A convenient and highly sensitive method for quantification of ceramide in tape-stripping samples by dot-blot analysis as a skin barrier assessment technique in murine macerated skin

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  • Mugita Yuko
    Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo
  • Minematsu Takeo
    Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo
  • Huang Lijuan
    Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo
  • Sugama Junko
    Department of Clinical Nursing, Graduate School of Medical Science, Kanazawa University
  • Ichikawa Yoshie
    Department of Clinical Nursing, Graduate School of Medical Science, Kanazawa University
  • Nakagami Gojiro
    Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo
  • Nagase Takashi
    Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo
  • Oe Makoto
    Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo
  • Noguchi Hiroshi
    Department of Life Support Technology（Molten）, Graduate School of Medicine, The University of Tokyo
  • Mori Taketoshi
    Department of Life Support Technology（Molten）, Graduate School of Medicine, The University of Tokyo
  • Sanada Hiromi
    Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo

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  • 浸軟皮膚における簡便かつ高感度な皮膚バリア機能評価法：テープストリッピングにより採取したマウス角質セラミドのドットブロット法による定量的解析

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Abstract

<p>　Accurate assessment of skin barrier function is important to provide appropriate care for barrier-impaired skin and to prevent skin diseases. Several methods of skin barrier assessment, including measurement of transepidermal water loss（TEWL）and quantification of ceramide collected by tape stripping, have been developed. However, TEWL measurement is highly susceptible to failure due to environmental factors, particularly moisture, and the present tape-stripping technique is too irritating for barrier-impaired skin. Thus, existing skin barrier assessment methods are not suitable, especially for maceration-induced barrier-impaired skin. We aimed to establish a minimally irritating, convenient, highly sensitive method of skin barrier assessment and to apply dot-blot analysis to the quantification of ceramide collected by a modified tape-stripping technique. Macerated skin conditions were reproduced in the dorsal skin of male C57BL/6 mice by the attachment of filter papers soaked with normal saline supplemented with Tween 20（0%, 0.1%, or 10%）. Thirty minutes after removal of the filter papers, skin barrier function was assessed in macerated and normal skin by TEWL measurement. Tape stripping was then performed on a smaller area than that used in the existing tape-stripping technique, and the amount of ceramide in the tape-stripping sample was quantified by dot-blot analysis and incubation using antibodies to ceramide. With skin barrier impairment by the skin maceration treatment, both TEWL measurements and ceramide signal intensities in our method were increased, and these two variables were significantly correlated. These results indicate that our ceramide quantification method is useful as a minimally irritating, convenient, highly sensitive skin barrier assessment technique.</p>

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