In vitro effects of matrix peptides on a cultured dermal-epidermal skin substitute

Composite dermal-epidermal skin substitutes rely on a firm attachment of human keratinocytes (HK) to the dermal substrate for graft survival on the wound. An in vitro study was performed assessing whether the addition of matrix peptides to the dermal substrate effected the epithelial thickness. Cultured grafts were made by attaching HK to the external surface of a collagen-chondroitin 6-sulfate (GAG) membrane and inoculating human fibroblasts (HF) internally. If the matrix peptide (RGD Peptide) was added to the collagen-GAG membrane prior to placement of the HK and HF, the resultant epithelial layer at the end of the normal 4-day culture period was significantly thicker (19.7 +/- 0.9 microns versus 13.5 +/- 1.0 microns). Subjectively, the HF content was also greater on the peptide-treated grafts. When HF were not placed on the cultured graft, i.e., only collagen-GAG membrane, RGD peptide, and HK, the resultant epithelial thickness was even greater (28.3 +/- 1.0 microns). These data suggest that addition of matrix peptides, which increase cell attachment to the dermal substrate, may prove effective in the improvement of this cultured composite dermal-epidermal skin substitute.