Thickness Of Lipid Film Research Articles

Optimization of Giant Unilamellar Vesicle Electroformation for Phosphatidylcholine/Sphingomyelin/Cholesterol Ternary Mixtures.

Artificial vesicles are important tools in membrane research because they enable studying membrane properties in controlled conditions. Giant unilamellar vesicles (GUVs) are specially interesting due to their similarity in size to eukaryotic cells. We focus on optimization of GUV production from phosphatidylcholine/sphingomyelin/cholesterol mixtures using the electroformation method. This mixture has been extensively researched lately due to its relevance for the formation of lipid rafts. We measured the effect of voltage, frequency, lipid film thickness, and cholesterol (Chol) concentration on electroformation successfulness using spin-coating for reproducible lipid film deposition. Special attention is given to the effect of Chol concentrations above the phospholipid bilayer saturation threshold. Such high concentrations are of interest to groups studying the role of Chol in the fiber cell plasma membranes of the eye lens or development of atherosclerosis. Utilizing atomic force and fluorescence microscopy, we found the optimal lipid film thickness to be around 30 nm, and the best frequency–voltage combinations in the range of 2–6 V and 10–100 Hz. Increasing the Chol content, we observed a decrease in GUV yield and size. However, the effect was much less pronounced when the optimal lipid film thickness was used. The results underline the need for simultaneous optimization of both electrical parameters and thickness in order to produce high-quality GUVs for experimental research.

Simultaneous monitoring of electroformation of phospholipid vesicles by quartz crystal microbalance and optical microscopy

The electroformation of giant vesicles from 1,2-Dimyristoyl-sn-Glycero-3-Phosphocholine (DMPC) was monitored using quartz crystal microbalance with dissipation monitoring (QCM-D) and optical microscopy, simultaneously using a novel sample cell design. A gold-coated QCM crystal was used as one of the electrodes and an Indium-tin-oxide (ITO)-coated glass slide was used as the second electrode for electroformation. Increases in the frequency and decreases in the dissipation were observed immediately upon voltage application between the two electrodes, indicating the loss of lipid from the QCM surface. Concurrently, we observed vesicles on the QCM electrode surface by differential interference contrast (DIC)-optical microscopy. The lipid-coated substrates were measured with AFM at various stages in the electroformation, and a significant change in the morphology of the lipid film was observed. Ellipsometry was used to find the average thickness of lipid film. The QCM data were fitted to a viscoelastic model to determine the viscoelastic properties and time dependence of the film thickness. All methods used to determine film thickness give values in reasonable quantitative agreement. Differences between the methods are consistent with what one might expect due to what is actually measured in the individual techniques. The comparison between mass loss and observed vesicles suggest that the vesicles formed are first localized to the substrate and then slowly released into the solution. By comparing the mass lost from the lipid film, to the total surface area of lipid vesicles observed, it is apparent that only a relatively small fraction of the lipid goes into the production of unilamellar vesicles with sizes detectable with optical microscopy.

Fabrication, Characterisation and Tribological Investigation of Artificial Skin Surface Lipid Films

This article deals with the tribology of lipid coatings that resemble those found on human skin. In order to simulate the lipidic surface chemistry of human skin, an artificial sebum formulation that closely resembles human sebum was spray-coated onto mechanical skin models in physiologically relevant concentrations (5–100 μg/cm2). Water contact angles and surface free energies (SFEs) showed that model surfaces with ≤25 μg/cm2 lipids appropriately mimic the physico-chemical properties of dry, sebum-poor skin regions. In friction experiments with a steel ball, lipid-coated model surfaces demonstrated lubrication effects over a wide range of sliding velocities and normal loads. In friction measurements on model surfaces as a function of lipid-film thickness, a clear minimum in the friction coefficient (COF) was observed in the case of hydrophilic, high-SFE materials (steel, glass), with the lowest COF (≈0.5) against skin model surfaces being found at 25 μg/cm2 lipids. For hydrophobic, low-SFE polymers, the COF was considerably lower (0.4 for PP, 0.16 for PTFE) and relatively independent of the lipid amount, indicating that both the mechanical and surface-chemical properties of the sliders strongly influence the friction behaviour of the skin-model surfaces. Lipid-coated skin models might be a valuable tool not only for tribologists but also for cosmetic chemists, in that they allow the objective study of friction, adhesion and wetting behaviour of liquids and emulsions on simulated skin-surface conditions.

