Studying the effects of suberythemal UV doses on the human stratum corneum by in vivo confocal Raman spectroscopy

Background: The stratum corneum (SC) plays an important role in skin barrier function. It acts as a protective barrier against water loss, eliminates foreign substances and micro-organisms and acts against harmful effects of UVR. Objectives: Our aim was to study the impact of suberythemal doses of UVA and UVB exposure on the molecular structure, organization and barrier function of the SC by following different Raman descriptors. Materials & Methods: Twenty female volunteers, aged 20-30 years, with healthy skin were enrolled. Doses of 95 mJ/cm² UVA and 15 mJ/cm² UVB were applied to volunteers' forearms. In vivo Raman measurements were performed at irradiated and control regions. Results: The impact of UVA and UVB irradiation was observed following several Raman descriptors, i.e. the ratio of vasymCH2/vsymCH2 (2885 cm-1/2850 cm-1) corresponding to the organizational order of the lipid bilayer. Water content and mobility descriptors were obtained by calculating vOH/vCH ratio. Finally, protein secondary structure was evaluated based on the 1670 cm-1/1650 cm-1 ratio related to ß sheets and α helices, respectively. Conclusion: UVA induced a loosening of the lateral packing of lipids immediately after irradiation. In contrast, delayed impact caused a tightening of the lipid barrier, an increase in water content -mainly in the unbound water fraction and a higher relative amount of ß sheets in SC proteins. Overall, these observations may explain the thickening of the SC observed in previous studies. A UVB dose of 15 mJ/cm² was apparently below the threshold necessary to induce significant changes despite the trends observed in this study.

Studies on stratum corneum metabolism: function, molecular mechanism and influencing factors.

BACKGROUND: Stratum corneum is located in the outermost layer of the skin and is the most important part of the skin barrier. Stratum corneum mainly contains keratinocytes, lipids, and desmosomes. Their normal metabolic process is closely related to the function of skin barrier. AIMS: This paper reviews the structure and function of stratum corneum, influencing factors, skin diseases, and common solutions. METHODS: An extensive literature search was conducted on the structure and function of stratum corneum, influencing factors, skin diseases, and common solutions. RESULTS: This paper reviews the structure and function of stratum corneum and the influence of various factors on stratum corneum metabolism. At the same time, the existing skin problems, skin diseases, and common solutions are summarized. CONCLUSIONS: This information will help to understand the function, molecular mechanism, and influencing factors of stratum corneum metabolism, and provide new ideas for stratum corneum health management and cosmetic research and development.

Virgin coconut oil as prophylactic therapy against alcohol damage on skin in COVID times.

BACKGROUND: Increased frequency of using alcohol-based hand sanitizers (ABHS) by consumers during COVID times have resulted in increased incidences of skin issues on palms. OBJECTIVE: (1) To quantify skin damage with increased usage frequency of ABHS by consumers and (2) To evaluate Virgin Coconut Oil (VCO) as natural prophylactic agent to counter the adverse effects. METHODS: In-home usage study was carried out with 60 volunteers for a 15-day intervention-Control Group: 6 applications per day of ABHS and Test Group: Overnight VCO use (6-8 drops) followed by 6× usage per day of ABHS. This leg included dermatological evaluation and WHO Self-Assessment Scale for skin health. Another leg of measurement included non-invasive instrumental study (Moisture & TEWL Probes, Tape Strip for protein content and IR spectroscopy for protein & lipid content) on forearm of 12 subjects (25-60 years age) with and without VCO application and repeated alcohol exposure. RESULTS: In-home usage study established consumer experiencing skin protective effect of VCO in the context of ABHS onslaught. 25% increase in perceived moisture content was recorded for VCO users, using WHO Self-Assessment Scale. Instrumental studies confirmed an increase in TEWL and decrease in lipids & protein content. Overnight VCO application resists the extraction which builds up with repeated application. CONCLUSIONS: Current work provides evidence of compromised hand skin barrier with ABHS daily usage. Overnight VCO application helps prepare the skin for next day alcohol use. Based on the findings, a regimen of overnight VCO application on hands as a natural prophylactic is recommended.

