A Quantitative Approach to Characterize the Surface Modification on Nanoparticles Based on Localized Dielectric Environments.

Nanoparticles (NPs) are utilized for the functionalization of composite materials and nanofluids. Although oxide NPs (e.g., silica (SiO2)) exhibit less dispersibility in organic solvents or polymers due to their hydrophilic surface, the surface modification using silane coupling agents can improve their dispersibility in media with low dielectric constants. Herein, SiO2 NPs were functionalized using octyltriethoxysilane (OTES, C8) and dodecyltriethoxysilane (DTES, C12), wherein the degrees of surface modification of SiO2@C8 and SiO2@C12 were quantitatively evaluated based on the ratio of modifier to surface silanol group (θ) and the volume fraction of organic modifier to total particle volume (ϕR). The variations of surface properties were revealed by analyzing the Hansen solubility parameters (HSP). Particularly, the surface modification using OTES or DTES significantly affected the polarity (δP) of NPs. The local dielectric environments of surface-modified SiO2 NPs were characterized using a solvatochromic dye, Laurdan. By analyzing the peak position of the steady-state emission spectrum of Laurdan in a NP suspension, the apparent dielectric environments surrounding NPs (εapp) were obtained. A good correlation between ϕR and εapp was observed, indicating that ϕR is a reliable quantity for understanding the properties of surface-modified NPs. Furthermore, the generalized polarization (GP) of NPs was investigated. The surface-modified SiO2 NPs with higher ϕR (≥0.15) exhibited GP > 0, suggesting that the modifiers are well-organized on the surface of NPs. The localized dielectric environment surrounding NPs could be predicted by analyzing the volume fraction of nonpolar moieties derived from modifiers. Alternatively, εapp and GP can be utilized for understanding the properties of inorganic-organic hybrid NPs.

Surface lattice resonance in three-dimensional plasmonic arrays fabricated via self-assembly of silica-coated gold nanoparticles.

HYPOTHESIS: Three-dimensional plasmonic nanoparticle arrays in which the nanoparticles are assembled with a certain distance apart are expected to exhibit unique optical properties attributed to surface lattice resonances because of the interactions between the nanoparticle layers. EXPERIMENTS: Multi-layered gold nanoparticle arrays were created to experimentally prove surface lattice resonances from three-dimensional arrays. Silica-coated gold nanoparticles were employed as building blocks for the array because the distance between the nanoparticles can be tuned by adjusting the thickness of the silica coating. Employing highly monodisperse building blocks enabled to fabricate both single-layered and multi-layered plasmonic arrays via a confined convective assembly method. FINDINGS: Multi-layering of monodisperse building blocks brought about some additional peaks corresponded to Bragg diffraction of gold nanoparticle periodic array and the interactions between layers in a hexagonal close-packed structure of the nanoparticles, respectively. Most importantly, the multi-layered arrays exhibited a distinctive extinction peak at the same wavelength as that observed from the single-layered array, proving the realization of surface lattice resonances from the three-dimensional plasmonic array.

Fluorescence Spectroscopic Analysis of Lateral and Transbilayer Fluidity of Exosome Membranes.

Exosomes are small extracellular vesicles (sEVs) involved in distal cell-cell communication and cancer migration by transferring functional cargo molecules. Membrane domains similar to lipid rafts are assumed to occur in exosome membranes and are involved in interactions with target cells. However, the bilayer membrane properties of these small vesicles have not been fully investigated. Therefore, we examined the fluidity, lateral domain separation, and transbilayer asymmetry of exosome membranes using fluorescence spectroscopy. Although there were some differences between the exosomes, TMA-DPH anisotropy showing moderate lipid chain order indicated that ordered phases comprised a significant proportion of exosome membranes. Selective TEMPO quenching of the TMA-DPH fluorescence in the liquid-disordered phase indicated that 40-50% of the exosome membrane area belonged to the ordered phase based on a phase-separated model. Furthermore, NBD-PC in the outer leaflet showed longer fluorescence lifetimes than those in the inner leaflets. Therefore, the exosome membranes maintained transbilayer asymmetry with a topology similar to that of the plasma membranes. In addition, the lateral and transbilayer orders of exosome membranes obtained from different cell lines varied, probably depending on the different membrane lipid components and compositions partially derived from donor cells. As these higher membrane orders and asymmetric topologies are similar to those of cell membranes with lipid rafts, raft-like functional domains are possibly enriched on exosome membranes. These domains likely play key roles in the biological functions and cellular uptake of exosomes by facilitating selective membrane interactions with target organs.

