Barrier Characteristics of Different Human Skin Types Investigated with X-Ray Diffraction, Lipid Analysis, and Electron Microscopy Imaging

Schreiner, Volker; Gooris, Gert S.; Pfeiffer, Stephan; Lanzendörfer, Ghita; Wenck, Horst; Diembeck, W.; Proksch, Ehrhardt; Bouwstra, Joke A.

doi:10.1046/j.1523-1747.2000.00941.x

Cited by 122 publications

(92 citation statements)

References 27 publications

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“…This suggests that both NLS-ethanol and magainin reduced cholesterol crystallinity, which reduces lipid bilayer order . [26] Small-angle XRD on untreated skin revealed a number of characteristic peaks, notably corresponding to ceramides (d = 6.13 nm) and crystalline cholesterol (d = 3.38 nm) [27]. Treatment with NLS-ethanol reduced both the ceramide and crystalline cholesterol peaks and addition of magainin reduced them still farther.…”

Section: X-ray Diffractionmentioning

confidence: 99%

Transdermal delivery enhanced by magainin pore-forming peptide

Kim

Ludovice

Prausnitz

2007

Journal of Controlled Release

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In this study we tested the hypothesis that magainin, a peptide known to form pores in bacterial cell membranes, can increase skin permeability by disrupting stratum corneum lipid structure. We further hypothesized that magainin's enhancement requires co-administration with a surfactant chemical enhancer to increase magainin penetration into the skin. In support of these hypotheses, exposure to a known surfactant chemical enhancer, N-lauroyl sarcosine (NLS), in 50% ethanol solution increased in vitro skin permeability to fluorescein 15 fold and the combination of magainin and NLS-ethanol synergistically increased skin permeability 47 fold. In contrast, skin permeability was unaffected by exposure to magainin without co-enhancement by NLS-ethanol. Furthermore, confocal microscopy showed that magainin in the presence of NLS-ethanol penetrated deeply and extensively into stratum corneum, whereas magainin alone penetrated poorly into the skin. Additional analysis by Fouriertransform infrared spectroscopy, X-ray diffraction, and differential scanning calorimetry showed that NLS-ethanol disrupted stratum corneum lipid structure and that the combination of magainin and NLS-ethanol disrupted stratum corneum lipids even further. Altogether, these data suggest that NLSethanol increased magainin penetration into stratum corneum, which further increased stratum corneum lipid disruption and skin permeability. We believe this is the first study to demonstrate the use of a pore-forming peptide to increase skin permeability. This study also introduces the novel concept of using a first chemical enhancer to increase penetration of a second chemical enhancer into the skin to synergistically increase skin permeability to a model drug.

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Section: X-ray Diffractionmentioning

confidence: 99%

Transdermal delivery enhanced by magainin pore-forming peptide

Kim

Ludovice

Prausnitz

2007

Journal of Controlled Release

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“…This model, however, does not explicitly address the conformation of the acylceramides, which are known to be necessary for proper arrangement of the SC barrier. Some authors have in fact proposed that the strength of the 4.5 nm scattering peak of SC is linked to the content of the acylceramide EOH ( 7 ). A number of other models of lipid organization also have been proposed earlier to explain scattering patterns (11)(12)(13)(14)(15).…”

Section: Lipid Structural Variations Across the Scmentioning

confidence: 99%

“…Most of the SAXS studies of human SC fi nd refl ections corresponding to ‫ف‬ 4.5 nm and ‫ف‬ 6.5 nm inter-bilayer spacings ( 4,6,7 ). Some studies have also detected a refl ection corresponding to a spacing of 11-13.5 nm ( 6,8 ), referred to as the "long periodicity phase" ( 6 ).…”

mentioning

confidence: 99%

Micron-scale assessment of molecular lipid organization in human stratum corneum using microprobe X-ray diffraction

