(a) Fig.1 Bricks and mortar model of stratum corneum X Fig.2(a) (b) 1,2) 2 1 (b) Fig.2 Lamellar structure in the intercellular lipid a
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1 Netsu Sokutei Structure and Phase Transitions of Intercellular Lipid Assembly in Stratum Corneum Ichiro Hatta, Kana Nakanishi, and Noboru Ohta (Received June 23, 2007; Accepted July 24, 2007) Stratum corneum which is composed of corneocytes and intercellular lipid matrix plays an important role in barrier function and maintenance of water in skin. The intercellular lipid matrix bears the main part in the function. Therefore, it is important to make clear the structure formed by the intercellular lipids. It is well-known from X-ray diffraction, electron diffraction, neutron scattering, etc. that the structure consists of long and short lamellar structures and in the orthogonal direction hexagonal and orthorhombic hydrocarbon-chain packings. However, until now the relation between each lamellar structure and each hydrocarbon-chain packing has been unresolved. From the thermal analysis and the temperature scanning small- and wide-angle X- ray diffraction analysis in stratum corneum, we revealed that there are two domains: One is the long lamellar structure with the hexagonal hydrocarbon-chain packing and the other the short lamellar structure with the orthorhombic hydrocarbon-chain packing. Based upon the structural evidence we can further study the functional mechanism of cosmetics, the drug penetration mechanism, etc. Keywords: stratum corneum; thermal analysis; X-ray diffraction; lipid; lamellar 20 µm Fig The Japan Society of Calorimetry and Thermal Analysis. 159
2 (a) Fig.1 Bricks and mortar model of stratum corneum X Fig.2(a) (b) 1,2) 2 1 (b) Fig.2 Lamellar structure in the intercellular lipid assembly of stratum corneum. (a) Long lamellar structure. (b) Short lamellar structure. Fig.2(b) 13 nm Fig nm 6nm 1 broad-narrow-broad 3) Fig.2(a) Broad-narrow-broad 1 1 Fig.3 hexagonal orthorhombic 0.42 nm 0.42 nm 0.37 nm Fig nm 0.37 nm 160
3 (a) (b) (a) Fig.3 Hydrocarbon-chain packing in intercellular lipid assembly of stratum corneum. (a) Hexagonal hydrocarbon-chain packing. (b) Orthorhombic hydrocarbon-chain packing. 1 X Fig.4(a) 20 S nm nm nm ) Fig.4(a) X Fig.4(b) 20 S nm nm 2.69 nm nm S nm nm 4) Fig.4(b) S nm 1 S nm 1 5) T / (b) T / S / nm 1 S / nm Fig.4 Simultaneous SAXD and WAXD measurements of the temperature dependence of intensity in hairless mouse stratum corneum. (a) SAXD. (b) WAXD (See Fig.1 in reference 4 in detail). 1 µm 4 25 wt% 6) Fig.4(a)1 2 3 Fig.2(b) 7,8) Fig.5(a) Intensity S 2 (arb.units) Intensity (arb.units)
4 Full Width at Half Maximum / nm 1 Spacing / nm Fig.5 (a) Water Content/ wt% (b) Water Content/ wt% (a) Dependence of spacing on the water content in the stratum corneum for the 2nd order diffraction of the long lamellar structure ( )and for the 1st order diffraction of the short lamellar structure ( ). (b) Full width at half maximum for the above X-ray diffraction peaks. spacing spacing 13.6 nm 2 2 Fig.4(a) 0.15 nm 2 Fig.4(a) Bouwstra 9,10) spacing 20 wt% 5 wt% 20 wt% 3 11) water pool water pool X 8) 5.7 nm 24 h6.2 nm 5.7 nm 6.2 nm Fig.5(a) X X Fig.5(b) X 7) Bouwstra 9,10) X wt% Fig.5(b) X wt% Bouwstra 9,10) X wt% 2) wt% X wt% Fig.5(b) 162
