Gel Domains Research Articles

Young modulus of supported lipid membranes containing milk sphingomyelin in the gel, fluid or liquid-ordered phase, determined using AFM force spectroscopy

The biological membrane surrounding milk fat globules (MFGM) exhibits lateral phase separation of lipids, interpreted as gel or liquid-ordered phase sphingomyelin-rich (milk SM) domains dispersed in a fluid continuous lipid phase. The objective of this study was to investigate whether changes in the phase state of milk SM-rich domains induced by temperature (T < Tm or T > Tm) or cholesterol affected the Young modulus of the lipid membrane. Supported lipid bilayers composed of MFGM polar lipids, milk SM or milk SM/cholesterol (50:50 mol) were investigated at 20 °C and 50 °C using atomic force microscopy (AFM) and force spectroscopy. At 20 °C, gel-phase SM-rich domains and the surrounding fluid phase of the MFGM polar lipids exhibited Young modulus values of 10–20 MPa and 4–6 MPa, respectively. Upon heating at 50 °C, the milk SM-rich domains in MFGM bilayers as well as pure milk SM bilayers melted, leading to the formation of a homogeneous membrane with similar Young modulus values to that of a fluid phase (0–5 MPa). Upon addition of cholesterol to the milk SM to reach 50:50 mol%, membranes in the liquid-ordered phase exhibited Young modulus values of a few MPa, at either 20 or 50 °C. This indicated that the presence of cholesterol fluidized milk SM membranes and that the Young modulus was weakly affected by the temperature. These results open perspectives for the development of milk polar lipid based vesicles with modulated mechanical properties.

Lateral Segregation of Palmitoyl Ceramide-1-Phosphate in Simple and Complex Bilayers

Ceramide-1-phosphate is a minor sphingolipid with important functions in cell signaling. In this study, we examined the propensity of palmitoyl ceramide-1-phosphate (Cer-1P) to segregate laterally into ordered domains in different bilayer compositions at 23 and 37°C and compared this with segregation of palmitoyl ceramide (PCer) and palmitoyl sphingomyelin (PSM). The ordered-domain formation in the fluid phosphatidylcholine bilayers was determined using the emission lifetime changes of trans-parinaric acid and from differential scanning calorimetry thermograms. The lateral segregation of Cer-1P was examined when hydrated to bilayers in Tris buffer (50 mM Tris, 140 mM NaCl (pH 7.4)). At this pH, Cer-1P was negatively charged. The lateral segregation propensity of Cer-1P in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) bilayers was intermediate between PCer and PSM. Based on differential scanning calorimetry analysis, we observed that the gel domains formed by Cer-1P in POPC bilayers (POPC:Cer-1P 70:30 by mol) were less stable (melting interval 16–37°C) than the corresponding POPC and PCer gel domains at equal composition (melting interval 20–55°C). The gel-phase melting enthalpy was also much lower in Cer-1P (1.5 kcal/mol) than in the PCer-containing POPC bilayers (9 kcal/mol). Cer-1P appeared to be at least partially miscible with PCer domains in POPC bilayers. Cer-1P domains were stabilized in the presence of PSM (POPC:PSM 85:15), similarly as seen with PCer-rich domains. In bilayers at 37°C, with an approximate outer-leaflet cell membrane composition (sphingomyelin and cholesterol enriched, aminophospholipid poor), Cer-1P segregation did not lead to the formation of ordered domains, at least when compared with PCer segregation. In bilayers with an approximate inner-leaflet composition (sphingomyelin poor, cholesterol and aminophospholipid enriched), Cer-1P also failed to form ordered domains. PCer segregated into ordered domains only after the PCer/cholesterol ratio exceeded an approximate equimolar ratio.