Preferential adsorption of L-tryptophan by L-phospholipid coated porous polymer particles.

Chiral selective adsorption of L-amino acid, tryptophan (Trp) was achieved using phospholipid membrane-coated porous polymer particles (PPPs). PPPs with numerous pores were prepared by in situ polymerization of divinylbenzene, and then coated with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC, L-phospholipid) via the impregnation method. Elemental mapping of energy dispersive X-ray (EDX) analysis revealed that DPPC molecules were distributed to the surface and the inner part of PPPs, where almost all the DPPC molecules applied for impregnation were deposited on PPPs. The phospholipid membrane properties of DPPC-PPPs were characterized using the fluorescence probe 6-lauroyl-2-dimethylaminonaphthalene (Laurdan). The results show that DPPC-PPPs possessed a lipid membrane-like environment similar to pure DPPC liposomes, especially at temperatures below 35 °C. DPPC-PPPs slightly adsorbed L-Trp and D-Trp at 45 °C, while DPPC-PPPs significantly adsorbed L-Trp but not D-Trp at 30 °C: enantio excess (e.e.) was 75.0%. The time course of Trp adsorption was investigated: for both enantiomers, similar adsorption behaviors were observed for 30 h, thus suggesting surface adsorption onto DPPC-PPPs. L-Trp adsorption continued after 30 h, suggesting that L-Trp could be distributed in the inner part of DPPC-PPPs. Interestingly, the reused DPPC-PPPs featured improved adsorption performance, suggesting that the deposited DPPC membranes on PPPs could act as chiral selectors for L-Trp. The optical resolution of L-/D-Trp was performed using DPPC-PPPs, resulting in the e.e. of D-Trp was > 60%. Thus, DPPC-PPPs have the potential of chiral selective adsorption of L-amino acid, which can be used as chiral separation materials.

Surface Characteristics of Antibacterial Polystyrene Nanoparticles Synthesized Using Cationic Initiator and Comonomers.

Polymer nanoparticles have attracted attention as antibacterial materials, but the function of the polymer itself has not yet been clarified sufficiently. To estimate the essential surface properties of antibacterial polymer nanoparticles, herein, we synthesized cationic polystyrene (PSt) nanoparticles via soap-free emulsion polymerization using 2,2'-azobis-[2-(1,3-dimethyl-4,5-dihydro-1H-imidazol-3-ium-2-yl)]propane triflate (ADIP) as initiator. The conversion of total monomers was drastically increased through the addition of the commoner (vinylbenzyl)trimethylammonium chloride (VBTMAC), where unimodal size distributions (Cv ≤ 10%) were obtained at comonomer molar ratios between 0.0083 and 0.0323. The adsorption behavior of a solvatochromic anionic fluorescent dye revealed the surface charge density (σ) and affinity with anionic molecules (K) of PSt nanoparticles. The PSt nanoparticles with increased K values exhibited antibacterial activity against Staphylococcus epidermidis, with a minimum inhibitory concentration of at least 0.69 mg/mL. To determine a plausible mechanism for the antibacterial activity, the membrane damage induced by PSt nanoparticles was evaluated using an assay utilizing polydiacetylene vesicles as the model for negatively charged bilayer membranes. The PSt nanoparticles exhibiting large K values disturbed the bilayer structure of the model membrane system, suggesting that the synthesized PSt nanoparticles could be utilized as a contact-killing antibacterial agent.

Insight into the Exosomal Membrane: From Viewpoints of Membrane Fluidity and Polarity.

Numerous research studies have been done for exosomes, particularly focusing on membrane proteins and included nucleic acids, and the volume of the knowledge about the lipids in the exosomal membrane has been increasing. However, the dynamic property of the exosomal membrane is hardly studied. By employing milk exosome as an example, herein the exosomal membrane was characterized focusing on the membrane fluidity and polarity. The lipid composition and phase state of milk exosome (exosome from bovine milk) were estimated. The milk exosome contained enriched Chol (43.6 mol % in total lipid extracts), which made the membrane in the liquid-ordered (lo) phase by interacting with phospholipids. To suggest a model of exosomal vesicle cargo, the liposome compositions that mimic milk exosome were studied: liposomes were made of cholesterol (Chol), milk sphingomyelin (milk SM), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). By using fluorescent probes 1,6-diphenyl-1,3,5-hexatriene and 6-dodecanoyl-2-dimethylaminonaphthalene, the microenvironments of submicron-sized membranes of exosome and model liposomes were investigated. The membrane fluidity of milk exosome was slightly higher than those of Chol/milk SM/POPC liposomes with a similar content of Chol, suggesting the presence of enriched unsaturated lipids. The most purposeful membrane property was obtained by the liposome composition of Chol/milk SM/POPC = 40/15/45. From the above, it is concluded that Chol is a fundamental component of the milk exosomal membrane to construct the enriched lo phase, which could increase the membrane rigidity and contribute to the function of exosome.

Correlation of Secondary Particle Number with the Debye-Hückel Parameter for Thickening Mesoporous Silica Shells Formed on Spherical Cores.

Mesoporous silica shells were formed on nonporous spherical silica cores during the sol-gel reaction to elucidate the mechanism for the generation of secondary particles that disturb the efficient growth of mesoporous shells on the cores. Sodium bromide (NaBr) was used as a typical electrolyte for the sol-gel reaction to increase the ionic strength of the reactant solution, which effectively suppressed the generation of secondary particles during the reaction wherein a uniform mesoporous shell was formed on the spherical core. The number of secondary particles (N 2nd) generated at an ethanol/water weight ratio of 0.53 was plotted against the Debye-Hückel parameter κ to quantitatively understand the Debye screening effect on secondary particle generation. Parameter κa, where a is the average radius of the secondary particles finally obtained in the silica coating, expresses the trend in N 2nd at different concentrations of ammonia and NaBr. N 2nd was much lower than that expected theoretically from the variation of secondary particle sizes at a constant Debye-Hückel parameter. A similar correlation with κa was observed at the high and low ethanol/water weight ratios of 0.63 and 0.53, respectively, with different hydrolysis rate constants. The good correlation between N 2nd and κa revealed that controlling the ionic strength of the silica coating is an effective approach to suppress the generation of secondary particles for designing mesoporous shells with thicknesses appropriate for their application as high-performance liquid chromatography column packing materials.

Modulation of the Belousov-Zhabotinsky Reaction with Lipid Bilayers: Effects of Lipid Head Groups and Membrane Properties.

The Belousov-Zhabotinsky (BZ) reaction is an oscillating reaction due to periodic oscillations that happen in the concentration of some intermediates. Such systems can be applied together with hydrophobic membranes to create an autonomous behavior in artificial systems. However, because of a complex set of reactions happening in such systems, the interferences caused by hydrophobic membranes are not easily understood. In this study, we tested lipid membranes composed of trimethylammonium-propane (TAP) and phosphate (PA) lipids in an attempt to break down how the polar region of phosphatidylcholine (PC) lipid membranes affect the BZ reaction. According to our findings, the trimethylammonium group and membrane fluidity are crucial to change the frequency of oscillations in the reaction. In addition, the results also indicate a possible complexation of cerium ions with membranes with a phosphate head group.

Systematic Characterization of Nanostructured Lipid Carriers from Cetyl Palmitate/Caprylic Triglyceride/Tween 80 Mixtures in an Aqueous Environment.

