Allogenic graft material and tissue engineering have recently shown promising results for the improvement of both esthetic and functional outcomes in the treatment of large skin defects. We chose human amniotic membrane as a cellular scaffold in order to develop a skin substitute for later in vivo uses. Various methods of de-epithelialization of the human amniotic membrane were evaluated by histological analysis including hematoxylin-eosin and laminin staining, optic coherence tomography, and scanning electron microscopy with 0.25/0.02% trypsin/ethylenediaminetetraacetic acid treatment and mechanical cell removal showing an almost complete loss of the epithelium and a mainly intact basement membrane. Novel examination of human amniotic membrane by optic coherence tomography 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 human amniotic membrane. We achieved a mostly keratinized surface on the epidermal side with a confluent fibroblast network on the chorion side.

中文翻译：

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基于人羊膜的组织工程皮肤替代品的开发。

近来，同种异体移植材料和组织工程学已显示出可喜的结果，可改善大皮肤缺损的美学和功能结局。我们选择人羊膜作为细胞支架，以开发出皮肤替代品以用于以后的体内用途。通过组织学分析评估了人类羊膜脱上皮的各种方法，包括苏木精-伊红和层粘连蛋白染色，光学相干断层扫描以及0.25 / 0.02％胰蛋白酶/乙二胺四乙酸处理的扫描电子显微镜和机械细胞去除，显示几乎完成上皮和基本完整的基底膜丢失。通过光学相干断层扫描对人羊膜进行新颖检查是可行的，但是由于没有可比较的数据，在处理和解释组织方面遇到了困难。随后，我们开发了一种气液界面细胞培养物，以在去上皮化的人羊膜上培养角质形成细胞和成纤维细胞。我们在表皮一侧实现了大部分角化表面，在绒毛膜一侧实现了融合的成纤维细胞网络。