Wound closure with EDC cross-linked cultured skin substitutes grafted to athymic mice

Collagen-glycosaminoglycan (C-GAG) sponges are commonly utilized as a substitute for the extracellular matrix of dermal tissue. Cultured skin substitutes (CSS) were assessed, after fabrication using sponges cross-linked with 1-ethyl-3-3-dimethylaminopropylcarbodiimide hydrochloride (EDC) at 0, 1, 5, or 50 mm, for development of viable, stratified skin tissue anatomy in vitro, and for wound contraction and cell viability in vivo. Cross-linking the C-GAG sponges with EDC reduced in vitro contraction of the CSS from a 39% reduction in area in the 0 mm CSS to 0% in the 50 mm group. Conversely, the wounds closed with 0, 1 and 5 mm EDC groups exhibited significantly less wound contraction than the 50 mm group. Engraftment of human cells occurred in 86%, 83%, and 83% of the wounds treated with CSS fabricated using 0, 1, and 5 mm EDC cross-linked sponges, respectively, which were significantly higher engraftment rates than the 50 mm group (17%). These data suggest that low concentrations of EDC can be used to improve the biochemical stability of the C-GAG component of CSS in vitro, and promote stable wound closure.