Nanostructured lipid carriers (NLCs) are gaining attention as the new generation of lipid vehicles. These carriers consist of saturated lipids with small drops of liquid oil dispersed into the inner lipid matrix and are stabilized by a surfactant. Conventionally, NLC-based drug delivery systems have been widely studied, and many researchers are looking into the composition of NLC properties to improve the performance of NLCs. The membrane fluidity and polarity of self-assembling lipids are also essential properties that must be affected by membrane compositions; however, such fundamental characteristics have not been studied yet. In this study, NLCs were prepared from cetyl palmitate (CP), caprylic triglyceride (CaTG), and Tween 80 (T80). Structural properties, such as particle size and ζ-potential of the CP/CaTG/T80 ternary mixtures, were investigated. Then, the systematic characterization of self-assembly properties using fluorescence-based analysis was applied for the first time to the NLC system. As a final step, the ternary diagram was developed based on the self-assembly properties to summarize the possible structures formed at different compositions. The results showed four states: micelle-like, oil-in-water (O/W) emulsion-like, solid lipid nanoparticle-like, and intermediate (solid-liquid coexistence). For the purpose of making the lipid matrix more liquified, the heterogeneous state and the disordered state of the O/W emulsion-like structure might fulfill the criteria of NLCs. Finally, the ternary diagram provides new information about the assembly state of NLC constituents that could become an important reference for developing high-performance NLCs.

Investigation of Quercetin interaction behaviors with lipid bilayers: Toward understanding its antioxidative effect within biomembrane.

Quercetin (QCT), existing in common dietary sources, is an abundant bioflavonoid with planar structure and exerts multiple pharmacological effects. Herein, four kinds of liposomes were prepared as model biomembranes, and then the partition coefficient, distribution in lipid membrane and influence of the QCT on the membrane properties were evaluated. The partition of QCT to lipid membranes was affected by both membrane phase state and the interference of QCT on membrane properties. The location of QCT in lipid membrane was related to the phase state of lipid membrane. In addition, influence of QCT on the compaction of the hydrocarbon tail in lipid membranes was dependent on the unsaturation degree of lipid molecules. Finally, about its antioxidant activity, from the results of 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay, it can be concluded that the interaction of QCT with lipid membrane greatly influences on QCT reductive activity in lipid membrane. Furthermore, mass spectrometry of DOPC molecule showed no lipid oxidation in the presence of QCT, indicating that in addition to the QCT ability toward radical scavenging, the ordering effect of QCT in unsaturated lipid membrane would be helpful to protect lipid membrane from oxidation by inhibiting radical diffusion (synergy effect). Based on lipid membrane analysis, our study made it clear that the effect of QCT on various lipid membrane and its relation with the antioxidant effect of QCT within lipid membrane. Therefore, our analytical method and findings would be also helpful for understanding the mechanism of other antioxidants effects on biomembrane.

Double-Inverse-Opal-Structured Particle Assembly as a Novel Immobilized Photocatalytic Material.

Immobilization of photocatalysts on supports is an important method of adding highly active photocatalysts to a continuous flowing system without the need for photocatalyst recovery. However, direct immobilization prevents exposure to all photocatalytically active surfaces. Therefore, to immobilize particulate photocatalysts, while exposing the photocatalytic surface to organic pollutant water in a continuous flowing system, in this study, we employed double-inverse-opal (DIO) with periodically arranged, interconnected macropores, each containing a single photocatalytic particle. Increasing the macropore size successfully enhanced the decomposition rate of organic dye due to the high diffusion rate of dye molecules in the macropores of thin DIOs. However, an excessive increase in macropore size lowered the decomposition rate of dye molecules because an increase in DIO thickness caused the attenuation of light used to excite the photocatalytic particles. This study presents novel, immobilized photocatalytic DIO-structured particles that can be employed in continuous flowing reaction systems by tuning the photocatalytic particle size, macropore size, and DIO thickness.