Coupling of Membrane Nanodomain Formation and Enhanced Electroporation near Phase Transition

Biological cells are enveloped by a heterogeneous lipid bilayer that prevents the uncontrolled exchange of substances between the cell interior and its environment. In particular, membranes act as a continuous barrier for salt and macromolecules to ensure proper physiological functions within the cell. However, it has been shown that membrane permeability strongly depends on temperature and, for phospholipid bilayers, displays a maximum at the transition between the gel and fluid phase. Here, extensive molecular dynamics simulations of dipalmitoylphosphatidylcholine bilayers were employed to characterize the membrane structure and dynamics close to phase transition, as well as its stability with respect to an external electric field. Atomistic simulations revealed the dynamic appearance and disappearance of spatially related nanometer-sized thick ordered and thin interdigitating domains in a fluid-like bilayer close to the phase transition temperature (Tm). These structures likely represent metastable precursors of the ripple phase that vanished at increased temperatures. Similarly, a two-phase bilayer with coexisting gel and fluid domains featured a thickness minimum at the interface because of splaying and interdigitating lipids. For all systems, application of an external electric field revealed a reduced bilayer stability with respect to pore formation for temperatures close to Tm. Pore formation occurred exclusively in thin interdigitating membrane nanodomains. These findings provide a link between the increased membrane permeability and the structural heterogeneity close to phase transition.

Nonlamellar-Phase-Promoting Colipids Enhance Segregation of Palmitoyl Ceramide in Fluid Bilayers

Ceramide is an important intermediate in sphingolipid homeostasis. We examined how colipids, with negative intrinsic curvature and which may induce curvature stress in the bilayers, affected the segregation of palmitoyl ceramide (PCer). Such colipids include 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and tetra-linoleoyl cardiolipin (CL). In 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) bilayers, PCer formed ordered, gel-like domains at concentrations above 10 mol% at 23°C, as evidenced by the change in the average lifetime of the trans-parinaric acid emission. When POPE or DOPE were included in the DOPC bilayer (at 20:80 or 40:60 POPE or DOPE to DOPC, by mol), the lateral segregation of PCer was facilitated in a concentration-dependent manner, and less PCer was required for the formation of the ordered ceramide-rich domains. Inclusion of CL in the DOPE bilayer (at 10:90 or 20:80 CL to PC, by mol) also caused a similar facilitation of the lateral segregation of PCer. The PCer-rich domains formed in the presence of POPE, DOPE, or CL in DOPC bilayers were slightly more thermostable (by 2–10°C) when compared to PCer-rich domains in DOPC-only bilayers. Nonlamellar phases were not present in bilayers in which the effects of POPE or DOPE on PCer segregation were the largest, as verified by 31P NMR. When palmitoyl sphingomyelin was added to the different bilayer compositions at 5 mol%, relative to the phospholipids, PCer segregated into gel domains at lower concentrations (2–3 mol% PCer), and the effect of POPE on PCer segregation was eliminated. We suggest that the effects of POPE, DOPE, and CL on PCer segregation was in part influenced by their effects on membrane curvature stress and in part because of unfavorable interactions with PCer due to their unsaturated acyl chains. These lipids are abundant in mitochondrial membranes and are likely to affect functional properties of saturated ceramides in them.

Core Scale Modeling of Polymer Gel Dehydration by Spontaneous Imbibition

SummaryCapillary spontaneous imbibition (SI) of solvent (water bound in gel) from formed polymer gel into an adjacent, oil-saturated porous medium was recently observed in laboratory experiments. Loss of water from the gel by SI might influence the blocking capacity of the gel residing in a fracture, by decreasing its volume, and might contribute to gel failure, often observed in water-wet oil fields.This work presents an original modeling approach to simulate and interpret spontaneous imbibition of water from Cr(III)-acetate-hydrolyzed-polyacrylamide (HPAM) gel into adjacent oil-saturated rock matrix. Simulations were compared to experiments on the core scale, using two different boundary conditions: all faces open (AFO) and two-ends-open free spontaneous imbibition (TEOFSI). Capillary forces enable water (used as gel solvent) to enter the rock matrix. The gel particle network itself is, however, inhibited from entering because of its structure, and remains on the surface of the rock matrix. We developed a theory that describes the gel as a compressible porous medium and describes the flow of water through gel. The polymer structure of the gel is proposed to constitute a gel matrix of constant solid volume. Gel porosity, defined by the volume fraction of solvent, is modeled as a function of pore pressure and gel compressibility. Gel permeability is modeled as function of gel porosity using a Kozeny-Carman approach. The flow equations for the gel and core domains were solved simultaneously by implementing the proposed description into the core-scale simulator IORCoreSim. The gel surrounding the core was discretized and included as a part of the total grid.The simulated flow of water through and from the gel occurred in a transient manner, driven by the coupled gradients in gel fluid pressure and gel porosity. Gel porosity initially decreased in a layer close to the core surface because of reduced aqueous pressure, and continued to decrease in layers away from the core surface. The propagation rate was controlled by two main gel parameters: First, gel compressibility controlled the pressure gradient within the gel network, and the amount of water transported from the outer part of the gel toward the core surface to balance the pore pressure; and, second, gel permeability limited how fast water could flow within the gel at a given pressure gradient, thus increasing the time scale of the overall imbibition process.

