Wagner Malina, Walter Peter, Salla Sabine, Johnen Sandra, Plange Niklas, Rütten Stephan, Goecke Tamme W, Fuest Matthias
Department of Ophthalmology, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany.
Department of Electron Microscopy, University Hospital RWTH Aachen, Aachen, Germany.
Graefes Arch Clin Exp Ophthalmol. 2018 Jun;256(6):1117-1126. doi: 10.1007/s00417-018-3973-1. Epub 2018 Apr 5.
Amniotic membrane (AM) is an essential tool in ocular surface reconstruction. In this study, we analyzed the differential effects of glycerol and straight storage at - 80 °C for up to 6 months on the structural, biological, and mechanical properties of amniotic membrane (AM).
Human placentae of 11 different subjects were analyzed. AMs were stored at - 80 °C, either with a 1:1 mixture of Dulbecco's modified Eagle medium and glycerol (glycerol) or without any medium or additives (straight). Histological image analysis, tensile strength, cell viability, and basic fibroblast growth factor (bFGF) secretion were evaluated at 0.5, 1, 3, and 6 months.
Histologically, neither glycerol nor straight storage significantly altered the epithelial or stromal structure of the AM. However, the cell number of the stroma was significantly reduced during the freezing process, independently of the storage method (p = 0.05-0.001). Tensile strength and Young's modulus were not influenced by the storage method, but longer storage periods significantly increased the tensile strength of the AMs (p = 0.028). Cell viability was higher in glycerol rather than straight AM samples for up to 3 months of storage (p = 0.047-0.03). Secretion of bFGF at 3 months of storage was significantly higher in glycerol versus straight frozen AM samples (p = 0.04).
Glycerol led to higher cell viability and higher bFGF secretion for up to 3 months of AM storage. However, no significant differences between the two methods were observed at 6 months of storage at - 80 °C.
羊膜（AM）是眼表重建的重要工具。在本研究中，我们分析了甘油处理以及在-80°C直接储存长达6个月对羊膜（AM）的结构、生物学和力学性能的不同影响。
分析了11名不同受试者的人胎盘。羊膜在-80°C下储存，一种是与杜氏改良 Eagle 培养基和甘油按1:1混合（甘油组），另一种是不添加任何培养基或添加剂（直接储存组）。在0.5、1、3和6个月时评估组织学图像分析、拉伸强度、细胞活力和碱性成纤维细胞生长因子（bFGF）分泌情况。
组织学上，甘油处理和直接储存均未显著改变羊膜的上皮或基质结构。然而，在冷冻过程中，无论储存方法如何，基质细胞数量均显著减少（p = 0.05 - 0.001）。拉伸强度和杨氏模量不受储存方法影响，但储存时间延长显著增加了羊膜的拉伸强度（p = 0.028）。在长达3个月的储存期内，甘油处理的羊膜样本中的细胞活力高于直接储存的样本（p = 0.047 - 0.03）。储存3个月时甘油处理的羊膜样本中bFGF的分泌显著高于直接冷冻的样本（p = 0.04）。
在羊膜储存长达3个月时，甘油处理可导致更高的细胞活力和更高的bFGF分泌。然而，在-80°C储存6个月时，两种方法之间未观察到显著差异。
