Jester J V, Maurer J K, Petroll W M, Wilkie D A, Parker R D, Cavanagh H D
University of Texas, Southwestern Medical Center, Dallas 75235-9057, USA.
Toxicol Pathol. 1996 Jul-Aug;24(4):412-28. doi: 10.1177/019262339602400404.
The purpose of this study was to assess the ability of in vivo confocal microscopy (CM) to provide noninvasively derived histopathologic correlates of surfactant-induced eye irritation from which specific pathologic mechanisms can be identified and later evaluated in alternative in vitro models. Rats and rabbits, divided into groups of 5, received 10 microliters of an anionic or cationic surfactant in one eye with the other eye used as a control. At specified times, eyes were examined and scored for ocular irritancy using a penlight and slit-lamp. Subsequently, corneas were evaluated by in vivo CM to evaluate epithelial layer thickness and surface epithelial cell area, corneal thickness, depth of necrosis, inflammation, fibrosis, and endothelial injury. At 3 hr, the anionic surfactant produced slight irritation with peak scores of 12.4 and 8.0 out of a possible 110 in the rats and rabbits, respectively. In vivo CM revealed changes limited to the corneal epithelium that decreased in thickness to 78% in rats and 81% in rabbits at 3 hr. This decrease in the thickness correlated with a significant decrease in surface epithelial cell area from 2,061 +/- 395 microns2 to 567 +/- 330 microns2 in the rats and 1,523 +/- 185 microns2 to 934 +/- 71 microns2 in the rabbits (p < 0.005 and 0.005, respectively). The cationic surfactant produced severe irritation in both the rats and rabbits with peak scores of 85.4 and 80.2 occurring at day 2, respectively. In vivo CM in the rats showed complete loss of corneal epithelium, lysis of keratocytes, and loss of corneal endothelium. In the rabbits, injury appeared limited to the anterior cornea with complete loss of epithelium and loss of keratocytes extending to 52% of the corneal thickness. These findings establish the application of noninvasive, in vivo CM to qualitatively and quantitatively characterize the pathobiology of ocular irritation in situ. This information will be important in the development and evaluation of mechanistically based in vitro alternatives for ocular irritancy testing.
本研究的目的是评估体内共聚焦显微镜（CM）提供表面活性剂诱导的眼部刺激的非侵入性组织病理学关联的能力，据此可确定特定的病理机制，并随后在其他体外模型中进行评估。将大鼠和兔子分成每组5只，在一只眼睛中滴入10微升阴离子或阳离子表面活性剂，另一只眼睛作为对照。在特定时间，使用手电筒和裂隙灯检查眼睛并对眼刺激性进行评分。随后，通过体内共聚焦显微镜评估角膜，以评估上皮层厚度和表面上皮细胞面积、角膜厚度、坏死深度、炎症、纤维化和内皮损伤。在3小时时，阴离子表面活性剂产生轻微刺激，大鼠和兔子的最高得分分别为12.4和8.0（满分110分）。体内共聚焦显微镜显示变化仅限于角膜上皮，在3小时时大鼠角膜上皮厚度降至78%，兔子降至81%。这种厚度的降低与表面上皮细胞面积的显著减少相关，大鼠从2,061±395平方微米降至567±330平方微米，兔子从1,523±185平方微米降至934±71平方微米（分别为p<0.005和0.005）。阳离子表面活性剂在大鼠和兔子中均产生严重刺激，最高得分分别在第2天达到85.4和80.2。大鼠的体内共聚焦显微镜显示角膜上皮完全丧失、角膜细胞溶解和角膜内皮丧失。在兔子中，损伤似乎仅限于前角膜，上皮完全丧失，角膜细胞丧失延伸至角膜厚度的52%。这些发现确立了非侵入性体内共聚焦显微镜在定性和定量表征原位眼部刺激病理生物学方面的应用。该信息对于基于机制的体外眼刺激性测试替代方法的开发和评估将具有重要意义。
