Influence of short chain ceramides and lipophilic penetration enhancers on the nano-structure of stratum corneum model membranes studied using neutron diffraction

Abstract

Oriented stratum corneum model lipid membranes were used to study the influence of the short chain ceramides (CER)[NP] and [AP] as well as the impact of the lipophilic penetration enhancer molecules oleic acid (OA) and isopropyl myristate (IPM) on the lipid nanostructure. The influence of the enhancer molecules were studied using specifically deuterated OA and IPM and neutron diffraction. ²H NMR spectroscopy was used to study the impact of the ceramides’ degree of order within the stratum corneum model lipid membranes. It was found that CER[NP] forms two very stable phases with high resistance against temperature increase. Phase B showed unusual hydration behavior as no water uptake of this phase was observed. The ²H NMR spectroscopic measurements showed that CER[NP] based ternary model system had a higher state of lamellar order in comparison to CER[AP] based lipid matrix.

The studies confirmed that the short chain ceramides, particularly CER[NP], have a very high impact on the integrity of the Stratum corneum lipid bilayers. The penetration enhancer OA has not influenced the repeat distance of the model membrane based on CER[AP], and was not able to induce a phase separation in the investigated lipid matrix. However, a disorder and a fluidisation of the model membranes were observed when OA was incorporated. IPM showed the same effect but two phases (assigned as phase A and B) appeared, when IPM was used as penetration enhancer and incorporated into the model membrane. Furthermore, two arrangements of IPM were identified in phase A using deuterated IPM. A model of the nanostructure of the Stratum corneum lipid membranes is presented.

Key words： nano-structure of the stratum corneum; ceramide; penetration enhancer; model membrane; neutron diffraction; ²H NMR spectroscopy

References

1 Kessner D, Kiselev M A, Dante S, Hauss T, Lersch P, Wartewig S, Neubert R H H. Arrangement of ceramide [EOS] in a stratum corneum lipid model matrix: new aspects revealed by neutron diffraction studies. European Biophysics Journal with Biophysics Letters, 2008, 37(6): 989–999

2 Kessner D, Kiselev M A, Hauss T, Dante S, Wartewig S, Neubert R H H. Localisation of partially deuterated cholesterol in quaternary SC lipid model membranes: a neutron diffraction study. European Biophysics Journal with Biophysics Letters, 2008, 37(6): 1051–1057

3 Kiselev M A. Conformation of ceramide 6 molecules and chain-flip transitions in the lipid matrix of the outermost layer of mammalian skin, the stratum corneum. Crystallography Reports, 2007, 52(3): 525–528

4 Kiselev M A, Ryabova N Y, Balagurov A M, Dante S, Hauss T, Zbytovska J, Wartewig S, Neubert R H H. New insights into the structure and hydration of a stratum corneum lipid model membrane by neutron diffraction. European Biophysics Journal, 2005, 34(8): 1030–1040

5 Wegener M, Neubert R H H, Rettig W, Wartewig S. Structure of stratum corneum lipids characterized by FT-Raman spectroscopy and DSC. III. Mixtures of ceramides and cholesterol. Chemistry and Physics of Lipids, 1997, 88(1): 73–82

6 Zbytovska J, Vavrova K, Kiselev M A, Lessieur P, Wartewig S, Neubert R H H. The effects of transdermal permeation enhancers on thermotropic phase behaviour of a stratum corneum lipid model. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 2009, 351(1-3): 30–37

7 Schroeter A, Kiselev M A, Hauss T, Dante S, Neubert R H H. Evidence of free fatty acid interdigitation in stratum corneum model membranes based on ceramide [AP] by deuterium labelling. Biochimica et Biophysca Acta. Biomembranes, 2009, 1788(10): 2194

8 Engelbrecht T N, Deme B, Dobner B, Neubert R H H. Study of influence of penetration enhancer isopropylmyristate on the nanostructure of stratum corneum lipid model membranes using neutron diffraction and deuterium labelling. Skin Pharmacology and Physiology, 2012, 25(4): 200–207

