title: Development of a one-step approach for the reconstruction of full thickness skin defects using minced split thickness skin grafts and biodegradable synthetic scaffolds as a dermal substitute.
abstract: BACKGROUND
  Tissue 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.
  
  
  OBJECTIVE
  To develop a one-step approach to wound reconstruction using finely minced split thickness skin and a biodegradable synthetic dermal substitute.
  
  
  METHODS
  This 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.
  
  
  RESULTS
  Skin 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.
  
  
  CONCLUSIONS
  We 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.
authors.affiliation: Department of Plastic and Reconstructive Surgery, Sheffield Teaching Hospitals, NHS Foundation Trust, United Kingdom.
authors.forename: Kavita
authors.initials: K
authors.lastname: Sharma
authors.affiliation: Department of Materials Science and Engineering, Kroto Research Institute, University of Sheffield, Sheffield S3 7HQ, United Kingdom.
authors.forename: Anthony
authors.initials: A
authors.lastname: Bullock
authors.affiliation: Department of Plastic and Reconstructive Surgery, Sheffield Teaching Hospitals, NHS Foundation Trust, United Kingdom.
authors.forename: David
authors.initials: D
authors.lastname: Ralston
authors.affiliation: Department of Materials Science and Engineering, Kroto Research Institute, University of Sheffield, Sheffield S3 7HQ, United Kingdom. Electronic address: s.macneil@sheffield.ac.uk.
authors.forename: Sheila
authors.initials: S
authors.lastname: MacNeil
mesh_terms: D020341:Absorbable Implants; D004847:Epithelial Cells; D006801:Humans; D007150:Immunohistochemistry; D008747:Methylcellulose; D008855:Microscopy, Electron, Scanning; D011091:Polyesters; D012867:Skin / Q000166:cytology*; D016038:Skin Transplantation / Q000379:methods*; D017768:Skin, Artificial; D017154:Stromal Cells; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D019737:Transplants / Q000166:cytology*
chemical_list: D011091:Polyesters; C033616:poly(lactide); D008747:Methylcellulose
keywords: Burn reconstruction; Electrospun scaffold; Minced human skin; Skin substitute; Wound reconstruction