title: Topical nutrients promote engraftment and inhibit wound contraction of cultured skin substitutes in athymic mice.
abstract: Routine treatment of burns with cultured skin substitutes (CSS) has been limited by poor engraftment and by scarring. Hypothetically, topical application of essential nutrients and/or growth factors may support epithelial survival temporarily during graft vascularization. CSS, composed of human epidermal keratinocytes and dermal fibroblasts attached to collagen-glycosaminoglycan substrates, were incubated for 19 d in media optimized for keratinocytes. CSS, human xenografts, murine autografts, or no grafts were applied orthotopically to full-thickness skin wounds (2 x 2 cm) in athymic mice. Wounds were irrigated for 14 d with 1 ml/d modified cell culture medium or with saline containing epidermal growth factor, or were treated with dry dressings. After 6 weeks, treated sites were scored for percentage original wound area (mean +/- SEM) and percentage HLA-ABC-positive healed wounds [(number positive/n) x 100], and tested for significance (analysis of variance, p < 0.0001; Tukey test, p < 0.05). The data showed that CSS irrigated with nutrient medium were not statistically different in wound area (67.8 +/- 5.1%) from murine autografts (63.3 +/- 2.9%) but were statistically larger than human xenograft, no graft, or CSS treated with saline irrigation or dry dressings. HLA-ABC expression was 100% in CSS with nutrient irrigation, 86% in CSS with saline irrigation, 83% in CSS without irrigation, and 75% in xenografts with nutrient irrigation. These findings suggest that availability of essential nutrients supports keratinocyte viability during graft vascularization of CSS.
authors.affiliation: Shriners Burns Institute, Cincinnati, Ohio 45229-3095.
authors.forename: S T
authors.initials: ST
authors.lastname: Boyce
authors.forename: A P
authors.initials: AP
authors.lastname: Supp
authors.forename: M D
authors.initials: MD
authors.lastname: Harriger
authors.forename: D G
authors.initials: DG
authors.lastname: Greenhalgh
authors.forename: G D
authors.initials: GD
authors.lastname: Warden
mesh_terms: D000284:Administration, Oral; D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D046508:Culture Techniques; D006025:Glycosaminoglycans / Q000494:pharmacology; D006133:Growth Substances / Q000494:pharmacology; D015234:HLA-A Antigens / Q000032:analysis; D015235:HLA-B Antigens / Q000032:analysis; D015236:HLA-C Antigens / Q000032:analysis; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D012867:Skin / Q000033:anatomy & histology / Q000187:drug effects / Q000276:immunology; D016038:Skin Transplantation / Q000473:pathology; D014945:Wound Healing / Q000502:physiology*
chemical_list: D006025:Glycosaminoglycans; D006133:Growth Substances; D015234:HLA-A Antigens; D015235:HLA-B Antigens; D015236:HLA-C Antigens; D003094:Collagen