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Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty

PURPOSE: To evaluate the effect of media composition and storage method on pre-prepared Descemet's membrane endothelial keratoplasty (DMEK) grafts. METHODS: 50 corneas were used. Endothelial wound healing and proliferation in different media were assessed using a standard injury model. DMEK gra...

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Autores principales: Bhogal, Maninder, Matter, Karl, Balda, Maria S, Allan, Bruce D
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BMJ Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5136687/
https://www.ncbi.nlm.nih.gov/pubmed/27543290
http://dx.doi.org/10.1136/bjophthalmol-2016-308855
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author Bhogal, Maninder
Matter, Karl
Balda, Maria S
Allan, Bruce D
author_facet Bhogal, Maninder
Matter, Karl
Balda, Maria S
Allan, Bruce D
author_sort Bhogal, Maninder
collection PubMed
description PURPOSE: To evaluate the effect of media composition and storage method on pre-prepared Descemet's membrane endothelial keratoplasty (DMEK) grafts. METHODS: 50 corneas were used. Endothelial wound healing and proliferation in different media were assessed using a standard injury model. DMEK grafts were stored using three methods: peeling with free scroll storage; partial peeling with storage on the stroma and fluid bubble separation with storage on the stroma. Endothelial cell (EC) phenotype and the extent of endothelial overgrowth were examined. Global cell viability was assessed for storage methods that maintained a normal cell phenotype. RESULTS: 1 mm wounds healed within 4 days. Enhanced media did not increase EC proliferation but may have increased EC migration into the wounded area. Grafts that had been trephined showed evidence of EC overgrowth, whereas preservation of a physical barrier in the bubble group prevented this. In grafts stored in enhanced media or reapposed to the stroma after trephination, endothelial migration occurred sooner and cells underwent endothelial-mesenchymal transformation. Ongoing cell loss, with new patterns of cell death, was observed after returning grafts to storage. Grafts stored as free scrolls retained more viable ECs than grafts prepared with the fluid bubble method (74.2± 3% vs 60.3±6%, p=0.04 (n=8). CONCLUSION: Free scroll storage is superior to liquid bubble and partial peeling techniques. Free scrolls only showed overgrowth of ECs after 4 days in organ culture, indicating a viable time window for the clinical use of pre-prepared DMEK donor material using this method. Methods for tissue preparation and storage media developed for whole corneas should not be used in pre-prepared DMEK grafts without prior evaluation.
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spelling pubmed-51366872016-12-08 Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty Bhogal, Maninder Matter, Karl Balda, Maria S Allan, Bruce D Br J Ophthalmol Laboratory Science PURPOSE: To evaluate the effect of media composition and storage method on pre-prepared Descemet's membrane endothelial keratoplasty (DMEK) grafts. METHODS: 50 corneas were used. Endothelial wound healing and proliferation in different media were assessed using a standard injury model. DMEK grafts were stored using three methods: peeling with free scroll storage; partial peeling with storage on the stroma and fluid bubble separation with storage on the stroma. Endothelial cell (EC) phenotype and the extent of endothelial overgrowth were examined. Global cell viability was assessed for storage methods that maintained a normal cell phenotype. RESULTS: 1 mm wounds healed within 4 days. Enhanced media did not increase EC proliferation but may have increased EC migration into the wounded area. Grafts that had been trephined showed evidence of EC overgrowth, whereas preservation of a physical barrier in the bubble group prevented this. In grafts stored in enhanced media or reapposed to the stroma after trephination, endothelial migration occurred sooner and cells underwent endothelial-mesenchymal transformation. Ongoing cell loss, with new patterns of cell death, was observed after returning grafts to storage. Grafts stored as free scrolls retained more viable ECs than grafts prepared with the fluid bubble method (74.2± 3% vs 60.3±6%, p=0.04 (n=8). CONCLUSION: Free scroll storage is superior to liquid bubble and partial peeling techniques. Free scrolls only showed overgrowth of ECs after 4 days in organ culture, indicating a viable time window for the clinical use of pre-prepared DMEK donor material using this method. Methods for tissue preparation and storage media developed for whole corneas should not be used in pre-prepared DMEK grafts without prior evaluation. BMJ Publishing Group 2016-11 2016-08-19 /pmc/articles/PMC5136687/ /pubmed/27543290 http://dx.doi.org/10.1136/bjophthalmol-2016-308855 Text en Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/ This is an Open Access article distributed in accordance with the terms of the Creative Commons Attribution (CC BY 4.0) license, which permits others to distribute, remix, adapt and build upon this work, for commercial use, provided the original work is properly cited. See: http://creativecommons.org/licenses/by/4.0/
spellingShingle Laboratory Science
Bhogal, Maninder
Matter, Karl
Balda, Maria S
Allan, Bruce D
Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty
title Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty
title_full Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty
title_fullStr Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty
title_full_unstemmed Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty
title_short Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty
title_sort organ culture storage of pre-prepared corneal donor material for descemet's membrane endothelial keratoplasty
topic Laboratory Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5136687/
https://www.ncbi.nlm.nih.gov/pubmed/27543290
http://dx.doi.org/10.1136/bjophthalmol-2016-308855
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