Cell-inspired, massive electromodulation of friction via transmembrane fields across lipid bilayers.

Transient electric fields across cell bilayer membranes can lead to electroporation and cell fusion, effects crucial to cell viability whose biological implications have been extensively studied. However, little is known about these behaviours in a materials context. Here we find that transmembrane electric fields can lead to a massive, reversible modulation of the sliding friction between surfaces coated with lipid-bilayer membranes-a 200-fold variation, up to two orders of magnitude greater than that achieved to date. Atomistic simulations reveal that the transverse fields, resembling those at cell membranes, lead to fully reversible electroporation of the confined bilayers and the formation of inter-bilayer bridges analogous to the stalks preceding intermembrane fusion. These increase the interfacial dissipation through reduced hydration at the slip plane, forcing it to revert in part from the low-dissipation, hydrated lipid-headgroup plane to the intra-bilayer, high-dissipation acyl tail interface. Our results demonstrate that lipid bilayers under transmembrane electric fields can have striking materials modification properties.

Direct measurement of the viscoelectric effect in water.

The viscoelectric effect concerns the increase in viscosity of a polar liquid in an electric field due to its interaction with the dipolar molecules and was first determined for polar organic liquids more than 80 y ago. For the case of water, however, the most common polar liquid, direct measurement of the viscoelectric effect is challenging and has not to date been carried out, despite its importance in a wide range of electrokinetic and flow effects. In consequence, estimates of its magnitude for water vary by more than three orders of magnitude. Here, we measure the viscoelectric effect in water directly using a surface force balance by measuring the dynamic approach of two molecularly smooth surfaces with a controlled, uniform electric field between them across highly purified water. As the water is squeezed out of the gap between the approaching surfaces, viscous damping dominates the approach dynamics; this is modulated by the viscoelectric effect under the uniform transverse electric field across the water, enabling its magnitude to be directly determined as a function of the field. We measured a value for this magnitude, which differs by one and by two orders of magnitude, respectively, from its highest and lowest previously estimated values.

Geometric tumor embolic budding characterizes inflammatory breast cancer.

PURPOSE: Inflammatory breast cancer (IBC) is characterized by numerous tumor emboli especially within dermal lymphatics. The explanation remains a mystery. METHODS: This study combines experimental studies with two different IBC xenografts with image algorithmic studies utilizing human tissue microarrays (TMAs) of IBC vs non-IBC cases to support a novel hypothesis to explain IBC's sina qua non signature of florid lymphovascular emboli. RESULTS: In the human TMAs, compared to tumor features like nuclear grade (size), mitosis and Ki-67 immunoreactivity which show that IBC is only modestly more proliferative with larger nuclei than non-IBC, what really sets IBC apart is the markedly greater number of tumor emboli and distinctly smaller emboli whose numbers indicate geometric or exponential differences between IBC and non-IBC. In the experimental xenograft studies, Mary-X gives rise to tight spheroids in vitro which exhibit dynamic budding into smaller daughter spheroids whereas Karen-X exhibits only loose non-budding aggregates. Furthermore Mary-X emboli also bud dramatically into smaller daughter emboli in vivo. The mechanism that regulates this involves the generation of E-cad/NTF1, a calpain-mediated cleavage 100 kDa product of 120 kDa full length membrane E-cadherin. Inhibiting this calpain-mediated cleavage of E-cadherin by blocking either the calpain site of cleavage (SC) or the site of binding (SB) with specific decapeptides that both penetrate the cell membrane and mimic either the cleavage site or the binding site on E-cadherin, inhibits the generation of E-cad/NTF1 in a dose-dependent manner, reduces spheroid compactness and decreases budding. CONCLUSION: Since E-cad/NFT1 retains the p120ctn binding site but loses the α-and ß-catenin sites, promoting its 360° distribution around the cell's membrane, the vacilating levels of this molecule trigger budding of both the spheroids as well as the emboli. Recurrent and geometric budding of parental emboli into daughter emboli then would account for the plethora of emboli seen in IBC.

Patient-Reported Prevalence, Characteristics, and Impact of Leg Cramps in an Urban Primary Care Clinic.