Doucet

Potter

Baltenneck

et al. 2014

Journal of Lipid Research

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This article is available online at http://www.jlr.org complex and specifi c architecture of the inter-corneocyte lipid matrix ( 2 ) composed of a mixture of ceramides, cholesterol, and long-chain fatty acids. Ultrastructural studies, using electron microscopy and diffraction techniques, have revealed that these lipids are organized in stacked bilayers that are predominantly parallel to the skin surface ( 3-5 ). Part of the lipids is covalently bound to the corneocytes whereas another part represents the "free lipids" removable by solvent extraction.X-ray diffraction is an excellent technique for studying the spatial organization of the intercellular matrix because the lipid molecules are highly ordered and oriented in a crystalline structure. This technique has provided a number of important results with regard to the SC structure. In particular, small-angle X-ray scattering (SAXS) gives information on the inter-bilayer distance in the multi-lamellar lipid structures, whereas the wide-angle X-ray scattering (WAXS) data contain information about the in-plane crystalline arrangement of lipids (lattice type).Most of the SAXS studies of human SC fi nd refl ections corresponding to ‫ف‬ 4.5 nm and ‫ف‬ 6.5 nm inter-bilayer spacings ( 4, 6, 7 ). Some studies have also detected a refl ection corresponding to a spacing of 11-13.5 nm ( 6, 8 ), referred to as the "long periodicity phase" ( 6 ). These data indicate that the SC lipid matrix has a complex organization into multiple sublayers or even phase-separated domains with different lamellar repeat distances.Similar complexity is found at the molecular level using WAXS, where two sets of peaks are found, corresponding to orthorhombic (peaks at 0.41 and 0.37 nm) and hexagonal (peak at 0.41 nm) packing of the lipid chains. In addition, a fraction of lipids might exist in a fl uid or amorphous state, which would then contribute to the broad band corresponding to an average distance of 0.46 nm. Abstract Lipid and protein components of the stratum corneum (SC) are organized in complex supramolecular arrangements. Exploring spatial relations between various possible substructures is important for understanding the barrier function of this uppermost layer of epidermis. Here, we report the fi rst study where micro-focus X-ray scattering was used for assessing fi ne structural variations of the human skin barrier with micrometer resolution. We found that the scattering profi les were unchanged when scanning in the direction parallel to the SC surface. Furthermore, small-angle scattering profi les did not change as a function of depth in the SC, confi rming that the lipid lamellar spacings remained the same throughout the SC. However, the wide-angle scattering data showed that the orthorhombic phase was more abundant in the middle layers of the SC, whereas the hexagonal phase dominated in the surface layers both at the external and the lowest part of the SC; i.e., the lipids were most tightly packed in the middle region of the SC. Taken together, our results demonstrate that micro...

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“…The predominant chain lengths of these -hydroxy fatty acids are 32-carbon saturated as well as 32-and 34-carbon monounsaturated species that are acylated with linoleic acid via the -OH position. This unique epidermal lipid has been shown to be important for both the lamellar organization of the barrier lipids (6,7) and for the formation of a lipid monolayer covalently bound to surface proteins of the corneocytes (3,8,9).…”

mentioning

confidence: 99%

Sphingolipid metabolism during epidermal barrier development in mice

Doering

Brade

Sandhoff

2002

Journal of Lipid Research

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In rodents, a competent skin barrier to water loss is formed within 2 or 3 days prior to birth. Acquisition of barrier function during rat gestation correlates with the formation of a stratum corneum enriched in ceramides, cholesterol, and fatty acids (Aszterbaum, M., G. K. Menon, K. R. Feingold, and M. L. Williams. 1992. Ontogeny of the epidermal barrier to water loss in the rat: correlation of function with stratum corneum structure and lipid content. Pediatr. Res. 31: 308-317). We analyzed the formation and epidermal localization of glucosylceramides during embryonic skin barrier development in Balb/c mice. Using immunohistochemistry, epidermal glucosylceramides were hardly detectable 3 days prior to birth. After further 24 h of gestation the level of glucosylceramides was maximal and decreased with increasing gestational age. In parallel, glucosylceramides were targeted to the apical side of the outermost granular keratinocyte layer. A spectrum of five distinct epidermal ceramides was present 2 days prior to birth. With ongoing gestation the composition of the ceramide fraction changed markedly. Most importantly, the level of -hydroxylated acylceramides decreased paralleled by the formation of the corneocyte lipid envelope. This structure consists of -hydroxylated ceramides and fatty acids bound to surface proteins of the corneocytes. The covalent attachment of ceramides or glucosylceramides correlated with the maturation of the stratum corneum and might contribute to its chemical and enzymatic resistance. -Doering, T., H. Brade, and K. Sandhoff. Sphingolipid metabolism during epidermal barrier development in mice.

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Barrier Characteristics of Different Human Skin Types Investigated with X-Ray Diffraction, Lipid Analysis, and Electron Microscopy Imaging

Cited by 122 publications

References 27 publications

Transdermal delivery enhanced by magainin pore-forming peptide

Transdermal delivery enhanced by magainin pore-forming peptide

Micron-scale assessment of molecular lipid organization in human stratum corneum using microprobe X-ray diffraction

Sphingolipid metabolism during epidermal barrier development in mice

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