Systematic investigation of factors, such as the impact of emulsifiers, which influence the measurement of skin barrier integrity by in-vitro trans-epidermal water loss (TEWL)

Trans-epidermal water loss (TEWL) has been the most widely used method to assess the integrity of the skin barrier and evaluate the irritation potential or the protective properties of topical products for many years. It detects the amount of water that diffuses across the stratum corneum (SC) to the external environment. As one of the most important functions of the skin is to keep water inside the body, an increase in TEWL is used to indicate the skin's impaired barrier function. So far, a variety of commercial instruments are available to measure the TEWL. Their applications mainly focus on the in-vivo TEWL measurements for dermatological examinations or formulation development. Recently, an in-vitro TEWL probe has also been commercially released enabling preliminary tests with excised skin samples. In our study, we first aimed to optimize the experimental procedures for detecting the in-vitro TEWL of porcine skin. Secondly, different kinds of emulsifiers were applied to the skin, including polyethylene glycol-containing emulsifiers (PEG-ylated emulsifiers), sorbitan esters, cholesterol, and lecithin. Sodium lauryl sulfate (SLS) was used as a positive control, and water as a negative control. Based on the findings, we established a protocol for accurately measuring the in-vitro TEWL values, emphasizing that the temperature of the skin sample should be constantly maintained at 32℃. Subsequently, the influences of emulsifiers on the in-vitro TEWL were analyzed. They indicated a significant skin barrier impairment of PEG-20 cetyl ether, PEG-20 stearyl ether, and SLS on in-vitro skin. Furthermore, we interestingly found that there consistently was an alteration of the TEWL values, even after the application of water to the skin. Our findings are of special interest, as the European Medicines Agency (EMA) recommends the use of in-vitro TEWL to determine skin barrier intactness during Franz cell experiments. Thus, this study provides a validated protocol for measuring the in-vitro TEWL and elucidates the impact of emulsifiers on the skin barrier. It also improves the understanding of tolerable variations of in-vitro TEWL and offers recommendations for its use in research.