Polymer Coating and Lipid Phases Regulate Semiconductor Nanorods' Interaction with Neuronal Membranes: A Modeling Approach.

The interplay between nanoparticles (NPs) and cell membranes is extremely important with regard to using NPs in biology applications. With the aim of unraveling the dominating factors on the molecular scale, we have studied the interaction between polymer-coated semiconductor nanorods (NRs) made of cadmium selenium/cadmium sulfur and model lipid membranes. The zeta potential (ζ) of the NRs was tuned from having a negative value (-24 mV) to having a positive one (+11 mV) by changing the amine content in the polymer coating. Supported lipid bilayers (SLBs) and lipid monolayers (LMs) were used as model membranes. Lipid mixtures containing anionic or cationic lipids were employed in order to change the membrane ζ from -77 to +49 mV; lipids with saturated hydrophobic chains were used to create phase-separated gel domains. NR adsorption to the SLBs was monitored by quartz crystal microbalance with dissipation monitoring; interactions with LMs with the same lipid composition were measured by surface pressure-area isotherms. The results showed that the NRs only interact with the model membrane if the mutual Δζ is higher than 70 mV; at the air-water interface, positively charged NRs remove lipids from the anionic lipid mixtures, and the negative ones penetrate the space between the polar heads in the cationic mixtures. However, the presence of gel domains in the membrane inhibits this interaction. The results of the Derjaguin-Landau-Verwey-Overbeek model frame indicate that the interaction occurs not only due to electrostatic and van der Waals forces, but also due to steric and/or hydration forces.

Regimes of Complex Lipid Bilayer Phases Induced by Cholesterol Concentration in MD Simulation

Cholesterol is essential to the formation of phase-separated lipid domains in membranes. Lipid domains can exist in different thermodynamic phases depending on the molecular composition and play significant roles in determining structure and function of membrane proteins. We investigate the role of cholesterol in the structure and dynamics of ternary lipid mixtures displaying phase separation using molecular dynamics simulations, employing a physiologically relevant span of cholesterol concentration. We find that cholesterol can induce formation of three regimes of phase behavior: 1) miscible liquid-disordered bulk, 2) phase-separated, domain-registered coexistence of liquid-disordered and liquid-ordered domains, and 3) phase-separated, domain-antiregistered coexistence of liquid-disordered and newly identified nanoscopic gel domains composed of cholesterol threads we name “cholesterolic gel” domains. These findings are validated and discussed in the context of current experimental knowledge, models of cholesterol spatial distributions, and models of ternary lipid-mixture phase separation.

Anti-icing Properties on Surfaces through a Functional Composite: Effect of Ionic Salts.

