Use of an in vitro model of tissue‐engineered human skin to study keratinocyte attachment and migration in the process of reepithelialization

Abstract

To produce a stable epidermis, keratinocytes need to be firmly attached to the basement membrane. However, following wounding, keratinocytes are required to develop a migratory phenotype in order to reepithelialize the wound. To investigate some of the issues underlying reepithelialization, we have developed a three-dimensional in vitro model of tissue-engineered skin, comprising sterilized human dermis seeded with human keratinocytes and dermal fibroblasts. Using this model, we have shown that the inclusion of fibroblasts within the model increases the stability of keratinocyte attachment. We have also demonstrated that keratinocyte migration occurs most effectively in the absence of a basement membrane and following the inclusion of fibroblasts in the model. In addition, subjecting the keratinocyte layer to mechanical trauma induces a migratory phenotype. We conclude that this three-dimensional in vitro wound model can be used to increase our understanding of the factors that enhance keratinocyte migration and hence wound healing in vivo.

Intravital insights in skin wound healing using the mouse dorsal skin fold chamber

“…More recently, a variety of in vivo (Breuing et al 1992;Rossio-Pasquier et al 1999;Escamez et al 2004;Geer et al 2004) and in vitro models (Kamamoto et al 2003;Harrison et al 2006) have been introduced to evaluate the process of wound Values are given as means ± sem; one-way repeated measures ANOVA with *P < 0.05; ANOVA with †P < 0.05 vs. functional density normal ; n.s., not significant. healing and skin regeneration.…”

“…For this purpose different models have been established to study skin regeneration in health and disease (Breuing et al 1992;Rossio-Pasquier et al 1999;Escamez et al 2004;Geer et al 2004;Harrison et al 2006). While most preparations used in those studies are not well suited for observing the microcirculatory process of wound healing, the skin fold chamber seems to be an ideal model, allowing repeated analysis of the regenerating skin as well as the skin microcirculation over a prolonged period of time.…”

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

“…5 Besides two-dimensional models, three-dimensional human skin equivalents have been developed for dermatological research. The dermal part of these models are generated using various scaffolds including decellularized dermis, 6 collagen hydrogels, 7,8 glycosaminoglycans 9 or synthetic materials. 10 Employing these skin equivalents, the role of epithelial-mesenchymal interactions 11 , the re-epithelialization, the cellular crosstalk between fibroblasts and keratinocytes and the influence of different growth factors can be studied.…”

Vascularization of Novel Porcine Acellular Dermal Matrix

“…In addition, implanting cultured autologous keratinocyte and FB in dermal substitute and constructing permanent wound coverage is an important subject in research on skin tissue engineering. At present, it has been successfully conducted in vitro, but no clinical validation is reported 10,11) . At the same time, preparation of functional dermal substitute containing epidermis, FB, VEC, Langhan's cell, sweat gland cell, hair follicle cell and pigment cell, and constructing ideal tissue-engineered dermal substitute have also be attempted 12,13) .…”