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Reepithelialization in Focus: Rapid Screening of Wound Closure Using Organotypic Skin Models

: Kiesewetter, Lisa; Littau, Laura; Boccaccini, Aldo R.; Walles, Heike; Groeber-Becker, Florian Kai


Wound repair and regeneration 27 (2019), No.3, pp.A31
ISSN: 1067-1927
ISSN: 1524-475X
Wound Healing Society (Annual Meeting) <31, 2019, San Antonio/Tex.>
Fraunhofer ISC ()
Wundheilung; Epidermis

Background: Up to today, animal studies are the gold standard for preclinical testing of new therapeutics for cutaneous wound healing. Alternative in vitro studies are mostly restricted to 2D cell cultures and thus only poorly reflect the complex physiological wound situation.
Methods: To overcome this limitation, we present two 3D models based on human primary cells for studying different stages of cutaneous wound healing in vitro. For observation of the reepithelialization, human epidermal models are wounded locally using a biopsy punch and wound healing is monitored. We introduce impedance spectroscopy as a method to determine the efficacy of wound healing non‐destructively via the skin barrier. Results are compared to phase‐contrast microscopy, laser scanning and histology images as well as metabolic activity. For allowing the integration and analysis of deeper wounds, we expanded the models to full thickness skin equivalents with a collagen‐based extracellular matrix including fibroblasts.
Results: Upon wounding, the impedance of epidermal models with initial values between 6.3 and 11.4kΩcm2 dropped significantly by ‐95%. During on‐line monitoring over fourteen days the barrier recovered, indicated by significant increases in the impedance back to levels between 7.9 and 14.8kΩcm2. For full thickness skin equivalents, too, this proved to be a promising screening technique with impedances starting at 276Ωcm2 and dropping by ‐63% following wounding. With ongoing wound closure, the barrier recovered to 310Ωcm2. Reepithelialization appeared to be donor dependent for both models, correlating well with results from imaging techniques and metabolic activity.
Conclusions: Altogether, impedance analysis allows for rapid screening and evaluation of test substances and wound therapies, whereby only a small set of models is needed, cutting the costs of preclinical studies. Finally, with the help of these in vitro models, the amount of animals used in preclinical studies could be reduced.