预加载后弹力层内皮角膜移植术中内皮细胞丢失的定量分析
Quantitative Analysis of Endothelial Cell Loss in Preloaded Descemet Membrane Endothelial Keratoplasty Grafts.
作者信息
Wolle Meraf A, DeMill David L, Johnson Lauren, Lentz Stephen I, Woodward Maria A, Mian Shahzad I
机构信息
*Kellogg Eye Center, University of Michigan, Ann Arbor, MI;†Wilmer Eye Institute, Johns Hopkins University, Baltimore, MD;‡Devers Eye Institute, Portland, OR;§Eversight Michigan, Ann Arbor, MI; and¶Department of Internal Medicine, Division of Metabolism, Endocrinology and Diabetes, University of Michigan, MI, USA.
出版信息
Cornea. 2017 Nov;36(11):1295-1301. doi: 10.1097/ICO.0000000000001301.
PURPOSE
Availability of preloaded Descemet membrane endothelial keratoplasty (pDMEK) tissue may increase acceptance of DMEK in surgical management of endothelial disease. The goal of this study was to determine the safety of pDMEK grafts for 24 hours before surgery by analyzing endothelial cell loss (ECL) using 2 image analysis software programs.
METHODS
A total of 18 cadaveric corneas were prepared for DMEK using a standardized technique and loaded in a modified Jones tube injector. Nine of the corneas were injected into Calcein AM vital dye after 1 minute (controls), and the remaining 9 corneas were left preloaded for 24 hours before injection into vital dye for staining. The stained corneas were imaged using an inverted confocal microscope. ECL was then analyzed and quantified by 2 different graders using 2 image analysis software programs.
RESULTS
The control DMEK tissue resulted in 22.0% ± 4.0% ECL compared with pDMEK tissue, which resulted in 19.2% ± 7.2% ECL (P = 0.31). Interobserver agreement was 0.93 for MetaMorph and 0.92 for Fiji. The average time required to process images with MetaMorph was 2 ± 1 minutes and with Fiji was 20 ± 10 minutes. Intraobserver agreement was 0.97 for MetaMorph and 0.93 for Fiji.
CONCLUSIONS
Preloading DMEK tissue is safe and may provide an alternative technique for tissue distribution and surgery for DMEK. The use of MetaMorph software for quantifying ECL is a novel and accurate imaging method with increased efficiency and reproducibility compared with the previously validated Fiji.
目的
预装载的Descemet膜内皮角膜移植术（pDMEK）组织的可用性可能会提高DMEK在角膜内皮疾病手术治疗中的接受度。本研究的目的是通过使用2种图像分析软件程序分析内皮细胞丢失（ECL），来确定手术前24小时pDMEK移植物的安全性。
方法
使用标准化技术为DMEK制备了总共18只尸体角膜，并装载到改良的Jones管注射器中。其中9只角膜在1分钟后注射钙黄绿素AM活性染料（对照组），其余9只角膜在注射活性染料染色前预装载24小时。使用倒置共聚焦显微镜对染色后的角膜进行成像。然后由2名不同的分级人员使用2种图像分析软件程序对ECL进行分析和量化。
结果
与pDMEK组织相比，对照DMEK组织的ECL为22.0%±4.0%，而pDMEK组织的ECL为19.2%±7.2%（P=0.31）。MetaMorph的观察者间一致性为0.93，Fiji为0.92。使用MetaMorph处理图像的平均时间为2±1分钟，使用Fiji为20±10分钟。MetaMorph的观察者内一致性为0.97，Fiji为0.93。
结论
预装载DMEK组织是安全的，可能为DMEK的组织分配和手术提供一种替代技术。与先前验证的Fiji相比，使用MetaMorph软件量化ECL是一种新颖且准确的成像方法，具有更高的效率和可重复性。
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