Comparison of dermal and eschar fibroblasts in engineered skin equivalents

Deep dermal injuries, like burns, often heal with (hypertrophic) scar formation. Dermal fibroblasts play an important role in the wound healing process. Fibroblasts derived from burnt tissue (eschar) have different characteristics then healthy dermal fibroblasts. In this study, human dermal fibroblasts were isolated from healthy skin and burnt tissue (eschar) from 10 patients. Fibroblasts were cultured to P2 and RNA was isolated for microarray analysis using an Agilent Human 44k array. Results of the array was analyzed using R to create a dataset which compared the expression between dermal and eschar derived fibroblasts. In addition, gene-expression data was coupled to the KEGG-pathway database to explore the differences and identify the significantly altered pathways. Furthermore, full-skin equivalents (FSE) were produced with human primary keratinocytes and dermal or eschar fibroblasts using collagen-elastin scaffolds. RNA was isolated from these FSE's and qPCR was performed on the synthesized cDNA. A volcano-plot was generated to reveal significantly up and downregulated genes from the micro-array data. For the FSE data, qPCR was performed on a selection of genes based on the microarray. A full description of the data and findings can be found in the corresponding manuscript.