Analysis of Cryopreservation Protocols and Their Harmful Effects on the Endothelial Integrity of Human Corneas

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Analysis of Cryopreservation Protocols and Their Harmful Effects on the Endothelial Integrity of Human CorneasAuthor(s)
Date
2021-11-22Citation
Rodríguez-Fernández S, Álvarez-Portela M, Rendal-Vázquez E, Piñeiro-Ramil M, Sanjurjo-Rodríguez C, Castro-Viñuelas R, Sánchez-Ibáñez J, Fuentes-Boquete I, Díaz-Prado S. Analysis of cryopreservation protocols and their harmful effects on the endothelial integrity of human corneas. Int J Mol Sci. 2021 Nov 22;22(22):12564.
Abstract
[Abstract] Corneal cryopreservation can partially solve the worldwide concern regarding donor cornea shortage for keratoplasties. In this study, human corneas were cryopreserved using two standard cryopreservation protocols that are employed in the Tissue Bank of the Teresa Herrera Hospital (Spain) to store corneas for tectonic keratoplasties (TK protocol) and aortic valves (AV protocol), and two vitrification protocols, VS55 and DP6. Endothelial viability and general corneal state were evaluated to determine the protocol that provides the best results. The potential corneal cryopreservation protocol was studied in detail taking into consideration some cryopreservation-related variables and the endothelial integrity and stroma arrangement of the resulting cryopreserved corneas. TK corneas showed mostly viable endothelial cells, while the others showed few (AV) or none (DP6 and VS55). The corneal structure was well maintained in TK and AV corneas. TK corneas showed endothelial acellular areas surrounded by injured cells and a normal-like stromal fiber arrangement. Cryoprotectant solutions of the TK protocol presented an increasing osmolality and a physiological pH value. Cooling temperature rate of TK protocol was of 1 °C/min to −40 °C and 3 °C/min to −120 °C, and almost all of dimethyl sulfoxide left the tissue after washing. Future studies should be done changing cryopreservation-related variables of the TK protocol to store corneas of optical grade.
Keywords
Corneal cryopreservation
Cryoprotective agent
Corneal endothelium
Corneal storage
Corneal transplantation
DMSO
Cryoprotective agent
Corneal endothelium
Corneal storage
Corneal transplantation
DMSO
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Creative Commons Attribution 4.0 International Licence (CC-BY 4.0)
ISSN
1422-0067