This study reports the potential of a unique functional composite for anti-icing applications. To date, various ionic salt formulations have been applied to prevent ice accumulation on surfaces. However, salt can be removed by external factors and large amounts must be used to attain anti-icing properties. Incorporating hydrophilic salts into hydrophobic mediums and controlled release of specific agents can provide effective solution to reduce ice accumulation on surfaces. Here, we developed functional polymer composites with salt pockets of altered ionic salts consisting of potassium formate (KCOOH), sodium chloride (NaCl), or magnesium chloride (MgCl2). We dissolved ionic salts in hydrophilic gel domains and dispersed in a hydrophobic styrene–butadiene–styrene polymer matrix. Na+ and Cl– ions delayed ice formation by 42.6 min at −2 °C compared to that for unmodified surfaces. Functional composites prepared with the NaCl ionic salt exhibited better anti-icing behavior at −2 °C because of their high concentration compared to that of the composites prepared with KCOOH and MgCl2 ionic salts. We also characterized the release of ionic salts from composite-modified hydrophobic medium separately up to 118 days. Furthermore, we monitored freezing of water on composite-incorporated or composite-coated hydrophobic surfaces in a camera-integrated cold chamber with a uniform temperature (−2 °C). The results demonstrated significant increases in the delay of freezing on composite-incorporated or composite-coated surfaces compared to that on controls. We observed altered effects of each ionic salt on the mechanical, morphological, and functional properties of the composite-incorporated or composite-coated hydrophobic surfaces. Our results suggested that the efficiency of a polymer composite to promote anti-icing behavior on a surface is directly related to the type and concentration of the particular ionic salt incorporation into the composite. This approach is promising and demonstrates significant potential of the ionic salt embedded within polymer composite-modified hydrophobic surfaces to attain delayed icing function.

Molecular Structure and Permeability at the Interface between Phase-Separated Membrane Domains.

Phase-separated membrane domains, also known as lipid rafts, are believed to play an important role in cell function. Although most rafts are sterol-enriched membrane regions, evidence suggests that living cells may also contain gel-like rafts. Interactions between gel and fluid domains have a large impact on membrane properties, as is the case with permeability. The membrane permeability may reach a peak at the main phase transition temperature, by far exceeding the values recorded at the fluid phase. It has been proposed that gel-fluid interfaces are leaky, but the effect has not yet been demonstrated at the molecular level. Here, we performed atomistic molecular dynamics simulations of phospholipid bilayers with coexisting gel-like and fluid domains. We found that the thickness mismatch between both phases, the membrane elasticity, and the lipid packing acted together to promote the formation of a thickness minimum at the gel-fluid interface. Free energy calculations showed that pore-mediated ionic permeation was strongly facilitated at the constriction region, whereas water permeation by simple diffusion was only marginally affected. Long-lived, peristaltic undulations were recorded at the bulk fluid phase near the main transition temperature. They gave rise to thickness minima that, although shallower than the interface constrictions, could also enhance permeability. Finally, we demonstrated that an interface constriction was also formed at the boundaries of regular, cholesterol-enriched lipid rafts. Our simulation results will hopefully contribute to a better understanding of biological processes such as transport, signaling, and cellular damage promoted by low temperature and dehydration.

Effective Bending Rigidity of Membranes with Rigid Inclusions

The ability of biological membranes to bend and deform is essential to a wide range of cell processes. Significant advances in understanding the elasticity of lipid bilayers underlying these deformations have been made by studying homogeneous fluid membranes. However, biological membranes are far from homogeneous, and less is known about how the heterogeneity affects the material properties of the membrane. Here we use neutron spin echo spectroscopy (NSE) to study the effective bending rigidity (κeff) of heterogeneous lipid membranes with coexisting rigid gel and soft fluid domains. Our data show that κeff scales directly, but not linearly, with the area faction of gel phase and are in excellent agreement with theoretical predictions for heterogeneous lipid membranes with rigid inclusions. While it is intuitively expected that κeff should in some way depend on the fraction of gel phase, the functional form of the scaling has important implications for understanding the effects of rigid inclusions, such as proteins or lipid domains, on the effective stiffness of biological membranes.

Complex Effects of 24:1 Sphingolipids in Membranes Containing Dioleoylphosphatidylcholine and Cholesterol

The effects of C24:1 sphingolipids have been tested in phospholipid bilayers containing cholesterol. Confocal microscopy, differential scanning calorimetry, and atomic force microscopy imaging and force curves have been used. More precisely, the effects of C24:1 ceramide (nervonoyl ceramide, nCer) were evaluated and compared to those of C16:0 ceramide (palmitoyl ceramide, pCer) in bilayers composed basically of dioleoylphosphatidylcholine, sphingomyelin (either C24:1, nSM or C16:0, pSM) and cholesterol. Combination of equimolecular amounts of C24:1 and C16:0 sphingolipids were also studied under the same conditions. Results show that both pCer and nCer are capable of forming segregated gel domains. Force spectroscopy data point to nCer having a lower stiffening effect than pCer, while the presence of nSM reduces the stiffness. DSC reveals Tm reduction by nSM in every case. Furthermore, pSM seems to better accommodate both ceramides in a single phase of intermediate properties, while nSM partial accommodat...

