Cutting Edge of the Pathogenesis of Atopic Dermatitis: Sphingomyelin Deacylase,the Enzyme Involved in Its Ceramide Deficiency,Plays a Pivotal Role

Atopic dermatitis (AD) is characterized clinically by severe dry skin and functionally by both a cutaneous barrier disruption and an impaired water-holding capacity in the stratum corneum (SC) even in the nonlesional skin. The combination of the disrupted barrier and water-holding functions in nonlesional skin is closely linked to the disease severity of AD, which suggests that the barrier abnormality as well as the water deficiency are elicited as a result of the induced dermatitis and subsequently trigger the recurrence of dermatitis. These functional abnormalities of the SC are mainly attributable to significantly decreased levels of total ceramides and the altered ceramide profile in the SC. Clinical studies using a synthetic pseudo-ceramide (pCer) that can function as a natural ceramide have indicated the superior clinical efficacy of pCer and, more importantly, have shown that the ceramide deficiency rather than changes in the ceramide profile in the SC of AD patients plays a central role in the pathogenesis of AD. Clinical studies of infants with AD have shown that the barrier disruption due to the ceramide deficiency is not inherent and is essentially dependent on postinflammatory events in those infants. Consistently, the recovery of trans-epidermal water loss after tape-stripping occurs at a significantly slower rate only at 1 day post-tape-stripping in AD skin compared with healthy control (HC) skin. This resembles the recovery pattern observed in Niemann–Pick disease, which is caused by an acid sphingomyelinase (aSMase) deficiency. Further, comparison of ceramide levels in the SC between before and after tape-stripping revealed that whereas ceramide levels in HC skin are significantly upregulated at 4 days post-tape-stripping, their ceramide levels remain substantially unchanged at 4 days post-tape-stripping. Taken together, the sum of these findings strongly suggests that an impaired homeostasis of a ceramide-generating process may be associated with these abnormalities. We have discovered a novel enzyme, sphingomyelin (SM) deacylase, which cleaves the N-acyl linkage of SM and glucosylceramide (GCer). The activity of SM deacylase is significantly increased in AD lesional epidermis as well as in the involved and uninvolved SC of AD skin, but not in the skin of patients with contact dermatitis or chronic eczema, compared with HC skin. SM deacylase competes with aSMase and β-glucocerebrosidase (BGCase) to hydrolyze their common substrates, SM and GCer, to yield their lysoforms sphingosylphosphorylcholine (SPC) and glucosylsphingosine (GSP), respectively, instead of ceramide. Consistently, those reaction products (SPC and GSP) accumulate to a greater extent in the involved and uninvolved SC of AD skin compared with chronic eczema or contact dermatitis skin as well as HC skin. Successive chromatographies were used to purify SM deacylase to homogeneity with a single band of ≈43 kDa and with an enrichment of >14,000-fold. Analysis of a protein spot with SM deacylase activity separated by 2D-SDS-PAGE using MALDI-TOF MS/MS allowed its amino acid sequence to be determined and to identify it as the β-subunit of acid ceramidase (aCDase), an enzyme consisting of α- and β-subunits linked by amino-bonds and a single S-S bond. Western blotting of samples treated with 2-mercaptoethanol revealed that whereas recombinant human aCDase was recognized by antibodies to the α-subunit at ≈56 and ≈13 kDa and the β-subunit at ≈43 kDa, the purified SM deacylase was detectable only by the antibody to the β-subunit at ≈43 kDa. Breaking the S-S bond of recombinant human aCDase with dithiothreitol elicited the activity of SM deacylase with an apparent size of ≈40 kDa upon gel chromatography in contrast to aCDase activity with an apparent size of ≈50 kDa in untreated recombinant human aCDase. These results provide new insights into the essential role of SM deacylase as the β-subunit aCDase that causes the ceramide deficiency in AD skin.     