5 Endothermic 2 mj s 1 T / Fig.6 Thermogram for stratum corneum of a hairless mouse on the 1st heating, 1st cooing, 2nd heating and 2nd cooling runs. The scan rate was 10 min 1. X spacing wt% wt% wt% Walkley DSC 6,12-14) DSC 0 Walkley 25 wt% 12) Inoue 29 wt% 13) Takenouchi 28 wt% 14) Imokawa 25 wt% 33.3 % 33.3 wt% [0.333/( )] 100 wt% 6) DSC wt% 3 0 DSC Van Duzee 15) Goldman 16) Goodman 17) Gay 18) Dreher 19) Rehfeld 20) Goldman 21) wt% Van Duzee 15) 40, 75, 85, Gay 18) min 1 22) Fig DSC 1 Fig , 39, 52, 73, DSC 163
6 1 48, min Fig J g J g J g J g ) X 4) 2 Fig.2(a) Fig.2(b) hexagonal orthorhombic Fig.3 X 4) 1 X 1 Fig.4(a) (b) , 39, 51, A. X S nm nm 1 51 Fig.4(a) A B nm 1 6 nm nm 1 5 nm Fig.4(a) B C. 32 S 2.24 nm nm 51 S 2.19 nm nm 32 Fig.6 Fig.4(b) C 22) D S 2.43 nm nm 71 S 2.39 nm nm Fig.4(b) D Fig.4(a) (b) A C Fig.4(a) (b) B D 4 X 120 9,10) 164
7 Intensity S 2 / arb. units S / nm 1 Fig.7 Change of X-ray diffraction profiles with temperature for the 2nd order, 3rd order and 4th order diffractions of the long lamellar structure and for the 1st order diffraction of the short lamellar structure. Fig Fig S nm nm Bouwstra 10 Fig.3 4) X 1) G. S. K. Pilgram and J. A. Bouwstra, Basic and Clinical Dermatology 26, 107 (2004). 2) I. Hatta and N. Ohta, Photon Factory Activity Report 2003 #21, 49 (2004). 3) D. C. Swartzendruber, P. W. Wertz, D. J. Kirko, K. C. Madison, and D. T. Downing, J. Invest. Dermatol. 92, 251 (1989). 4) I. Hatta, N. Ohta, K. Inoue, and N. Yagi, Biochim. Biophys. Acta 1758, 1830 (2006). 5) G. S. K. Pilgram, A. M. Engelsma-van Pelt, J. A. Bouwstra, and H. K. Koerten, J. Invest. Dermatol. 113, 403 (1999). 6) G. Imokawa, H. Kuno, and M. Kawai, J. Invest. Dermatol. 96, 845 (1991). 7) N. Ohta, S. Ban, H. Tanaka, S. Nakata, and I. Hatta, Chem. Phys. Lipids 123, 1 (2003). 8) G. Ch. Charalambopoulou, T. A. Sterotis, Th. Hauss, A. K. Stubos, and N. K. Kanellopoulos, Physica B 165
8 350, e603 (2004). 9) J. A. Bouwstra, G. S. Gooris, J. A. van der Spek, and W. Bras, J. Invest. Dermatol. 97, 1005 (1991). 10) J. A. Bouwstra, G. S. Gooris, J. A. van der Spek, S. Lavrijsen, and W. Bras, Biochim. Biophys. Acta 1212, 183 (1994). 11) J. A. Bouwstra, A. de Graaff, G. S. Gooris, J. Nijsee, J. W. Wiechers, and A. C. van Aelst, J. Invest. Dermatol. 120, 750 (2003). 12) K. Walkeley, J. Invest. Dermatol. 59, 225 (1972). 13) T. Inoue, K. Tujii, K. Okamoto, and K. Toda, J. Invest. Dermatol. 86, 689 (1986). 14) M. Takenouchi, H. Suzuki, and H. Tagami, J. Invest. Dermatol. 87, 574 (1986). 15) B. F. Van Duzee, J. Invest. Dermatol. 65, 404 (1975). 16) G. M. Goldman, D. B. Guzek, R. R. Harris, J. E. Mckie, and R. O. Potts, J. Invest. Dermatol. 86, 255 (1986). 17) M. Goodman and B. W. Barry, Analytical Proceedings 23, 397 (1986). 18) C. L. Gay, R. H. Guy, G. M. Golden, V. H. W. Mak, and M. L. Francoeur, J. Invest. Dermatol. 103, 233 (1994). 19) F. Dreher, P. Walde, P. Walther, and E. Wehrli, J. Control. Release 45, 131 (1997). 20) S. J. Rehfeld and P. M. Elias, J. Invest. Dermatol. 79, 1 (1982). 21) G. M. Goldman, D. B. Guzek, A. H. Kennedy, J. E. McKie, and R. O. Potts, Biochemistry 26, 2382 (1987). 22) I. Hatta, K. Nakanishi, and K. Ishikiriyama, Thermochim. Acta 431, 94 (2005). X Ichiro Hatta, Industrial Application Div., Japan Synchrotron Radiation Research Institute, TEL , FAX , hatta@spring8.or.jp Kana Nakanishi, TORAY Research Center Inc., TEL , FAX , Kana_Nakanishi@trc.toray.co.jp Noboru Ohta, Industrial Application Div., Japan Synchrotron Radiation Research Institute, noboru_o@spring8.or.jp X 166
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