9 Engelbrecht T N, Schroeter A, Hauss T, Neubert R H H. Lipophilic penetration enhancers and their impact to the bilayer structure of stratum corneum lipid model membranes: neutron diffraction studies based on the example oleic acid. Biochimica et Biophysica Acta. Biomembranes, 2011, 1808(12): 2798–2806

10 Holbrook K A, Odland G F. Regional differences in the thickness (cell layers) of the human stratum corneum: an ultrastructural analysis. Journal of Investigative Dermatology, 1974, 62(4): 415–422

11 Scheuplein R J, Morgan L J. “Bound water” in keratin membranes measured by a microbalance technique. Nature, 1967, 214(5087): 456–458

12 Loomans M E, Hannon D P. An electron microscopic study of the effects of subtilisin and detergents on human stratum corneum. Journal of Investigative Dermatology, 1970, 55(2): 101–114

13 Blank I H, Scheuplein R J. Transport into and within skin. British Journal of Dermatology, 1969, S 81: 4–10

14 Mackenzie J C. Ordered structure of the stratum corneum of mammalian skin. Nature, 1969, 222(5196): 882

15 Candi E, Schmidt R, Melino G. The cornified envelope: a model of cell death in the skin. Nature Reviews. Molecular Cell Biology, 2005, 6(4): 328–340

16 Elias P M. Epidermal lipids, barrier function, and desquamation. Journal of Investigative Dermatology, 1983, 80(1s Suppl): 44s–49s

17 Elias P M. Structure and function of the stratum-corneum permeability barrier. Drug Development Research, 1988, 13(2-3): 97–105

18 Elias P M, Friend D S. The permeability barrier in mammalian epidermis. Journal of Cell Biology, 1975, 65(1): 180–191

19 Elias P M, Goerke J, Friend D S. Mammalian epidermal barrier layer lipids–composition and influence on structure. Journal of Investigative Dermatology, 1977, 69(6): 535–546

20 Landmann L. The epidermal permeability barrier. Anatomy and Embryology, 1988, 178(1): 1–13

21 El Maghraby G M, Barry B W, Williams A C. Liposomes and skin: from drug delivery to model membranes. European Journal of Pharmaceutical Sciences, 2008, 34(4-5): 203–222

22 Gray G, Yardley H. Different populations of pig epidermal-cells–isolation and lipid composition. Journal of Lipid Research, 1975, 16(6): 441–447

23 Gray G, Yardley H. Lipid compositions of cells isolated from pig, human, and rat epidermis. Journal of Lipid Research, 1975, 16(6): 434–440

24 Wertz P W, van den Bergh B. The physical, chemical and functional properties of lipids in the skin and other biological barriers. Chemistry and Physics of Lipids, 1998, 91(2): 85–96

25 Law S L, Wertz P W, Swartzendruber S C A, Squier C A. Regional variation in content, composition and organization of porcine epithelial barrier lipids revealed by thin-layer chromatography and transmission electron microscopy. Archives of Oral Biology, 1995, 40(12): 1085–1091

26 Di Nardo A, Wertz P, Giannetti A, Seidenari S. Ceramide and cholesterol composition of the skin of patients with atopic dermatitis. Acta Dermato-Venereologica, 1998, 78(1): 27–30

27 Motta S, Monti M, Sesana S, Mellesi L, Ghidoni R, Caputo R. Abnormality of water barrier function in psoriasis. Role of ceramide fractions. Archives of Dermatology, 1994, 130(4): 452–456

28 Madison K C, Swartzendruber D C, Wertz P W, Downing D T. Presence of intact intercellular lipid lamellae in the upper layers of the stratum corneum. Journal of Investigative Dermatology, 1987, 88(6): 714–718

29 Swartzendruber D C, Wertz P W, Kitko D J, Madison K C, Downing D T. Molecular models of the intercellular lipid lamellae in mammalian stratum corneum. Journal of Investigative Dermatology, 1989, 92(2): 251–257

30 Forslind B. A domain mosaic model of the skin barrier. Acta Dermato-Venereologica, 1994, 74(1): 1–6

31 Bouwstra J A, Gooris G S, van der Spek J A, Bras W. Structural investigations of human stratum corneum by small-angle X-ray scattering. Journal of Investigative Dermatology, 1991, 97(6): 1005–1012

