation of hydrogen bonds between the guanidinium group and its oxygen atoms, and (ii) induces bulk ß-sheet and α-helix conformations of the respective polypeptides. The working hypothesis is that such different interactions may be used to tune the film properties when compressed to form extended structures (ESs). Neutron reflectometry reveals that application of a high compression ratio (4.5 : 1) results in the nanoscale self-assembly of ESs containing up to two PLL-wrapped SDS bilayers. Brewster angle microscopy provides images of the PLL/SDS ESs as discrete regions on the micrometre scale while additional linear regions of PLA/SDS ESs mark macroscopic film folding. Ellipsometry demonstrates high stability of the different ESs formed. The collapse of PLL/SDS films upon compression to a very high ratio (10 : 1) is irreversible due to the formation of solid domains that remain embedded in the film upon expansion while that of PLA/SDS films is reversible. These findings demonstrate that differences in the side group of a polypeptide can have a major influence on controlling the film properties, marking a key step in the development of this new film formation mechanism for the design of biocompatible and/or biodegradable films with tailored properties for applications in tissue engineering, biosensors and antimicrobial coatings.

Developing advanced models of biological membranes with hydrogenous and deuterated natural glycerophospholipid mixtures.

Cellular membranes are complex systems that consist of hundreds of different lipid species. Their investigation often relies on simple bilayer models including few synthetic lipid species. Glycerophospholipids (GPLs) extracted from cells are a valuable resource to produce advanced models of biological membranes. Here, we present the optimisation of a method previously reported by our team for the extraction and purification of various GPL mixtures from Pichia pastoris. The implementation of an additional purification step by High Performance Liquid Chromatography-Evaporative Light Scattering Detector (HPLC-ELSD) enabled for a better separation of the GPL mixtures from the neutral lipid fraction that includes sterols, and also allowed for the GPLs to be purified according to their different polar headgroups. Pure GPL mixtures at significantly high yields were produced through this approach. For this study, we utilised phoshatidylcholine (PC), phosphatidylserine (PS) and phosphatidylglycerol (PG) mixtures. These exhibit a single composition of the polar head, i.e., PC, PS or PG, but contain several molecular species consisting of acyl chains of varying length and unsaturation, which were determined by Gas Chromatography (GC). The lipid mixtures were produced both in their hydrogenous (H) and deuterated (D) versions and were used to form lipid bilayers both on solid substrates and as vesicles in solution. The supported lipid bilayers were characterised by quartz crystal microbalance with dissipation monitoring (QCM-D) and neutron reflectometry (NR), whereas the vesicles by small angle X-ray (SAXS) and neutron scattering (SANS). Our results show that despite differences in the acyl chain composition, the hydrogenous and deuterated extracts produced bilayers with very comparable structures, which makes them valuable to design experiments involving selective deuteration with techniques such as NMR, neutron scattering or infrared spectroscopy.

Engineering phosphatidylinositol-4,5-bisphosphate model membranes enriched in endocytic cargo: A neutron reflectometry, AFM and QCM-D structural study.

The combination of in vitro models of biological membranes based on solid-supported lipid bilayers (SLBs) and of surface sensitive techniques, such as neutron reflectometry (NR), atomic force microscopy (AFM) and quartz crystal microbalance with dissipation monitoring (QCM-D), is well suited to provide quantitative information about molecular level interactions and lipid spatial distributions. In this work, cellular plasma membranes have been mimicked by designing complex SLB, containing phosphatidylinositol 4,5-bisphosphate (PtdIns4,5P2) lipids as well as incorporating synthetic lipo-peptides that simulate the cytoplasmic tails of transmembrane proteins. The QCM-D results revealed that the adsorption and fusion kinetics of PtdIns4,5P2 are highly dependent of Mg2+. Additionally, it was shown that increasing concentrations of PtdIns4,5P2 leads to the formation of SLBs with higher homogeneity. The presence of PtdIns4,5P2 clusters was visualized by AFM. NR provided important insights about the structural organization of the various components within the SLB, highlighting that the leaflet symmetry of these SLBs is broken by the presence of CD4-derived cargo peptides. Finally, we foresee our study to be a starting point for more sophisticated in vitro models of biological membranes with the incorporation of inositol phospholipids and synthetic endocytic motifs.

Soft Colloidal Particles at Fluid Interfaces.

The assembly of soft colloidal particles at fluid interfaces is reviewed in the present paper, with emphasis on the particular case of microgels formed by cross-linked polymer networks. The dual polymer/colloid character as well as the stimulus responsiveness of microgel particles pose a challenge in their experimental characterization and theoretical description when adsorbed to fluid interfaces. This has led to a controversial and, in some cases, contradictory picture that cannot be rationalized by considering microgels as simple colloids. Therefore, it is necessary to take into consideration the microgel polymer/colloid duality for a physically reliable description of the behavior of the microgel-laden interface. In fact, different aspects related to the above-mentioned duality control the organization of microgels at the fluid interface, and the properties and responsiveness of the obtained microgel-laden interfaces. This works present a critical revision of different physicochemical aspects involving the behavior of individual microgels confined at fluid interfaces, as well as the collective behaviors emerging in dense microgel assemblies.

In-situ study of the impact of temperature and architecture on the interfacial structure of microgels.

The structural characterization of microgels at interfaces is fundamental to understand both their 2D phase behavior and their role as stabilizers that enable emulsions to be broken on demand. However, this characterization is usually limited by available experimental techniques, which do not allow a direct investigation at interfaces. To overcome this difficulty, here we employ neutron reflectometry, which allows us to probe the structure and responsiveness of the microgels in-situ at the air-water interface. We investigate two types of microgels with different cross-link density, thus having different softness and deformability, both below and above their volume phase transition temperature, by combining experiments with computer simulations of in silico synthesized microgels. We find that temperature only affects the portion of microgels in water, while the strongest effect of the microgels softness is observed in their ability to protrude into the air. In particular, standard microgels have an apparent contact angle of few degrees, while ultra-low cross-linked microgels form a flat polymeric layer with zero contact angle. Altogether, this study provides an in-depth microscopic description of how different microgel architectures affect their arrangements at interfaces, and will be the foundation for a better understanding of their phase behavior and assembly.

A broad perspective to particle-laden fluid interfaces systems: from chemically homogeneous particles to active colloids.

Particles adsorbed to fluid interfaces are ubiquitous in industry, nature or life. The wide range of properties arising from the assembly of particles at fluid interface has stimulated an intense research activity on shed light to the most fundamental physico-chemical aspects of these systems. These include the mechanisms driving the equilibration of the interfacial layers, trapping energy, specific inter-particle interactions and the response of the particle-laden interface to mechanical perturbations and flows. The understanding of the physico-chemistry of particle-laden interfaces becomes essential for taking advantage of the particle capacity to stabilize interfaces for the preparation of different dispersed systems (emulsions, foams or colloidosomes) and the fabrication of new reconfigurable interface-dominated devices. This review presents a detailed overview of the physico-chemical aspects that determine the behavior of particles trapped at fluid interfaces. This has been combined with some examples of real and potential applications of these systems in technological and industrial fields. It is expected that this information can provide a general perspective of the topic that can be exploited for researchers and technologist non-specialized in the study of particle-laden interfaces, or for experienced researcher seeking new questions to solve.