【作者】 赵峰；

【导师】 徐建江；

【作者基本信息】 复旦大学 ， 眼科学， 2009， 硕士

【摘要】 第一部分兔角膜后弹力层撕除内皮损伤模型的实验研究目的建立兔角膜后弹力层撕除内皮损伤模型,观察角膜内皮修复过程。方法健康成年雌性新西兰兔9只,左眼用于实验。建立兔角膜后弹力层撕除内皮损伤模型。术后分别于1天,1周,1月,2月,3月裂隙灯下观察角膜情况并拍摄数码照片。术后1月,2月,3月分别取兔角膜行台盼蓝-茜素红联合染色、苏木素-伊红(HE)染色组织病理学观察及扫描电镜检查,观察角膜内皮修复过程。结果兔角膜后弹力层撕除角膜内皮损伤术后1天,裂隙灯下可见损伤区域角膜水肿浑浊。术后1月,角膜水肿程度减轻,范围缩小。台盼蓝-纤素红染色见缺损区为粗糙的大片蓝紫色着染区,无明显角膜内皮细胞形态可见。HE染色和扫描电镜提示角膜内皮细胞开始生长。扫描电镜下见损伤区已被角膜内皮细胞覆盖,缺损区域周围的内皮细胞延展伸长,类似于成纤维细胞,呈纺锤形。术后2月,角膜水肿明显减轻,台盼蓝-茜素红染色显示损伤区域蓝紫色染色区范围减小,其间可见增大异形的角膜内皮细胞。HE染色和扫描电镜提示内皮细胞数目增多,形态趋于规则。术后3月,角膜无明显水肿,台盼蓝-茜素红染色,HE染色和扫描电镜提示损伤区域角膜内皮细胞数量和形态基本恢复正常。呈紧密排列的规则的六边形结构。结论角膜后弹力层撕除可成功建立兔角膜内皮损伤模型,其损伤区域角膜内皮于术后3月基本恢复正常。第二部分角膜神经破坏对兔角膜后弹力层撕除内皮损伤修复影响的实验研究目的在兔角膜后弹力层撕除内皮损伤模型的基础上建立兔角膜神经损伤模型,观察角膜神经破坏对角膜后弹力层撕除内皮损伤修复的影响。方法健康成年雌性新西兰兔27只,左眼用于实验。随机分成3组,每组9只。A组行单纯角膜后弹力层撕除内皮损伤术;B组行角膜后弹力层撕除内皮损伤联合角膜放射状深板层切开术;C组行角膜后弹力层撕除内皮损伤联合角膜环行深板层切开术。分别于术后1天,1周,1月,2月,3月裂隙灯下观察角膜情况并拍摄数码照片,角膜水肿浑浊程度分级。术后1月,2月,3月分别取兔角膜行台盼蓝-茜素红联合染色,组织病理HE染色及扫描电镜观察,并比较各组内皮缺损面积、角膜内皮细胞密度以及内皮细胞异形性等参数。结果术后1周各组角膜均呈明显水肿,组间无统计学差异,术后3月C组水肿评分则明显高于A组(P＜0.05)。术后3月,A组角膜内皮缺损区完全修复,内皮细胞密度达1016.59±456.18个/mm~2;B组角膜内皮少量缺损区存在,内皮细胞密度511.06±143.89个/m~2;C组角膜内皮散在多处缺损区,内皮细胞形体大,异形性高,细胞密度275.29±41.66个/mm~2,明显低于A组(P＜0.01)结论兔角膜神经破坏可能影响角膜后弹力层撕除内皮损伤的修复过程。神经损伤程度越重,修复过程越缓慢。更多还原

【Abstract】 PartⅠThe Study of Descement’s Stripping with Endothelial Wound Healing in Rabbit CorneaPURPOSE.To observe morphologieal in corneal endothelium wound healing after Descement’s Stripping in rabbit eyes.METHODS.9 healthy New Zealand white rabbies under Descement’s Stripping with endothelial wound on left eyes.Endothelial wound healing after Descement’s Stripping was studied in rabbit eyes observed by slit microscope,histopathology and scanning electron microscope. RESULTS.At 1 day PO,corneal edema in injured zone was observed by slit-lamp microscope.At 1 month PO,severity of corneal edema decreased and its zone diminished.It was found by trypan bleu and alizarin red S staining that blue-purple fibrous membrane covered injured zone in corneal posterior surface without endothelial cells.Corneal endothelial cells were indicated by HE staining.And scanning electromicroscope showed that injured zone was covered by spindle-shaped corneal endothelial cells resembling fibroblasts.These endothelial cells contained actin filaments in agreement with cultured endothelial cells in vitro.At 2 month PO,corneal edema decreased clearly. It was found by trypan bleu and alizarin red S staining that the injured zone diminished,some increasing and obform corneal endothelial resurfaced these area.And at 3 month PO,corneal wound had been resurfaced by a normal corneal endothelial surface layer.Conclusion Our study demonstrated that Descemet’s stripping in Rabbit could feasibly obtain a valuable model to observe the process of endothelial healing. Corneal endothelium recovered completely by 3 month later.PartⅡThe Experimental Study of Influences of Corneal Nerve Damage with Endothelial Wound Healing in Rabbit CorneaPurpose To establish a model of corneal nerve damage based on the model of Descemet’s stripping with corneal endothelial injury in Rabbit.And to investigate the influences of corneal nerve damage on corneal endothelial healing.Methods A total of 27 Female healthy adult New Zealand white rabbits were randomly divided into three groups:control group A,control group B,and experimental group. The left eye of each rabbit was used in this study.Only Descemet’s stripping was performed in control group A;Descemet’s stripping combined with radial deep lamellar keratotomy was performed in control group B;Descemet’s stripping combined with circular deep lamellar keratotomy was performed in experimental group.At 1 day,1 week,1 month,2 month,and 3 month post-operation(PO),the severity of corneal edema was graded according to observation by slit-lamp microscope.At 1 week,1 month,2 month,and 3 month PO,the operated corneas were excised for trypan blue alizarin red vital staining and HE staining,in addition to scanning electromicroscopy,in order to observe the process of endothelial healing.The evaluations by vital staining included count,size of endothelial defect area,density of regenerated endothelial cell and frequency of hexagonal endothelial cell.Results At 1 day PO,although corneal edema happened in each group,the experimental group was most serious,and control group B was significantly worse than control group A. The corneal nerve disappeared around the incision without regenerated nerve fibers in the experimental group.At 3 month PO,evaluations by vital staining showed that control group A was better than control group B while the latter was better than experimental group,and differences between groups were all significant.更多还原