Rate of change in central corneal thickness: A viability indicator for conventional drainage tissues in organ culture
Rate of change in central corneal thickness: A viability indicator for conventional drainage tissues in organ culture
复制标题
DOI：
10.1016/j.exer.2005.10.027
复制
发表时间：
2006-06-01
影响因子：
3.4
通讯作者：
Stamer, W. D.
中科院分区：
文献类型：
--
作者：
Wan, Z.;Brigatti, L.;Stamer, W. D.
人体前段器官培养是了解小梁网络生物学的有力工具。然而，很大比例的培养前节段的数据是不可用的，因为组织不能满足质量控制要求，例如具有足够的小梁网组织学。本研究的目的是评估一种新的、实时的方法来评估常规引流组织在人体前段灌注模型中的生存能力。采用标准技术对22个人角膜前段进行灌注培养1周，同时测量角膜流出设施和角膜中央厚度（CCT）。灌注固定后，对来自每个前段所有四个象限的引流组织的甲苯胺蓝染色组织学切片进行分级，角膜中心的内皮细胞核用4′，6-二氨基-2-苯基吲哚染色并计数。我们发现，大多数具有稳定流出设施的前节段的CCT随时间而下降，而具有不稳定流出设施的前节段的CCT测量未能随时间下降（P < 0.01）。当比较CCT测量值与流出组织的组织学外观时，我们发现在11/11的病例中，具有可接受组织学评分的前节段CCT斜率为负（P < 0.01）。相反，在3/4的病例中，组织学评分不可接受的前节段的CCT斜率为正。最后，我们观察到CCT测量值与角膜内皮密度之间的显著关系（P < 0.01）。因此，在灌注过程中测量CCT的简单方法为评估灌注过程中前段的活力提供了第二个有用的方法。(c) 2006 Elsevier Ltd.版权所有。
Organ culture of human anterior segments is a powerful tool for understanding trabecular meshwork biology. However, data from a significant percentage of cultured anterior segments are unusable because tissues fail to meet quality control requirements, such as having adequate trabecular meshwork histology. The purpose of the present study was to evaluate a novel, real time method for assessing the viability of conventional drainage tissues in the human anterior segment perfusion model. Twenty-two human anterior segments were perfusion cultured using standard techniques for one week while measuring outflow facility and central corneal thickness (CCT). After perfusion-fixation, toludine blue-stained histological sections of drainage tissues from all four quadrants of each anterior segment were graded and endothelial cell nuclei from cornea centers were stained with 4',6-diamidino-2-phenylindole and counted. We found that most anterior segments with a stable outflow facility had a CCT that decreased over time, while anterior segments with an unstable outflow facility had CCT measurements that failed to decrease over time (P < 0.01). When comparing CCT measurements to histological appearance of outflow tissues, we found that in 11/11 cases, anterior segments with an acceptable histological score had a negative CCT slope (P < 0.01). Conversely in 3/4 instances, anterior segments with an unacceptable histological score had a positive CCT slope. Lastly, we observed a significant relationship between CCT measurements and corneal endothelial density (P < 0.01). Thus, the simple procedure of measuring CCT during anterior segment perfusion provides a second useful measure to assess the viability of the anterior segment during the perfusion process. (c) 2006 Elsevier Ltd. All rights reserved.