Anthony V. Rawlings, Ph.D. Journal of Investigative Dermatology

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Stratum Corneum Moisturization at the Molecular Level: An Update in Relation to the Dry Skin Cycle Anthony V. Rawlings, Ph.D. Journal of Investigative Dermatology Volume 124, Issue 6, Pages 1099-1110 (June 2005) DOI: 10.1111/j.1523-1747.2005.23726.x Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 – (Left) Visible light macrograph of dry skin on the outer lower leg (approx. 50x), showing lifting squame. (Right) SEM micrograph of carbon tape applied to dry outer lower leg skin (500x); note compacted corneocytes in disarray. Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Typical structure of the epidermis and critical steps in formation of the stratum corneum. Modified from Rawlings, AV; Scott, IR; Harding, CR & Bowser, PA. Stratum Corneum Moisturization at the Molecular Level. J. Invest. Dermatol. 103 (5) 731-740 (1994) and Rawlings, AV & Harding CR. Moisturization & skin barrier function. Dermatologic Therapy. 17: 43-48 (2004). Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Water profile averaged over a single rectangular region of a cryosection obtained from an individual with normal skin, grade 0.5 (A). The horizontal axis is distance across the SC with the SC/granulosum junction indicated by a vertical line. Water profile averaged over a single rectangular region of a cryosection obtained from an individual with dry skin, grade 4 (B). From Warner, RR & Lilly, NA. Correlation of water content with ultrastructure in the stratum corneum. In: Bioengineering of the skin: Water & the stratum corneum. Pp 3-12. Eds: Elsner, P, Berardesca, E & Maibach HI. CRC Press Inc. (1994) Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Structures of human stratum corneum ceramides. Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5a ‘Sandwich model’, the characteristics of which are: (1) the liquid sublattice is located in the central lipid layer of this phase, in this layer mainly unsaturated linoleic acid and cholesterol are present; (2) in the sublattice adjacent to the central layer a gradual change in lipid mobility occurs due to the presence of less mobile long saturated hydrocarbon chains; (3) only a small fraction of lipids forms a fluid phase in the SC, therefore one can assume that this central lipid layer is not a continuous phase. 5b. The liquid phase parallel to the basal layers of the lamellae facilitates transport and therefore communication between the desmosomes. Modified from Bouwstra J, Pilgram G, Gooris G, Koerten H and Ponec M. New aspects of the skin barrier organization. Skin Pharm Appl Skin Physiol 14: 52-62 (2001). Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Organisation of stratum corneum lipids in tape-strippings of individuals with clinically normal skin. Transmission electron micrographs of tape-strippings. Ultrastructural changes in lipid organisation towards the surface of the stratum corneum: A. First strip; absence of bilayers and presence of amorphous lipidic material. B. Second strip; disruption of lipid lamellae. C. Third strip; normal lipid lamellae. (x200,000). From: Rawlings, AV, Watkinson, A, Rogers, J, Mayo, AM, Hope J and IR Scott, Abnormalities in stratum corneum structure, lipid composition and desmosomal degradation in soap-induced winter xerosis, J Soc Cosmet Chem 45 203-220 (1994) Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 7 Electron micrographs of tape strippings of normal skin (grade 1). Degradation of corneodesmosomes (CD) toward the surface of the stratum corneum: A. First strip; CD fully degraded. B. Second strip; CD partially degraded and encapsulated by lipid lamellae. C. Third strip; CD partially degraded, vaculation of structure. D. Third strip, normal CD in contact with lamellar lipids. From: Rawlings, AV, Watkinson, A, Rogers, J, Mayo, AM, Hope J and IR Scott, Abnormalities in stratum corneum structure, lipid composition and desmosomal degradation in soap-induced winter xerosis, J Soc Cosmet Chem 45 203-220 (1994) Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 8 Fluorescence & Normaski phase contrast microscopy of TRIC stained cornified envelopes demonstrating increased fluorescence labelling of Cer compared with CEf. Harding CR, Long S, Richardson J, Rogers J, Zhang Z, Bush A & Rawlings AV. The cornified cell envelope: an important marker of stratum corneum maturation in healthy and dry skin. Int. J. Cosmet Sci. 25:1-11 (2003). Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 9 Double Staining of CE’s with Nile red and anti-involucrin. A. Face. B. Upper Arm. Modified from Hirao T, Denda M and Takahashi M. Identification of immature cornfield envelopes in the barrier-impaired epidermis by characterization of their hydrophobicity and antigenicities of the components. Exp. Dermatol. 10: 35-44, (2001). Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 10 Distribution profile of the maximal compressional forces (uN) of individual CE’s. Top panel shows the force range for Cer and the bottom for CEf. The maximal compression force was significantly different between the corneocytes. Harding CR, Long S, Richardson J, Rogers J, Zhang Z, Bush A & Rawlings AV. The cornified cell envelope: an important marker of stratum corneum maturation in healthy and dry skin. Int. J. Cosmet. Sci. 25:1-11 (2003). Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 11 Schematic representation of profilaggrin catabolism and filaggrin hydrolysis to NMF within the SC. Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 12 Semiquantitative in vivo concentration profiles of NMF and sweat constituents in the stratum corneum of the thenar as determined by Raman spectroscopy. From Caspers PJ, Lucassen GW, Carter EA, Bruining HA and Puppels GJ. In vivo confocal raman microspectroscopy of the skin: non-invasive determination of molecular concentration profiles. J. Invest. Dermatol. 116: 434-442 (2001). Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 13 Organisation of stratum corneum lipids in tape-stripping of subjects with winter xerosis. Transmission electron micrographs of tape strippings of individuals with severe xerosis. Perturbation in lipid organisation towards the surface of the stratum corneum. D. First strip; disorganised lipid lamellae. E. Second strip; disorganisd lipid lamellae. F. Third strip; normal lipid lamellae (x200,000). From: Rawlings, AV, Watkinson, A, Rogers, J, Mayo, AM, Hope J and IR Scott, Abnormalities in stratum corneum structure, lipid composition and desmosomal degradation in soap-induced winter xerosis, J Soc Cosmet Chem 45 203-220 (1994) Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 14 Percentage distribution of Cer and CEf in normal and soap dried dry skin. *p<0.05. Harding CR, Long S, Richardson J, Rogers J, Zhang Z, Bush A & Rawlings AV. The cornified cell envelope: an important marker of stratum corneum maturation in healthy and dry skin. Int. J. Cosmet. Sci. 25:1-11 (2003). Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 15 Schematic diagram showng pivotal events within the "dry skin cycle". Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 16 Results from the treatment phase of a Kligman-type regression study (products applied twice-daily at 2mg/cm2 to randomised sites on the outer, lower leg of female subjects [n=36] with inclusion of a no-treament control). Products represented high-efficacy commercial moisturisers with ingredients of differing dry skin relief mechanism. Key: NT = no treatment control; N = niacinamide-containing lotion; A = lactic acid-containing moisturiser; other product codes represent commercial products with high loadings of traditional humectants and emollients (including glycerin and petrolatum). Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 17 Summary of stratum corneum maturation & corneodesmolysis in normal (A) and dry skin (B). Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 18 Semiquantitative in vivo concentration profiles of water on different body sites. Courtesy of Gerwin Puppels & Peter Caspers at River Diagnostics. Journal of Investigative Dermatology 2005 124, 1099-1110DOI: (10.1111/j.1523-1747.2005.23726.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions