Oxygen-Carrying Capacity of Perfluorohexyloctane, a Novel Eye Drop for Dry Eye Disease.

Objective: One-hundred percent perfluorohexyloctane (PFHO) is a water-free, preservative-free eye drop approved by the Food and Drug Administration in the United States for the treatment of dry eye disease. PFHO has shown relief of dry eye signs and symptoms in clinical trials and has potent antievaporative action in vitro. The objective of this study was to measure the level of oxygen in PFHO. Methods: T1 relaxation times (time taken for proton spins to translate from a random alignment to an alignment with the main magnetic field) for fluorine-19 in perfluorohexyloctane were measured using fluorine-19 nuclear magnetic resonance spectroscopy. The level of oxygen was interpolated from published data. Results: The hydrogen-1 and fluorine-19 nuclear magnetic resonance spectra of PFHO were well resolved and the resonance assignments and intensities were as expected. The T1 values calculated for the CF3 group resonance in the current study was 0.901 seconds and 1.12 seconds at 25 °C and 37 °C, respectively. The T1 values for the CF2 group resonances increased by 17% to 24% with an increase in temperature from 25 °C to 37 °C. The mean (SD) partial pressure of oxygen in PFHO was calculated to be 257 (36) mm Hg and 270 (38) mm Hg at 25 °C and 37 °C, respectively. Conclusions: The current study confirms that PFHO contains a significant amount of oxygen, more so than that calculated for tears in equilibrium with air. Once instilled on the eye, PFHO is not expected to be a barrier to the oxygen necessary for a healthy cornea and may in fact deliver nonreactive oxygen to the cornea to facilitate healing in patients with dry eye disease.

Relationship Between Human Meibum Lipid Composition and the Severity of Meibomian Gland Dysfunction: A Spectroscopic Analysis.

Purpose: Information on the relationship between meibum lipid composition and severity of meibomian gland dysfunction (MGD) is limited. The purpose of this study was to analyze the molecular components of meibum collected from individuals with no MGD, mild-to-moderate MGD, and severe MGD. Methods: Adults with and without MGD were enrolled in a prospective, multicenter, exploratory clinical trial (ClinicalTrials.gov Identifier: NCT01979887). Molar ratios of cholesteryl ester to wax ester (RCE/WE) and aldehyde to wax ester (Rald/WE) in meibum samples were measured with 1H-NMR spectroscopy. Results were evaluated for participants grouped by MGD disease status and severity (non-MGD, mild-to-moderate MGD, and severe MGD), as defined by maximum meibum quality scores, Schirmer test results, and Subject Ocular Symptom Questionnaire responses. Results: Sixty-nine meibum samples from 69 individuals were included in the analysis: 24 non-MGD, 24 mild-to-moderate MGD, and 21 severe MGD. Mean RCE/WE was 0.29 in non-MGD, 0.14 in mild-to-moderate MGD (P = 0.038 vs. non-MGD, 51% lower), and 0.07 in severe MGD (P = 0.16 vs. mild-to-moderate MGD, 52% lower; P = 0.002 vs. non-MGD, 76% lower). Mean Rald/WE was 0.00022 in non-MGD, 0.00083 in mild-to-moderate MGD (P = 0.07 vs. non-MGD, 277% higher), and 0.0024 in severe MGD (P = 0.003 vs. mild-to-moderate MGD, 190% higher; P < 0.001 vs. non-MGD, 992% higher). Conclusions: RCE/WE was lowest and Rald/WE was highest in the severe MGD cohort, suggesting that these meibum constituent molar ratios may result from the pathophysiology associated with MGD and can impact ocular surface lipid and tear film homeostasis. These findings may potentially help identify targets for MGD treatment.

In Vitro Inhibition of Evaporation with Perfluorohexyloctane, an Eye Drop for Dry Eye Disease.