32 Bouwstra J A, Dubbelaar F E, Gooris G S, Ponec M. The lipid organisation in the skin barrier. Acta Dermato-Venereologica. Supplementum, 2000, 208(Suppl): 23–30

33 White S H, Mirejovsky D, King G I. Structure of lamellar lipid domains and corneocyte envelopes of murine stratum corneum. An X-ray diffraction study. Biochemistry, 1988, 27(10): 3725–3732

34 Iwai I, Han H, Hollander L D, Svensson S, Ofverstedt L G, Anwar J, Brewer J, Bloksgaard M, Laloeuf A, Nosek D, Masich S, Bagatolli L A, Skoglund U, Norlen L. The human skin barrier is organized as stacked bilayers of fully extended ceramides with cholesterol molecules associated with the ceramide sphingoid moiety. Journal of Investigative Dermatology, 2012, 132(9): 2215–2225

35 Trommer H, Neubert R H H. Overcoming the stratum corneum: The modulation of skin penetration–a review. Skin Pharmacology and Physiology, 2006, 19(2): 106–121

36 Barry B W. Mode of action of penetration enhancers in human skin. Journal of Controlled Release, 1991, 15(3): 237–248

37 Nagle J F, Tristram-Nagle S. Structure of lipid bilayers. Biochimica et Biophysica Acta, 2000, 1469(3): 159–195

38 Seul M, Sammon M J. Reparation of surfactant multilayer films on solid substrates by deposition from organic solution. Thin Solid Films, 1990, 185(2): 287–305

39 Rowat A C, Kitson N, Thewalt J L. Interactions of oleic acid and model stratum corneum membranes as seen by ²H NMR. International Journal of Pharmaceutics, 2006, 307(2): 225–231

40 Sternin E, Bloom M, Mackay A L. De-pakeing of NMR spectra. Journal of Magnetic Resonance (San Diego, Calif.), 1983, 55(2): 274–282

41 Davis J H, Jeffrey K R, Bloom M, Valic M I, Higgs T P. Quadrupolar echo deuteron magnetic resonance spectroscopy in ordered hydrocarbon chains. Chemical Physics Letters, 1976, 42(2): 390–394

42 Davis J H, Jeffrey K R. Temperature dependence of chain disorder in potassium palmitate-water –A deuterium NMR study. Chemistry and Physics of Lipids, 1977, 20(2): 87–104

43 Takeuchi Y, Yamaoka Y, Fukushima S, Miyawaki K, Taguchi K, Yasukawa H, Kishimoto S, Suzuki M. Skin penetration enhancing action of cis-unsaturated fatty acids with omega-9, and omega-12-chain lengths. Biological & Pharmaceutical Bulletin, 1998, 21(5): 484–491

44 Tonegawa A, Michiue A, Masuda T, Ohno T, Matsuura H, Yamada K, Okuda T. Deuterium NMR and Raman spectroscopic studies on conformational behavior of lipophilic chains in the C12E3/decane/water system. Zeitschrift für Naturforschung A–A Journal of Physical Sciences, 2002, 57(6-7): 320–326

45 Engelbrecht T N, Schroeter A, Hauss T, Deme B, Scheidt H A, Huster D, Neubert R H H. The impact of ceramides NP and AP to nanostructure of stratum corneum lipid model membranes. Part I: Neutron diffraction and ²H NMR studies on multilamellar models based on ceramides with symmetric alkyl chain length distribution. Soft Matter, 2012, 8(24): 6599–6607

46 Engelbrecht T N, Hauss T, Suss K, Vogel A, Roark M, Feller S E, Neubert R H H, Dobner B. Characterisation of a new ceramide EOS species: Synthesis, investigation of the thermotropicphase behaviour and influence on the bilayer architecture of stratum corneum lipid model membranes. Soft Matter, 2011, 7(19): 8998–9011

47 Mitriaikina S, Muller-Goymann C C. Synergetic effects of isopropyl alcohol (IPA) and isopropyl myristate (IPM) on the permeation of betamethasone-17-valerate from semisolid Pharmacopoeia bases. Journal of Drug Delivery Science and Technology, 2007, 17(5): 339–346