Role of ceramides 2 and 5 in the structure of the stratum corneum lipid barrier

We utilized Fourier transform infrared (FTIR) spectroscopy to investigate headgroup and chain interactions in model SC lipid barriers containing equimolar amounts of deuterated hexadecanoic acid, cholesterol, and ceramide 2 (non-hydroxy sphingosine) or ceramide 5 (alpha-hydroxy sphingosine). In the ceramide 2 model the thermotropic response of the CD _ 2 and CH _ 2 stretching modes indicates that hexadecanoic acid begins to disorder at 42 degrees C while ceramide 2 remains ordered until 52 degrees C. Additionally, splitting of the CD _ 2 bending and CH _ 2 rocking modes provides evidence for separate orthorhombic hexadecanoic acid and ceramide domains. The ceramide amide I mode (1650 cm ; -1) is split into two components indicating strong intermolecular hydrogen bonding between headgroups. In the ceramide 5 model, the CH _ 2 and CD _ 2 stretching frequencies again reveal highly conformationally ordered ceramide 5 and hexadecanoic acid chains. Splitting of both the CD _ 2 bending and CH _ 2 rocking modes is observed. However, the CH _ 2 rocking frequencies indicate distorted packing of the ceramide. The collapse of these highly ordered phases, and the onset of conformational disorder, occurs at 50 degrees C for both ceramide 5 and hexadecanoic acid. The amide I and II frequencies of ceramide 5 indicate strong H-bonding, although neither mode is split. Our results demonstrate that model SC lipid systems have quite different physical properties depending on whether they contain ceramide 2 or 5. From this we infer that ceramide 2 and 5 make distinct contributions to the structural biophysics of the SC lipid barrier. Our observation of ordered lipid domains is also consistent with the recently proposed domain mosaic model of the skin barrier.     

The chemistry,function and (patho)physiology of stratum corneum barrier ceramides

Research on understanding of the chemistry, function and (patho)physiology of stratum corneum (SC) lipids and especially ceramides has evolved over the last two decades. This has been made successful through the application of separation techniques that have become increasingly more sophisticated, and it has become increasingly evident that our understanding of these molecules remains in its infancy. Thirteen classes of ceramides with over 300 and possibly up to 1000 distinct ceramide species have been identified suggesting an exquisitely subtle relationship between the types of ceramides and physical and chemical behaviour. Nevertheless, research has demonstrated the importance of the correct SC lipid lamellar architecture with conformationally‐ordered lipid bilayers, the presence of long‐chain ceramides, as either free or covalently bound lipids, greater quantities of phytosphingosine‐containing ceramides and a high SC lipid/protein ratio is essential for optimal barrier function. These features are known to change in a variety of physiological and pathophysiological conditions. Clearly, there is more to be learned but as we further decipher the complexity of SC lipids and understand their individual roles in the SC, we will learn how to better treat the disorders of cornification.     

Superior effect of isostearyl isostearate on improvement in stratum corneum water permeability barrier function as examined by the plastic occlusion stress test

Dry skin is a major dermatological problem and consumer research indicates that although current moisturizers are effective they are not completely meeting consumer expectation. Several technological approaches have been taken but influencing stratum corneum (SC) lipid phase behaviour as a novel water permeability barrier‐enhancing and moisturizing mechanism has only been started to be investigated recently. Both the long periodicity SC lipid lamellar phase and the orthorhombic lipid packing state have been proposed to define optimal SC water permeability barrier properties. Several lipophillic moisturizers have been tested for their ability to modify SC lipid lateral packing namely glyceryl monoisostearate (GMIS), isopropyl isostearate (IPIS) and isostearyl isostearate (ISIS) of which IPIS and ISIS are reported to induce the orthorhombic phase. Despite the improvements in the lateral packing of SC lipids, these ingredients have been shown not to improve transepidermal water loss. However, using a novel skin surface water loss method we have observed for the first time significant improvements in SC water permeability barrier function for ISIS compared with IPIS, GMIS and petrolatum. However, using synthetic membranes and measuring water vapour transport rates we showed that the isostearyl esters were not occlusive like petrolatum. As the effects of ISIS were not because of what would be considered as true occlusion, we propose that the differences in the SC water permeability barrier properties from use of ISIS to the other ingredients tested are because of its reported effects on SC lipid phase behaviour. Further studies probably using spectroscopic approaches, however, will be needed to specifically test this hypothesis in vivo.     