Objective: Perfluorohexyloctane (PFHO) MIEBOTM, formerly (NOV03) is a single component, water-free eye drop approved by the Food and Drug Administration in the United States for the treatment of dry eye disease. We evaluated the in vitro inhibitory effect of PFHO on the evaporation rate (Revap) of saline. Methods: Evaporation rates were measured gravimetrically at 25°C or 35°C. The evaporation rate (Revap) of phosphate-buffered saline (PBS) was measured following the application of 11-200 µL PFHO or 100 µL artificial tears (Soothe XP [Bausch + Lomb, Bridgewater, New Jersey], Systane Balance [Alcon, Fort Worth, Texas], and Systane Ultra [Alcon]). The effect of PFHO on the Revap of PBS was further evaluated following the addition of 50 mg/mL mucin to PBS and compared with that of meibum lipid collected from a 68 year-old White volunteer. Results: At 25°C the mean (SEM) Revap of PBS alone or PFHO alone was 4.06 (0.06) and 0.137 (0.004) µm/min, respectively. Layering 100 µL PFHO over PBS inhibited the Revap of PBS by 81% (P < 0.0001), whereas artificial tears had no effect. The presence of mucin attenuated the inhibition of the Revap of PBS by PFHO by 17% (P < 0.0001). At 35°C, the Revap of PBS was inhibited by 88% when layering 100 µL PFHO over PBS and 28% when applying a single 11 µL drop of PFHO (P value < 0.0001 for both). Meibum lipid inhibited the Revap of PBS by 8% at this temperature, whereas the combination of a drop of PFHO plus meibum inhibited the Revap of PBS by 34%. Conclusions: PFHO significantly inhibited the Revap of saline in this in vitro model. The data support the idea that PHFO may form an antievaporative layer on the tear film surface and may be a functional substitute for the native tear-film lipid layer in patients with dry eye disease.

Correlations between bulk and surface properties of meibomian lipids with alteration of wax-to-sterol esters content.

In a recent study (Ewurum et al., 2021), wax (WE) and sterol esters (CE) from human meibum secretions (MGS) were separated and reconstituted with controlled WE/CE ratios (0%, 20%, 30%, 40%, 50%, 75% and 100% CE weight fractions). It was found that the alterations in the CE content of WE/CE mixtures modified the hydrocarbon chain conformation and packing of the mixture. A major question that emerges is whether the spectroscopic packing parameters determined for bulk meibum translate to a change in the performance of meibomian layers at the air/water interface, as it is the surface film functionality that is crucial for the performance of MGS at the ocular surface. The study of human meibum films with Langmuir surface balance was performed to access the surface properties at blink-like deformations of the film area. Surface pressure (π)-area (A) isocycles and stress relaxations were used to assess the layer's reorganization during area cycling and dilatational elasticity, respectively. The morphology of the films was monitored by Brewster angle microscopy. It was found that the increased order and chain melting temperature of the bulk samples correlated with a raise in the maximum surface pressure attained at minimal surface area and in the transient dilatational modulus of the meibomian layers. Such correlations may allow for development of an improved understanding between the bulk and surface properties of human meibum and of other natural and synthetic tear lipid films.

Meibum Lipid Composition and Conformation in Parkinsonism.

PURPOSE: Patients with Parkinson's disease (PD) exhibit unstable tear films. Tear film lipid composition and structure are related to tear film stability and dry eye and tear lipids have not been characterized in people with PD. The aim of this study is to characterize Meibum tear lipids in donors with PD using 1H-NMR and infrared spectroscopy. METHODS: Three cohorts were compared: meibum from donors with PD (Mp) n = 10, meibum from donors with PD and dry eye (Mpd) n = 3, meibum from donors without PD (Mn) n = 29. RESULTS: There were no significant differences, P > 0.05, in hydrocarbon branching for Mp compared with Mn. Mn contained twice as much cholesteryl esters compared with Mp, P < 0.0001. The cooperativity of the phase transition was significantly 37% lower for Mp compared with Mn, P < 0.0001. Mpd was much more ordered (stiffer) with compared with Mp and Mn, P < 0.0001. CONCLUSION: Changes in meibum lipid composition and structure could be a marker for and/or contribute to increase the susceptibility of dry eye in patients with PD. A less cooperative phase transition for Mp compared with Mn indicates that Mp was more heterogeneous and/or contained more contaminants than Mn. The data support the idea that more ordered lipid contributes to dry eye.

A spectroscopic approach to measuring meibum lipid composition and conformation in donors with SjÓ§gren's syndrome.

