Lipid and ultrastructural characterization of reconstructed skin models.
The study aimed at evaluating tissue architecture and quality of the permeability barrier in commercially available reconstructed human skin models; EpiDerm, SkinEthic and Episkin in comparison to native tissue. For this purpose, tissue architecture was examined by electron microscopy and epidermal lipid composition was analyzed by HPTLC. Stratum corneum lipid organization was investigated by electron microscopy in combination with RuO(4) post-fixation and by SAXD. Ultrastructurally, the overall tissue architecture showed high similarities with native epidermis. In the stratum corneum extracellular space, lipid lamellae consisting of multiple alternating electron-dense and electron-lucent bands were present. This regular pattern was not seen throughout the whole stratum corneum probably due to the observed irregular lamellar body extrusion in some areas. Lipid analyses revealed the presence of all major epidermal lipid classes. Compared with native epidermis the content of polar ceramides 5 and 6 was lower, ceramide 7 was absent, and the content of free fatty acids was very low. These differences in lipid composition may account for differences observed in SAXD pattern of Episkin and EpiDerm penetration models. In the latter only the long-distance periodicity unit of about 12 nm was observed and the short periodicity unit was missing. In conclusion, all three skin models provide a promising means for studying the effects of topically applied chemicals, although the observed deviations in tissue homeostasis and barrier properties need to be optimized.
Ponec
M
M
Sylvius Laboratory, Department of Dermatology, Leiden University Medical Center, P.O. Box 9503, 2300 RA, Leiden, The Netherlands. ponec@mail.medfac.leidenuniv.nl
Boelsma
E
E
Weerheim
A
A
Mulder
A
A
Bouwstra
J
J
Mommaas
M
M
D001485:Basement Membrane / Q000648:ultrastructure; D002518:Ceramides / Q000032:analysis; D005230:Fatty Acids, Nonesterified / Q000032:analysis; D006801:Humans; D008055:Lipids / Q000032:analysis*; D010743:Phospholipids / Q000032:analysis; D012867:Skin / Q000737:chemistry / Q000648:ultrastructure*; D014280:Triglycerides / Q000032:analysis; D014961:X-Ray Diffraction
D002518:Ceramides; D005230:Fatty Acids, Nonesterified; D008055:Lipids; D010743:Phospholipids; D014280:Triglycerides