Objective: Bioengineered skin substitutes were created to address wound healing problems. Skin substitutes contains live human cells seeded onto a matrix to provide cytokine, growth factor and other proteins from ECM required to decrease healing time. These products are classified based on their durability, the cells seeded on them and their originality. In this study, we aimed to investigate fibrin-collagen hydrogel as a new scaffold to design a bilayer temporary skin equivalent Methods: Fibrin gel was prepared by crosslinking fibrinogen with thrombin and mixing it with collagen type 1. Human fibroblasts and keratinocytes were isolated from skin biopsies of healthy donors and foreskin and the cells were seeded onto 3D hydrogel layer by layer. Morphological assessment, histological analysis, immunocytochemistry were performed to characterize the scaffold properties Result: The results of scaffold characterization demonstrated good porosity, cell viability and biocompatibility of the scaffold Conclusion: Fibrin-collagen as a natural material in organotypic cell culture modeling demonstrates that hydrogel scaffolds can be properly designed to generate bilayer or composite temporary skin grafts