Changes Caused by Liposomes to the Belousov-Zhabotinsky Reaction.

The Belousov-Zhabotinsky (BZ) reaction has been applied to give autonomous dynamic behaviors to artificial systems. This reaction is conducted in an aqueous system, but it produces some hydrophobic intermediates, such as bromine. On the basis of previous works about reactions in the lipid bilayer, we investigated how liposome membranes (water-oil interface) affect the BZ reaction. Herein diacylglycerophosphocholine (PC) molecules with a variety of hydrocarbon tails were selected as components of liposomes, and the BZ reaction in the presence of the liposomes was characterized. As a result, membrane fluidity was the main characteristic leading to changes in the reaction behavior. The decrease of the frequency of oscillations was relevant to membrane fluidity, suggesting the interaction of bromine species in the hydrophobic site of the liposomes. In addition, the heterogeneous membrane (so+ld) of DMPC showed a fast decrease in the amplitude of oscillations. Conclusively, characteristics of the hydrophobic environment play a role in the reaction.

Evaluation of Molecular Ordering in Bicelle Bilayer Membranes Based on Induced Circular Dichroism Spectra.

Bicelles are submicrometer-sized disc-shaped molecular self-assemblies that can be obtained in aqueous solution by dispersing mixtures of certain amphiphiles. Although phospholipid bicelle and phospholipid vesicle assemblies adopt similar lipid bilayer structures, the differences in bilayer characteristics, especially physicochemical properties such as bilayer fluidity, are not clearly understood. Herein, we report the lipid ordering properties of bicelle bilayer membranes based on induced circular dichroism (ICD) and fluorescence polarization analyses using 1,6-diphenyl-1,3,5-hexatriene (DPH) as a probe. Bicelles were prepared by using 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC), while pure DMPC vesicles and pure DHPC micelles were used as references. At temperatures below the phase transition temperature of DMPC, the bicelles showed lower membrane fluidities, whereas DHPC micelles showed higher membrane fluidity, suggesting no significant differences in bilayer fluidity between the bicelle and vesicle assemblies. The ICD signals of DPH were induced only when the membrane was in ordered (solid-ordered or ripple-gel) phases. In the bicelle systems, the ICD of DPH was more significant than that of the DMPC vesicle. The induced chirality of DPH was dependent on the chirality of the bilayer lipid. Compared to that of the DMPC/DHPC bicelle, the ICD of the 1,2-dipalmitoyl-sn-glycero-3-phosphocholine/DHPC bicelle was higher, while that of the bovine sphingomyelin/DHPC bicelle was lower. Because the lipids are tightly packed in the ordered phase, the ICD intensity reflects the molecular ordering state of the lipids in the bicelle bilayer.

Enzymatic hydrolysis of cellulose recovered from ionic liquid-salt aqueous two-phase system.

Regenerated cellulose can be prepared by treatment with an ionic liquid (IL) and an anti-solvent such as water, which significantly enhances the enzymatic hydrolysis in comparison to crystalline cellulose. The IL-aqueous two-phase system (IL-ATPS) is consisted of IL-condensed top phase and salt-condensed bottom phase, which could be suitable to produce regenerated cellulose with smaller amount of IL. Using IL-ATPS with different pH, the enzymatic saccharification efficiency of crystalline cellulose was determined. The use of 1-allyl-3-methylimidazolium chloride resulted in relatively higher yield of glucose production as compared to 1-butyl-3-methylimidazolium chloride. The IL-ATPS showing optimal pH for cellulase was prepared with mixed salt (NaH2PO4/Na2HPO4 = 5/1 (wt/wt)), which provide a regenerated cellulose with the pH range of 4.8-4.9 in enzymatic reaction mixture. Using such regenerated cellulose as feed of saccharification, the final yield of glucose was about 70%.

Membrane Surface-Enhanced Raman Spectroscopy for Cholesterol-Modified Lipid Systems: Effect of Gold Nanoparticle Size.