Dry skin and impairment of barrier function associated with itch - new insights

Itch is a common symptom in dry skin related to inflammatory skin diseases, normal aging, and systemic diseases such as chronic renal failure, and HIV. However, correlations between itch and objective measures of barrier function and skin dryness such as skin hydration and transepidermal water loss have been rarely found. Recent experimental evidence indicates that damage to the stratum corneum with acetone/ether and water elicits a scratching response in mice and rats. These responses correlate to the number of PGP 9.5 immunoreactive fibers in the epidermis and to FOS-like immunoreactivity in the spinal cord. Other neuromediators involved in the pathogenesis of itch in dry skin are nerve growth factor (NGF), muscarinic acetylcholine receptors, and opiates. Serine proteases such as tryptase and their respective proteinase-activating receptor 2 (PAR2), recently found in both skin and nerves of patients with atopic eczema, suggest that these molecules may have a role in itch in dry skin. This has also been exemplified in the itchy and hyperkeratotic phenotype of the stratum corneum chymotryptic enzyme (SCCE) transgenic mouse model, which is over-expressing a serine protease. Developing inhibitors to these neuropeptides and mediators may be an attractive strategy for anti-itch treatment. The significant progress made in development of moisturizers may have an additional benefit in reducing the itch associated with dry skin. Formulating topical combination therapies containing moisturizers and anti-pruritics can significantly reduce the itch associated with dry skin. This paper will review the current clinical knowledge on the association between dry skin and itch and the recent advances in understanding the pathophysiology of this problem.     

Non-invasive assessment of stratum corneum structure and function

Methods for the study of the stratum corneum in situ are described and their application to clinical problems and cosmetic science are discussed. In vivo measurements of the thickness of skin and the components of skin can be made using high frequency pulsed ultrasound. Improved resolution of the device should allow measurement of epidermal thickness, and even that of the stratum corneum, which may vary with hydration. The protective ability of the stratum corneum can be estimated in a number of ways, the most familiar technique being transepidermal water loss measurements. Mechanical properties of the stratum corneum have been widely investigated in vivo by numerous researchers. A recent technique devised by us to deform the stratum corneum using an extendable metal frame has been used on patients with ichthyotic disorders. The deformation or compliance of the skin surface has been measured using profilometric methods, and compared to normal individuals. The results appear to be in agreement with the loss of flexural ability experienced by ichthyotic patients. Other mechanical methods more usually involve the measurement of the forces generated by skin while undergoing extension. Three techniques are described which have been used to investigate hydrational change with emollient application. Finally, the phenomenon of desquamation is discussed and methods presented for its evaluation. Passive collection of corneocytes using chambers is useful for estimation of the rates of desquamation, but are laborious and inconvenient. Forced desquamation using the desquamator enables rapid controlled harvesting of corneocytes. In addition to this, the dansyl chloride fluorescence technique has been adopted for estimation of stratum corneum kinetics using visual comparison and fluorimetric techniques. These techniques require further refinement and validation, but the tests available now are nonetheless important in understanding the biology and pharmacology of the stratum corneum.     

The role of sebum and epidermal lipids in the cosmetic properties of skin

Lipids found in the skin are derived both from the keratinized epidermis and from the sebaceous glands. The composition of human sebum and epidermal lipids has only been fully elucidated during the past few years, and the role of these lipids is still being evaluated. This presentation reviews sebaceous gland structure and function composition of human sebum, the effects of aging and hormones on sebum secretion and the role of sebum in acne and in dry skin. In addition, a review of the role of epidermal lipids in the properties of skin includes consideration of the structure and function of the epidermis, the composition of the epidermal lipids, and the function of lipids in epidermal differentia-tion and water barrier properties.
Le rle du sébum et des lipides de I'épiderme dans les propriétes cosmétiques de la peau     

