Abstract
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.
