title: An ex vivo human skin model for studying skin barrier repair.
abstract: In the studies described in this study, we introduce a novel ex vivo human skin barrier repair model. To develop this, we removed the upper layer of the skin, the stratum corneum (SC) by a reproducible cyanoacrylate stripping technique. After stripping the explants, they were cultured in vitro to allow the regeneration of the SC. We selected two culture temperatures 32 °C and 37 °C and a period of either 4 or 8 days. After 8 days of culture, the explant generated SC at a similar thickness compared to native human SC. At 37 °C, the early and late epidermal differentiation programmes were executed comparably to native human skin with the exception of the barrier protein involucrin. At 32 °C, early differentiation was delayed, but the terminal differentiation proteins were expressed as in stripped explants cultured at 37 °C. Regarding the barrier properties, the SC lateral lipid organization was mainly hexagonal in the regenerated SC, whereas the lipids in native human SC adopt a more dense orthorhombic organization. In addition, the ceramide levels were higher in the cultured explants at 32 °C and 37 °C than in native human SC. In conclusion, we selected the stripped ex vivo skin model cultured at 37 °C as a candidate model to study skin barrier repair because epidermal and SC characteristics mimic more closely the native human skin than the ex vivo skin model cultured at 32 °C. Potentially, this model can be used for testing formulations for skin barrier repair.
authors.affiliation: Department of Drug Delivery Technology, Leiden Academic Centre for Drug Research, Leiden University, Leiden, The Netherlands.
authors.forename: Mogbekeloluwa O
authors.initials: MO
authors.lastname: Danso
authors.forename: Tineke
authors.initials: T
authors.lastname: Berkers
authors.forename: Arnout
authors.initials: A
authors.lastname: Mieremet
authors.forename: Farzia
authors.initials: F
authors.lastname: Hausil
authors.forename: Joke A
authors.initials: JA
authors.lastname: Bouwstra
mesh_terms: D001706:Biopsy; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D003487:Cyanoacrylates / Q000737:chemistry; D004817:Epidermis / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D008055:Lipids / Q000737:chemistry*; D009924:Organ Culture Techniques / Q000379:methods*; D010539:Permeability; D010619:Phenazines / Q000737:chemistry; D012867:Skin / Q000378:metabolism* / Q000473:pathology; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D014961:X-Ray Diffraction
chemical_list: D003487:Cyanoacrylates; D008055:Lipids; D010619:Phenazines; C009195:safranine T
keywords: epidermal differentiation; lipid composition; lipid organization; skin barrier repair; stratum corneum