Allogeneic bilayered tissue-engineered skin promotes full-thickness wound healing in ovine model

Skin injuries can be treated via grafting. However, since both autologous and allogeneic skin grafts have their own limitations, skin tissue engineering was developed as an alternative approach to promote healing. Autologous tissue-engineered skin requires a few weeks for cell culture and cannot be used for acute treatment. In this study, healing potential of allogeneic bilayered tissue-engineered skin (BTES) was evaluated in sheep. Isolated allogeneic skin cells from the sheep were cultured using a combined medium of DKSFM:F12:DMEM in the ratio 2: 1: 1 supplemented with 5% FBS. Differential trypsinization was later carried out to separate the fibroblasts and keratinocytes. The allogeneic BTES was fabricated using allogeneic fibrin as biomaterial by polymerizing the fibrin-keratinocyte layer with calcium chloride followed by fibrin-fibroblast layer. Half of the wounds were isolated with PVC rings to prevent cell migration. The wounds were treated with allogeneic BTES and silk, as control. Wounds were observed at days 7, 14 and 21 to determine the rate of re-epithelialization. After 3 weeks, the sheep were euthanized. Histological evaluation with H & E, elastin van Gieson and Masson's trichrome staining showed that allogeneic BTES treated wound healed faster compared to the control group. In conclusion, allogeneic BTES has the potential to be developed as off the shelf product for rapid treatment of full thickness wound.