Optimal configuration of confocal Raman spectroscopy for precisely determining stratum corneum thickness: Evaluation of the effects of polyoxyethylene stearyl ethers on skin.

Properties regarding stratum corneum (SC), the outermost membrane of the skin, remain an active area in dermatologic and cosmetic research. The reduced thickness of SC is associated with varied adverse statuses such as skin lipid deficiency, skin barrier dysfunctions and skin deceases, etc. Emulsifiers with existing irritative effects on skin components also face the risk of decreasing SC thickness. We have been focusing on the effects of PEGylated emulsifiers on the skin and have an interest in finding the role of their polyethylene glycol (PEG)-chain length in tuning skin irritations. With this aim, PEG-stearyl ethers with different numbers of hydrophilic chains were applied on the skin, and their influence on skin thickness was discovered to determine their skin barrier effect. Confocal Raman spectroscopy (CRS) with extensive application in skin research was used here. To obtain the precise determination of skin thickness, our secondary aim was to find the optimal CRS configuration referring to varied objectives and pinhole sizes where further study is still in demand. Therefore, SC thickness measured via eddy current approach served as reference. The applied PEG-stearyl ethers formed the system to achieve varied thicknesses. Results confirmed that the skin interactions rose with increasing PEG-chain length, however only up to a certain limit, with decreasing effects recorded from PEG-40 stearyl ether and no effects observed from PEG-100 stearyl ether. Simultaneously, CRS combined with water immersion objective and 50 µm pinhole presented the most consistent values to the references and exhibited better spectral intensity and signal-to-noise ratio. Correlation plots involving different cases of configurations were calculated for error corrections. Taken together, this work helps to identify the potential mechanisms governing the interactions between PEG-stearyl ethers and skin and offers powerful evidence of using CRS as a reliable alternative to obtain accurate thickness values.

The grease trap: uncovering the mechanism of the hydrophobic lid in Cutibacterium acnes lipase.

Acne is one of the most common dermatological conditions, but the details of its pathology are unclear, and current management regimens often have adverse effects. Cutibacterium acnes is known as a major acne-associated bacterium that derives energy from lipase-mediated sebum lipid degradation. C. acnes is commensal, but lipase activity has been observed to differ among C. acnes types. For example, higher populations of the type IA strains are present in acne lesions with higher lipase activity. In the present study, we examined a conserved lipase in types IB and II that was truncated in type IA C. acnes strains. Closed, blocked, and open structures of C. acnes ATCC11828 lipases were elucidated by X-ray crystallography at 1.6-2.4 Å. The closed crystal structure, which is the most common form in aqueous solution, revealed that a hydrophobic lid domain shields the active site. By comparing closed, blocked, and open structures, we found that the lid domain-opening mechanisms of C. acnes lipases (CAlipases) involve the lid-opening residues, Phe-179 and Phe-211. To the best of our knowledge, this is the first structure-function study of CAlipases, which may help to shed light on the mechanisms involved in acne development and may aid in future drug design.

Prediction of neonatal acne based on maternal lipidomic profiling.

BACKGROUND: Neonatal acne occurs in the first few weeks after birth. Some lesions are more serious and leave scars. Maternal surface skin lipids (SSL) have a strong correlation with SSL of infants. The establishment of prediction rank model based on maternal SSL is essential to the prevention and treatment of neonatal acne. METHOD: Surface skin lipids samples were collected from the mothers (M) of 56 neonatal acne patients and the mothers (HM) of 19 healthy infants. Surface skin lipids from the right forehead were collected using a noninvasive method. UPLC-QTOF-MS was applied to detect SSL. Partial least squares discriminant analysis and receiver operating characteristic (ROC) analysis were performed to screen and validate potential lipids. Random forest (RF) and ROC analysis were used to establish a prediction model and evaluate its accuracy. RESULTS: Sixteen altered potential lipids belonging to fatty acids, sphingomyelins, and glycerides were associated with M. M had less lipids than HM. Spearman's correlation of 16 lipids revealed 9 with high correlation. They were chosen as characteristic values of the RF prediction model. And the model showed an average accuracy of 98% in the validation set. CONCLUSION: We have established an RF model for predicting neonatal acne and have shown that high skin barrier-related lipids were markers for predicting neonatal acne.