29Si{27Al}, 27Al{29Si} and 27Al{1H} double-resonance NMR spectroscopy study of cementitious sodium aluminosilicate gels (geopolymers) and gel-zeolite composites.

The influence of starting materials and synthesis route on the properties and the structure of cementitious sodium aluminosilicate gels is not fully understood, partly due their amorphous nature and the fact that they often contain residual reactants, which can make the results of single-pulse NMR spectroscopy applied to these materials difficult to interpret or ambiguous. To overcome some of these limitations, 29Si{27Al} TRAPDOR NMR as well as 27Al{29Si} and 27Al{1H} REDOR NMR spectroscopy were applied to materials synthesized by the one-part alkali-activation route from three different amorphous silica starting materials, including rice husk ash. The latter led to formation of a fully amorphous sodium aluminosilicate gel (geopolymer), while the materials produced from the other silicas contained amorphous phase and crystalline zeolites. Application of the double-resonance NMR methods allowed to identify hydrous alumina gel domains in the rice husk ash-based material as well as significantly differing amounts of residual silica in the three cured materials. Four-coordinated Al existed not only in the aluminosilicate gel framework but also in a water-rich chemical environment with only a small amount of Si in proximity, likely in the alumina gel or possibly present as extra-framework Al in the aluminosilicate gel. The results demonstrate how the employment of different silica starting materials determines the phase assemblage of one-part alkali-activated materials, which in turn influences their engineering properties such as the resistance against chemically/biologically aggressive media.

Preparation and Membrane Properties of Oxidized Ceramide Derivatives.

Ceramide is a bioactive lipid with important roles in several biological processes including cell proliferation and apoptosis. Although 3-ketoceramides that contain a keto group in place of the 3-OH group of ceramide occur naturally, ceramide derivatives oxidized at the primary 1-OH group have not been identified to date. To evaluate how the oxidative state of the 1-OH group affects the physical properties of membranes, we prepared novel ceramide derivatives in which the 1-OH group was oxidized to a carboxylic acid (PCerCOOH) or methylester (PCerCOOMe) and examined the rigidity of their monolayers and the formation of gel domains in palmitoyloleoylphosphatidylcholine (POPC) or sphingomyelin (SM) bilayers. As a result, PCerCOOH and PCerCOOMe exhibited membrane properties similar to those of native ceramide, although the deprotonated form of PCerCOOH, PCerCOO-, exhibited markedly lower rigidity and higher miscibility with POPC and SM. This was attributed to the electrostatic repulsion of the negative charge, which hampered the formation of the ceramide-enriched gel domain. The similarities in the properties of PCerCOOMe and ceramide revealed the potential to introduce various functional groups onto PCerCOOH via ester or amide linkages; therefore, these derivatives will also provide a new strategy for developing molecular probes, such as fluorescent ceramides, and inhibitors of ceramide-related enzymes.

Investigation of Nanoscopic Phase Separations in Lipid Membranes Using Inverse FCS

Measurement of the sizes of nanoscopic particles is a difficult challenge, especially in two-dimensional systems such as cell membranes. We have extended inverse fluorescence correlation spectroscopy (iFCS) to endow it with unique advantages for measuring particle size from the nano- to the microscale. We have augmented iFCS with an analysis of moments of fluorescence fluctuations and used it to measure stages of phase separation in model lipid bilayer membranes. We observed two different pathways for the growth of phase domains. In one, nanoscopic gel domains appeared first and then gradually grew to micrometer size. In the other, the domains reached micrometer size quickly, and their number gradually increased. These measurements demonstrate the value of iFCS measurements through their ability, to our knowledge, to provide new information about the mechanism of lipid phase separation and potentially about the physical basis of naturally occurring nanodomains such as lipid rafts.

