Comparison of rat epidermal keratinocyte organotypic culture (ROC) with intact human skin: Lipid composition and thermal phase behavior of the stratum corneum

The present report is a part of our continuing efforts to explore the utility of the rat epidermal keratinocyte organotypic culture (ROC) as an alternative model to human skin in transdermal drug delivery and skin irritation studies of new chemical entities and formulations. The aim of the present s...

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Published in:	Biochimica et biophysica acta Vol. 1778; no. 4; pp. 824 - 834
Main Authors:	Pappinen, Sari, Hermansson, Martin, Kuntsche, Judith, Somerharju, Pentti, Wertz, Philip, Urtti, Arto, Suhonen, Marjukka
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 01.04.2008
Subjects:	Animals Calorimetry, Differential Scanning Ceramides - analysis Ceramides - chemistry Ceramides - classification Chromatography, Thin Layer DSC Epidermis - chemistry Epidermis - cytology Humans Keratinocytes - chemistry Keratinocytes - cytology Lipid composition Lipids - analysis Lipids - chemistry Mass Spectrometry Models, Biological Organ Culture Techniques Rats Reconstructed epidermis ROC Stratum corneum Temperature TLC FFA NP Reconstructed epidermis Lipid composition Stratum corneum NS AH EOP ROC EOS TLC AP AS DSC REK NH EOH
ISSN:	0005-2736, 0006-3002, 1879-2642
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Abstract	The present report is a part of our continuing efforts to explore the utility of the rat epidermal keratinocyte organotypic culture (ROC) as an alternative model to human skin in transdermal drug delivery and skin irritation studies of new chemical entities and formulations. The aim of the present study was to compare the stratum corneum lipid content of ROC with the corresponding material from human skin. The lipid composition was determined by thin-layer chromatography (TLC) and mass-spectrometry, and the thermal phase transitions of stratum corneum were studied by differential scanning calorimetry (DSC). All major lipid classes of the stratum corneum were present in ROC in a similar ratio as found in human stratum corneum. Compared to human skin, the level of non-hydroxyacid-sphingosine ceramide (NS) was increased in ROC, while α-hydroxyacid-phytosphingosine ceramide (AP) and non-hydroxyacid-phytosphingosine ceramides (NP) were absent. Also some alterations in fatty acid profiles of ROC ceramides were noted, e.g., esterified ω-hydroxyacid-sphingosine contained increased levels of oleic acid instead of linoleic acid. The fraction of lipids covalently bound to corneocyte proteins was distinctly lower in ROC compared to human skin, in agreement with the results from DSC. ROC underwent a lipid lamellar order to disorder transition ( T 2) at a slightly lower temperature (68 °C) than human skin (74 °C). These differences in stratum corneum lipid composition and the thermal phase transitions may explain the minor differences previously observed in drug permeation between ROC and human skin.
AbstractList	The present report is a part of our continuing efforts to explore the utility of the rat epidermal keratinocyte organotypic culture (ROC) as an alternative model to human skin in transdermal drug delivery and skin irritation studies of new chemical entities and formulations. The aim of the present study was to compare the stratum corneum lipid content of ROC with the corresponding material from human skin. The lipid composition was determined by thin-layer chromatography (TLC) and mass-spectrometry, and the thermal phase transitions of stratum corneum were studied by differential scanning calorimetry (DSC). All major lipid classes of the stratum corneum were present in ROC in a similar ratio as found in human stratum corneum. Compared to human skin, the level of non-hydroxyacid-sphingosine ceramide (NS) was increased in ROC, while alpha-hydroxyacid-phytosphingosine ceramide (AP) and non-hydroxyacid-phytosphingosine ceramides (NP) were absent. Also some alterations in fatty acid profiles of ROC ceramides were noted, e.g., esterified omega-hydroxyacid-sphingosine contained increased levels of oleic acid instead of linoleic acid. The fraction of lipids covalently bound to corneocyte proteins was distinctly lower in ROC compared to human skin, in agreement with the results from DSC. ROC underwent a lipid lamellar order to disorder transition (T2) at a slightly lower temperature (68 degrees C) than human skin (74 degrees C). These differences in stratum corneum lipid composition and the thermal phase transitions may explain the minor differences previously observed in drug permeation between ROC and human skin.The present report is a part of our continuing efforts to explore the utility of the rat epidermal keratinocyte organotypic culture (ROC) as an alternative model to human skin in transdermal drug delivery and skin irritation studies of new chemical entities and formulations. The aim of the present study was to compare the stratum corneum lipid content of ROC with the corresponding material from human skin. The lipid composition was determined by thin-layer chromatography (TLC) and mass-spectrometry, and the thermal phase transitions of stratum corneum were studied by differential scanning calorimetry (DSC). All major lipid classes of the stratum corneum were present in ROC in a similar ratio as found in human stratum corneum. Compared to human skin, the level of non-hydroxyacid-sphingosine ceramide (NS) was increased in ROC, while alpha-hydroxyacid-phytosphingosine ceramide (AP) and non-hydroxyacid-phytosphingosine ceramides (NP) were absent. Also some alterations in fatty acid profiles of ROC ceramides were noted, e.g., esterified omega-hydroxyacid-sphingosine contained increased levels of oleic acid instead of linoleic acid. The fraction of lipids covalently bound to corneocyte proteins was distinctly lower in ROC compared to human skin, in agreement with the results from DSC. ROC underwent a lipid lamellar order to disorder transition (T2) at a slightly lower temperature (68 degrees C) than human skin (74 degrees C). These differences in stratum corneum lipid composition and the thermal phase transitions may explain the minor differences previously observed in drug permeation between ROC and human skin. The present report is a part of our continuing efforts to explore the utility of the rat epidermal keratinocyte organotypic culture (ROC) as an alternative model to human skin in transdermal drug delivery and skin irritation studies of new chemical entities and formulations. The aim of the present study was to compare the stratum corneum lipid content of ROC with the corresponding material from human skin. The lipid composition was determined by thin-layer chromatography (TLC) and mass-spectrometry, and the thermal phase transitions of stratum corneum were studied by differential scanning calorimetry (DSC). All major lipid classes of the stratum corneum were present in ROC in a similar ratio as found in human stratum corneum. Compared to human skin, the level of non-hydroxyacid-sphingosine ceramide (NS) was increased in ROC, while alpha-hydroxyacid-phytosphingosine ceramide (AP) and non-hydroxyacid-phytosphingosine ceramides (NP) were absent. Also some alterations in fatty acid profiles of ROC ceramides were noted, e.g., esterified omega-hydroxyacid-sphingosine contained increased levels of oleic acid instead of linoleic acid. The fraction of lipids covalently bound to corneocyte proteins was distinctly lower in ROC compared to human skin, in agreement with the results from DSC. ROC underwent a lipid lamellar order to disorder transition (T2) at a slightly lower temperature (68 degrees C) than human skin (74 degrees C). These differences in stratum corneum lipid composition and the thermal phase transitions may explain the minor differences previously observed in drug permeation between ROC and human skin. The present report is a part of our continuing efforts to explore the utility of the rat epidermal keratinocyte organotypic culture (ROC) as an alternative model to human skin in transdermal drug delivery and skin irritation studies of new chemical entities and formulations. The aim of the present study was to compare the stratum corneum lipid content of ROC with the corresponding material from human skin. The lipid composition was determined by thin-layer chromatography (TLC) and mass-spectrometry, and the thermal phase transitions of stratum corneum were studied by differential scanning calorimetry (DSC). All major lipid classes of the stratum corneum were present in ROC in a similar ratio as found in human stratum corneum. Compared to human skin, the level of non-hydroxyacid-sphingosine ceramide (NS) was increased in ROC, while α-hydroxyacid-phytosphingosine ceramide (AP) and non-hydroxyacid-phytosphingosine ceramides (NP) were absent. Also some alterations in fatty acid profiles of ROC ceramides were noted, e.g., esterified ω-hydroxyacid-sphingosine contained increased levels of oleic acid instead of linoleic acid. The fraction of lipids covalently bound to corneocyte proteins was distinctly lower in ROC compared to human skin, in agreement with the results from DSC. ROC underwent a lipid lamellar order to disorder transition ( T 2) at a slightly lower temperature (68 °C) than human skin (74 °C). These differences in stratum corneum lipid composition and the thermal phase transitions may explain the minor differences previously observed in drug permeation between ROC and human skin.
Author	Kuntsche, Judith Wertz, Philip Suhonen, Marjukka Urtti, Arto Hermansson, Martin Pappinen, Sari Somerharju, Pentti
Author_xml	– sequence: 1 givenname: Sari surname: Pappinen fullname: Pappinen, Sari organization: Department of Pharmaceutics, University of Kuopio, P.O. Box 1627, 70211 Kuopio, Finland – sequence: 2 givenname: Martin surname: Hermansson fullname: Hermansson, Martin organization: Institute of Biomedicine, Department of Biochemistry, University of Helsinki, P.O. Box 63, 00014 Helsinki, Finland – sequence: 3 givenname: Judith surname: Kuntsche fullname: Kuntsche, Judith organization: Department of Pharmaceutics, University of Kuopio, P.O. Box 1627, 70211 Kuopio, Finland – sequence: 4 givenname: Pentti surname: Somerharju fullname: Somerharju, Pentti organization: Institute of Biomedicine, Department of Biochemistry, University of Helsinki, P.O. Box 63, 00014 Helsinki, Finland – sequence: 5 givenname: Philip surname: Wertz fullname: Wertz, Philip organization: Dows Institute, University of Iowa, Iowa City IA 52242, USA – sequence: 6 givenname: Arto surname: Urtti fullname: Urtti, Arto organization: Department of Pharmaceutics, University of Kuopio, P.O. Box 1627, 70211 Kuopio, Finland – sequence: 7 givenname: Marjukka surname: Suhonen fullname: Suhonen, Marjukka email: Marjukka.Suhonen@uku.fi organization: Department of Pharmaceutics, University of Kuopio, P.O. Box 1627, 70211 Kuopio, Finland
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Keywords	FFA NP Reconstructed epidermis Lipid composition Stratum corneum NS AH EOP ROC EOS TLC AP AS DSC REK NH EOH
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Snippet	The present report is a part of our continuing efforts to explore the utility of the rat epidermal keratinocyte organotypic culture (ROC) as an alternative...
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SubjectTerms	Animals Calorimetry, Differential Scanning Ceramides - analysis Ceramides - chemistry Ceramides - classification Chromatography, Thin Layer DSC Epidermis - chemistry Epidermis - cytology Humans Keratinocytes - chemistry Keratinocytes - cytology Lipid composition Lipids - analysis Lipids - chemistry Mass Spectrometry Models, Biological Organ Culture Techniques Rats Reconstructed epidermis ROC Stratum corneum Temperature TLC
Title	Comparison of rat epidermal keratinocyte organotypic culture (ROC) with intact human skin: Lipid composition and thermal phase behavior of the stratum corneum
URI	https://dx.doi.org/10.1016/j.bbamem.2007.12.019 https://www.ncbi.nlm.nih.gov/pubmed/18211819 https://www.proquest.com/docview/70438255
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