Patients with SjÓ§gren's syndrome (SS) have dry eye associated with meibomian gland dysfunction (MGD). The meibum from donors with dry eye due to MGD but without SS (MMGD) presents with lower levels of cholesteryl ester, less straight chains, and more ordered hydrocarbon chains compared with meibum from donors without MGD (Mn). The aim of the current study was to compare the composition and hydrocarbon chain conformation of meibum from donors with Sjögren's syndrome (Mss) to Mn and MMGD. Meibum was expressed from patients with SS using an ILUX instrument (Alcon Inc., Fort Worth TX). All of the nine meibum donors with SS were female. Meibum composition was characterized using 1H-NMR and meibum hydrocarbon chain conformation was measured using fourier transform infrared spectroscopy. Meibum from every donor with SS measured contained a significantly (P < 0.01) higher cholesteryl ester/wax ester ratio and more straight chains compared with donors without SS or dry eye. None of the nine phase transitional parameters were significantly different, P > 0.05, for Mss compared with Mn. Nor was the CH3/CH2 band height ratio used to estimate the number of hydrocarbon CH3 and CH2 moieties different, P = 0.22, for Mss compared with Mn. In conclusion, the compositional differences between Mss compared with Mn did not result in differences in any of the nine meibum lipid phase transitional parameters measured. The compositional differences observed between Mss and Mn could be markers for or contribute to SS as the differences could lead to tear film lipid packing differences other than conformational differences.

A spectroscopic study of the composition and conformation of cholesteryl and wax esters purified from meibum.

Elucidating wax ester (WE) and cholesteryl ester (CE) compositional, structural and functional relationships is key to our understanding of how these lipids are involved in natural and pathological processes. Little is known about how CE and WE interact with one another. The focus of the present study is to bridge this gap of knowledge. CE and WE were collected from human meibum as a source of esters with complex hydrocarbon chains. MgO column chromatography was used to separate WE and CE. The esters were characterized using 1H-NMR and Fourier transform infrared spectroscopy. The complexity of the hydrocarbon chains of native WE and CE influenced how changes in the ratio of WE and CE ester influenced some lipid phase transitional parameters but not others. Changes in CE content of WE/CE mixtures undoubtedly modifies the hydrocarbon chain conformation and packing of the mixture. The nature of the change depends on the conformation of the WE and CE. Differences in the complexity of the hydrocarbon chains are likely not to be a major influence on alterations in the order or phase transition temperature when more ordered WE is added to less ordered CE.

Hyaluronic acid-lipid binding.

BACKGROUND: Phospholipid (PL)-hyaluronic acid (HA) interactions are relevant to aging-associated vitreous humor liquefaction, therapies for dry eye disease, skin-care products and synovial joint lubrication. Phosphatidyl choline-HA interactions have been well characterized. However, other major lipids found in tears, vitreous humor and synovial joints have not. The purpose of this study was to bridge this gap of knowledge. METHODS: HA (1600 kDa) at 5 mg/mL, was mixed with various lipids ranging in concentration from 0.1 to 10 mg/mL in D2O. HA-PL binding was measured from the decrease in HA proton resonance intensity with binding using a nuclear magnetic resonance spectrometer. RESULTS: Cholesterol weakly bound to HA, followed by monoglyceride and palmitoyl palmitate < phosphatidyl choline, phosphatidic acid and sphingomyelin. The maximum amount of PL bound was 14 ± 1 µmoles inferring a 1 to 1 molar ratio of bound PL to HA dimer. Monoglyceride and palmitoyl palmitate required two to three times more lipid to achieve 100% bound HA compared to PL. CONCLUSIONS: Physiological levels of HA, phosphatidyl choline and sphingomyelin would result in only 4% of the hydrophobic hydrogens of HA to be bound. HA-PL binding interactions could be important for therapeutic use of HA in eye drops in future studies to treat dry eye and to trap PL entering the VH to keep them from forming light scattering micelles. HA-lipid binding may also be relevant to the therapeutic effects of topical skin-care products. Both head group and hydrocarbon chain moieties influence HA-lipid interactions.

Lipid conformational order and the etiology of cataract and dry eye.

