摘要
目的探讨不同乙醇作用时间对角膜上皮瓣活性的影响，并确定角膜上皮瓣的解剖分离层面。方法按标准准分子激光角膜上皮瓣下磨镶术（LASEK）方法制备7只尸体眼角膜上皮瓣，其中对照组1只眼，其余6只分为A、B、C3个组，每组2只眼，乙醇浸润时间分别为20、30和40s。对7只眼进行HE、增殖细胞核抗原（PCNA）、层黏连蛋白免疫组化及Ⅶ型胶原免疫荧光染色，评价角膜上皮瓣细胞活性，确定LASEK手术中角膜上皮瓣的解剖分离层面，采用X^2检验分析。结果角膜上皮瓣为复层上皮细胞组织，没有Bowman’s层和角膜基质成分，PCNA阳性细胞率为A组〉B组〉C组，其laminin染色在角膜床和上皮瓣基底膜一侧均有呈片段样棕色条纹状，其collagen Ⅶ染色阳性反应呈一条绿色线形，而且只存在于角膜床，上皮瓣基底膜侧不着色。结论角膜上皮瓣活性随乙醇浸润时间延长而降低；乙醇浸润法制备角膜上皮瓣的组织分离层面位于角膜上皮基底膜内，并且定位于基底膜的透明层和致密层之间。
Objective To evaluate the viability of corneal epithelial cells and to determine the anatomic cleavage on the epithelial basement membrane after various exposure times to 20% ethanol during epithelial flap preparation in laser-assisted subepithelial keratectomy （LASEK） in cadaver eyes. Methods Six human cadaver eyes were exposed to 20% ethanol for 20,30 and 40 seconds （2 eyes for each group）, and another one eye was used as the control. PCNA staining was performed to determine the viability of corneal epithelial cells. Immunofluorescence staining using monoclonal antibodies against collagen Ⅶ, and immunohistological staining using monoclonal antibodies against laminin were performed to detect the anatomic location of the cleavage plane on the corneal epithelial flaps created by 20 seconds exposure to 20% ethanol in cadaver eyes. Results Hematoxylin and eosin staining of epithelial flaps revealed a coherent stratified epithelium. The PCNA positive rates of the epithelial ceils in the flap decreased in the 20-second group,30-second group and 40-second group successively. Immunohistological staining to laminin was patchy in the lifted flap and the remaining corneal basement membrane. Immunofluorescence to collagen Ⅶ, the main component of anchoring fibrils remained exclusively in the corneal bed. Conclusions Viability of the epithelial flap decreased with longer time exposure to ethanol. The cleavage plane of the ethanol-treated corneal epithelial flap is located between the lamina lucida and the lamina densa of the basement membrane where laminin forms hemidesmosome.
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2008年第12期1088-1092,共5页
Chinese Journal of Ophthalmology
关键词
角膜切削术
上皮下
激光
上皮
角膜
基膜
增殖细胞核抗原
层粘连蛋白
乙醇
Keratectomy, subepithelial, laeser-assisted
Epithelium, corneal
Basement membrane
Proliferating cell nuclear antigen
Laminin
Ethanol
