BACKGROUND: Extended full thickness skin defects still represent a considerable therapeutic challenge as ideal strategies for definitive autologous coverage are still not available. Tissue engineering of whole skin represents an equally attractive and ambitious novel approach. We have recently shown that laboratory-grown human skin analogues with near normal skin anatomy can be successfully transplanted on immuno-incompetent rats. The goal of the present study was to engineer autologous porcine skin grafts for transplantation in a large animal model (pig study = intended preclinical study). MATERIALS AND METHODS: Skin biopsies were taken from the pig's abdomen. Epidermal keratinocytes and dermal fibroblasts were isolated and then expanded on culture dishes. Subsequently, highly concentrated collagen hydrogels and collagen/fibrin hydrogels respectively, both containing dermal fibroblasts, were prepared. Fibroblast survival, proliferation, and morphology were monitored using fluorescent labelling and laser scanning confocal microscopy. Finally, keratinocytes were seeded onto this dermal construct and allowed to proliferate. The resulting in vitro generated porcine skin substitutes were analysed by H&E staining and immunofluorescence. RESULTS: Dermal fibroblast proliferation and survival in pure collagen hydrogels was poor. Also, the cells were mainly round-shaped and they did not develop 3D-networks. In collagen/fibrin hydrogels, dermal fibroblast survival was significantly higher. The cells proliferated well, were spindle-shaped, and formed 3D-networks. When these latter dermal constructs were seeded with keratinocytes, a multilayered and partly stratified epidermis readily developed. CONCLUSION: This study provides compelling evidence that pig cell-derived skin analogues with near normal skin anatomy can be engineered in vitro. These tissue-engineered skin substitutes are needed to develop a large animal model to establish standardized autologous transplantation procedures for those studies that must be conducted before "skingineering" can eventually be clinically applied.
BACKGROUND: Extended full thickness skin defects still represent a considerable therapeutic challenge as ideal strategies for definitive autologous coverage are still not available. Tissue engineering of whole skin represents an equally attractive and ambitious novel approach. We have recently shown that laboratory-grown human skin analogues with near normal skin anatomy can be successfully transplanted on immuno-incompetent rats. The goal of the present study was to engineer autologous porcine skin grafts for transplantation in a large animal model (pig study = intended preclinical study). MATERIALS AND METHODS: Skin biopsies were taken from the pig's abdomen. Epidermal keratinocytes and dermal fibroblasts were isolated and then expanded on culture dishes. Subsequently, highly concentrated collagen hydrogels and collagen/fibrin hydrogels respectively, both containing dermal fibroblasts, were prepared. Fibroblast survival, proliferation, and morphology were monitored using fluorescent labelling and laser scanning confocal microscopy. Finally, keratinocytes were seeded onto this dermal construct and allowed to proliferate. The resulting in vitro generated porcine skin substitutes were analysed by H&E staining and immunofluorescence. RESULTS: Dermal fibroblast proliferation and survival in pure collagen hydrogels was poor. Also, the cells were mainly round-shaped and they did not develop 3D-networks. In collagen/fibrin hydrogels, dermal fibroblast survival was significantly higher. The cells proliferated well, were spindle-shaped, and formed 3D-networks. When these latter dermal constructs were seeded with keratinocytes, a multilayered and partly stratified epidermis readily developed. CONCLUSION: This study provides compelling evidence that pig cell-derived skin analogues with near normal skin anatomy can be engineered in vitro. These tissue-engineered skin substitutes are needed to develop a large animal model to establish standardized autologous transplantation procedures for those studies that must be conducted before "skingineering" can eventually be clinically applied.
Authors: Irene Montaño; Clemens Schiestl; Jörg Schneider; Luca Pontiggia; Joachim Luginbühl; Thomas Biedermann; Sophie Böttcher-Haberzeth; Erik Braziulis; Martin Meuli; Ernst Reichmann Journal: Tissue Eng Part A Date: 2010-01 Impact factor: 3.845
Authors: M Raghunath; T Bächi; M Meuli; S Altermatt; R Gobet; L Bruckner-Tuderman; B Steinmann Journal: J Invest Dermatol Date: 1996-05 Impact factor: 8.551
Authors: Sophie Böttcher-Haberzeth; Thomas Biedermann; Agnieszka S Klar; Luca Pontiggia; Jürgen Rac; David Nadal; Clemens Schiestl; Ernst Reichmann; Martin Meuli Journal: Pediatr Surg Int Date: 2014-02 Impact factor: 1.827
Authors: Clemens Schiestl; Thomas Biedermann; Erik Braziulis; Fabienne Hartmann-Fritsch; Sophie Böttcher-Haberzeth; Margarete Arras; Nikola Cesarovic; Flora Nicolls; Carsten Linti; Ernst Reichmann; Martin Meuli Journal: Pediatr Surg Int Date: 2011-03 Impact factor: 1.827
Authors: Sophie Böttcher-Haberzeth; Agnieszka S Klar; Thomas Biedermann; Clemens Schiestl; Claudia Meuli-Simmen; Ernst Reichmann; Martin Meuli Journal: Pediatr Surg Int Date: 2012-11-30 Impact factor: 1.827
Authors: Marketa Bacakova; Julia Pajorova; Denisa Stranska; Daniel Hadraba; Frantisek Lopot; Tomas Riedel; Eduard Brynda; Margit Zaloudkova; Lucie Bacakova Journal: Int J Nanomedicine Date: 2017-02-09