Lens and tear film lipids are as unique as the systems they reside in. The major lipid of the human lens is dihydrosphingomylein, found in quantity only in the lens. The lens contains a cholesterol to phospholipid molar ratio as high as 10:1, more than anywhere else in the body. Lens lipids contribute to maintaining lens clarity, and alterations in lens lipid composition due to age are likely to contribute to cataract. Lens lipid composition reflects adaptations to the unique characteristics of the lens: no turnover of lens lipids or proteins; the lowest amount of oxygen of any tissue; and contains almost no intracellular organelles. The tear film lipid layer (TFLL) is also unique. The TFLL is a thin (100 nm) layer of lipid on the surface of tears covering the cornea that contributes to tear film stability. The major lipids of the TFLL are wax esters and cholesterol esters that are not found in the lens. The hydrocarbon chains associated with the esters are longer than those found anywhere else in the body (as long as 32 carbons), and many are branched. Changes in the composition and structure of the 30,000 different moieties of TFLL contribute to the instability of tears. The focus of the current review is how spectroscopy has been used to elucidate the relationships between lipid composition, conformational order and function, and the etiology of cataract and dry eye.

Changes in meibum composition following plaque bachytherapy for choroidal melanoma.

OBJECTIVES: Dry eye is common when external beam radiation is used for the treatment of choroidal melanoma (CM). As meibum structure and composition have been related to dry eye, we determined if plaque bachytherapy for CM alters meibum composition. DESIGN: 1H-NMR spectroscopy was used to measure the lipid composition of meibum. SETTING: The University of Louisville, Kentucky, USA. PARTICIPANTS: All 13 participants had CM and one participant had iris melanoma. MAIN OUTCOME MEASURES: Cholesteryl ester (CE) to wax ester (WE) ratio, amount of meibum esters (ME) and meibum lipid saturation were measured. RESULTS: ME decreased by 80%±18% (±99% CI) in 11 eyes that were treated compared with the contralateral untreated eye. ME increased by 181% in two eyes that were treated compared with the contralateral untreated eye. The mole % CE/WE for meibum was significantly (p<0.0001) 67% lower in eyes that were irradiated compared with control eyes from donors without CM and were not treated. Plaque brachytherapy induced the de-esterification of CE. The intensity of the meibum cis double bond resonances did not change significantly (p>0.05). CONCLUSION: Eyes that had plaque brachytherapy had a lower amount of expressible meibum and a lower CE/WE ratio compared with meibum from the contralateral eye that received no treatment and eyes that did not have uveal melanoma. Both the quality and quantity of meibum should be considered in designing a therapy for dry eye after plaque brachytherapy.

Heterozygous Loss of Yap1 in Mice Causes Progressive Cataracts.

Purpose: Yap1 encodes an evolutionarily conserved transcriptional coactivator and functions as a down-stream effector of the Hippo signaling pathway that controls tissue size and cell growth. Yap1 contributes to lens epithelial development. However, the effect of Yap1 haplodeficiency on the lens epithelium and its role in the development of cataracts has not been reported. The aim of the current study is to investigate Yap1 function and its regulatory mechanisms in lens epithelial cells (LECs). Methods: Lens phenotypes were investigated in Yap1 heterozygous mutant mice by visual observation and histological and biochemical methods. Primary LEC cultures were used to study regulatory molecular mechanism. Results: The heterozygous inactivation of Yap1 in mice caused cataracts during adulthood with defective LEC phenotypes. Despite a normal early development of the eye including the lens, the majority of Yap1 heterozygotes developed cataracts in the first six months of age. Cataract was preceded by multiple morphological defects in the lens epithelium, including decreased cell density and abnormal cell junctions. The low LEC density was coincident with reduced LEC proliferation. In addition, expression of the Yap1 target gene Crim1 was reduced in the Yap1+/- LEC, and overexpression of Crim1 restored Yap1+/- LEC cell proliferation in vitro. Conclusions: Homozygosity of the Yap1 gene was critical for adequate Crim1 expression needed to maintain the constant proliferation of LEC and to maintain a normal-sized lens. Yap1 haplodeficiency leads to cataracts.

In-vitro and ex-situ regional mass spectral analysis of phospholipids and glucose in the vitreous humor from diabetic and non-diabetic human donors.

