Malina Wagner1, Peter Walter1, Sabine Salla1, Sandra Johnen1, Niklas Plange1, Stephan Rütten2, Tamme W Goecke3, Matthias Fuest4. 1. Department of Ophthalmology, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany. 2. Department of Electron Microscopy, University Hospital RWTH Aachen, Aachen, Germany. 3. Department of Gynecology and Obstetrics, RWTH Aachen University, Aachen, Germany. 4. Department of Ophthalmology, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany. mfuest@ukaachen.de.
Abstract
PURPOSE: 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). METHODS: 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. RESULTS: 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). DISCUSSION: 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.
PURPOSE: 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). METHODS: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. RESULTS: 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). DISCUSSION: 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.
Authors: H Thomasen; M Pauklin; B Noelle; G Geerling; J Vetter; P Steven; K-P Steuhl; D Meller Journal: Curr Eye Res Date: 2011-01-28 Impact factor: 2.424
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Authors: Beau J Fenner; Nur Zahirah B M Yusoff; Matthias Fuest; Lei Zhou; Francisco Bandeira; Howard Y Cajucom-Uy; H K Tan; Jodhbir S Mehta; Gary H F Yam Journal: Eye Vis (Lond) Date: 2019-10-12