5 nm Silver Nanoparticles Amplify Clinical Features of Atopic Dermatitis in Mice by Activating Mast Cells

The triggering effect of silver nanoparticles (NPs) on the induction of allergic reactions is evaluated, by studying the activation of mast cells and the clinical features of atopic dermatitis in a mouse model. Granule release is induced in RBL‐2H3 mast cells by 5 nm, but not 100 nm silver NPs. Increases in the levels of reactive oxygen species (hydrogen peroxide and mitochondrial superoxide) and intracellular Ca⁺⁺ in mast cells are induced by 5 nm silver NPs. In a mouse model of atopic dermatitis induced by a mite allergen, the skin lesions are more severe and appear earlier in mice treated simultaneously with 5 nm silver NPs and allergen compared with mice treated with allergen alone or 100 nm silver NPs and allergen. The histological findings reveal that number of tryptase‐positive mast cells and total IgE levels in the serum increase in mice treated with 5 nm silver NPs and allergen. The results in this study indicate that cotreatment with 5 nm silver NPs stimulates mast cell degranulation and induces earlier and more severe clinical alterations in allergy‐prone individuals.     

Development of Phospholipids Vesicular Nanocarrier for Topical Delivery of Tea Tree Oil in Management of Atopic Dermatitis Using BALB/c Mice Model

This work aims to develop and evaluate the efficacy of tea tree oil (TTO) ethosomal cream with improved deposition in skin layers for treatment of atopic dermatitis (AD). Ethosomes of TTO are developed using phosphatidylcholine (2% and 3% w/v) and ethanol (20%, 30%, and 40% w/v). Ethosomes are evaluated for percent entrapment efficiency (%EE), vesicle size, zeta potential, and in vitro drug diffusion. Ethosomal creams with optimized ethosomal dispersion are developed and evaluated for physicochemical parameters, thermal stability, ex vivo permeation, skin retention, and in vitro cytotoxicity using HaCat skin cell lines in comparison to conventional creams of TTO. In vivo investigations of optimized creams are performed using BALB/c mice model. The %EE, vesicle size, and zeta potential for optimized ethosomes are found to be 76.19 ± 3.26%, 333.6 nm, and –35.3 mV, respectively. Ethosomal creams showed higher deposition in the epidermis and dermis. The optimized creams are non-cytotoxic to HaCat cell lines. The creams significantly reduce the inflammatory response by decreasing the clinical score and infiltration of white blood cells, eosinophils, and IgE antibodies. Overall efficacy of ethosomal cream is higher than conventional cream. In conclusion, optimized ethosomal cream of TTO shows good efficacy for treatment of AD. Practical applications : The method used for the formulation of ethosomes is simple and can be easily scaled up on the industrial level. The loading of TTO within ethosomes can increase the efficiency by enhanced drug deposition in the epidermis and might also improve its stability against oxidative degradation. Topical ethosomal cream of TTO can improve patient compliance by avoidance of adverse effects linked with corticosteroids and could be a possible complementary or alternative therapy in management of AD.     

Mesenchymal stem cells and cell-free preparations for treating atopic dermatitis

Atopic dermatitis (AD) is a chronic cutaneous inflammatory disease caused by an interaction between genetic, immune and epidermal barrier factors. Several treatments can be used to treat this disease but there are patients that do not respond to actual drugs. So, there is a need to develop effective therapies for AD. Mesenchymal stem cells (MSCs) are non-hematopoietic multipotent adult progenitor cells with immunomodulatory power and self-regenerating capacity to repair tissue damage, so they could be a potential effective treatment for AD. MSCs-Conditioned Medium (CM) and MSCs-exosomes are cell-free preparation with molecules secreted by stem cells that could be also beneficial for AD. This viewpoint reviews the actual development of MSCs, MSCs-CM and MSCs-exosomes for treating patients with AD.     

‘Nickel Allergy’ arising from decorative nickel plated and alloyed articles: prevention at source