The causes of vitreous humor (VH) liquefaction remain unclear. Diabetes accelerates this process and other ocular diseases. The weakening of the blood-retina barrier observed with diabetes could enhance the rate of transfer of relatively small molecules such as glucose (Glu) and phospholipids (PLs) from the retina to the VH. Glucose and PLs have been detected previously in VH but their regional distributions are not known. The mapping of Glu and PLs in VHs from subjects with and without diabetes could reveal the roles of these molecules in VH liquefaction. Diabetic and non-diabetic human eyes were acquired from the Kentucky Lions Eye Bank and frozen immediately. Each VH was removed and halved along the sagittal plane. One half was stamped on a matrix assisted laser desorption ionization (MALDI) plate. Either p-Nitroanaline (26 mg/mL MeOH:CHCl3) or 2,5-dihydroxybenzoic acid (20 mg/mL H2O:acetonitrile) was used as matrix. Glu and PLs were extracted from the remaining sections and analyzed. Data were acquired using a MALDI-mass spectrometer. The levels of Glu and PLs were significantly greater in VH from diabetics (VHd) compared with VH from non-diabetics (VHnd). VHds showed the highest relative levels of PLs in the posterior VH, followed by the anterior and central regions. Throughout the entire VH, the most abundant PLs were phosphatidylcholines followed by sphingomyelins. For Glu, the relative intensities were ~3 times higher in the posterior region of VHd (12 ± 1.3) compared with VHnd (6.5 ± 0.7) VHs. Regional studies showed that relative to the posterior VHd, the Glu levels were lower in the anterior (8.1 ± 1.0) and central (6.7 ± 0.8) regions. For the VHnds, the values for the central and anterior regions were 5.9 ± 1.2 and 4.7 ± 0.9, respectively. PLs and Glu are most abundant in the posterior region relative to the central and anterior zones of VHs. This trend was observed in VHd and VHnd, but PLs and Glu levels were significantly higher in VHds. These results support the possibility that higher levels of Glu and PLs accelerate VH liquefaction in diabetic patients. As liquefaction begins in the posterior region, the higher abundance of PLs and Glu in this zone also suggests that they may play a role in liquefaction. The specific molecular interactions affected by Glu and PLs in the collagen/hyaluronan/water network need to be examined.

Meibum lipid hydrocarbon chain branching and rheology after hematopoietic stem cell transplantation.

PURPOSE: Meibum from donors who have had hematological stem cell transplantations (MHSCT) are susceptible to severe dry eye symptoms and exhibit very high lipid order (stiffness) compared with meibum from donors without dry eye (Mn). Since lipid order could have functional consequences, we compared the rheology and composition of Mn and MHSCT to measure meibum compositional, structural and functional relationships. METHODS: The rheology and composition was measured using Langmuir trough and 1H NMR spectroscopy, respectively. RESULTS: MHSCT and Mn was studied from 16 to 43 donors, respectively, using NMR spectroscopy. MHSCT contained significantly 16% more straight chain and 24% less iso-chain hydrocarbons compared with Mn. The cholesteryl ester to wax ester molar ratio, and hydrocarbon chain unsaturation were not significantly different, for MHSCT compared with Mn.Surface pressure-area isotherms of meibum from 30 donors without dry-eye were grouped into 4 pools (PC) and meibum from 32 donors with dry eye who had hematopoietic stem cell transplantation (PT) were grouped into 3 pools. Above 15 years of age the Пmax and (Cs -1)max increased with age for both the PC and the PT cohorts. (Cs -1)max values were higher for PT samples compared with age matched PC samples, indicating they had higher elasticity and stiffness. A more ordered lipid could contribute to the formation of a discontinuous patchy tear film lipid layer, which in turn results in deteriorated spreading, and decreased surface elasticity. CONCLUSIONS: The composition and rheology of meibum from donors with dry eye and who have had HSCT support the idea from other studies that more ordered meibum may contribute to or be a marker of dry eye.

Concentration dependent cholesteryl-ester and wax-ester structural relationships and meibomian gland dysfunction.