This study addresses the prevalence and characteristics of leg cramps in 294 primary care patients (mean age = 46.5 years), with 51.7% reporting leg cramps. Patients who experience resting or exercise-induced leg cramps were more likely to be older and female. Cramp severity averaged 5.6 on a scale of 1-10 and disturbed sleep "sometimes" or "often" in 55% of patients. Most patients did not discuss cramps with their clinician. Our study reveals a possible shift in patients who experience leg cramps to younger age and chronicity. Resting leg cramps should be reviewed by clinicians as a symptom of declining health and advancing aging.

Effect of cholesterol on the mechanical stability of gel-phase phospholipid bilayers studied by AFM force spectroscopy.

The remarkably low sliding friction of articular cartilage in the major joints such as hips and knees, which is crucial for its homeostasis and joint health, has been attributed to lipid bilayers forming lubricious boundary layers at its surface. The robustness of such layers, and thus their lubrication efficiency at joint pressures, depends on the lipids forming them, including cholesterol which is a ubiquitous component, and which may act to strengthen of weaken the bilayer. In this work, a systematic study using an atomic force microscope (AFM) was carried out to understand the effect of cholesterol on the nanomechanical stability of two saturated phospholipids, DSPC (1,2-distearoyl-sn-glycero-3-phosphatidlycholine) and DPPC (1,2-dipalmitoyl-sn-glycero- phosphatidylcholine), that differ in acyl chain lengths. Measurements were carried out both in water and in phosphate buffer solution (PBS). The nanomechanical stability of the lipid bilayers was quantitatively evaluated by measuring the breakthrough force needed to puncture the bilayer by the AFM tip. The molar fractions of cholesterol incorporated in the bilayers were 10% and 40%. We found that for both DSPC and DPPC, cholesterol significantly decreases the mechanical stability of the bilayers in solid-ordered (SO) phase. In accordance with the literature, the strengthening effect of salt on the lipid bilayers was also observed. For DPPC with 10 mol % cholesterol, the effect of tip properties and the experimental procedure parameters on the breakthrough forces were also studied. Tip radius (2-42 nm), material (Si, Si3N4, Au) and loading rate (40-1000 nm/s) were varied systematically. The values of the breakthrough forces measured were not significantly affected by any of these parameters, showing that the weakening effect of cholesterol does not result from such changes in experimental conditions. As we have previously demonstrated that mechanical robustness improves the tribological performance of lipid layers, this study helps to shed light on the mechanism of physiological lubrication. Nanoindentation of SDPC bilayers.

Organohydrogels with High-Speed Lubrication by Confining Polymer Chain Mobility by an Interpenetrated Heteronetwork.

Hydrogels with pure hydrophilic network have received much attention due to their excellent low frictional behavior. However, the lubrication performance of hydrogels is not satisfied under high-speed condition due to the energy dissipation caused by adsorbed polymer chains as well as the failure of lubricating mechanisms accompanied by the transition of lubrication regime. In this work, interpenetrating double-network organohydrogels were constructed by combining hydrophilic and oleophilic polymer networks to modify the physiochemical properties of surface polymer chains, especially the chain mobility. The oleophilic polymer network spatially restricting the mobility of the swollen hydrophilic network in water, resulted in a low coefficient of friction (ca. 0.01) compared with conventional hydrogels at high speed (0.1 m s-1 ). Meanwhile, the organohydrogels had superior wear resistance, with almost no wear observed on the sliding track after 5 k cycles of rubbing at high speed. The design concept of organohydrogels can be extended to a variety of low-wear, highly-lubricating materials.

Lipid-Bilayer Assemblies on Polymer-Bearing Surfaces: The Nature of the Slip Plane in Asymmetric Boundary Lubrication.

