University Eye Clinic, Maastricht University Medical Center, Maastricht, The Netherlands.
Department of Cell Biology-Inspired Tissue Engineering, MERLN Institute for Technology-Inspired Regenerative Medicine, Maastricht, The Netherlands; and.
Cornea. 2020 Nov;39(11):1407-1414. doi: 10.1097/ICO.0000000000002391.
This study compares the effect of the transport of conventionally prestripped Descemet membrane endothelial keratoplasty (DMEK) tissue with the DMEK revolutionary advanced Preloadable Injection Device (RAPID) preloaded transport system from Geuder AG (Heidelberg, Germany). Endothelial cell loss, tissue integrity, endothelial cell phenotype, and viability were assessed and compared.
Twelve DMEK grafts were prestripped by the cornea bank and transported using the following 2 conditions: conventional flask (n = 6) or a preloaded transport cartridge (DMEK RAPID, n = 6). After transport, tissues were analyzed for cell density; denuded areas; immunolocalization of corneal endothelial markers, such as ZO-1, CD166, and Na/K ATPase; histology analysis; and cell viability staining with Hoechst, calcein AM, and ethidium homodimer.
Endothelial cell loss (10.35% vs. 9.15%) did not differ between transport conditions. Histological analysis confirmed the integrity of the Descemet membrane and endothelial cell layer with both transport conditions. Similarly, the corneal endothelial cell mosaic was conserved in both conditions. The ZO-1 tight junctions confirmed the integrity of the confluent corneal endothelial cell monolayer. CD166 and Na/K ATPase detection with immunofluorescence was also comparable. A similar percentage of dead cells was reported in both conditions (18.1% vs. 16.73%). Moreover, the surface covered with calcein-positive cells (59.02% vs. 61.95%) did not differ between transport conditions.
Our results suggest that DMEK grafts can be prestripped or preloaded into a novel transport cartridge and shipped to the clinic with comparable endothelial cell loss, phenotypical marker expression, and viability to the conventional prestripped donor tissue.
本研究比较了传统预撕迪斯美膜内皮角膜移植术(DMEK)组织的运输效果与 Geuder AG(德国海德堡)的革命性先进预加载注射装置(RAPID)预加载运输系统的效果。评估和比较了内皮细胞损失、组织完整性、内皮细胞表型和活力。
12 个 DMEK 移植物由角膜库预撕,使用以下 2 种条件进行运输:传统培养瓶(n=6)或预加载运输盒(DMEK RAPID,n=6)。运输后,分析组织的细胞密度;裸区;角膜内皮标志物如 ZO-1、CD166 和 Na/K ATPase 的免疫定位;组织学分析;以及用 Hoechst、钙黄绿素 AM 和 ethidium homodimer 进行细胞活力染色。
两种运输条件下,内皮细胞损失(10.35%对 9.15%)没有差异。组织学分析证实了两种运输条件下的迪斯美膜和内皮细胞层的完整性。同样,两种条件下的角膜内皮细胞镶嵌都得到了保存。ZO-1 紧密连接证实了连续的角膜内皮细胞单层的完整性。免疫荧光检测 CD166 和 Na/K ATPase 的结果也相似。两种条件下报告的死亡细胞比例相似(18.1%对 16.73%)。此外,用 calcein 阳性细胞覆盖的表面(59.02%对 61.95%)在两种运输条件之间没有差异。
我们的结果表明,DMEK 移植物可以预撕或预加载到新型运输盒中,并运往临床,与传统预撕供体组织相比,内皮细胞损失、表型标志物表达和活力相当。