Abstract
The main barrier of the skin is formed by the lipids in the apical skin layer, the stratum corneum (SC). In SC mainly ceramides (CER), free fatty acids (FFA) and cholesterol (CHOL) are present. The CER are composed of at least six different fractions. CER 1 has an exceptional molecular structure as it contains a linoleic acid linked to a long-chain ω-hydroxy acid (C > 30). The SC lipids are organized in two lamellar phases with periodicities of approximately 6 and 13 nm, respectively. Recent studies revealed that ceramides isolated from pig SC mixed with cholesterol in confined ratios mimic stratum corneum lipid phase behavior closely (Bouwstra, J.A., et al. 1996. J. Lipid Res. 37: 999–1011). In this paper the role of CER 1 for the SC lipid lamellar organization was studied. For this purpose lipid phase behavior of mixtures of CHOL and total ceramide fraction was compared with that of mixtures of CHOL and a ceramide mixture lacking CER 1. These studies showed that in the absence of CER 1 almost no long periodicity phase was formed over a wide CHOL/CER molar ratio. A model is proposed for the molecular arrangement of the two lamellar phases. This model is based on the dominant role CER 1 plays in the formation of the long periodicity phase, electron density distribution calculations, and observations, such as i) the bimodal distribution of the fatty acid chain lengths of the ceramides, ii) the phase separation between long-chain ceramides and short-chain ceramides in a monolayer approach, and iii) the absence of swelling of the lamellae upon increasing the water content organization in SC. In this molecular model the short periodicity phase is composed of only two high electron density regions indicating the presence of only one bilayer, similar to that often found in phospholipid membranes. The molecular arrangement in the long periodicity phase is very exceptional. This phase most probably consists of two broad and one narrow low electron density regions. The two broad regions are formed by partly interdigitating ceramides with long-chain fatty acids of approximately 24–26 C atoms, while the narrow low-electron density region is formed by fully interdigitating ceramides with a short free fatty acid chain of approximately 16 to 18 C atoms.—Bouwstra, J. A., G. S. Gooris, F. E. R. Dubbelaar, A. M. Weerheim, A. P. IJzerman, and M. Ponec. Role of ceramide 1 in the molecular organization of the stratum corneum lipids. J. Lipid Res. 1998. 39: 186–196.
Highlights
The main barrier of the skin is formed by the lipids in the apical skin layer, the stratum corneum (SC)
The CER(2–6) or CER(1–6) mixtures were subsequently mixed with CHOL at varying molar ratios or in equimolar ratios with both CHOL and free fatty acids (FFA)
In a recent study in which the phase behavior of mixtures prepared from CHOL and CER(1–6) was examined (15), we found that over a large range of CHOL/ CER(1–6) molar ratios, two lamellar phases were present with repeat distances of 5.2 and 12.2 nm, respectively
Summary
Fresh pig skin was obtained from a slaughter house and SC was isolated from the skin as described before (15). Epidermal lipids were extracted using the method of Bligh and Dyer (20). The extracted lipids were applied on a silica gel 60 (Merck) column with a diameter of 2 cm and a length of 33 cm. The various lipid classes were eluted sequentially using various solvent mixtures as published recently (15). The lipid composition of collected fractions was established by one-dimensional high performance thin-layer chromatography, as described before (21). Authentic standards (Sigma) were run in parallel. The quantification was performed after charring using a photodensitometer with automatic peak integration (Desaga, Germany). Isolated fractions were mixed to achieve mixtures of CER(2–6) or mixtures of CER(1–6)
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