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

Sari Pappinen, Martin Hermansson, Judith Kuntsche, Pentti Somerharju, Philip Wertz, Arto Urtti, Marjukka Suhonen

Research output: Contribution to journalJournal articleResearchpeer-review

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 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.
Original languageEnglish
JournalBBA General Subjects
Volume1778
Issue number4
Pages (from-to)824-34
Number of pages11
ISSN0304-4165
DOIs
Publication statusPublished - 2008

Fingerprint

Keratinocytes
Lipids
Skin
Linoleic Acid
Oleic Acid
Thin Layer Chromatography
Pharmaceutical Preparations
Fatty Acids
Proteins

Keywords

  • Animals
  • Calorimetry, Differential Scanning
  • Ceramides
  • Chromatography, Thin Layer
  • Epidermis
  • Humans
  • Keratinocytes
  • Lipids
  • Mass Spectrometry
  • Models, Biological
  • Organ Culture Techniques
  • Rats
  • Temperature

Cite this

Pappinen, Sari ; Hermansson, Martin ; Kuntsche, Judith ; Somerharju, Pentti ; Wertz, Philip ; Urtti, Arto ; Suhonen, Marjukka. / Comparison of rat epidermal keratinocyte organotypic culture (ROC) with intact human skin : lipid composition and thermal phase behavior of the stratum corneum. In: BBA General Subjects. 2008 ; Vol. 1778, No. 4. pp. 824-34.
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Comparison of rat epidermal keratinocyte organotypic culture (ROC) with intact human skin : lipid composition and thermal phase behavior of the stratum corneum. / Pappinen, Sari; Hermansson, Martin; Kuntsche, Judith; Somerharju, Pentti; Wertz, Philip; Urtti, Arto; Suhonen, Marjukka.

In: BBA General Subjects, Vol. 1778, No. 4, 2008, p. 824-34.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Comparison of rat epidermal keratinocyte organotypic culture (ROC) with intact human skin

T2 - lipid composition and thermal phase behavior of the stratum corneum

AU - Pappinen, Sari

AU - Hermansson, Martin

AU - Kuntsche, Judith

AU - Somerharju, Pentti

AU - Wertz, Philip

AU - Urtti, Arto

AU - Suhonen, Marjukka

PY - 2008

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N2 - 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.

AB - 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.

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KW - Calorimetry, Differential Scanning

KW - Ceramides

KW - Chromatography, Thin Layer

KW - Epidermis

KW - Humans

KW - Keratinocytes

KW - Lipids

KW - Mass Spectrometry

KW - Models, Biological

KW - Organ Culture Techniques

KW - Rats

KW - Temperature

U2 - 10.1016/j.bbamem.2007.12.019

DO - 10.1016/j.bbamem.2007.12.019

M3 - Journal article

C2 - 18211819

VL - 1778

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EP - 834

JO - B B A - General Subjects

JF - B B A - General Subjects

SN - 0304-4165

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ER -