l-Ascorbic acid alkyl esters action on stratum corneum model membranes: An insight into the mechanism for enhanced skin permeation.

L-ascorbic acid alkyl esters (ASCn) are lipophilic forms of vitamin C, which act as skin permeation enhancers. We investigated the physical changes induced by incorporating ASCn into stratum corneum (SC) lipid membranes and correlated this with the mechanism proposed in the literature for skin permeation enhancement phenomena. We used lipid monolayers to explore the 2D structure and elasticity of the lipid-enhancer systems. As a comparison, the classic permeation enhancer, oleic acid (OA) and the non-enhancer analogue stearic acid (SA) were analysed. The incorporation of ASCn or OA into SC membranes resulted in more liquid-like films, with a dose-dependent lowering of the compressibility modulus. Brewster angle microscopy (BAM) evidenced partial miscibility of the enhancer with SC lipid components, stabilising the liquid-expanded phase. At the nanoscale, AFM showed that SC lipids form heterogeneous membranes, which underwent structural alterations after incorporating ASCn and fatty acids, such as SA and OA. The lower, cholesterol-enriched phase appears to concentrate the enhancers, whilst the higher ceramide-enriched phase concentrated the non-enhancer SA. Our results and previously reported pieces of evidence indicate a strong pattern in which the rheological properties of SC lipid films are determinant for skin permeation phenomena.

Hyperalphalipoproteinemic scavenger receptor BI knockout mice exhibit a disrupted epidermal lipid barrier.

Scavenger receptor class B type I (SR-BI) mediates the selective uptake of cholesteryl esters (CE) from high-density lipoproteins (HDL). An impaired SR-BI function leads to hyperalphalipoproteinemia with elevated levels of cholesterol transported in the HDL fraction. Accumulation of cholesterol in apolipoprotein B (apoB)-containing lipoproteins has been shown to alter skin lipid composition and barrier function in mice. To investigate whether these hypercholesterolemic effects on the skin also occur in hyperalphalipoproteinemia, we compared skins of wild-type and SR-BI knockout (SR-BI-/-) mice. SR-BI deficiency did not affect the epidermal cholesterol content and induced only minor changes in the ceramide subclasses. The epidermal free fatty acid (FFA) pool was, however, enriched in short and unsaturated chains. Plasma CE levels strongly correlated with epidermal FFA C18:1 content. The increase in epidermal FFA coincided with downregulation of cholesterol and FFA synthesis genes, suggesting a compensatory response to increased flux of plasma cholesterol and FFAs into the skin. Importantly, the SR-BI-/- epidermal lipid barrier showed increased permeability to ethyl-paraminobenzoic acid, indicating an impairment of the barrier function. In conclusion, increased HDL-cholesterol levels in SR-BI-/- mice can alter the epidermal lipid composition and lipid barrier function similarly as observed in hypercholesterolemia due to elevated levels of apoB-containing lipoproteins.

Development of new 3D human ex vivo models to study sebaceous gland lipid metabolism and modulations.

