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Updated: Jul 14, 2026

Isolation and Culture of Primary Mouse Keratinocytes from Neonatal and Adult Mouse Skin
Published on: July 14, 2017
Cytokine and chemokine release upon prolonged mechanical loading of the epidermis
Debbie Bronneberg1, Sander W Spiekstra, Lisette H Cornelissen
1Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands. d.bronneberg@tue.nl
Abstract:
At this moment, pressure ulcer risk assessment is dominated by subjective measures and does not predict pressure ulcer development satisfactorily. Objective measures are, therefore, needed for an early detection of these ulcers. The current in vitro study evaluates cytokines and chemokines [interleukin 1alpha (IL-1alpha), interleukin 1 receptor antagonist (IL-1RA), tumor necrosis factor alpha (TNF-alpha) and interleukin 8 (CXCL8/IL-8)] as early markers for mechanically-induced epidermal damage. Various degrees of epidermal damage were induced by subjecting commercially available epidermal equivalents (EpiDerm) to increasing pressures (0, 50, 75, 100, 150, and 200 mmHg) for 24 h, using a loading device. At the end of the loading experiment, tissue damage was assessed by histological examination and by evaluation of the cell membrane integrity. Cytokines and chemokines were determined in the culture supernatant. Sustained epidermal loading resulted in an increased release of IL-1alpha, IL-1RA, TNF-alpha and CXCL8/IL-8. This was first observed at 75 mmHg, when the tissue was only slightly damaged. Swollen cells, vacuoles, necrosis and affected cell membranes were observed at pressures higher than 75 mmHg. Furthermore, at 150 and 200 mmHg, the cells in the lower part of the epidermis were severely compressed. In conclusion, IL-1alpha, IL-1RA, TNF-alpha and CXCL8/IL-8 are released in vitro as a result of sustained mechanical loading of the epidermis. The first increase in cytokines and chemokines was observed when the epidermal tissue was only slightly damaged. Therefore, these cytokines and chemokines are potential markers for the objective, early detection of mechanically-induced skin damage, such as pressure ulcers.
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