This work builds on papers published in Transactions during 2015 and 2018 reporting research into low-cost commercial methods for the prevention of nickel release from decorative nickel plated articles, rendering them suitable for placement on the European market in accordance with the requirements of REACH. ‘Nickel Allergy’ sometimes occurs when nickel-containing articles are in direct and prolonged contact with the skin, leading to corrosion of elemental nickel by sweat, liberating sufficient nickel ions to be absorbed through the skin and initiate an allergenic effect. The EU ‘Nickel Restrictions’ impose limits on the amount of nickel released from articles intended for use in this application, but permits a non-nickel surface coating that can ensure the rate of nickel release does not exceed 0.5?µg?cm^?2 week^?1 after 2 years of normal use. The official tests for coated items are simulated wear and corrosion under EN 12472 followed by determination of nickel release under EN 1811. Earlier work concluded that suitable barrier coatings over bright electrodeposited nickel are regular chromium deposited from a hexavalent electrolyte, microporous trivalent chromium from a chloride electrolyte and UV cured PU electrophoretic coatings. Further tests reported here focused on nickel release from examples of wearable articles such as costume jewellery and watch cases. A typical flash coating of gold over bright nickel is thin and porous and being more noble, causes the rate of nickel release to be accelerated; but this can be prevented by an intermediate barrier coating of electrodeposited palladium. To round out the relevance of this study on wearable articles, nickel release tests were also conducted on nickel-containing Grades 304 (UNS S30400) and 316 (UNS S31600) austenitic stainless steels, plus a typical gold alloy containing nickel. All passed the nickel release tests satisfactorily.     

Mild and severe udder cleft dermatitis—Prevalence and risk factors in Swedish dairy herds

Udder cleft dermatitis (UCD) is an inflammatory skin condition affecting the anterior parts of the udder of dairy cows. The lesions may present as mild or severe skin lesions and have been associated with mastitis and digital dermatitis. The full etiology and pathogenesis are not understood and no large-scale studies have investigated prevalence and risk factors. Therefore, the main objectives of the study were to investigate the prevalence of mild and severe UCD in Swedish dairy herds and to identify risk factors associated with such lesions. We also wanted to investigate risk factors for all cases of UCD and to determine whether UCD increases the risk for mastitis and culling. A random sample of 100 freestall dairy herds were included in the study, and each herd was visited once. Cows were registered as having no, mild, or severe UCD. Additional cow and herd data were obtained via observations, interviews, and the Swedish Official Milk Recording Scheme. The data were analyzed using logistic regression models to identify risk factors for mild and severe UCD. In total, data from 3,479 cows in 99 herds were analyzed. The prevalence of mild and severe UCD was 19 and 9%, respectively. Lesions were found in 98 of 99 herds but the within-herd prevalence of mild (0–43%) and severe (0–33%) UCD varied notably between herds. Breed (Swedish Red compared with Swedish Holstein), certain udder conformation traits, and higher parity were risk factors associated with increased risk of UCD. In addition, cows with hock lesions and cows in herds with high incidence of culling due to hoof and leg diseases had a higher risk for mild UCD. More days in milk and high milk yield were cow-related risk factors associated with severe UCD. Three housing-related factors (shorter cubicles, mattress as cubicle base, and cubicles installed before 2001 compared with 2001–2005), a high incidence of veterinary-treated clinical mastitis and culling due to udder diseases, and a low incidence of culling of first-parity cows in early lactation were herd-related risk factors associated with increased risk for severe UCD. In addition, cows in herds with a high proportion of heifers older than 17 mo that were not inseminated were associated with lower risk of all UCD. Finally, UCD was not associated with the outcomes milk somatic cell count, veterinary-treated clinical mastitis, or culling in the multivariable analyses. The etiology of UCD is most likely multifactorial, involving udder conformation traits and other cow-related risk factors as well as herd-related risk factors. The high prevalence of severe UCD lesions in Swedish dairy cows emphasizes the need for preventive measures and efficient treatments.     

Short Communication: Scoring of Digital Dermatitis During Milking as an Alternative to Scoring in a Hoof Trimming Chute

Digital dermatitis is a serious problem in dairy production in many countries. In many settings, it is important to evaluate the digital dermatitis status of individual cows or an entire dairy herd. Such an evaluation has traditionally been done in a hoof trimming chute. An evaluation in the milking parlor can take place without disturbing the cows to a large extent, it can be done using less labor compared with an evaluation in a hoof trimming chute, and is cheaper than using a chute. The objective was to evaluate the sensitivity and specificity of a rapid screening method for digital dermatitis in the milking parlor, without using any specialized tools and taking approximately 15 s/cow. All lactating cows in 3 commercial Danish dairy herds were included. Cows were first scored for the presence of digital dermatitis during milking and the next day all cows were scored during hoof trimming. A 6-point nominal scoring system based on a visual inspection of the digital dermatitis lesions was used. For the analysis, the scores were dichotomized (digital dermatitis positive or digital dermatitis negative). Additionally, lesions were classified as small (diameter ≤ 2 cm) or large (diameter >2 cm). Sensitivity and specificity were calculated using observations from the hoof trimming chute as the “gold standard” and observations during milking as the diagnostic test. Relatively large variation was found between herds with an overall sensitivity of 0.65 (95% confidence interval: 0.59 to 0.72) and a specificity of 0.84 (0.81 to 0.87). The sensitivity increased to 0.69 (0.62 to 0.76), when only large lesions were assessed. The method has several advantages compared with evaluation in a chute and may be a useful tool in the daily hoof health management in dairy herds.     