OBJECTIVES: Sebaceous glands maintain skin homeostasis by producing sebum. Low production can induce hair loss and fragile skin. Overproduction provokes seborrhoea and may lead to acne and inflammatory events. To better study sebaceous gland maintenance, sebocyte maturation, lipid production and ageing or inflammatory processes, we developed innovative 3D ex vivo models for human sebaceous glands. MATERIALS AND METHODS: Culture conditions and analytical methods optimized on sebocyte monolayers were validated on extracted sebaceous glands and allowed the development of two 3D models: (a) "air-liquid" interface and (b) human fibronectin-coated "sandwich" method. Lipid production was assessed with microscopy, fluorometry or flow cytometry analysis after Nile Red staining. Specific lipids (particularly squalene and peroxidized squalene) were measured by Gas or liquid Chromatography and Mass spectrometry. RESULTS: This study allowed us to select appropriate conditions and design Seb4Gln culture medium inducing sebocyte proliferation and neutral lipid production. The "air-liquid" model was appropriate to induce sebocyte isolation. The "sandwich" model enabled sebaceous gland maintenance up to 42 days. A treatment with Insulin Growth Factor-1 allowed validation of the model as we succeeded in mimicking dynamic lipid overproduction. CONCLUSION: Functional sebocyte maturation and physiological maintenance were preserved up to 6 weeks in our models. Associated with functional assays, they provide a powerful platform to mimic physiological skin lipid metabolism and to screen for active ingredients modulating sebum production.

Supplemental feeding of phospholipid-enriched alkyl phospholipid from krill relieves spontaneous atopic dermatitis and strengthens skin intercellular lipid barriers in NC/Nga mice.

Plasmalogen (Pls) is a glycerophospholipid derived from alkyl phospholipid (Alk) with antioxidant functions in vivo. The present study investigated the effects of ether phospholipids, such as Pls and Alk, on intercellular lipid barriers in the skin of NC/Nga mice, a model of atopic dermatitis (AD). NC/Nga mice fed Alk showed increased plasma levels of Alk and Pls. The AD-related changes in ceramide composition in the skin were abrogated by oral administration of Alk. Moreover, Alk suppressed skin inflammation in AD mice. These results indicate that Alk partially fortifies the stratum corneum lipid barrier and may be an effective treatment for AD. Abbreviations: Pls: plasmalogen; PlsCho: choline plasmalogen; PlsEtn: ethanolamine plasmalogen; Alk: alkyl phospholipid; TJ: tight junction; FA: fatty acid; AD: atopic dermatitis; SO: soybean oil; FO: fish oil; DHA: docosahexaenoic acid; EPA: eicosapentaenoic acid; TG: triglyceride; PL: phospholipid; RF: retention factor; AlkCho: choline-type alkyl phospholipid; AlkEtn: ethanolamine-type alkyl phospholipid; LC-MS/MS: liquid chromatography-tandem mass spectrometry; FAR1: fatty acyl-coenzyme (Co)A reductase 1.

The mechanism of skin lipids influencing skin status.

Skin lipids, compose of sebocyte-, keratinocyte-, and microbe- derived lipids, dramatically influence skin status by different mechanisms. (I) Physical chemistry function: They are "mortar" to establish the physico-chemical barrier function of skin; (II) Biochemistry function: They function as signals in the complex signaling network originating at the epidermal level; (III) Microecology function: Sebocyte- and keratinocyte-derived lipids vary the composition of microbial skin flora, and microorganisms metabolize them to produce lipids as signal starting signaling transduction. Importantly, further research needs lipidiomics, more powerful analytical ability and high-throughput manner, to identify skin lipid components into individual species. The validation of lipid structure and function to research the process that lipid species involved in. Additional, the integration of lipidomics data with other omics strategies can develop the power to study the mechanism of skin lipids influencing skin status.

Lipidomic platform for structural identification of skin ceramides with α-hydroxyacyl chains.