Complex Effects of 24:1 Sphingolipids in Membranes Containing Dioleoylphosphatidylcholine and Cholesterol.

The effects of C24:1 sphingolipids have been tested in phospholipid bilayers containing cholesterol. Confocal microscopy, differential scanning calorimetry, and atomic force microscopy imaging and force curves have been used. More precisely, the effects of C24:1 ceramide (nervonoyl ceramide, nCer) were evaluated and compared to those of C16:0 ceramide (palmitoyl ceramide, pCer) in bilayers composed basically of dioleoylphosphatidylcholine, sphingomyelin (either C24:1, nSM or C16:0, pSM) and cholesterol. Combination of equimolecular amounts of C24:1 and C16:0 sphingolipids were also studied under the same conditions. Results show that both pCer and nCer are capable of forming segregated gel domains. Force spectroscopy data point to nCer having a lower stiffening effect than pCer, while the presence of nSM reduces the stiffness. DSC reveals Tm reduction by nSM in every case. Furthermore, pSM seems to better accommodate both ceramides in a single phase of intermediate properties, while nSM partial accommodation of ceramides generates different gel phases with higher stiffnesses caused by interceramide cooperation. If both pSM and nSM are present, a clear preference of both ceramides toward pSM is observed. These findings show the sharp increase in complexity when membranes exhibit different sphingolipids of varying N-acyl chains, which should be a common issue in an actual cell membrane environment.

Phase behavior of supported lipid bilayers: A systematic study by coarse-grained molecular dynamics simulations.

Solid-supported lipid bilayers are utilized by experimental scientists as models for biological membranes because of their stability. However, compared to free standing bilayers, their close proximity to the substrate may affect their phase behavior. As this is still poorly understood, and few computational studies have been performed on such systems thus far, here we present the results from a systematic study based on molecular dynamics simulations of an implicit-solvent model for solid-supported lipid bilayers with varying lipid-substrate interactions. The attractive interaction between the substrate and the lipid head groups that are closest to the substrate leads to an increased translocation of the lipids from the distal to the proximal bilayer-leaflet. This thereby leads to a transbilayer imbalance of the lipid density, with the lipid density of the proximal leaflet higher than that of the distal leaflet. Consequently, the order parameter of the proximal leaflet is found to be higher than that of the distal leaflet, the higher the strength of lipid interaction is, the stronger the effect. The proximal leaflet exhibits gel and fluid phases with an abrupt melting transition between the two phases. In contrast, below the melting temperature of the proximal leaflet, the distal leaflet is inhomogeneous with coexisting gel and fluid domains. The size of the fluid domains increases with increasing the strength of the lipid interaction. At low temperatures, the inhomogeneity of the distal leaflet is due to its reduced lipid density.

The molecular mechanism of Nystatin action is dependent on the membrane biophysical properties and lipid composition.

Nystatin (Nys) is a pore forming broad-spectrum and efficient antifungal drug with significant toxicity in mammalian organisms. In order to develop a non-toxic and more effective Nys formulation, its molecular mechanism of action at the cell membrane needs to be better understood. It is widely accepted that Nys activity and toxicity depend on the presence and type of membrane sterols. Taking advantage of multiple biophysical methodologies, we now show that the formation and stabilization of Nys aqueous pores, which are associated with Nys cytotoxicity, occur in the absence of membrane sterols. Our results suggest that the Nys mechanism of action is driven by the presence of highly ordered membrane domains capable of stabilizing the Nys oligomers. Moreover, Nys pore formation is accompanied by strong Nys-induced membrane reorganization that depends on membrane lipid composition and seems to underlie the Nys cytotoxic effect. Accordingly, in membranes enriched in a gel-phase forming phospholipid, Nys incorporates within the phospholipid-enriched gel domains, where it forms pores able to expand the gel domains. In contrast, in membranes enriched in gel domain forming sphingolipids, Nys-induced pore formation occurs through the destabilization of the gel phase. These results show that the Nys mechanism of action is complex and not only dependent on membrane sterols, and provide further insight into the molecular details governing Nys activity and toxicity.

