摘要目的 从影像学及组织学角度,分析兔眼角膜损伤部位在不同时间点的细胞构成及细胞外基质混浊程度的变化.方法 建立兔眼自体板层角膜移植模型,按术后1周,2周,1月和3月时间点分为4组,每组5只.进行共焦显微镜检查,应用曲线拟合和边缘检测的方法 分析图像,确定角膜基质层的混浊程度及细胞数量;在对应时间点以CD34和α-SMA标记角膜基质细胞和肌成纤维细胞,免疫荧光法观察角膜损伤部位的表达变化.结果 (1)术后2周的Confocal图像显示损伤处存在激活的角膜基质细胞,术后一月数量最多,此时细胞外基质的混浊程度也为明显.(2)术后两周和一月时损伤处α-SMA染色呈现阳性,CD34染色呈弱阳性.结论 (1)Confocal检查可以良好地反应角膜损伤后修复过程中细胞数量及细胞外基质混浊的程度.(2)角膜损伤后角膜基质细胞向肌成纤维细胞转化,肌成纤维细胞的数量与细胞外基质的混浊程度有关.

abstractsObjective To evaluate the confocal and histological microscopic features of corneal interface in rabbit after auto lamellar keratoplasty. Methods Auto lamellar keratoplasty was performed on 20 rabbit eyes. Confocal examination of lamellar interface was performed 1 week, 2 weeks, 1 month and 3 months postoperatively. Five corneal specimens were studied by immunohistochemistry using monoclonal antibodies against CD34 and α -SMA on each time point. Results Confocal microscopy revealed numerous highly reflective keratocytes at the interface 2 weeks postoperatively. Keratocyte density and extracellular matrix reflectivity peaked 1 month postoperatively. The lamellar interface showed positive staining for α -SMA 2 weeks and 1 month postoperatively, while the staining for CD34 weakened. Conclusions Substantial keratocyte activation and corneal opacity occur from 2 weeks to 1 month after auto lamellar keratoplasty. The keratocyte transform into myofibroblast, which may account for the increased corneal refloctivity.