Functional Characterization of Cultured Keratinocytes after Acute Cutaneous Burn Injury

Gerd G Gauglitz,Guido Henkel V Donnersmarck,Siegfried Zedler,Eugen Faist,Jasmin Fuhr,Felix V Spiegel,Jürgen Schauber

doi:10.1371/journal.pone.0029942

Gerd G Gauglitz, Guido Henkel V Donnersmarck + Show 5 more

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https://doi.org/10.1371/journal.pone.0029942

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Abstract

BackgroundIn addition to forming the epithelial barrier against the outside environment keratinocytes are immunologically active cells. In the treatment of severely burned skin, cryoconserved keratinocyte allografts gain in importance. It has been proposed that these allografts accelerate wound healing also due to the expression of a favourable - keratinocyte-derived - cytokine and growth factor milieu.MethodsIn this study the morphology and cytokine expression profile of keratinocytes from skin after acute burn injury was compared to non-burned skin. Skin samples were obtained from patients after severe burn injury and healthy controls. Cells were cultured and secretion of selected inflammatory mediators was quantified using Bioplex Immunoassays. Immunohistochemistry was performed to analyse further functional and morphologic parameters.ResultsHistology revealed increased terminal differentiation of keratinocytes (CK10, CK11) in allografts from non-burned skin compared to a higher portion of proliferative cells (CK5, vimentin) in acute burn injury. Increased levels of IL-1α, IL-2, IL-4, IL-10, IFN-γ and TNFα could be detected in culture media of burn injury skin cultures. Both culture groups contained large amounts of IL-1RA. IL-6 and GM-CSF were increased during the first 15 days of culture of burned skin compared to control skin. Levels of VEGF, FGF-basic, TGF-ß und G-CSF were high in both but not significantly different. Cryoconservation led to a diminished mediator synthesis except for higher levels of intracellular IL-1α and IL-1ß.ConclusionSkin allografts from non-burned skin show a different secretion pattern of keratinocyte-derived cytokines and inflammatory mediators compared to keratinocytes after burn injury. As these secreted molecules exert auto- and paracrine effects and subsequently contribute to healing and barrier restoration after acute burn injury therapies affecting this specific cytokine/growth factor micromilieu could be beneficial in burned patients.

Highlights

Loss of the integrity of large portions of the skin as a result of burn injury may lead to major disability or even death
As the skin forms an active barrier protecting our organism from the outside environment rapid restoration of the epidermal barrier is of vital relevance after acute burn injury
Inclusion criteria included: Admission within 24 hours post burn injury, burns covering more than 25% of the total body surface area (TBSA),70 years of age, written consent to the experimental protocol

Summary

Introduction

Loss of the integrity of large portions of the skin as a result of burn injury may lead to major disability or even death. As the skin forms an active barrier protecting our organism from the outside environment rapid restoration of the epidermal barrier is of vital relevance after acute burn injury. Appropriate wound care is mandatory and various treatment modalities have been utilized to improve and accelerate wound healing. Growth factors and cytokines play major roles in the well-orchestrated integration of the complex biological and molecular events underlying cutaneous wound healing, including cell migration and proliferation, extracellular matrix deposition, angiogenesis and tissue remodeling [9]. In addition to forming the epithelial barrier against the outside environment keratinocytes are immunologically active cells. It has been proposed that these allografts accelerate wound healing due to the expression of a favourable - keratinocyte-derived - cytokine and growth factor milieu

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