Disrupted skin barrier, one of the severe attributes of inflammatory skin diseases, is caused by lower content and pathological changes of lipids in the uppermost skin layer-stratum corneum (SC). Restoring skin barrier with native skin lipids, especially ceramides (Cers), appears to be a promising therapy with minimum side effects. For testing the efficiency of these formulations, suitable in vitro models of the skin with disrupted barriers are needed. For the similarity with the human tissue, our models were based on the pig ear skin. Three different ways of skin barrier disruption were tested and compared: tape stripping, lipid extraction with organic solvents, and barrier disruption by sodium lauryl sulfate. The level of barrier disruption was investigated by permeation studies, and parameters of each method were modified to reach significant changes between the non-disrupted skin and our model. Fourier transform infrared (FTIR) spectroscopy was employed to elucidate the changes of the skin permeability on the molecular scale. Further, the potential of the developed models to be restored by skin barrier repairing agents was evaluated by the same techniques. We observed a significant decrease in permeation characteristics through our in vitro models treated with the lipid mixtures compared to the untreated damaged skin, which implied that the skin barrier was substantially restored. Taken together, the results suggest that our in vitro models are suitable for the screening of potential barrier repairing agents.

• Klíčová slova
• FTIR spectroscopy, ceramides, liposomes, permeation experiment, skin barrier disruption models, stratum corneum lipids, topical treatment,
• MeSH
• ceramidy * MeSH
• epidermis MeSH
• kůže * MeSH
• lipidy MeSH
• permeabilita MeSH
• prasata MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ceramidy * MeSH
• lipidy MeSH

Mutations in the filaggrin (FLG) gene are strongly associated with common dermatological disorders such as atopic dermatitis. However, the exact underlying pathomechanism is still ambiguous. Here, we investigated the impact of FLG on skin lipid composition, organization, and skin acidification using a FLG knockdown (FLG-) skin construct. Initially, sodium/hydrogen antiporter (NHE-1) activity was sufficient to maintain the acidic pH (5.5) of the reconstructed skin. At day 7, the FLG degradation products urocanic (UCA) and pyrrolidone-5-carboxylic acid (PCA) were significantly decreased in FLG- constructs, but the skin surface pH was still physiological owing to an upregulation of NHE-1. At day 14, secretory phospholipase A2 (sPLA2) IIA, which converts phospholipids to fatty acids, was significantly more activated in FLG- than in FLG+. Although NHE-1 and sPLA2 were able to compensate the FLG deficiency, maintain the skin surface pH, and ensured ceramide processing (no differences detected), an accumulation of free fatty acids (2-fold increase) led to less ordered intercellular lipid lamellae and higher permeability of the FLG- constructs. The interplay of the UCA/PCA and the sPLA2/NHE-1 acidification pathways of the skin and the impact of FLG insufficiency on skin lipid composition and organization in reconstructed skin are described.

• MeSH
• atopická dermatitida metabolismus   patologie   MeSH
• filagriny MeSH
• fosfolipasy A2, skupina II metabolismus   MeSH
• genový knockdown MeSH
• koncentrace vodíkových iontů MeSH
• kůže cytologie   metabolismus   MeSH
• kyselina pyrrolidonkarboxylová metabolismus   MeSH
• kyselina urokanová metabolismus   MeSH
• kyseliny mastné neesterifikované metabolismus   MeSH
• kyseliny metabolismus   MeSH
• lidé MeSH
• metabolismus lipidů fyziologie   MeSH
• Na(+)-H(+) antiport metabolismus   MeSH
• permeabilita MeSH
• proteiny intermediálních filament nedostatek   genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• filagriny MeSH
• FLG protein, human MeSH Prohlížeč
• fosfolipasy A2, skupina II MeSH
• growth factor-activatable Na-H exchanger NHE-1 MeSH Prohlížeč
• kyselina pyrrolidonkarboxylová MeSH
• kyselina urokanová MeSH
• kyseliny mastné neesterifikované MeSH
• kyseliny MeSH
• Na(+)-H(+) antiport MeSH
• proteiny intermediálních filament MeSH