The Reconstructed Human Epidermis Models in Fundamental Research
Large skin trauma allows pathogens to invade, and causes water, electrolyte and protein loss, inducing adverse events often incompatible with sustained life if the wound is not rapidly covered and the lost tissue replaced. This last point is often critica
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A. Coquette, Y. Poumay
Contents 67.1
Introduction . . . . . . . . . . . . . . . . . . . . . . . . 967
67.2
Key Developments in In Vitro Human Epidermis Reconstruction . . . . . . . . . . . . . 967
67.3
The Culture and Morphology of Reconstructed Human Epidermis (RHE) Models . . . . . . . . . . . . . . . . . . . . . . . . . . . . 968
67.4
Reconstructed Human Epidermis Models and Skin Physiopathology . . . . . . . . . . . . . 969
67.5
Uses of RHE as Alternatives to Animal/ Human Testing . . . . . . . . . . . . . . . . . . . . . . 971
67.5.1 In Vitro Skin Irritation . . . . . . . . . . . . . . . . 971 67.5.2 In Vitro Skin Sensitization . . . . . . . . . . . . . 971 67.5.3 In Vitro Skin Permeation and Metabolization . . . . . . . . . . . . . . . . . . . . . . 972 References . . . . . . . . . . . . . . . . . . . . . . . . . 973
67.1
the development of several commercial skin substitutes. Although they represent significant advances in wound treatment, they are of relative efficacy and can induce adverse events such as infectious diseases and allergic reactions in patients with known sensitivity to materials of bovine origin [46]. Moreover, a risk of prior diseases is also present even if serum and/or collagen are sourced from countries theoretically free from bovine spongiform encephalitis [44]. Therefore, a more efficient and safe means of cultivation are needed in order to ensure the safety of the patient, such as the use of serum-free culture media. Moreover, for significant progresses in clinical efficacy, the understanding of the complex process of wound healing and development of the novel growth factor and gene therapies would benefit from research models that mimic closely the human skin epidermis/dermis structure, and in which studies of the cell-cell and cell-matrix interactions could be possible.
Introduction
67.2
Large skin trauma allows pathogens to invade, and causes water, electrolyte and protein loss, inducing adverse events often incompatible with sustained life if the wound is not rapidly covered and the lost tissue replaced. This last point is often critical in wound management because of the poor availability of skin samples. The recent advances in the in vitro techniques of keratinocyte culture have led to
The first stratified colonies of human keratinocytes from single cells were obtained by Rheinwald and Green [72], and these cultures were even used for grafting in patients with extensive burns or for treatment of giant naevi [29, 30]. Although this model remains as a reference for human therapy because of
Key Developments in In Vitro Human Epidermis Reconstruction
968
the selection of cells of high clonogenic potential, it is limited for fundamental and applied research. In fact, differentiating keratinocytes exhibit a strong involucrin expression in the first suprabasal cell layer, a situation that differs from in vivo differentiation where involucrin is mainly induced in upper spinous and granular layers [2]. Moreover, the simultaneous
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