Stratum corneum keratin structure, function and formation - a comprehensive review

A comprehensive review on stratum corneum keratin organization, largely based on the recently published cubic rod-packing and membrane templating model [J. Invest. Dermatol., 123, 2004, 715], is presented. Keratin is the major non-aqueous component (wt/wt) of stratum corneum. As 90-100% of the stratum corneum water is thought to be located intracellularly one may presume that keratin also is a major factor (together with filaggrin-derived free amino acids) determining stratum corneum hydration level and water holding capacity. This water holding capacity depends in turn on the structural organization of the corneocyte keratin intermediate filament network. The cubic rod-packing model for the structure and function of the stratum corneum cell matrix postulates that corneocyte keratin filaments are arranged according to a cubic-like rod-packing symmetry. It is in accordance with the cryo-electron density pattern of the native corneocyte keratin matrix and could account for the swelling behaviour and the mechanical properties of mammalian stratum corneum. The membrane templating model for keratin dynamics and for the formation of the stratum corneum cell matrix postulates the presence in viable epidermal cellular space of a highly dynamic small lattice parameter (<30 nm) membrane structure with cubic-like symmetry, to which keratin is associated. It further proposes that membrane templating, rather than spontaneous self-assembly, is responsible for keratin intermediate filament formation and dynamics. It is in accordance with the cryo-electron density patterns of the native keratinocyte cytoplasmic space and could account for the characteristic features of the keratin network formation process, the dynamic properties of keratin intermediate filaments, the close lipid association of keratin, the insolubility in non-denaturating buffers and pronounced polymorphism of keratin assembled in vitro, and the measured reduction in cell-volume and hydration level between stratum granulosum and stratum corneum.     

甘薯渣多糖提取、结构鉴定及体外功能研究

利用水提醇沉法提取甘薯渣多糖（SPRP）,分析多糖基本结构,测定其抗氧化能力、降血糖能力及清除亚硝酸盐阻断亚硝胺合成作用。结果表明：最佳工艺条件为料液比1:30 g/mL、提取时间5.5 h、提取温度80℃,多糖得率5.614%。经测定,SPRP的相对分子质量为2.04×105 g/mol。单糖组成的物质的量比为甘露糖：鼠李糖：葡萄糖：半乳糖：木糖=7.34:1.08:11.56:15.14:1,是α-D-吡喃型多糖。SPRP的体外抗氧化活性和降血糖能力较好,其清除ABTS＋·的IC50为0.881mg/mL,抑制α-葡萄糖苷酶和α-淀粉酶的IC50分别为1.116 mg/mL、1.326 mg/mL。当质量浓度为2.0mg/mL时,亚硝酸盐清除率为81.02%,亚硝胺阻断率为81.63%。     

Purification, characterization, and primary structure of a novel N-acyl-d-amino acid amidohydrolase from Microbacterium natoriense TNJL143-2

A novel N-acyl-d-amino acid amidohydrolase (DAA) was purified from the cells of a novel species of the genus Microbacterium. The purified enzyme, termed AcyM, was a monomeric protein with an apparent molecular weight of 56,000. It acted on N-acylated hydrophobic d-amino acids with the highest preference for N-acetyl-d-phenylalanine (NADF). Optimum temperature and pH for the hydrolysis of NADF were 45°C and pH 8.5, respectively. The k(cat) and K(m) values for NADF were 41?s(-1) and 2.5?mM at 37°C and pH 8.0, although the enzyme activity was inhibited by high concentrations of NADF. Although many known DAAs are inhibited by 1?mM EDTA, AcyM displayed a 65% level of its full activity even in the presence of 20?mM EDTA. Based on partial amino acid sequences of the purified enzyme, the full-length AcyM gene was cloned and sequenced. It encoded a protein of 495 amino acids with a relatively low sequence similarity to a DAA from Alcaligenes faecalis DA1 (termed AFD), a binuclear zinc enzyme of the α/β-barrel amidohydrolase superfamily. The unique cysteine residue that serves as a ligand to the active-site zinc ions in AFD and other DAAs was not conserved in AcyM and was replaced by alanine. AcyM was the most closely related to a DAA of Gluconobacter oxydans (termed Gox1177) and phylogenetically distant from AFD and all other DAAs that have been biochemically characterized thus far. AcyM, along with Gox1177, appears to represent a new phylogenetic subcluster of DAAs.     