Three-Phase Coexistence in Lipid Membranes

Phospholipid ternary systems are useful model systems for understanding lipid-lipid interactions and their influence on biological properties such as cell signaling and protein translocation. Despite extensive studies, there are still open questions relating to membrane phase behavior, particularly relating to a proposed state of three-phase coexistence, due to the difficulty in clearly distinguishing the three phases. We look in and around the region of the phase diagram where three phases are expected and use a combination of different atomic force microscopy (AFM) modes to present the first images of three coexisting lipid phases in biomimetic cell lipid membranes. Domains form through either nucleation or spinodal decomposition dependent upon composition, with some exhibiting both mechanisms in different domains simultaneously. Slow cooling rates are necessary to sufficiently separate mixtures with high proportions of lo and lβ phase. We probe domain heights and mechanical properties and demonstrate that the gel (lβ) domains have unusually low structural integrity in the three-phase region. This finding supports the hypothesis of a “disordered gel” state that has been proposed from NMR studies of similar systems, where the addition of small amounts of cholesterol was shown to disrupt the regular packing of the lβ state. We use NMR data from the literature on chain disorder in different mixtures and estimate an expected step height that is in excellent agreement with the AFM data. Alternatively, the disordered solid phase observed here and in the wider literature could be explained by the lβ phase being out of equilibrium, in a surface kinetically trapped state. This view is supported by the observation of unusual growth of nucleated domains, which we term “tree-ring growth,” reflecting compositional heterogeneity in large disordered lβ phase domains.

Spatially-resolved soft materials for controlled release - hybrid hydrogels combining a robust photo-activated polymer gel with an interactive supramolecular gel.

Hybrid hydrogels based on self-assembling low-molecular-weight gelator (LMWG) DBS-CONHNH2 (DBS = 1,3;2,4-dibenzylidene-d-sorbitol) and crosslinked polymer gelator (PG) PEGDM (poly(ethyleneglycol) dimethacrylate) are reported, and an active pharmaceutical ingredient (naproxen, NPX) is incorporated. The use of PEGDM as PG enhances the mechanical stiffness of the hybrid gel (G' increases from 400 to 4500 Pa) - the LMWG enhances its stability to very high frequency. Use of DBS-CONHNH2 as LMWG enables interactions with NPX and hence allows pH-mediated NPX release - the PG network is largely orthogonal and only interferes to a limited extent. Use of photo-activated PEGDM as PG enables spatially-resolved photo-patterning of robust hybrid gel domains within a preformed LMWG network - the presence of the LMWG enhances the spatial resolution. The photo-patterned multi-domain gel retains pH-mediated NPX release properties and directionally releases NPX into a compartment of higher pH. The two components within these hybrid PG/LMWG hydrogels therefore act largely independently of one another, although they do modify each others properties in subtle ways. Hybrid hydrogels capable of spatially controlled unidirectional release have potential applications in tissue engineering and drug-delivery.

Epithelial tethering of MUC5AC-rich mucus impairs mucociliary transport in asthma.

The development of pathologic mucus, which is not readily cleared from the airways, is an important contributor to the morbidity and mortality associated with asthma. It is not clear how the major airway mucins MUC5AC and MUC5B are organized within the mucus gel or how this gel contributes to airway obstruction in asthma. Here, we demonstrated that mucus plugs from individuals with fatal asthma are heterogeneous gels with distinct MUC5AC- and MUC5B-containing domains. Stimulation of cultured human bronchial epithelial cells with IL-13, a key mediator in asthma, induced the formation of heterogeneous mucus gels and dramatically impaired mucociliary transport. Impaired transport was not associated with defects in ciliary function but instead was related to tethering of MUC5AC-containing mucus gel domains to mucus-producing cells in the epithelium. Replacement of tethered mucus with untethered mucus restored mucociliary transport. Together, our results indicate that tethering of MUC5AC-containing domains to the epithelium causes mucostasis and likely represents a major cause of mucus plugging in asthma.