A gold nanoparticle (AuNP) has a localized surface plasmon resonance peak depending on its size, which is often utilized for surface-enhanced Raman scattering (SERS). To obtain information on the cholesterol (Chol)-incorporated lipid membranes by SERS, AuNPs (5, 100 nm) were first functionalized by 1-octanethiol and then modified by lipids (AuNP@lipid). In membrane surface-enhanced Raman spectroscopy (MSERS), both signals from 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and Chol molecules were enhanced, depending on preparation conditions (size of AuNPs and lipid/AuNP ratio). The enhancement factors (EFs) were calculated to estimate the efficiency of AuNPs on Raman enhancement. The size of AuNP100nm@lipid was 152.0 ± 12.8 nm, which showed an surface enhancement Raman spectrum with an EF2850 value of 111 ± 9. The size of AuNP5nm@lipid prepared with a lipid/AuNP ratio of 1.38 × 104 (lipid molecule/particle) was 275.3 ± 20.2 nm, which showed the highest enhancement with an EF2850 value of 131 ± 21. On the basis of fluorescent probe analyses, the membrane fluidity and polarity of AuNP@lipid were almost similar to DOPC/Chol liposome, indicating an intact membrane of DOPC/Chol after modification with AuNPs. Finally, the membrane properties of AuNP@lipid systems were also discussed on the basis of the obtained MSERS signals.

Ergosterol-Induced Ordered Phase in Ternary Lipid Mixture Systems of Unsaturated and Saturated Phospholipid Membranes.

The sterol ergosterol (Erg) is ubiquitous in the membranes of lower eukaryotes such as fungi. To investigate the interactions between Erg and phosphocholine (PC) molecules, we studied ternary lipid mixture systems composed of unsaturated 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), saturated 1,2-dipalmitoyl-sn-glycero-3-phophocholine (DPPC), and Erg. Bilayer membrane fluidity and polarity were systematically analyzed using fluorescent probes. The presence of ≥30 mol % of Erg exhibited a significant ordering effect and stabilized membrane properties when temperatures increased, suggesting that Erg has a similar function to cholesterol (Chol) in comparable lipid systems that form a liquid-ordered phase. Erg was also observed to have a significant condensing effect at the monolayer level in saturated PC-enriched systems. The phase behavior of Erg in bilayer systems was compared with that of Chol, with the data suggesting that Erg behaves in a similar manner to Chol in membranes enriched with saturated lipids.

Melting-Temperature-Dependent Interactions of Ergosterol with Unsaturated and Saturated Lipids in Model Membranes.

Sterols such as cholesterol (Chol) and ergosterol (Erg) are known to regulate membrane properties in higher eukaryotes and in lower eukaryotes, respectively. To better understand the modulation of membrane properties by Erg, binary lipid membranes composed of Erg and diacylglycerophosphocholine (PC) were studied in Langmuir monolayer and bilayer vesicle systems. From the excess area measured by pressure-area isotherms, attractive interactions between Erg and saturated PC were significant above the melting temperature (Tm) of PC. Conversely, repulsive interactions were observed at temperatures below Tm. From the analyses of membrane fluidity and polarity using fluorescence probes, similar trends were observed for bilayer systems where Erg had an ordering effect on saturated PC vesicles in the fluid state. However, Chol had a stronger ordering effect than Erg. In unsaturated PC systems, Erg did not alter membrane ordering. These findings demonstrate that the interaction of Erg with the fluid-state PC lipids will maintain lower-eukaryote membranes in a more ordered state, similar to the effect of cholesterol in higher eukaryotes.

Characterization of Ionic Liquid Aqueous Two-Phase Systems: Phase Separation Behaviors and the Hydrophobicity Index between the Two Phases.

