Picture Overviews - Skingraft on base of hAM

Allogenic graft material and tissue engineering have recently shown promising results for the improvement of both aesthetic and functional outcomes in the treatment of large skin defects. We chose human amniotic membranes (hAM) as a cellular scaffold in order to develop a skin substitute for later in-vivo uses. Various methods of de-epithelialization of the hAM were evaluated by histological analysis including hematoxylin-eosin and laminin staining, optic coherence tomography (OCT), and scanning electron microscopy with 0.25/0.02% trypsin/EDTA treatment and mechanical cell removal showing a complete loss of the epithelium and a mainly intact basement membrane. Novel examination of hAM by OCT was feasible, but difficulties were experienced in handling and interpretation of the tissue as no comparable data exist. Subsequently, we developed an air-liquid-interface cell culture to cultivate keratinocytes and fibroblasts on the de-epithelialized hAM. When comparing various air-liquid cell-culture media and resulting cell viability, we found that a mainly keratinocyte-based medium containing a low amount of serum provided good keratinocyte and fibroblast viability. We achieved a mostly keratinized surface on the epidermal side with a confluent fibroblast network on the chorion side.