• Kavita Sharma, Anthony Bullock, David Ralston, and Sheila MacNeil.
• Department of Plastic and Reconstructive Surgery, Sheffield Teaching Hospitals, NHS Foundation Trust, United Kingdom.
• Burns. 2014 Aug 1;40(5):957-65.

IntroductionTissue engineering has progressed in delivering laboratory-expanded keratinocytes to the clinic; however the production of a suitable alternative to a skin graft, containing both epidermis and dermis still remains a challenge.AimTo develop a one-step approach to wound reconstruction using finely minced split thickness skin and a biodegradable synthetic dermal substitute.MethodsThis was explored in vitro using scalpel diced pieces of split thickness human skin combined with synthetic electrospun polylactide (PLA) scaffolds. To aid the spreading of tissue, 1% methylcellulose was used and platelet releasate was examined for its effect on cellular outgrowth from tissue explants. The outcome parameters included the metabolic activity of the migrating cells and their ability to produce collagen. Cell presence and migration on the scaffolds were assessed using fluorescence microscopy and SEM. Cells were identified as keratinocytes by immunostaining for pan-cytokeratin. Collagen deposition was quantified by using Sirius red.ResultsSkin cells migrated along the fibers of the scaffold and formed new collagen. 1% methylcellulose improved the tissue handling properties of the minced skin. Platelet releasate did not stimulate the migration of skin cells along scaffold fibers. Immunohistochemistry and SEM confirmed the presence of both epithelial and stromal cells in the new tissue.ConclusionWe describe the first key steps in the production of a skin substitute to be assembled in theatre eliminating the need for cell culture. Whilst further experiments are needed to develop this technique it can be a useful addition to armamentarium of the reconstructive surgeon.Copyright © 2013 Elsevier Ltd and ISBI. All rights reserved.

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