Facial skin mapping: from single point bio‐instrumental evaluation to continuous visualization of skin hydration,barrier function,skin surface pH,and sebum in different ethnic skin types

Dry skin is one of the most important concerns of consumers worldwide. Despite huge efforts over several decades, the personal care industry still does not offer a perfect solution to satisfy the unmet needs of consumers for moisturising treatments in different ethnic groups. The paucity of data for the underlying cellular and biochemical problems in, and the effects of moisturisers on photodamaged facial skin may partly explain this. Mainly, single point measurements are used to understand the effects of products on skin physiology even on surrogate skin sites such as the non‐photodamaged volar forearm. Some groups have developed discontinuous facial maps of skin biophysical properties, however, in 2014 a continuous facial analysis of bio‐instrumental evaluations was developed using a heat map approach. These maps enabled a continuous visualization of features that not only revealed an unexpected complexity of facial skin but also indicated that use of surrogate skin sites for facial skin is inappropriate. We have demonstrated that remarkable gradients of skin hydration, TEWL, skin surface pH and sebum exist within short distances across the face and the gradients are distinctive among different ethnic groups. In addition, these studies have demonstrated that darkly‐pigmented individuals do not necessarily have a better skin barrier function than their less‐pigmented counterparts and that Caucasians have a lower facial skin surface pH compared with more pigmented subjects. Overall, there are no correlations between capacitance, TEWL and skin surface pH including individual topology angle values. Novel 3D camera approaches have also been used to facilitate a more precise assignment of measurement sites and visualisation. The 3D facial colour mappings illustrated precisely the local moisturising effects of a moisturising cream. There were subtle ethnic differences in efficacy that may be related to underlying skin biochemistry and/or ethnic differences in product application. A placebo‐controlled study using conductance measurements in Chinese subjects is also reported. Finally, a new whole face statistical approach has been taken to prove differences in skin parameters but also of moisturiser treatment that adds further to our understanding of the ethnic differences in skin physiology and product application. This paper reviews the background of the development and application of this methodology.     

Lemon (Citrus limon, Burm.f.) essential oil enhances the trans-epidermal release of lipid-(A, E) and water-(B6, C) soluble vitamins from topical emulsions in reconstructed human epidermis

Topical bioavailability of lipid‐ and water‐soluble vitamins is a critical issue for protecting or anti‐ageing formulations. Using 17‐day‐old SkinEthic^® reconstructed human epidermis, we investigated (at 34°C) the role of lemon EO in enhancing the penetration of α‐tocopherol (E) and retinyl acetate (A), pyridoxine (B₆) and ascorbic acid (C), released from O/W or W/O emulsions. D‐limonene, α‐pinene and p‐cymene (65.9, 2.2 and 0.5%w/w of the oil) had skin permeability coefficients Ps (10^?3 cm h^?1) of 0.56 ± 0.03 (or 0.73 ± 0.02), 0.72 ± 0.05 (or 0.98 ± 0.05) and 0.84 ± 0.04 (or 1.14 ± 0.04), respectively, when incorporated in a W/O (or O/W) emulsion. Vitamins B6, C and A had Ps values of (3.0 ± 0.4) × 10^?3, (7.9 ± 0.6) × 10^?3 and (0.37 ± 0.02) × 10^?5 cm h^?1, respectively, and their flux through the skin was enhanced by a factor of 4.1, 3.4 and 5.8, respectively, in the presence of lemon EO. The penetration of vitamin E was nine‐fold enhanced. Lemon EO produced only reversible modification of TEWL, and it is a safe and effective penetration enhancer for topical administration of lipid‐ and water‐soluble vitamins.