Phospholipid-macromolecule complexes have been proposed to form highly efficient, lubricating boundary layers at artificial soft surfaces or at biological surfaces such as articular cartilage, where the friction reduction is attributed to the hydration lubrication mechanism acting at the exposed, hydrated head groups of the lipids. Here we measure, using a surface force balance, the normal and frictional interactions between model mica substrates across several different configurations of phosphatidylcholine (PC) lipid aggregates and adsorbed polymer (PEO) layers, to provide insight into the nature of such lubricating boundary layers in both symmetric and especially asymmetric configurations. Our results reveal that, irrespective of the configuration, the slip plane between the sliding surfaces reverts wherever possible to a bilayer-bilayer interface where hydration lubrication reduces the friction strongly. Where such an interface is not available, the sliding friction remains high. These findings may account for the low friction observed between both biological and synthetic hydrogel surfaces which may be asymmetrically coated with lipid-based boundary layers and fully support the hydration lubrication mechanism attributed to act at such boundary layers.

Effects of Hyaluronan Molecular Weight on the Lubrication of Cartilage-Emulating Boundary Layers.

Osteoarthritic joints contain lower-molecular-weight (MW) hyaluronan (hyaluronic acid, HA) than healthy joints. To understand the relevance of this HA size effect for joint lubrication, the friction and surface structure of cartilage-emulating surfaces with HA of different MWs were studied using a surface force balance (SFB) and atomic force microscopy (AFM). Gelatin (gel)-covered mica surfaces were coated with high-MW HA (HHA), medium-MW HA (MHA), or low-MW HA (LHA), and lipids of hydrogenated soy l-α-phosphatidylcholine (HSPC) in the form of small unilamellar vesicles, using a layer-by-layer assembly method. SFB results indicate that the gel-HHA-HSPC boundary layer provides very efficient lubrication, attributed to hydration lubrication at the phosphocholine headgroups exposed by the HA-attached lipids, with friction coefficients (COF) as low as 10-3-10-4 at contact stresses at least up to P = 120 atm. However, for the gel-MHA-HSPC and gel-LHA-HSPC surfaces, the friction, initially low, increases sharply at much lower pressures (up to 30-60 atm at most). This higher friction with the shorter chains may be due to their weaker total adhesion energy to the gelatin, where the attraction between the negatively charged HA and the weakly positively charged gelatin is attributed largely to counterion-release entropy. Thus, the complexes of LHA and MHA with the lubricating HSPC lipids are more easily removed by shear during sliding, especially at high stresses, than the HHA-HSPC complex, which is strongly adhered to gelatin. This is ultimately the reason for lower-pressure lubrication breakdown with the shorter polysaccharides. Our results provide molecular-level insight into why the decrease in HA molecular weight in osteoarthritic joints may be associated with higher friction at the articular cartilage surface, and may have relevance for treatments of osteoarthritis involving intra-articular HA injections.

Normal and shear forces between boundary sphingomyelin layers under aqueous conditions.

Sphingomyelin is one of the predominant phospholipid groups in synovial joints, where lipids have been strongly implicated in the boundary lubrication of articular cartilage; however, little attention has been paid to its lubrication behavior. In this study, we demonstrate that sphingomyelin is an excellent boundary lubricant by measuring the normal and shear forces between sphingomyelin-layer-coated surfaces with a surface force balance under aqueous conditions. Slightly negatively-charged egg sphingomyelin vesicles were adsorbed on mica either by calcium bridging or by charge screening with high concentration monovalent salt. The normal force profiles between opposing egg sphingomyelin layers (vesicles or bilayers) show long-ranged weak repulsion and short-ranged strong repulsion on approaching. Friction coefficients, calculated from the highest load, were (7.2 ± 1.7) × 10-4 at contact stresses of 9.1 ± 0.7 MPa across 0.3 mM liposome dispersion in 0.03 mM Ca2+, and (0.8-3.5) × 10-3 at contact stresses of 7.6 ± 0.8 MPa across 0.3 mM liposome dispersion in 150 mM NaNO3. Similar or slightly lower friction coefficients of (5.3 ± 0.8) × 10-4 at 9.8 ± 0.2 MPa were obtained by replacing the liposome dispersion in 0.03 mM Ca2+ by water. Such low friction coefficients, attributed to the hydration lubrication mechanism, are comparable to those of phosphatidylcholine lipids, which have been widely recognized as excellent aqueous biolubricants. Therefore, we believe that sphingomyelin, in parallel with phosphatidylcholine, contributes to the remarkably good boundary lubrication in synovial joints.

