Skin age is one of the biological factors affecting percutaneous absorption. 1,2) To regulate the therapeutic efficacy of transdermal drug delivery, the difference in skin permeability at various ages is a subject of considerable importance. We previously reported that the permeability through the skin decreases with aging, 3,4) and that the extent of this decrease is higher for lipophilic permeants than for hydrophilic permeants. 4) We clarified that the thickness of skin strata is an important factor for limited percutaneous absorption in aging. However, other physiological changes in the skin with age, e.g., moisture content in the skin, lipid composition, lipid content, dermis thickness, and density of skin appendages, have been also reported. [5][6][7][8] Therefore the age-related differences in the skin barrier function must be further investigated based on physiological alterations in the skin to elucidate the mechanisms of the age dependency of transdermal absorption of drugs.There have been many reports on age-related changes in the transdermal permeation of drugs, 3,4,[6][7][8][9] however, age-related changes in skin permeability have not been fully interpreted because of large interindividual variance and the limited number of model permeants. Recently, it was reported that impedance parameters can sensitively reflect the properties of skin permeation pathways. [10][11][12] The impedance technique enables us to determine rapidly changes in skin physical properties, and one measurement provides information on pathways for both hydrophilic and lipophilic permeants. Therefore skin impedance analysis may be a useful method to elucidate the skin barrier function at different ages.In the present study, we attempted to apply impedance analysis to investigate age-dependent changes in permeation pathways of rat skin. In vitro skin impedance data of rats at various ages were compared. Physiological characteristics of the skin, e.g., the water and lipid content of the stratum corneum and the thickness of the skin strata, were also evaluated. Finally, we demonstrated the age dependency of relations between these electrical and physiological properties.
MATERIALS AND METHODS
AnimalsMale STD: Wistar rats at the age of 5, 10, 21, 90, and 180 d (Japan SLC Inc., Hamamatsu, Japan) were used. Rats were fed commercial food pellets and tap water ad libitum. Under sodium pentobarbital anesthesia (50 mg/kg, i.p.), a round section of the abdominal skin was excised, and then the subcutaneous fat and other extraneous tissue were trimmed off carefully. The skin sample was immediately used in the following skin impedance experiments.Skin Impedance Measurement and Analysis A sideby-side, two-chamber electrochemical cell with four electrodes, two for supplying current and two for recording signals, was designed according to the procedure of Burnett and Bagniefski.11) The rat skin was mounted in an O-ring seal between the two chambers, each having 23 ml volume and 1 cm 2 exposed area. The current-supplying electrode consisted of 0....