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

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 co...

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Autores principales:	Silvia Rodríguez-Fernández, Marcelino Álvarez-Portela, Esther Rendal-Vázquez, María Piñeiro-Ramil, Clara Sanjurjo-Rodríguez, Rocío Castro-Viñuelas, Jacinto Sánchez-Ibáñez, Isaac Fuentes-Boquete, Silvia Díaz-Prado
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	corneal cryopreservation cryoprotective agent corneal endothelium corneal storage corneal transplantation DMSO Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/73704223f4204e479a85704d8c8bb305
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spelling	oai:doaj.org-article:73704223f4204e479a85704d8c8bb3052021-11-25T17:57:55ZAnalysis of Cryopreservation Protocols and Their Harmful Effects on the Endothelial Integrity of Human Corneas10.3390/ijms2222125641422-00671661-6596https://doaj.org/article/73704223f4204e479a85704d8c8bb3052021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/22/12564https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Corneal 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.Silvia Rodríguez-FernándezMarcelino Álvarez-PortelaEsther Rendal-VázquezMaría Piñeiro-RamilClara Sanjurjo-RodríguezRocío Castro-ViñuelasJacinto Sánchez-IbáñezIsaac Fuentes-BoqueteSilvia Díaz-PradoMDPI AGarticlecorneal cryopreservationcryoprotective agentcorneal endotheliumcorneal storagecorneal transplantationDMSOBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12564, p 12564 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	corneal cryopreservation cryoprotective agent corneal endothelium corneal storage corneal transplantation DMSO Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	corneal cryopreservation cryoprotective agent corneal endothelium corneal storage corneal transplantation DMSO Biology (General) QH301-705.5 Chemistry QD1-999 Silvia Rodríguez-Fernández Marcelino Álvarez-Portela Esther Rendal-Vázquez María Piñeiro-Ramil Clara Sanjurjo-Rodríguez Rocío Castro-Viñuelas Jacinto Sánchez-Ibáñez Isaac Fuentes-Boquete Silvia Díaz-Prado Analysis of Cryopreservation Protocols and Their Harmful Effects on the Endothelial Integrity of Human Corneas
description	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.
format	article
author	Silvia Rodríguez-Fernández Marcelino Álvarez-Portela Esther Rendal-Vázquez María Piñeiro-Ramil Clara Sanjurjo-Rodríguez Rocío Castro-Viñuelas Jacinto Sánchez-Ibáñez Isaac Fuentes-Boquete Silvia Díaz-Prado
author_facet	Silvia Rodríguez-Fernández Marcelino Álvarez-Portela Esther Rendal-Vázquez María Piñeiro-Ramil Clara Sanjurjo-Rodríguez Rocío Castro-Viñuelas Jacinto Sánchez-Ibáñez Isaac Fuentes-Boquete Silvia Díaz-Prado
author_sort	Silvia Rodríguez-Fernández
title	Analysis of Cryopreservation Protocols and Their Harmful Effects on the Endothelial Integrity of Human Corneas
title_short	Analysis of Cryopreservation Protocols and Their Harmful Effects on the Endothelial Integrity of Human Corneas
title_full	Analysis of Cryopreservation Protocols and Their Harmful Effects on the Endothelial Integrity of Human Corneas
title_fullStr	Analysis of Cryopreservation Protocols and Their Harmful Effects on the Endothelial Integrity of Human Corneas
title_full_unstemmed	Analysis of Cryopreservation Protocols and Their Harmful Effects on the Endothelial Integrity of Human Corneas
title_sort	analysis of cryopreservation protocols and their harmful effects on the endothelial integrity of human corneas
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/73704223f4204e479a85704d8c8bb305
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Analysis of Cryopreservation Protocols and Their Harmful Effects on the Endothelial Integrity of Human Corneas

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