Molecular aspects of the main phase transition in lipid systems as a weak first-order phase transition: 2. Fluctuation phenomena and the role of molecule conformation in the behavior of lipid films

The influence of molecule “softness” on the thermodynamic behavior of membranes, on the character of the main phase transition, and on an anomalous increase in the distance between bilayers in the film (swelling) above the phase-transition temperature is studied. The formation of the so-called ripple phase in multilayer films (modulation in the membrane plane) below the phase-transition temperature and the role of such factors as the molecule parameters and the lipid-film thickness are considered. It is shown that swelling of a lipid film is caused by the fluctuation-induced tilt of the molecules and the corresponding decrease in the number of “soft” molecules, whereas the formation of the ripple phase is caused by fluctuations of the order parameter in the gel phase and the corresponding increase in the number of soft molecules.

Increased Tear Film Lipid Layer Thickness with the use of two Novel Lubricating Eyedrops

The lipid tear film stabilizes the tears by lowering the surface tension and preventing aqueous tear evaporation and may be analyzed by kinetic analysis of the tear interference images. This study investigated changes in the lipid film after application of a new emulsion-based eye drop of Refresh Endura (Allergan, Irvine, CA) in normal subjects and patients with dry eyes.Comparative, nonrandomized interventional study.Five normal subjects and 10 aqueous tear deficiency (ATD) patients with or without lipid tear deficiency were enrolled prospectively.A complete eye examination was performed, including symptom score, tear break-up time, dye staining, and fluorescein clearance test. One eye received a single dose of emulsion eye drop (EED), whereas the other eye received nonpreserved saline as a control. Kinetic analysis of tear interference images taken by DR-1 (Kowa, Japan) was performed before and at various times after instillation of the drops.Symptoms, pattern, thickness, and spread time of the tear film.Compared with the control, all eyes receiving EED showed rapid restructuring of the preexisting lipid film. In normal persons, restructuring resulted in a thick lipid band across the cornea and a significant increase of the lipid film thickness. For ATD patients, EED application resulted in either a scant, irregular tear film, which became covered by the lipid film, or a modified lipid film changed without forming a band or irregularities. Emulsion eye drops tended to be more comfortable in patients with more severe ATD. Kinetic analysis showed that the mean spread time improved for all patients after EED when compared with baseline or with controls (P<0.05).Emulsion eye drop produces significant changes in the tear film of normal and dry eye patients. Further investigation with a prolonged period of treatment is warranted to establish whether EED may help improve the spread and thickness of the lipid tear film in moderate and severe dry eyes.

Evaluation of Wettability of Diseased Conjunctiva in Superior Limbic Keratoconjunctivitis

The lipid tear film stabilizes the tears by lowering the surface tension and preventing aqueous tear evaporation and may be analyzed by kinetic analysis of the tear interference images. This study investigated changes in the lipid film after application of a new emulsion-based eye drop of Refresh Endura (Allergan, Irvine, CA) in normal subjects and patients with dry eyes.Comparative, nonrandomized interventional study.Five normal subjects and 10 aqueous tear deficiency (ATD) patients with or without lipid tear deficiency were enrolled prospectively.A complete eye examination was performed, including symptom score, tear break-up time, dye staining, and fluorescein clearance test. One eye received a single dose of emulsion eye drop (EED), whereas the other eye received nonpreserved saline as a control. Kinetic analysis of tear interference images taken by DR-1 (Kowa, Japan) was performed before and at various times after instillation of the drops.Symptoms, pattern, thickness, and spread time of the tear film.Compared with the control, all eyes receiving EED showed rapid restructuring of the preexisting lipid film. In normal persons, restructuring resulted in a thick lipid band across the cornea and a significant increase of the lipid film thickness. For ATD patients, EED application resulted in either a scant, irregular tear film, which became covered by the lipid film, or a modified lipid film changed without forming a band or irregularities. Emulsion eye drops tended to be more comfortable in patients with more severe ATD. Kinetic analysis showed that the mean spread time improved for all patients after EED when compared with baseline or with controls (P<0.05).Emulsion eye drop produces significant changes in the tear film of normal and dry eye patients. Further investigation with a prolonged period of treatment is warranted to establish whether EED may help improve the spread and thickness of the lipid tear film in moderate and severe dry eyes.