Surface Interactions between Boundary Layers of Poly(ethylene oxide)-Liposome Complexes: Lubrication, Bridging, and Selective Ligation †.

Poly(ethylene oxide), PEO, is widely exploited in biomedical applications, while phosphatidylcholine (PC) lipids (in the form of bilayers or liposomes) have been identified as very efficient boundary lubricants in aqueous media. Here we examine, using a surface force balance (SFB), the interactions between surface-adsorbed layers of PEO complexed with small unilamellar vesicles (SUVs, i.e. liposomes) or with bilayers of PC lipids, both well below and a little above their main gel-to-liquid phase-transition temperatures TM. The morphology of PEO layers (adsorbed onto mica), to which liposomes were added, was examined using atomic force microscopy (AFM) and cryo-scanning electron microscopy (cryo-SEM). Our results reveal that the PC lipids could attach to the PEO either as vesicles or as bilayers, depending on whether they were above or below TM. Under water (no added salt), excellent lubrication, with friction coefficients down to 10-3-10-4, up to contact stresses of 6.5 MPa (comparable to those in the major joints) was observed between two surfaces bearing such PEO-PC complexes. At 0.1 M KNO3 salt concentration (comparable to physiological salt levels), the friction between such surfaces was considerably higher, attributed to bridging by the polymer chains. Remarkably, such bridging could be suppressed and the friction could be restored to its previous low value if the KNO3 was replaced with NaNO3, as a result of the different PEO-mica ligation properties of Na+ compared to those of K+. Our results provide insight into the properties of PEO-PC complexes in potential applications, and large interfacial effects that can result from the seemingly innocuous replacement of K+ by Na+ ions.

Boundary Lubrication, Hemifusion, and Self-Healing of Binary Saturated and Monounsaturated Phosphatidylcholine Mixtures ⧫.

A wide range of phosphatidylcholine (PC) lipids with different degrees of unsaturation has been identified in the human synovial fluid and on the cartilage surface. The outstanding lubricity of the articular cartilage surface has been attributed to boundary layers comprising complexes of such lipids, though to date, only lubrication by single-component PC-lipid-based boundary layers has been investigated. As distinguishable lubrication behavior has been found to be related to the PC structures, we herein examined the surface morphology (on mica) and the lubrication ability of binary PC lipid mixtures, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), using atomic force microscopy (AFM) and a surface force balance (SFB). These two PC lipids are among the most abundant saturated and unsaturated PC components in synovial joints. Small unilamellar vesicles (SUVs) prepared from DPPC-POPC mixtures (8:2, 5:5, and 2:8, molar ratios) ruptured and formed bilayers on mica. The normal and shear forces between two DPPC-POPC bilayer-coated mica surfaces across the corresponding SUV dispersions show good boundary lubrication (friction coefficients ≤ ca. 10-4) up to contact stresses of 8.3 ± 2.2 MPa for 8:2 DPPC-POPC and 5.0 ± 1.7 MPa for the others. Hemifusion induced at high normal pressures was observed, probably because of the height mismatch of two components. Reproducible successive approaches after hemifusion indicate rapid self-healing of the mica-supported bilayers in the presence of the SUVs reservoir. This work is a first step to provide insight concerning the lubrication, wear, and healing of the PC-based boundary layers, which must consist of multicomponent lipid mixtures, on the articular cartilage surface.

Poly-phosphocholinated Liposomes Form Stable Superlubrication Vectors.

We have prepared phosphatidylcholine (PC) vesicles (liposomes) incorporating a novel lipid/poly-phosphocholine conjugate. This both stabilizes the liposomes against aggregation (for example, during storage or when being delivered) and allows them to act as very efficient lubricating elements readily attaining superlubric performance (defined as coefficient of friction µ < 10-2) via hydration lubrication at physiological salt concentrations and pressures. In contrast, vesicles sterically protected by poly(ethylene glycol) chains (PEGylation), which is the general method of choice, while being equally stable to aggregation are much poorer lubricants under these conditions, which is attributed to the relatively poor hydration of the PEG. Our approach enables the use of PC liposomes as stable superlubrication vectors in potential biomedical applications.