1-Allyl-3-methylimidazolium chloride [Amim][Cl] and 1-butyl-3-methylimidazolium chloride [Bmim][Cl] are water-soluble ionic liquids (ILs) that can from an aqueous two-phase system (ATPS) when mixed with specific salts. Herein, we prepared [Amim][Cl]- and [Bmim][Cl]-ATPSs by adding the salts (K2CO3, K2HPO4). To investigate the phase separation behavior of the IL-ATPSs, binodal curves were drawn at different temperatures and the length and slope of the tie lines were analyzed. The [Bmim][Cl]/K2HPO4 system underwent two-phase separation at lower temperature conditions, suggesting that the phase separation might depend on the salting-out effect in the bottom phase. Using the IL-ATPS, the distribution coefficients, Kaa, of amino acids were determined and used to characterize the hydrophobicity index (HF) between the top and bottom phases, which is a good indicator to understand the molecular partitioning behaviors in conventional ATPSs. The HF values of the IL-ATPSs were in the range 0.13-0.41 mol/kJ; these values were almost the same as the HF values reported for an ATPS composed of poly(ethylene glycol) and salt.

Lipid-Surrounding Water Molecules Probed by Time-Resolved Emission Spectra of Laurdan.

The hydration states of the interfacial region of lipid bilayers were investigated on the basis of the time-resolved emission spectra (TRES) analysis of 6-lauroyl-2-dimethylamino naphthalene (Laurdan), a common fluorescence probe used to analyze membrane hydration. TRES derived from long and short lifetime components were extracted from samples of different lipid species: 1,2-dipalmitoyl- sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl- sn-glycero-3-phosphocholine (DOPC), d- erythro- N-palmitoyl-sphingosylphosphorylcholine (PSM), and a DOPC/PSM binary bilayer system. Neither lifetime component (short or long) corresponded with the hydration properties; the short lifetime component of DOPC (1.97 ns) exhibited a peak at 440 nm, and the long lifetime components of DPPC and PSM (7.76 and 7.77 ns, respectively) exhibited peaks at the same wavelength. This similarity arose from the competition between the collisional quenching and the hydration effects of water molecules. Herein, this phenomenon was investigated using a plot of the lifetime τ and the peak position λ (τ vs λ plot), simultaneously visualizing both effects by deconvoluting the TRES. On the basis of collisional quenching theory, the distribution of the water population per lipid (water map) was generated. According to this theory, the τ vs λ plot was applied to the water map and the calculation of the number of water molecules per lipid, which is consistent with previous reports. This approach provides novel insights for the analysis of molecular hydration states using the fluorescence of Laurdan.

Solvatochromic Modeling of Laurdan for Multiple Polarity Analysis of Dihydrosphingomyelin Bilayer.

The hydration properties of the interface between lipid bilayers and bulk water are important for determining membrane characteristics. Here, the emission properties of a solvent-sensitive fluorescence probe, 6-lauroyl-2-dimethylamino naphthalene (Laurdan), were evaluated in lipid bilayer systems composed of the sphingolipids D-erythro-N-palmitoyl-sphingosylphosphorylcholine (PSM) and D-erythro-N-palmitoyl-dihydrosphingomyelin (DHPSM). The glycerophospholipids 1-palmitoyl-2-palmitoyl-sn-glycero-3-phosphocholine and 1-oleoyl-2-oleoyl-sn-glycero-3-phosphocholine were used as controls. The fluorescence properties of Laurdan in sphingolipid bilayers indicated multiple excited states according to the results obtained from the emission spectra, fluorescence anisotropy, and the center-of-mass spectra during the decay time. Deconvolution of the Laurdan emission spectra into four components based on the solvent model enabled us to identify the varieties of hydration and the configurational states derived from intermolecular hydrogen bonding in sphingolipids. Sphingolipids showed specific, interfacial hydration properties stemming from their intra- and intermolecular hydrogen bonds. Particularly, the Laurdan in DHPSM revealed more hydrated properties compared to PSM, even though DHPSM has a higher Tm than PSM. Because DHPSM forms hydrogen bonds with water molecules (in 2NH configurational functional groups), the interfacial region of the DHPSM bilayer was expected to be in a highly polar environment. The careful analysis of Laurdan emission spectra through the four-component deconvolution in this study provides important insights for understanding the multiple polarity in the lipid membrane.