Sequential changes of lipid tear film after the instillation of a single drop of a new emulsion eye drop in dry eye patients

Purpose The lipid tear film stabilizes the tears by lowering the surface tension and preventing aqueous tear evaporation and may be analyzed by kinetic analysis of the tear interference images. This study investigated changes in the lipid film after application of a new emulsion-based eye drop of Refresh Endura (Allergan, Irvine, CA) in normal subjects and patients with dry eyes. Design Comparative, nonrandomized interventional study. Participants Five normal subjects and 10 aqueous tear deficiency (ATD) patients with or without lipid tear deficiency were enrolled prospectively. Methods A complete eye examination was performed, including symptom score, tear break-up time, dye staining, and fluorescein clearance test. One eye received a single dose of emulsion eye drop (EED), whereas the other eye received nonpreserved saline as a control. Kinetic analysis of tear interference images taken by DR-1 (Kowa, Japan) was performed before and at various times after instillation of the drops. Main outcome measures Symptoms, pattern, thickness, and spread time of the tear film. Results Compared with the control, all eyes receiving EED showed rapid restructuring of the preexisting lipid film. In normal persons, restructuring resulted in a thick lipid band across the cornea and a significant increase of the lipid film thickness. For ATD patients, EED application resulted in either a scant, irregular tear film, which became covered by the lipid film, or a modified lipid film changed without forming a band or irregularities. Emulsion eye drops tended to be more comfortable in patients with more severe ATD. Kinetic analysis showed that the mean spread time improved for all patients after EED when compared with baseline or with controls ( P<0.05). Conclusions Emulsion eye drop produces significant changes in the tear film of normal and dry eye patients. Further investigation with a prolonged period of treatment is warranted to establish whether EED may help improve the spread and thickness of the lipid tear film in moderate and severe dry eyes.

Computer-synthesis of an interference color chart of human tear lipid layer, by a colorimetric approach.

To synthesize an interference color chart for the specific tear lipid layer interference camera, DR-1, for the conversion of the tear film lipid layer thickness into color graphic information--that is, for the quantification of the tear interference image--by a colorimetric approach. Because the color of the tear lipid layer interference image is visualized by a white light source interference phenomena, to produce gamma-corrected red, green, and blue (RGB) values of a specific interference color at a certain tear lipid film thickness, XYZ tristimulus values of the Commission Internationale de l'Eclairage (CIE) were obtained. XYZ tristimulus values were calculated from the light source spectrum of the DR-1 camera, the color-matching function of CIE, and the reflectance of the tear interference image in wavelengths ranging from 380 to 780 nm. These calculated interference colors were synthesized ranging from 0 nm to 1000 nm of lipid film thickness to produce a color chart. The applicability of the new color chart in the analysis of the lipid layer thickness was tested on a healthy control subject with normal tear function and a patient with dry eye who had aqueous tear deficiency and meibomian gland obstruction. The specific tear interference color chart for the DR-1 camera was obtained with RGB and XYZ tristimulus values. The interference chart ranged from 0 to approximately the 5th interference order. The interference colors from clinical DR-1 images could be converted to lipid thickness data by using the color chart system. A new tear interference color chart was developed in this study, which may be of benefit in converting tear interference color information to data describing the thickness of the tear film lipid layer.

Differentiation of lipid tear deficiency dry eye by kinetic analysis of tear interference images.

To use kinetic changes to characterize tear interference images in patients with lipid tear deficiency (LTD) dry eye. We used a DR-1 camera to digitize and analyze sequential images of tear interference on the central 8 mm of the cornea at the start of complete blinking in 11 healthy volunteers and 8 patients with LTD and noninflamed meibomian gland dysfunction. We studied tear lipid spread time and pattern, stability of the lipid after spread, and distribution of thickness in a prospective, case-control study. On complete lid closure, the lipid spread was horizontal in healthy eyes but vertical in LTD (P<.001). Mean +/- SD lipid spread time was 0.36 +/- 0.22 seconds in healthy eyes but 3.54 +/- 1.86 seconds in LTD (P<.001). Conventional DR-1 grading could not distinguish these groups (P =.32). Mean +/- SD lipid film thickness in healthy eyes was 74.5 +/- 6.9 nm, thicker than the 43.8 +/- 10.6 nm in LTD (P<.001), and this result was confirmed by qualification with intensity histogram (P<.001). Kinetic analysis of the tear interference revealed distinctive differences in the time and pattern of lipid spread and the distribution and stability of resultant lipid thickness between healthy subjects and patients with LTD. This method can be coupled with others for formulating effective therapies for patients with dry eye.