Introduction
Timely wound closure in massive burns remains an unmet clinical need. Engineered skin substitutes can provide an alternative treatment to split skin grafting. However, since the development of Cultured Epithelial Autograft (Green’s method), almost 50 years ago, limited alternatives have been introduced.
Methodology
Here we describe the production of an autologous human skin equivalent (HSE) for the successful treatment of a paediatric patient with a severe burn injury. The HSE was comprised of dermal and epidermal components, including a basement membrane. The epidermis was stratified and semi-matured, providing essential components for a definitive graft. A platelet-derived hydrogel, combined with autologous fibroblasts, acted as a niche for the stem and progenitor keratinocytes, isolated from a small piece of donor skin. The survival of the stem and progenitor keratinocytes in HSE was confirmed by immunofluorescence using Keratin 5, Integrin alpha 5 and beta 1 specific antibodies. Over 500 cm2 HSE was produced under good manufacturing practice (GMP) guidelines and tested clear of endotoxin, mycoplasma, bacteria and yeast pathogens.
Results
The HSE was grafted on a full thickness wound temporised with NovoSorb ® Biodegradable Temporising Matrix (BTM) and graft take was estimated to be 95% on day 14 post grafting. Histology, at 2 weeks and 6 weeks post grafting, confirmed persistence of a complete and continuous, although hyperproliferative, epidermis. Graft quality has remained superior to the patient’s autologous grafts at 6 months
Conclusion
This study is a proof of concept for the application of HSE as a treatment option in paediatric burn injuries with limited donor skin.