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Title: An investigation into the use of fibrin glue and cultured cells with tissue engineered skin replacements
Author: Currie, Lachlan James
Awarding Body: University of London
Current Institution: University College London (University of London)
Date of Award: 2003
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Cultured autologous keratinocytes have been used as an alternative to split skin grafts for patients with major full thickness burns. We have examined the use of cultured autologous keratinocytes with commercially available Integra® artificial skin and Tisseel fibrin glue. The Large White Pig was used as an animal model. Full thickness wounds were isolated in PTFE chambers to prevent wound contraction and marginal re-epithelialisation. Wounds were grafted with Integra® artificial skin seeded with cultured autologous keratinocytes with or without fibrin glue. The reconstitution of an epidermal cell layer was demonstrated both with or without the use of fibrin glue. There was a trend towards an increased percentage of epidermal cover when fibrin glue was used to seed Integra® (42.1% verses 20.4%) though this was not significant (P=0.097). However, fibrin glue did improve haemostasis and made Integra® application technically easier. Non-cultured autologous keratinocytes produced less epithelium than cultured autologous keratinocytes (6.7% verses 24.6%, P=0.009) and there was evidence of significant differentiated cyst formation within the matrix with the use of non-cultured cells. Cultured fibroblasts seeded into Integra® were shown to persist in-vivo for at least two weeks. This was true for both allogenic and autologous fibroblasts. However, the number of fibroblasts seeded into the matrix was insignificant in comparison to the number of fibroblasts derived from local mesenchymal cells. Spray delivery of cultured keratinocytes is also gaining widespread popularity as an alternative to the use of fragile sheets of keratinocytes. Chambered wounds sprayed with cultured autologous keratinocytes in a suspension of culture medium (without the use of a dermal replacement) were shown to produce a 25.8% epithelial cover by three weeks. When fibrin glue was used as the delivery vehicle for the sprayed cultured autologous cells there was no improvement in the percentage of epithelium at three weeks (P=0.802). In conclusion, autologous and allogenic fibroblasts seeded into Integra remain viable within the wound for at least two weeks. The use of fibrin glue to seed Integra with cultured autologous keratinocytes produces a good quality surface epithelium. When non-cultured autologous keratinocytes are seeded into Integra, cysts form within the matrix, preventing epithelial formation at the wound surface. The use of fibrin glue for spray delivery of cultured keratinocytes does not increase the quantity of epithelium formed in this model.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available