Fibroblasts improve performance of cultured composite skin substitutes on athymic mice.
BACKGROUND
This study investigated the impact of adding human fibroblasts to a cultured composite skin substitute model of cultured human keratinocytes and acellular human dermis.


METHODS
Skin substitutes were prepared by seeding human keratinocytes on the papillary side of acellular dermis with or without seeding fibroblasts on the reticular side. Performance of the grafts was compared both in vitro by histology and in vivo on surgically created full-thickness wounds on athymic mice. Graft size and contraction were measured and immunohistochemical stains were done to reveal vascularization.


RESULTS
Skin substitutes with fibroblasts formed thicker epidermis than skin substitutes without fibroblasts. When transplanted onto athymic mice, skin substitutes with fibroblasts maintained their original size with only 2% contraction. In contrast, skin substitutes without fibroblasts showed 29% contraction. Vascular basement membrane specific mouse CD31staining and endothelial cell specific mouse collagen type IV staining revealed vascularization as early as 1 week posttransplant in grafts with fibroblasts, and was significantly higher than grafts without fibroblasts at 2 weeks.


CONCLUSIONS
Addition of fibroblasts to keratinocyte based composite skin substitutes improves epidermis formation, enhances vascularization and reduces contraction.
Erdag
Gulsun
G
Shriners Hospital for Children, Massachusetts General Hospital, 51 Blossom Street, Boston, MA 02114, USA. erdag@helix.mgh.harvard.edu
Sheridan
Robert L
RL
D000818:Animals; D001485:Basement Membrane / Q000166:cytology; D002450:Cell Communication; D002455:Cell Division; D018920:Coculture Techniques; D003286:Contracture / Q000473:pathology; D004730:Endothelium, Vascular / Q000166:cytology; D000078404:Epidermal Cells; D005260:Female; D005347:Fibroblasts / Q000166:cytology*; D006085:Graft Survival; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D051379:Mice; D008819:Mice, Nude; D012039:Regional Blood Flow; D012867:Skin / Q000098:blood supply; D016038:Skin Transplantation / Q000379:methods*; D017768:Skin, Artificial