BACKGROUND: With dry eye, the ratio of cholesteryl ester (CE) to wax ester (WE) decreases substantially in meibum, but the functional and structural consequences of this change are speculative. The aim of this study is to confirm this finding and to bridge this gap in knowledge by investigating the effect of varying CE/WE ratios on lipid structure and thermodynamics. METHODS: Infrared spectroscopy was use to quantify CE and WE in human meibum and to measure hydrocarbon chain conformation and thermodynamics in a cholesteryl behenate, stearyl stearate model system. RESULTS: The CE/WE molar ratio was 36% lower for meibum from donors with dry eye due to meibomian gland dysfunction compared with meibum from donors without dry eye. CE (5 mol %) dramatically increased the phase transition temperature of pure WE from -0.12 °C to 63 °C in the mixture. Above 5 mol % CB, the phase transition temperature increased linearly, from 68.5 °C to 85 °C. In the ordered state, CE caused an increase in lipid order from about 72% trans rotamers to about 86% trans rotamers. Above 10% CE, the hydrocarbon chains were arranged in a monoclinic geometry. CONCLUSIONS: The CE/WE is lower in meibum from donors with dry eye due to meibomian-gland dysfunction. Major conformational changes in the hydrocarbon chains of wax and cholesteryl ester mixtures begin to occur with just 5% CB and above. GENERAL SIGNIFICANCE: CE-WE interactions may be important for in understanding lipid layer structure and functional relationships on the surface of tears, skin and plants.

Structural Differences in Meibum From Teenagers Without and With Dry Eye and Allogeneic Hematologic Stem Cell Transplantations.

Although dry eye occurs mostly in adults, dry eye may be induced in teens receiving allogeneic hematologic stem cell transplantations (AHSCT). Changes in meibum composition and structure has been associated with dry eye. The structure of meibum from teens with dye eye or teens with dry eye and AHSCT has not been studied, so in this study, we compared the structure of meibum from teens receiving AHSCT that had a dry eye with meibum from teens without AHSCT and without dry eye symptoms.

Differences in Meibum and Tear Lipid Composition and Conformation.

PURPOSE: The compositional, structural, and functional relationships of meibum may provide insights into the loss of tear film stability. Although the conformation of meibum lipids has been studied rigorously, that of tear lipids has not. METHODS: Tear lipids (TLHSCT) and meibum (MHSCT) from patients who had hematopoietic stem cell transplantation were pooled prospectively. The infrared spectra of meibum from donors with (MMGD) and without (Mn) meibomian gland dysfunction were retrospectively analyzed to measure the lipid composition and structure. The infrared CH stretching region was used to measure the relative content of CH3 and CH2 moieties in the meibum. RESULTS: The 3 major findings of the current study are as follows: 1) compared with Mn, MHSCT and MMGD had 18% fewer CH3 moieties; 2) compared with MHSCT, the phase transition temperature, cooperativity, and order were approximately 20% greater for TLHSCT; and 3) compared with Mn and MMGD, MHSCT and TLHSCT contained fewer double bonds. CONCLUSIONS: Tear lipids are more ordered than meibum lipids, which could have functional consequences. The human meibum peak height ratio of the CH3/CH2 bands is not a factor related to tear film stability with age or sex. The amount of CH3 moieties relative to CH2 moieties and saturation could contribute to a higher meibum lipid order associated with a younger age, meibomian gland dysfunction, and dry eye from hematopoietic stem cell transplantation. Therefore, the hydrocarbon order may be a marker of or contribute to an unstable tear film layer.

Lipid Saturation and the Rheology of Human Tear Lipids.

Elevated levels of acyl chain saturation of meibomian lipids are associated with enhanced tear film (TF) stability in infants to shortened TF breakup time with meibomian gland dysfunction. Thus, the effect of saturation on the surface properties of human TF lipids (TFLs) using a Langmuir surface balance and Brewster angle microscopy was studied. Lipid phase transitions were measured using infrared spectroscopy. The raise in the % of saturation resulted in thicker, and more elastic films at π = 12 mN/m, with the effects being proportional to the saturation level. At the same time, at lower (≤10 mN/m) π, the raise in saturation resulted in an altered spreading and modified structure of TFL layers. The strong impact of saturation on TFL surface properties correlated with a saturation induced increase of the TFL acyl chain order, phase transition temperature, and lipid-lipid interactions. The native TFL order and πmax were significantly greater, compared with native meibum collected from the same individual. Aggregation of lipids on the tear surface due to saturation was not as significant as it was for meibum. Although the surface pressure/area isotherms for TFL were similar for meibum, differences in rheology and phase transition parameters warrant the study of both.

Structural Differences in Meibum From Donors After Hematopoietic Stem Cell Transplantations.

