Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
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ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
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Advances in Clinical and Experimental Medicine

2018, vol. 27, nr 8, August, p. 1033–1036

doi: 10.17219/acem/73713

Publication type: original article

Language: English

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Structural changes arising from different thawing protocols on cryopreserved human allograft’s aortic valve leaflets

Robert Novotný1,2,3,A,B,C,D,E,F, Dáša Slížová4,C, Jaroslav Hlubocký1,2,E,F, Otakar Krs4,B,C, Jaroslav Špatenka5,6,A,E, Jan Burkert5,6,B,C, Radovan Fiala6,7,B, Petr Mitáš1,2,D,E, Pavel Mĕricka8,E,F, Miroslav Špaček1,2,B, Zuzana Hlubocká2,8,9,E, Jaroslav Lindner1,2,A,F

1 2nd Department of Cardiovascular Surgery, General University Hospital, Prague, Czech Republic

2 1st Faculty of Medicine, Charles University, Prague, Czech Republic

3 Transplant Surgery Department, Institute for Clinical and Experimental Medicine, Prague, Czech Republic

4 Department of Anatomy, Faculty of Medicine, Charles University, Hradec Králové, Czech Republic

5 Transplant Center, University Hospital Motol, Prague, Czech Republic

6 Department of Cardiovascular Surgery, Faculty Hospital Motol, Prague, Czech Republic

7 2nd Faculty of Medicine, Charles University, Prague, Czech Republic

8 Tissue Bank, Faculty Hospital Hradec Králové, Faculty of Medicine, Charles University, Czech Republic

9 2nd Internal Department of Cardiology and Angiology, General University Hospital, Prague, Czech Republic

Abstract

Background. The aim of our experimental work was to assess the impact and morphological changes that arise during different thawing protocols on human aortic valve (AV) leaflets resected from cryopreserved aortic root allografts (CARAs).
Objectives. Two thawing protocols were tested: 1. CARAs were thawed at a room temperature (23°C); 2. CARAs were placed directly into a water bath at a temperature of 37°C. After all the samples were thawed, non-coronary AV leaflets were sampled from each specimen and fixed in a 4% formaldehyde solution before they were sent for morphological analysis.
Material and Methods. All the samples were washed in distilled water for 5 min and dehydrated in a graded ethanol series (70%, 85%, 95%, and 100%) for 5 min at each level. The tissue samples were then immersed in 100% hexamethyldisilazane (HMDS) for 10 min, and then air-dried in an exhaust hood at room temperature. Processed samples were mounted on stainless steel stubs and coated with gold. Histological analysis was performed with the use of an electron microscope on a scanning mode operating at 25 kV – BS 301.
Results. Thawing protocol 1 (room temperature at 23°C): 6 (100%) samples showed loss of the endothelial covering of the basal membrane with no damage to the basal lamina. Thawing protocol 2 (water bath at 37°C): 5 (83%) samples showed loss of the endothelial covering of the basal membrane with no damage to the basal lamina. One (17%) sample showed loss of the endothelial covering the basal membrane with significant damage to the basal membrane.
Conclusion. Based on our experimental work, we can clearly conclude that cryopreserved AV leaflet allografts show identical structural changes at different rates of thawing.

Key words

aortic valve, allograft, thawing, cryopreserved, structural changes

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