Zhu Wen-qing, Xu Jian-jiang, Sun Xing-huai, Qiu Ting, Hong Jia-xu, Wang Yan, Wang Wen-tao
Department of Ophthalmology, Eye Ear Nose and Throat Hospital of Fudan University, Shanghai 200031, China.
Zhonghua Yan Ke Za Zhi. 2010 Jan;46(1):18-24.
To analyze the morphology on the ocular surface of severe alkali burns patients by in vivo laser scanning confocal microscopy.
This research was a retrospective observation case series. From February to November 2008 in Eye Ear Nose and Throat Hospital of Fudan University, 39 alkali burns patients who classified as III or IV according to Roper-Hall classification were enrolled in this study. They were divided into four groups according to the course of disease: A (less than 3 months), B (3 - 6 months), C (6 - 12 months) and D (over 12 months). In vivo laser scanning confocal microscopic examinations were performed on the injured cornea, the limbus and the bulbar conjunctiva and the images were recorded. The morphology of the injured cornea, the limbus and the bulbar conjunctiva was analyzed and the densities of the inflammatory cells and dendritic cells in the limbus were calculated. One-way analysis of variance was used to compare the means of the inflammatory cells and dendritic cells. Subsequently the data between two groups were analyzed by least significant difference.
The corneal epitheliums of the patients in Group A manifested large irregular features with hyperreflective cytoplasm and hyporeflective nuclei, sometimes losing cell features. There were numerous small hyperreflective inflammatory cells in groups beneath the superficial epitheliums. Shallow corneal stroma was edema, and it was hard to discriminate the morphology of the stromal cells. Deep stromal cells were in the activated state. The images of the endothelial layer were unclear. In Group B and Group C, there were the same manifestation of the superficial epitheliums as the group A and it disappeared in Group D. The inflammatory cells beneath the superficial epitheliums reduced and some residual basal epitheliums and hyperreflective conjunctiva-like epitheliums were visible in Group B and Group C. In Group D, there were small oval tight-arranged cells with punctiform hyperreflective nuclei instead of normal corneal basal epitheliums. In Group B, it was still hard to discriminate the morphology of the shallow stroma cells. Deep stromal cells were still in the activated state. In Group C and Group D, corneal stroma was replaced by the fibrous tissues. The images of the endothelial layer were still unclear in the other groups. The Vogt palisades in the limbus of the severe alkali burns patients were destroyed in all groups. There were rich vascular nets in the limbus. The densities of the limbal inflammatory cells in four groups were (4023 +/- 343), (2975 +/- 246), (2652 +/- 375), (2679 +/- 299) cells/mm(2), respectively. Significant difference in inflammatory cell density was found among groups (F = 40.001, P = 0.000). The densities of the limbal dendritic cells in four groups were (106 +/- 19), (132 +/- 35), (141 +/- 26), (98 +/- 24) cells/mm(2), respectively. Significant difference in dendritic cell density was found among groups (F = 8.053, P = 0.000). When the injured area of the conjunctiva was limited, it was hard to discriminate the morphology of the conjunctival epitheliums in both Group A and Group B. Numerous inflammatory cells infiltrated in the conjunctival lamina propria and goblet cells were invisible. In Group C and Group D, the conjunctival epitheliums were almost normal. There were still some inflammatory cells and dendritic cells in the conjunctival lamina propria, and there were residual goblet cells visible in parts of the patients. When the injured area of the conjunctiva was large, the conjunctivas in four groups displayed hyperreflective stripe fibrous tissues instead of normal conjunctival epitheliums.
The application of laser scanning confocal microscopy indicates that there is much difference on the cellular morphology of the ocular surface of severe alkali burns patients among diverse courses of the disease. The technique is a useful tool to the observation on the ocular surface of severe alkali burns patients.
应用活体激光扫描共聚焦显微镜分析重度碱烧伤患者眼表形态。
本研究为回顾性观察病例系列。2008年2月至11月，复旦大学附属眼耳鼻喉科医院收治的39例按Roper-Hall分类法为Ⅲ或Ⅳ级的碱烧伤患者纳入本研究。根据病程将其分为四组：A组（病程小于3个月）、B组（病程3 - 6个月）、C组（病程6 - 12个月）和D组（病程超过12个月）。对损伤的角膜、角膜缘和球结膜进行活体激光扫描共聚焦显微镜检查并记录图像。分析损伤的角膜、角膜缘和球结膜的形态，计算角膜缘炎症细胞和树突状细胞密度。采用单因素方差分析比较炎症细胞和树突状细胞的均值，随后两组间数据采用最小显著差法分析。
A组患者角膜上皮表现为大的不规则形态，细胞质高反射，细胞核低反射，有时失去细胞特征。浅表上皮下各层有大量小的高反射炎症细胞。角膜浅基质水肿，难以辨别基质细胞形态。深层基质细胞处于活化状态。内皮层图像不清。B组和C组浅表上皮表现与A组相同，D组消失。浅表上皮下炎症细胞减少，B组和C组可见一些残留的基底上皮和高反射的结膜样上皮。D组可见小的椭圆形紧密排列细胞，细胞核呈点状高反射，而非正常角膜基底上皮。B组仍难以辨别浅基质细胞形态。深层基质细胞仍处于活化状态。C组和D组角膜基质被纤维组织替代。其他组内皮层图像仍不清。所有组重度碱烧伤患者角膜缘的Vogt栅栏均被破坏。角膜缘有丰富的血管网。四组角膜缘炎症细胞密度分别为(4023±343)、(2975±246)、(2652±375)、(2679±299)个/mm²。组间炎症细胞密度差异有统计学意义（F = 40.001，P = 0.000）。四组角膜缘树突状细胞密度分别为(106±19)、(132±35)、(141±26)、(98±24)个/mm²。组间树突状细胞密度差异有统计学意义（F = 8.053，P = 0.000）。当结膜损伤面积局限时，A组和B组难以辨别结膜上皮形态。结膜固有层有大量炎症细胞浸润，杯状细胞不可见。C组和D组结膜上皮基本正常。结膜固有层仍有一些炎症细胞和树突状细胞，部分患者可见残留杯状细胞。当结膜损伤面积大时，四组结膜均表现为高反射条纹状纤维组织，而非正常结膜上皮。
激光扫描共聚焦显微镜检查显示，重度碱烧伤患者不同病程阶段眼表细胞形态差异较大。该技术是观察重度碱烧伤患者眼表的有用工具。
