Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty.
Organ culture storage of pre-prepared corneal donor material for Descemet's membrane endothelial keratoplasty.
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DOI：
10.1136/bjophthalmol-2016-308855
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发表时间：
2016-11
期刊：
影响因子：
--
通讯作者：
Allan BD
中科院分区：
文献类型：
--
作者：
Bhogal M;Matter K;Balda MS;Allan BD
评价培养基成分和储存方法对预制后弹力层内皮角膜移植术（DMEK）移植物的影响。使用了50个角膜。使用标准损伤模型评估不同介质中的内皮伤口愈合和增殖。使用三种方法储存 DMEK 移植物：自由滚动储存剥离；部分剥离并在基质上储存，并且流体气泡分离并在基质上储存。检查内皮细胞（EC）表型和内皮过度生长的程度。针对维持正常细胞表型的储存方法评估了整体细胞活力。 1 毫米的伤口在4天内愈合。增强培养基不会增加 EC 增殖，但可能会增加 EC 向受伤区域的迁移。环钻的移植物显示出 EC 过度生长的证据，而气泡组中物理屏障的保留阻止了这种情况。在储存在增强培养基中或环钻后重新放置到基质中的移植物中，内皮迁移发生得更快并且细胞经历内皮间质转化。将移植物返回储存后，观察到持续的细胞损失和新的细胞死亡模式。以自由卷轴形式保存的移植物比用流体气泡法制备的移植物保留了更多的存活 EC（74.2± 3% vs 60.3±6%，p=0.04 (n=8)）。自由卷轴存储优于液体气泡和部分剥离技术。自由卷轴仅在器官培养 4 天后才显示出 EC 过度生长，这表明使用此方法预制 DMEK 供体材料的临床使用的可行时间窗口。未经事先评估，为整个角膜开发的组织制备和存储介质不应用于预先准备的 DMEK 移植物。
To evaluate the effect of media composition and storage method on pre-prepared Descemet's membrane endothelial keratoplasty (DMEK) grafts. 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. 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). 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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影响因子：
4.2
作者：
Heindl, Ludwig M.;Schloetzer-Schrehardt, Ursula;Kruse, Friedrich E.
通讯作者：
Kruse, Friedrich E.
影响因子：
8.1
作者：
Monnereau, Claire;Quilendrino, Ruth;Melles, Gerrit R. J.
通讯作者：
Melles, Gerrit R. J.
影响因子：
2.8
作者：
Lie, Jessica T.;Birbal, Renuka;Melles, Gerrit R. J.
通讯作者：
Melles, Gerrit R. J.
DOI：
10.1034/j.1600-0420.1999.770306.x
发表时间：
1999-06-01
期刊：
ACTA OPHTHALMOLOGICA SCANDINAVICA
影响因子：
--
作者：
Ehlers, H;Ehlers, N;Hjortdal, JO
通讯作者：
Hjortdal, JO
DOI：
10.1007/s004170100354
发表时间：
2001-10-01
影响因子：
2.7
作者：
Moller-Pedersen, T;Hartmann, U;Engelmann, K
通讯作者：
Engelmann, K