title: Use of an in vitro model of tissue-engineered skin to investigate the mechanism of skin graft contraction.
abstract: Skin graft contraction leading to loss of joint mobility and cosmetic deformity remains a major clinical problem. In this study we used a tissue-engineered model of human skin, based on sterilized human adult dermis seeded with keratinocytes and fibroblasts, which contracts by up to 60% over 28 days in vitro, as a model to investigate the mechanism of skin contraction. Pharmacologic agents modifying collagen synthesis, degradation, and cross-linking were examined for their effect on contraction. Collagen synthesis and degradation were determined using immunoassay techniques. The results show that skin contraction was not dependent on inhibition of collagen synthesis or stimulation of collagen degradation, but was related to collagen remodelling. Thus, reducing dermal pliability with glutaraldehyde inhibited the ability of cells to contract the dermis. So did inhibition of matrix metalloproteinases and inhibition of lysyl oxidase-mediated collagen cross-linking, but not transglutaminase-mediated cross-linking. In summary, this in vitro model of human skin has allowed us to identify specific cross-linking pathways as possible pharmacologic targets for prevention of graft contracture in vivo.
authors.affiliation: Department of Tissue Engineering, Kroto Institute, University of Sheffield, UK. caroline.a.harrison@btinternet.com
authors.forename: Caroline A
authors.initials: CA
authors.lastname: Harrison
authors.forename: Fatma
authors.initials: F
authors.lastname: Gossiel
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Layton
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Bullock
authors.forename: Timothy
authors.initials: T
authors.lastname: Johnson
authors.forename: Aubrey
authors.initials: A
authors.lastname: Blumsohn
authors.forename: Sheila
authors.initials: S
authors.lastname: MacNeil
mesh_terms: D000328:Adult; D000629:Aminopropionitrile / Q000494:pharmacology; D002392:Catechin / Q000494:pharmacology; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000378:metabolism; D003286:Contracture / Q000473:pathology / Q000503:physiopathology*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D003470:Culture Media; D020405:Dermis / Q000166:cytology; D004305:Dose-Response Relationship, Drug; D004970:Estrone / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D019084:Fluorescent Antibody Technique, Indirect; D005976:Glutaral / Q000494:pharmacology; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007118:Immunoassay; D007150:Immunohistochemistry; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D007633:Keratins / Q000378:metabolism; D010781:Photography / Q000379:methods; D016038:Skin Transplantation; D017768:Skin, Artificial; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*; D000511:alpha-Macroglobulins / Q000494:pharmacology
chemical_list: D024141:Collagen Type IV; D003432:Cross-Linking Reagents; D003470:Culture Media; D000511:alpha-Macroglobulins; D000629:Aminopropionitrile; D004970:Estrone; D007633:Keratins; D002392:Catechin; D005976:Glutaral