Anomalous viscosity-time behavior of polysaccharide dispersions.

Using viscosity and dynamic light scattering (DLS) measurements, we monitored the changes in the properties of dispersions of chitosan (a cationic polysaccharide) in acidic solution over a period of up to 700 h. Different polymer concentrations, weight average molecular weights, and degrees of deacetylation were examined. We found that the solution rheology and chitosan aggregates continue to change even up to 700 h. It was observed, remarkably, using both capillary and cone and plate viscometry that the viscosity decreased significantly during the storage period of the chitosan dispersions, with a rapid initial decrease and a slow approach to the steady state value. DLS measurements over this period could be interpreted in terms of a gradual decrease in the size of the chitosan aggregates in the dispersion. This behavior is puzzling, insofar as one expects the dissolution of compact polymer aggregates with time into individual polymer chains to increase the viscosity rather than decrease it as observed: We attribute this apparently anomalous behavior to the fact that the chitosan aggregates are rigid crystalline rod-like entities, which dissolved with time from dispersion of overlapping rods (with high viscosity) into solution of individual random coils (with lower viscosity). A detailed model comparing the hydrodynamic behavior of the initial overlapping rod-like aggregates with the subsequent free coils in solution is in semi-quantitative agreement with our observation.

On the question of whether lubricants fluidize in stick-slip friction.

Intermittent sliding (stick-slip motion) between solids is commonplace (e.g., squeaking hinges), even in the presence of lubricants, and is believed to occur by shear-induced fluidization of the lubricant film (slip), followed by its resolidification (stick). Using a surface force balance, we measure how the thickness of molecularly thin, model lubricant films (octamethylcyclotetrasiloxane) varies in stick-slip sliding between atomically smooth surfaces during the fleeting (ca. 20 ms) individual slip events. Shear fluidization of a film of five to six molecular layers during an individual slip event should result in film dilation of 0.4-0.5 nm, but our results show that, within our resolution of ca. 0.1 nm, slip of the surfaces is not correlated with any dilation of the intersurface gap. This reveals that, unlike what is commonly supposed, slip does not occur by such shear melting, and indicates that other mechanisms, such as intralayer slip within the lubricant film, or at its interface with the confining surfaces, may be the dominant dissipation modes.

Lubrication of Articular Cartilage.

The major synovial joints such as hips and knees are uniquely efficient tribological systems, able to articulate over a wide range of shear rates with a friction coefficient between the sliding cartilage surfaces as low as 0.001 up to pressures of more than 100 atm. No human-made material can match this. The means by which such surfaces maintain their very low friction has been intensively studied for decades and has been attributed to fluid-film and boundary lubrication. Here, we focus especially on the latter: the reduction of friction by molecular layers at the sliding cartilage surfaces. In particular, we discuss such lubrication in the light of very recent advances in our understanding of boundary effects in aqueous media based on the paradigms of hydration lubrication and of the synergism between different molecular components of the synovial joints (namely hyaluronan, lubricin, and phospholipids) in enabling this lubrication.

Effect of Cholesterol on the Stability and Lubrication Efficiency of Phosphatidylcholine Surface Layers.

