Chen Conan, Solar Steven Jared, Lohmeier John, Terrin Staci, Baliga Satya, Wiener Batya Gold, Lewis Daniel Schouten, Chiang Eric, Barnes Kali Alexandra, Chaurasia Akash, Eghrari Allen O
University of Colorado Denver School of Medicine, Denver, Colorado, USA.
Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland, USA.
BMJ Open Ophthalmol. 2021 May 20;6(1):e000679. doi: 10.1136/bmjophth-2020-000679. eCollection 2021.
To assess feasibility and compare the effects of 96-hour shipment of Descemet membrane endothelial keratoplasty (DMEK) grafts as a scroll or a tri-fold on cell viability.
DMEK grafts were prepared at the Rocky Mountain Lions Eye Bank. Twenty pre-stripped DMEK grafts, paired from 10 donors, were either tri-folded in an endothelium-in configuration using microforceps and loaded into a plastic Treyetech cartridge, or suctioned in a scrolled endothelium-out configuration into a modified Jones Tube. Grafts were shipped via FedEx to a secondary location and back for 48 hours each way, resulting in a total shipping time of 96 hours. After shipping, grafts were removed from inserters onto glass slides and unfolded using viscoelastic with endothelium facing upwards. Calcein-AM stained grafts were imaged with a fluorescent microscope and endothelial cell loss (ECL) was measured using trainable segmentation in Fiji by a masked grader.
A total of 20 grafts were shipped for 96 hours, split between preloaded tri-folded (n=10) and preloaded scrolled (n=10) tissues. No significant difference in ECL was observed across groups after prolonged shipping (14.8% vs 13.7% ECL respectively, p=0.68).
For preloaded DMEK after 96 hours, both scrolled and tri-folded tissue demonstrated clinically acceptable levels of ECL. The data suggest a wider window of time for endothelial cell viability and is promising for the prospect of international shipment of preloaded grafts.
评估将Descemet膜内皮角膜移植术（DMEK）移植物以卷轴形式或三折形式进行96小时运输的可行性，并比较其对细胞活力的影响。
DMEK移植物在落基山狮子眼库制备。从10名供体获取的20片预剥离DMEK移植物，成对处理，其中一组使用显微镊以内皮朝内的方式三折后装入塑料Treyetech盒中，另一组以卷轴形式内皮朝外吸入改良的琼斯管。移植物通过联邦快递运往第二个地点，往返各48小时，总运输时间为96小时。运输后，将移植物从插入器中取出置于载玻片上，使用粘弹性物质展开，使内皮面朝上。用荧光显微镜对钙黄绿素-AM染色的移植物进行成像，并由经过训练的分级人员使用Fiji中的可训练分割功能测量内皮细胞损失（ECL）。
总共20片移植物运输96小时，分为预装载三折组（n = 10）和预装载卷轴组（n = 10）。长时间运输后，两组之间的ECL无显著差异（分别为14.8%和13.7%，p = 0.68）。
对于预装载的DMEK移植物，在运输96小时后，卷轴形式和三折形式的组织均显示出临床上可接受的ECL水平。数据表明内皮细胞活力的时间窗口更宽，这对于预装载移植物的国际运输前景很有希望。
