Search

Background The stratum corneum (SC) is a well-known structure responsible for the cutaneous barrier. Tight junctions (TJs) function as a paracellular barrier beneath the SC and are involved in the cutaneous barrier. It remains unclear how TJs are involved in the cutaneous barrier. Objective In order to clarify the role of TJs in the cutaneous barrier, we investigated skin equivalent models with disrupted TJ barriers focusing on the SC. Methods Skin equivalents with disrupted TJ barriers were established using GST-C-CPE, a peptide with specific inhibitory action against specific claudins. The changes of the SC barrier in the skin equivalents with disrupted TJ barriers were investigated and compared with control skin equivalents. Results An outside-to-inside skin barrier assay revealed a defective SC barrier in skin equivalents with disrupted TJ barriers. A detailed examination of the SC revealed an increase in the pH of the SC in the skin equivalent with disrupted TJ barriers. An electron microscopy showed the failure of lamellar structures to mature and the failure of keratohyalin granules to degrade in the skin equivalents with disrupted TJ barriers. A thin layer chromatography analysis showed an increase in polar lipids and a decrease in non-polar lipids. A western blot analysis showed an increase in filaggrin dimer and trimer and a decrease in filaggrin monomer. Conclusion We found that disrupted TJs obstructed the SC formation responsible for the cutaneous barrier. Our study indicates the possibility that impaired TJ barriers affect polar lipids and profilaggrin processing by disturbing the pH condition of the SC.

It has not been confirmed whether tight junctions (TJs) function as a paracellular permeability barrier in adult human skin. To clarify this issue, we performed a TJ permeability assay using human skin obtained from abdominal plastic surgery. Occludin, a marker protein of TJs, was expressed in the granular layer, in which a subcutaneously injected paracellular tracer, Sulfo-NHS-LC-Biotin (556.59 Da), was halted. Incubation with ochratoxin A decreased the expression of claudin-4, an integral membrane protein of TJs, and the diffusion of paracellular tracer was no longer prevented at the TJs. These results demonstrate that human epidermis possesses TJs that function as an intercellular permeability barrier at least against small molecules (∼550 Da). UVB irradiation of human skin xenografts and human skin equivalents (HSEs) resulted in functional deterioration of TJs. Immunocytochemical staining of cultured keratinocytes showed that occludin was localized into dot-like shapes and formed a discontinuous network when exposed to UVB irradiation. Furthermore, UVB irradiation downregulated the active forms of Rac1 and atypical protein kinase C, suggesting that their inactivation caused functional deterioration of TJs. In conclusion, TJs function as a paracellular barrier against small molecules (∼550 Da) in human epidermis and are functionally deteriorated by UVB irradiation.

Natural moisturizing factor (NMF) of the stratum corneum (SC) has been established to play important roles in the physical properties of the SC. Few studies, however, have investigated the specific influences of NMF components other than the amino acids. In this study, therefore, we focus on the relationship between the ion content and physical properties of the SC in 40 healthy subjects. Changes in the physical properties of the SC induced by the extraction of NMF were equivalent to the changes that took place from summer to winter, demonstrating the important role of NMF in the physical properties of the SC in healthy subjects. The seasonal changes in the physical properties of the SC from summer to winter were accompanied by significant decreases in the levels of lactate, potassium, sodium, and chloride in the SC. Lactate and potassium were the only components found to correlate significantly with the state of hydration, stiffness, and pH in the SC. Interestingly, the levels of lactate and potassium in the SC were also significantly correlated. Moreover, potassium lactate restored the SC hydration state decreased by extraction of NMF. These results suggest that lactate and potassium may play roles in maintaining the physical properties of the SC in healthy subjects.