The lubrication properties of saturated PC lipid vesicles containing high cholesterol content under high loads were examined by detailed surface force balance measurements of normal and shear forces between two surface-attached lipid layers. Forces between two opposing mica surfaces bearing distearoylphosphatidylcholine (PC) (DSPC) small unilamellar vesicles (SUVs, or liposomes), or bilayers, with varying cholesterol content were measured across water, whereas dimyristoyl PC (DMPC), dipalmitoyl PC (DPPC), and DSPC SUVs containing 40% cholesterol were measured across liposome dispersions of SUVs of the same lipid composition as in the adsorbed layers. The results clearly demonstrate decreased stability and resistance to normal load with the increase in cholesterol content of DSPC SUVs. Friction coefficients between two 10% cholesterol PC-bilayers were in the same range as for 40% cholesterol bilayers (µ ≈ 10-3), indicating that cholesterol has a more substantial effect on the mechanical properties of a bilayer than on its lubrication performance. We further find that the lubrication efficiency of DMPC and DPPC with 40% cholesterol is superior to that of DSPC 40% cholesterol, most likely because of enhanced hydration-lubrication in these systems. We previously found that when experiments are performed in the presence of a lipid reservoir, layers can self-heal and therefore their robustness is less important under such conditions. We conclude that the effect of cholesterol in decreasing the stability is more pronounced than its effect on hydration, but the stability is, in turn, less important when a lipid reservoir is present. This study complements our previous work and sheds light on the effect of cholesterol, a prominent and important physiological lipid, on the mechanical and lubrication properties of gel-phase lipid layers.

Modifying surface forces through control of surface potentials.

Combining direct surface force measurements with in situ regulation of surface potential provides an exceptional opportunity for investigating and manipulating interfacial phenomena. Recently, we studied the interaction between gold and mica surfaces in water with no added salt, while controlling the metal potential, and found that the surface charge at the metal may vary, and possibly even change its sign, as it progressively approaches the (constant-charge) mica surface [Langmuir, 2015, 31(47), 12845-12849]. Such a variation was found to directly affect the nature of the contact and adhesion between them due to exclusion of all mobile counterions from the intersurface gap. In this work, we extend this to examine the potential-dependent response of the adhesion and interaction between gold and mica to externally applied voltages and in electrolyte solution. Using a surface force balance (SFB) combined with a three-electrode electrochemical cell, we measured the normal interaction between gold and mica under surface potential regulation, revealing three interaction regimes - pure attraction, non-monotonic interaction from electrostatic repulsion to attraction (owing to charge inversion) and pure repulsion. Accordingly, the adhesion energy between the surfaces was found to vary both in no added salt water and, more strongly, in electrolyte solution. We justify this potential-dependent variation of adhesion energy in terms of the interplay between electrostatic energy and van der Waals (vdW) interaction at contact, and attribute the difference between the two cases to the weaker vdW interaction in electrolyte solution. Finally, we showed that through abruptly altering the gold surface potential from negative to positive and vice versa, the adhesion between gold and mica can be reversibly switched on and off. We surmise that the process of bringing the surface into contact is associated with the formation of a strong electric field O (108 V m-1) in the intersurface gap.

Probing the Surface Properties of Gold at Low Electrolyte Concentration.

Using the surface force balance (SFB), we studied the surface properties of gold in aqueous solution with low electrolyte concentration (∼10(-5) M and pH = 5.8), i.e., water with no added salt, by directly measuring the interaction between an ultrasmooth gold surface (ca. 0.2 nm rms roughness) and a mica surface. Under these conditions, specific adsorption of ions is minimized and its influence on the surface charge and surface potential of gold is markedly reduced. At open circuit potential, the electrostatic interaction between gold and mica was purely attractive and gold was found to be positively charged. This was further confirmed by force measurements against a positively charged surface, poly-l-lysine coated mica. Successive force measurements unambiguously showed that once gold and mica reach contact all counterions are expelled from the gap, confirming that at contact the surface charge of gold is equal and opposite in charge to that of mica. Further analysis of adhesion energy between the surfaces indicated that adhesion is mostly governed by vdW dispersion force and to a lesser extent by electrostatic interaction. Force measurements under external applied potentials showed that the gold-mica interaction can be regulated as a function of applied potential even at low electrolyte concentration. The gold-mica interaction was described very precisely by the nonlinearized Poisson-Boltzmann (PB) equation, where one of the surfaces is at constant charge, i.e., mica, and the other, i.e., gold, is at constant potential. Consequently, the gold surface potential could be determined accurately both at open circuit potential (OCP) and under different applied potentials. Using the obtained surface potentials, we were able to derive fundamental characteristics of the gold surface, e.g., its surface charge density and potential of zero charge (PZC), at very low electrolyte concentration.