Department of Ophthalmology, Saarland University Medical Center, Homburg, Germany.
Department of Ophthalmology, Saarland University Medical Center, Homburg, Germany.
Am J Ophthalmol. 2021 Jan;221:17-26. doi: 10.1016/j.ajo.2020.08.001. Epub 2020 Aug 11.
To present a novel, reproducible, and noninvasive method to quantify endothelial cell loss (ECL) of pre-stripped endothelial Descemet membrane lamellae (EDML) caused by its preparation and storage for 5 days.
Prospective laboratory investigation.
Thirty EDML were stripped from corneoscleral discs and placed in a well plate containing organ culture medium 1 without dextran. An additional 5 corneoscleral discs were also placed in the same medium and served as a control group. Endothelial cell density (ECD) was measured without any additional manipulation by using spectral microscopy following an extensive protocol by which 3 clear images from the center and periphery were used for each measurement, and each measurement was repeated 5 times. ECD was measured before and directly after preparation and on days 1, 2, and 5 of storage.
The average ECD of the 30 corneoscleral discs, which later underwent stripping, was 2,292 ± 308 cells/mm vs 2,129 ± 222 cells/mm for the 5 corneoscleral discs of the control group. The ECL of the control group was significantly lower than that of the EDML group (P < .0001), reaching ±2% versus 11 ± 5%, respectively, on day 0; 3% ± 4% versus 19 ± 10%, respectively, on day 1; 2% ± 2% versus 22% ± 11%, respectively on day 2; and 4% ± 3% versus 23% ± 9%, respectively, on day 5. Reproducibility of the results on all measurement days was good, with Cronbach alpha values ranging from 0.85 to 0.98.
A highly reproducible, noninvasive method was presented for measuring the ECD of the EDML. Prestripped EDML lose a significant amount of cells, up to 11%, due to the preparation process and up to 23% after 5 days of storage. Therefore, shipping them after several days of storage to be used in a DMEK surgery cannot be recommended.
介绍一种新颖、可重复且非侵入性的方法，以量化经过预处理和 5 天储存的预剥离内皮 Descemet 膜瓣（EDML）中内皮细胞的损失（ECL）。
前瞻性实验室研究。
从角膜巩膜瓣中剥离 30 个 EDML，并将其放置在含有不含葡聚糖的器官培养物 1 的培养皿中。另外 5 个角膜巩膜瓣也放置在相同的培养基中作为对照组。在没有任何额外操作的情况下，使用光谱显微镜测量内皮细胞密度（ECD），该测量采用了广泛的方案，即每个测量使用中心和外围的 3 个清晰图像，每个测量重复 5 次。在准备之前和之后以及储存的第 1、2 和 5 天直接测量 ECD。
30 个角膜巩膜瓣在进行剥离后，平均 ECD 为 2292±308 个细胞/mm，而 5 个角膜巩膜瓣的对照组为 2129±222 个细胞/mm。对照组的 ECL 明显低于 EDML 组（P<0.0001），在第 0 天分别达到±2%对 11±5%，在第 1 天分别达到±3%对 19±10%，在第 2 天分别达到±2%对 22%±11%，在第 5 天分别达到±4%对 23%±9%。所有测量日的结果均具有良好的可重复性，Cronbach alpha 值范围为 0.85 至 0.98。
提出了一种高度可重复、非侵入性的方法来测量 EDML 的 ECD。预剥离的 EDML 由于制备过程会损失大量细胞，最多可达 11%，在储存 5 天后损失细胞最多可达 23%。因此，不建议在储存几天后运送它们，以用于 DMEK 手术。
