212 Chapter 7 processes of skin inflammation, because they lack essential immune components [26,27]. To make in vitro models more appropriate, relevant immune cells and inflammatory mediators need to be incorporated. In this study, we generated a human full skin equivalent (FSE) model based on the collagen-elastin matrix MatriDerm® [28,29], as we described previously [30]. MatriDerm is a clinically applied matrix that provides a durable extracellular matrix architecture supporting epithelization in skin defects [31–34]. To study the effect of burn injury on cells from the innate and the adaptive immune system, monocytes and T cells isolated from human buffy coats were cultured in this human FSE. We studied the effect of burn injury on the phenotype and cytokine secretion of monocytes and T cells. By incorporating immune cells into the FSE, we set the first steps in the development of a more relevant skin model for the study of inflammatory reactions that occur after burn injury, while supporting the replacement and reduction of animal experiments. RESULTS Human full skin equivalents to study burn injury in vitro FSEs were produced by seeding human keratinocytes and fibroblasts into MatriDerm matrices containing collagen-elastin (see Figure 6A for procedure), as we previously described [30]. After 3 weeks of culture, the FSEs contained a properly developed epidermis and dermis. Burn injury was inflicted on the FSE and subsequently visualized by microscopy (Figure 1). Three days after injury, the burn was visible as the epidermis was detached from the injured area of the dermis. Figure 1. Histology of uninjured and burn-injured FSEs. H&E staining. (A) FSE after 3 weeks of culture. (B) FSE after 3 weeks of culture and 3 days after burn injury. The detached epidermis caused by the burn is clearly visible. Black scale bar = 100 µm; black arrows indicate burn injured area.
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