Skin ceramides are sphingolipids consisting of sphingoid bases, which are linked to fatty acids via an amide bond. Typical fatty acid acyl chains are composed of α-hydroxy fatty acid (A), esterified ω-hydroxy fatty acid (EO), non-hydroxy fatty acid (N), and ω-hydroxy fatty acid (O). We recently established a lipidomic platform to identify skin ceramides with non-hydroxyacyl chains using tandem mass spectrometry. We expanded our study to establish a lipidomic platform to identify skin ceramides with α-hydroxyacyl chains. Tandem mass spectrometry analysis of A-type ceramides using chip-based direct infusion nanoelectrospray-mass spectrometry showed the characteristic fragmentation pattern of both acyl and sphingoid units, which can be applied for structural identification of ceramides. Based on the tandem mass spectrometry fragmentation patterns of A-type ceramides, comprehensive fragmentation schemes were proposed. Our results may be useful for identifying A-type ceramides in the stratum corneum of human skin.

Contribution of human-related sources to indoor volatile organic compounds in a university classroom.

Although significant progress has been made in understanding the sources and chemistry of indoor volatile organic compounds (VOCs) during the past decades, much is unknown about the role of humans in indoor air chemistry. In the spring of 2014, we conducted continuous measurements of VOCs using a proton transfer reaction mass spectrometer (PTR-MS) in a university classroom. Positive matrix factorization (PMF) of the measured VOCs revealed a 'human influence' component, which likely represented VOCs produced from human breath and ozonolysis of human skin lipids. The concentration of the human influence component increased with the number of occupants and decreased with ventilation rate in a similar way to CO2 , with an average contribution of 40% to the measured daytime VOC concentration. In addition, the human skin lipid ozonolysis products were observed to correlate with CO2 and anticorrelate with O3 , suggesting that reactions on human surfaces may be important sources of indoor VOCs and sinks for indoor O3 . Our study suggests that humans can substantially affect VOC composition and oxidative capacity in indoor environments.

Epidermal hydration and skin surface lipids in patients with long-term complications of sulfur mustard poisoning.

BACKGROUND: Despite almost the three decades passed since the chemical attacks of Iraqi's army against the Iranian troops, some veterans are still suffering from long-term complications of sulfur mustard (SM) poisoning, including certain skin complaints specially dryness, burning, and pruritus. We thus aimed to evaluate the skin's water and lipid content in patients with a disability of >25% due to complications of SM poisoning and compare them with a matched control group. MATERIALS AND METHODS: Sixty-nine male participants were included in this study; 43 SM-exposed patients, and 26 normal controls from their close relatives. The water and lipid content was measured in four different locations: Extensor and flexor sides of forearms and lateral and medial sides of legs by the Corneometer CM 820/Sebumeter SM 810. Collected data was analyzed and P ≤ 0.05 was considered as statistically significant. RESULTS: The mean age of the patients and controls was 49.53 ± 11.34 (ranges: 40-71) and 29.08 ± 8.836 (ranges: 15-49 years), respectively. In the veterans group, the main cutaneous complaint was itching and skin dryness. Cherry angioma, dry skin, and pruritus were significantly more common in the SM-exposed cases than in the controls. (P = 0.01, 0.05, and 0.04, respectively). The moisture and lipid content of all areas were lower in the SM-exposed group, but it was only significant in skin sebum of lateral sides of legs (P = 0.02). CONCLUSION: Exposure to SM could decrease the function of stratum corneum and lipid production as a barrier, even after several years of its exposure.

Lipid Vesicles for the Skin Delivery of Diclofenac: Cerosomes vs. Other Lipid Suspensions.

PURPOSE: Lipid suspensions as drug carriers, including conventional liposomes, ethosomes, transferosomes, proniosomes, niosomes, PEG-PPG-PEG niosomes and stratum corneum liposomes (cerosomes), were formulated and compared. METHODS: Lipid vesicles were formulated and assessed with regards to enhancement of skin permeation of diclofenac and stability profiles of the formulations. Formulation-induced changes of the biophysical structure of excised human skin were monitored using the Fourier transform infrared spectroscopy. RESULTS: The stability profiles of these suspensions over 12 weeks did not show any significant drug leakage from the vesicles of interest (p > 0.05). FTIR observations indicated that the vesicles increased stratum corneum (SC) lipid fluidization and altered protein conformation. Skin permeability experiments showed that the free unencapsulated drug in the cerosomal formulations caused significant increase in drug permeation across the skin (p < 0.01). Low skin permeability of drug from the other lipid suspensions could be due to the entrapment of diclofenac within these vesicles which decreased the solubility of the hydrophilic drug in the skin lipids and the partition coefficient of the drug from these vesicles into the SC. CONCLUSION: Optimal drug entrapment in vesicles or alteration of the skin structure may not necessarily enhance the permeation of hydrophilic drugs across the human skin. These lipid vesicles may be further developed into carriers of both hydrophilic and hydrophobic drugs for topical and transdermal delivery, respectively.

