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[活体激光扫描共聚焦显微镜观察春季角结膜炎患者角膜的形态学特征]

[Morphological characteristics of cornea in patients with vernal keratoconjunctivitis by in vivo laser scanning confocal microscopy].

作者信息

Le Qi-hua, Hong Jia-xu, Zhu Wen-qing, Sun Xing-huai, Xu Jian-jiang

机构信息

Department of Ophthalmology, Eye Ear Nose and Throat Hospital of Fudan University, Shanghai 200031, China.

出版信息

Zhonghua Yan Ke Za Zhi. 2011 May;47(5):416-22.

Abstract

OBJECTIVE

To explore the morphological characteristics on cornea in patients with vernal keratoconjunctivitis (VKC) by the application of in vivo laser scanning confocal microscopy (LSCM).

METHODS

The experimental design was retrospective observation case series (case control study). Twenty-six patients, each diagnosed as bilateral VKC, were enrolled in the study, among which 13 were tarsal form, 5 were bulbar form and the rest were mixed form. Nine patients had the clinical course less than one year, eight subjects longer than three years, and the rest between them. Another twenty-six healthy volunteers with matching age and gender were selected as normal control. All participants had their right eyes examined with the in vivo confocal microscopy (HRT II/RCM). Central cornea and superior peripheral cornea were chosen as the examination points. The images were recorded automatically and cellular density of each layer was analyzed by installed software. Software ImageJ was utilized to analyze the density, diameter, branch number and tortuosity of subbasal nerve fiber in VKC patients. Independent t test was performed to assess the differences on cellular density between VKC patients and normal control, as well as those between central and peripheral cornea in VKC patients. Fisher chi-square test was used to compare the infiltration rate of Langerhans cells in corneal epithelium between VKC patients and controls. ANOVA was applied to assess the differences in cellular density among three subtypes, as well as among different duration of VKC. Independent t-test and chi-square test were applied to analyze the parameters of subbasal nerve fiber.

RESULTS

The morphological changes in cornea included the absence of superficial hyperreflective polygonal epithelial cells, infiltration of Langerhans cells in and(or) underneath corneal epithelium and activation of keratocytes in anterior stroma. Corneal epithelium conjunctivalization and stromal neovascularization could be identified in patients with corneal neovascular epithelium. Longitudinal or oblique dark striae could be found in the posterior stroma in patients with complicated keratoconus. The density of epithelial cells at central and peripheral cornea in healthy controls were (6033.1 ± 998.7) cells/mm(2) and (6098.4 ± 298.3) cells/mm(2), while that in VKC patients were (5972.2 ± 1148.2) cells/mm(2) and (6178.5 ± 318.9) cells/mm(2) respectively, the differences being no statistical significant between them (t = 1.191, 1.011; P = 0.238, 0.318). However, it's found in VKC patients that cellular density at peripheral cornea was significantly higher than that at central area (t = 2.249, P = 0.03). The density of anterior stromal cells at central and peripheral cornea in healthy controls was (1001.4 ± 125.3) cells/mm(2) and (924.6 ± 201.4) cells/mm(2), while that in VKC patients was (1184.5 ± 115.3) cells/mm(2) and (1101.4 ± 151.1) cells/mm(2), the difference bearing no statistical significance (t = 6.617, 3.439; P = 0.001). The density of posterior stromal cells in normal subjects and VKC patients was (537.7 ± 42.6) cells/mm(2) and (548.7 ± 79.8) cells/mm(2), that of endothelial cells was (2985.7 ± 401.2) cells/mm(2) and (3021.5 ± 383.3) cells/mm(2), respectively, neither difference had statistical significance (t = 0.174, 1.112; P = 0.864, 0.282). Langerhans cell infiltration could be identified in 61.5% (16 cases) VKC patients, which was significantly higher than normal control (2 cases, 7.7%) (χ(2) = 12.49, P = 0.001). Furthermore, much intense Langerhans cells infiltration was found in bulbar form and mix form than tarsal form. (t = 6.617, P = 0.001). The density and diameter of subbasal nerve fiber in VKC patients decreased significantly than those in normal subjects, whereas the tortuosity increased significantly.

CONCLUSIONS

The morphological changes of cornea in VKC patients mainly involve corneal epithelium, subbasal nerve fiber and anterior stroma. In vivo LSCM is helpful in discriminating the subtypes of VKC.

摘要

目的

应用活体激光扫描共聚焦显微镜(LSCM)探讨春季角结膜炎(VKC)患者角膜的形态学特征。

方法

实验设计为回顾性观察病例系列(病例对照研究)。纳入26例双侧诊断为VKC的患者,其中睑结膜型13例,球结膜型5例,其余为混合型。9例患者病程小于1年,8例患者病程大于3年,其余患者病程介于两者之间。另选26名年龄和性别匹配的健康志愿者作为正常对照。所有参与者均用活体共聚焦显微镜(HRT II/RCM)检查右眼。选择中央角膜和周边上角膜作为检查点。图像自动记录,并用安装的软件分析各层细胞密度。利用软件ImageJ分析VKC患者基底膜下神经纤维的密度、直径、分支数和迂曲度。采用独立t检验评估VKC患者与正常对照之间以及VKC患者中央角膜与周边角膜之间细胞密度的差异。采用Fisher卡方检验比较VKC患者与对照者角膜上皮中朗格汉斯细胞的浸润率。应用方差分析评估三种亚型之间以及不同病程的VKC患者之间细胞密度的差异。采用独立t检验和卡方检验分析基底膜下神经纤维的参数。

结果

角膜的形态学改变包括浅表高反射多边形上皮细胞缺失、角膜上皮内和(或)其下朗格汉斯细胞浸润以及前基质中角膜细胞活化。角膜新生血管上皮患者可出现角膜上皮结膜化和基质新生血管形成。圆锥角膜并发症患者后基质中可发现纵向或斜向的暗条纹。健康对照者中央角膜和周边角膜上皮细胞密度分别为(6033.1±998.7)个/mm²和(6098.4±298.3)个/mm²,而VKC患者分别为(5972.2±1148.2)个/mm²和(6178.5±318.9)个/mm²,两者差异无统计学意义(t = 1.191,1.011;P = 0.238,0.318)。然而,发现VKC患者周边角膜细胞密度显著高于中央区域(t = 2.249,P = 0.03)。健康对照者中央角膜和周边角膜前基质细胞密度分别为(1001.4±125.3)个/mm²和(924.6±201.4)个/mm²,而VKC患者分别为(1184.5±115.3)个/mm²和(1101.4±151.1)个/mm²,差异无统计学意义(t = 6.617,3.439;P = 0.001)。正常受试者和VKC患者后基质细胞密度分别为(537.7±42.6)个/mm²和(

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