PURPOSE: Meibum is considered to be a key component of tears that serve to protect the eye, and conformational changes in meibum have not been studied extensively within the population of patients who had hematopoietic stem cell transplantation (HSCT). The aim of this study was to determine possible lipid conformational changes in the meibum of patients who had HSCT. METHODS: Participants who had HSCT were randomly sampled for this prospective comparative study. Control participants did not have dry eye or had not undergone allogeneic or autologous stem cell transplantation. Fourier-transform infrared spectroscopy was used to measure meibum phase transition. RESULTS: Meibum was collected from both eyes of 36 donors without dry eye (Mc) and from 22 patients who had undergone HSCT (MHSCT). There were no significant differences between the phase transition parameters based on gender or race. The following were the significant differences (P < 0.0001) between the parameters for Mc compared with MHSCT : lipid order (% trans) at 33.4°C increased from 40 (1) to 54 (2), cooperativity decreased from 7.9 (0.4) to 5.4 (0.3), the phase transition temperature (C) increased from 30.3 (0.4) to 34.2 (0.9), and the magnitude of the phase transition (cm) increased from 4.0 (0.1) to 4.7 (0.5) (standard error of the mean). CONCLUSIONS: Conformational and thermodynamic differences were observed between Mc and MHSCT. The changes observed in the lipid conformation of meibum from patients receiving HSCTs suggest that meibum composition changes after stem cell transplantation, and clinicians should consider treating the meibomian glands to improve the ocular surface.

Human Meibum Cholesteryl and Wax Ester Variability With Age, Sex, and Meibomian Gland Dysfunction.

Purpose: Relationships between tear film lipid (TFL) layer composition, structure, and function could provide insight into the etiology of dry eye. The molar ratio of cholesteryl ester (CE)/wax ester (WE) was measured in meibum from normal donors (Mn) and compared with meibum from donors with meibomian gland dysfunction (MMGD). Methods: CE/WE was measured using nuclear magnetic resonance spectroscopy. Results: CE/WE was distributed into two populations with 81% distributed near 0.55 and 19% near 0.3. CE/WE were higher in donors 13 to 19 years old compared with donors 1 to 12 years old and 20 to 88 years old. CE/WE for MMGD was 30% lower, 0.34 ± 0.04, compared with Mn, 0.49 ± 0.04. There were no sex differences in CE/WE. There were no significant racial differences between the CE/WE ratios for Asians and Caucasians. The CE/WE ratio was higher for blacks and lower for Hispanics compared to Caucasians. Due to the small number sampled, confirmation of the later racial results is needed. The packing of CE and WE in the TFL layer was proposed. Conclusions: Although MMGD contains much less CE than Mn, factors other than the CE content, such as the levels of saturation and/or proteins, may be responsible for the higher order of MMGD. In addition to saturation, CE could contribute to the increase in order of Mn between 0 and 20 years of age. Observed changes in the meibum content of CE alone is not likely to influence tear film stability.

The optimum temperature for the heat therapy for meibomian gland dysfunction.

PURPOSE: Numerous devices have been developed to warm the eyelid as a therapy for Meibomian gland dysfunction. The optimum temperature for such therapy was determined. METHODS: Meibum lipid disorder versus temperature was calculated from previously published phase transition parameters measured using infrared spectroscopy. Phase transitions parameters were calculated from meibum obtained from donors with Meibomian gland dysfunction (MMGD), donors who were susceptible to dry eye after hematopoietic stem cell transplantations (MHSCT) and meibum from donors without dry eye (Mn). RESULTS: Heating Mn to 40 °C increases the lipid disorder by 20.4%-90%. Heating the meibum another 4° increases the disorder to 95%. MMGD is 73.2% disordered with no heating. Heating MMGD to 41.5 °C increases the disorder by 26.8%-90% disordered. Heating MMGD another 5.1 °C increases the disorder to 95%. As MHSCT is much more ordered, 56.9% disordered compared with Mn and MMGD, a higher temperature above safety limits, 52 and 59 °C is need to disorder MHSCT to 90 and 95% disorder. CONCLUSIONS: Heating the eye lid above the phase transition temperature of meibum increases the disorder of meibum lipid which could ameliorate dry eye symptoms. The optimum temperature for disordering Mn and MMGD to 90% maximum disorder is 40 and 41.5 °C, respectively. Safety issues and discomfort should be considered in obtaining an optimal level of disorder, especially for severe cases of dry eye.