Dermatitis-inducing furanocoumarins on leaf surfaces of eight species of Rutaceous and Umbelliferous plants

Eight species of Rutaceae or Umbelliferae, known to cause or suspected of causing photophytodermatitis, had the linear furanocoumarins psoralen, bergapten, and xanthotoxin on their leaf surfaces, in concentrations varying from 0.014 to 1800 /gmg/g fresh weight, equivalent to 0.17–56% of the total leaf concentration. The higher percentage generally observed for spring leaves compared to autumn leaves suggests a higher rate of transfer of these furanocoumarins to the surface in the younger leaves. Among the plants studied,Ruta graveolens had the highest surface concentrations of all three furanocoumarins. The relatively high effectiveness in causing dermatitis of some species with low surface concentrations may be explained by a more effective mechanism of transfer of the furanocoumarins to the skin. A role in the defense of the plant is suggested by their accumulation on the plant surface.A paper based on the work reported here was presented at the Groupe Polyphénols conference, Brock University, St. Catharines, Ontario, Canada, August 19–19, 1988.     

Allergic and toxic reactions caused by cream bases in dermatological patients

Irritant and allergic reactions from cream bases observed in eczema patients are reviewed. The frequency of allergic reactions to the ingredients of vehicles in eczema patients is an indication of allergic potential for normal skin. Against this background, hypoallergenic fragrances are still very difficult to produce and a proper mixture for epicutaneous testing is lacking. Among preservatives, parabens are obviously safer and less allergenic than chlorocresol and sorbic acid. Propylene glycol (PG) is very popular for its many beneficial properties. However, it is also an irritant and a sensitiser. The exact number of allergic delayed type reactions is difficult to establish because primary irritant epicutaneous test reactions often closely resemble allergic reactions. By doing epicutaneous tests with serial dilutions of PG and by making peroral challenge tests, the number of eczema patients allergic to PG has been estimated to be as high as 1%. Glycerol is much less active in producing toxic and allergic skin reactions. Common non-ionic emulsifiers and higher fatty alcohols cause allergic contact dermatitis occasionally. The allergenic properties of these substances have not yet been examined experimentally. It seems, however, that there are only small differences between the various emulsifying agents. Les allergies et réactions de toxicité causées par les constituants des crèmes sur les sujets atteints de dermatoses     

Chlorella vulgaris Attenuates Dermatophagoides Farinae-Induced Atopic Dermatitis-Like Symptoms in NC/Nga Mice

Atopic dermatitis (AD) is a chronic and inflammatory skin disease that can place a significant burden on quality of life for patients. AD most frequently appears under the age of six and although its prevalence is increasing worldwide, therapeutic treatment options are limited. Chlorella vulgaris (CV) is a species of the freshwater green algae genus chlorella, and has been reported to modulate allergy-inducible factors when ingested. Here, we examined the effect of CV supplementation on AD-like symptoms in NC/Nga mice. CV was orally administrated for six weeks while AD-like symptoms were induced via topical application of Dermatophagoides farinae extract (DFE). CV treatment reduced dermatitis scores, epidermal thickness, and skin hydration. Histological analysis also revealed that CV treatment reduced DFE-induced eosinophil and mast cell infiltration into the skin, while analysis of serum chemokine levels indicated that CV treatment downregulated thymus- and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC) levels. In addition, CV treatment downregulated mRNA expression levels of IL-4 and IFN-γ. Taken together, these results suggest that CV extract may have potential as a nutraceutical ingredient for the prevention of AD.