Human amniotic membrane (HAM) is a biomaterial with biological properties beneficial to tissue repair, serving as a substrate for cell cultivation. Irradiation is used for tissue sterilization, but can damage the HAM structure. The objective of this paper was to construct a skin substitute, composed of human keratinocytes cultured on glycerolated HAMs, and to evaluate the influence radiation on subsequent cell culture growth. Four batches of HAMs were glycerolated, and half of them were radio-sterilzed with 25 kGy. Non-irradiated glycerolated HAM (ni-HAM) and irradiated glycerolated HAM (i-HAM) samples were then de-epithelized and analyzed using optical microscopy (Picrossirius staining), immunofluorescence and electron microscopy. Subsequently, keratinocytes were cultured on ni- and i-HAMs, and either immersed or positioned at the air-liquid interface. The basement membranes of the ni-HAM group remained intact following de-epithelialization, whereas the i-HAM group displayed no evidence or remnant presence of these membranes. Concerning the keratinocyte cultures, the ni-HAM substrate promoted the growth of multi-layered and differentiated epithelia. Keratinocytes cultured on i-HAM formed epithelium composed of three layers of stratification and discrete cell differentiation. The glycerolated HAM was compatible with cultured epithelia, demonstrating its potential as a skin substitute. Irradiation at 25 kGy caused structural damage to the amnion.

中文翻译：

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放射灭菌的甘油化羊膜作为培养的人类上皮细胞的底物的评价。

人羊膜（HAM）是一种具有有益于组织修复的生物学特性的生物材料，可作为细胞培养的基质。辐照用于组织灭菌，但会损坏 HAM 结构。本文的目的是构建一种皮肤替代品，由在甘油化 HAM 上培养的人角质形成细胞组成，并评估辐射对后续细胞培养物生长的影响。四批 HAM 被甘油化，其中一半用 25 kGy 进行放射灭菌。然后对未辐照的甘油化 HAM (ni-HAM) 和辐照的甘油化 HAM (i-HAM) 样品进行去上皮并使用光学显微镜（Picrossirius 染色）、免疫荧光和电子显微镜进行分析。随后，角质形成细胞在 ni- 和 i-HAM 上培养，并且浸入或定位在气液界面处。ni-HAM 组的基底膜在去上皮后保持完整，而 i-HAM 组没有显示出这些膜的证据或残留。关于角质形成细胞培养物，ni-HAM 基质促进了多层分化上皮细胞的生长。在 i-HAM 上培养的角质形成细胞形成由三层分层和离散细胞分化组成的上皮。甘油化的 HAM 与培养的上皮细胞相容，证明了其作为皮肤替代品的潜力。25 kGy 的辐照对羊膜造成结构性损坏。关于角质形成细胞培养物，ni-HAM 基质促进了多层分化上皮细胞的生长。在 i-HAM 上培养的角质形成细胞形成由三层分层和离散细胞分化组成的上皮。甘油化的 HAM 与培养的上皮细胞相容，证明了其作为皮肤替代品的潜力。25 kGy 的辐照对羊膜造成结构性损坏。关于角质形成细胞培养物，ni-HAM 基质促进了多层分化上皮细胞的生长。在 i-HAM 上培养的角质形成细胞形成由三层分层和离散细胞分化组成的上皮。甘油化的 HAM 与培养的上皮细胞相容，证明了其作为皮肤替代品的潜力。25 kGy 的辐照对羊膜造成结构性损坏。