Construction of composite organotypic co-culture wound healing model.

A) Schematic drawing of stratified keratinocyte sheet growing on contracted collagen matrix containing fibroblasts (Layer II). This skin equivalent culture rests on a semi-permeable membrane and is nourished with medium from below, thus being exposed to an air-liquid interface. A wound is formed by punching a hole through the epithelium and collagen matrix. B) the wounded culture is transferred onto a second collagen matrix that has undergone contraction (Layer I). C) the resultant composite co-culture consists of two layers of contracted matrix and one layer of epithelium. Wound edges are seen at the transition zone from layer I to layer II and are noted with arrows. D) reepithelialization occurs as wounded keratinocytes migrate onto the collagen in layer II and is then followed by stratification of the tissue to reconstitute a fully stratified epithelium that covers the wound bed. E is a picture of the wound as depicted in C. on top of the wound a drop of self-assembling peptide containing or not the protein is added (scheme F). G is a picture of the wound covered by the drop of peptide stained with comassie blue.