1H NMR-based lipidomics of rodent fur: species-specific lipid profiles and SCD1 inhibitor-related dermal toxicity.

A method is described that allows noninvasive identification and quantitative assessment of lipid classes present in sebaceous excretions in rodents. The method relies on direct high-field proton NMR analysis of common group lipid protons in deuterated organic solvent extracts of fur. Extracts from as little as 15 mg of fur from rat, mouse, and hamster provided acceptable results on a 600 MHz NMR equipped with a cryogenically cooled proton-observe probe. In rats, sex- and age-related differences in lipid composition are larger than differences in fur collected from various body regions within an individual and much larger than interanimal differences in age- and sex-matched specimens. The utility of this method to noninvasively monitor drug-induced sebaceous gland atrophy in rodents is demonstrated in rats dosed with a stearoyl-CoA desaturase 1 (SCD1) inhibitor. In this model, a 35% reduction in sebum lipids, extracted from fur, was observed. Finally, structural elucidation of cholesta-7,24-dien-3ß-ol ester as the most prominent, previously unidentified sebum sterol ester in male Syrian hamsters is described. The utility of this method for drug and cosmetic safety and efficacy assessment is discussed.

Enhancement of skin radical scavenging activity and stratum corneum lipids after the application of a hyperforin-rich cream.

Hyperforin is well-known for its anti-inflammatory, anti-tumor, anti-bacterial, and antioxidant properties. The application of a hyperforin-rich verum cream could strengthen the skin barrier function by reducing radical formation and stabilizing stratum corneum lipids. Here, it was investigated whether topical treatment with a hyperforin-rich cream increases the radical protection of the skin during VIS/NIR irradiation. Skin lipid profile was investigated applying HPTLC on skin lipid extracts. Furthermore, the absorption- and scattering coefficients, which influence radical formation, were determined. 11 volunteers were included in this study. After a single cream application, VIS/NIR-induced radical formation could be completely inhibited by both verum and placebo showing an immediate protection. After an application period of 4weeks, radical formation could be significantly reduced by 45% following placebo application and 78% after verum application showing a long-term protection. Furthermore, the skin lipids in both verum and placebo groups increased directly after a single cream application but only significantly for ceramide [AP], [NP1], and squalene. After long-term cream application, concentration of cholesterol and the ceramides increased, but no significance was observed. These results indicate that regular application of the hyperforin-rich cream can reduce radical formation and can stabilize skin lipids, which are responsible for the barrier function.

Four characteristics and a model of an effective tear film lipid layer (TFLL).

It is proposed that a normal, effective tear film lipid layer (TFLL) should have the following four characteristics: 1) high evaporation resistance to prevent water loss and consequent hyperosmolarity; 2) respreadability, so it will return to its original state after the compression-expansion cycle of the blink; 3) fluidity sufficient to avoid blocking secretion from meibomian glands; 4) gel-like and incompressible structure that can resist forces that may tend to disrupt it. These characteristics tend to be incompatible; for example, lipids that form good evaporation barriers tend to be disrupted by compression-expansion cycles. It is noted that clues about the function and organization of the TFLL can be obtained by comparison with other biological lipid layers, such as lung surfactant and the lipid evaporation barrier of the skin. In an attempt to satisfy the conflicting characteristics, a "multilamellar sandwich model" of the TFLL is proposed, having features in common with the skin evaporation barrier.