血管内皮细胞替代角膜内皮细胞的可行性

目的:探讨培养的人脐静脉内皮细胞行猫眼角膜内皮细胞移植的可行性。方法:取第三代体外培养人脐静脉内皮细胞,种植在处理过的人羊膜上,待完全形成单层后用于细胞移植手术。正常健康家猫20只,分为A人脐静脉内皮细胞移植组:将载有完全融合的人脐静脉内皮细胞的羊膜移植到去除自体角膜内皮细胞及后弹力层的猫角膜上;B单纯羊膜移植对照组:将保存人羊膜移植到去除自体角膜内皮细胞及后弹力层的猫角膜上;C去除内皮细胞对照组:去除猫眼自体角膜内皮细胞及后弹力层后,既不移植羊膜,也不移植血管内皮细胞。术后1,2,4,8,12,24wk以裂隙灯观察角膜的透明度、眼前节炎症情况,同时行大体及裂隙灯照相。并以角膜厚度仪测量角膜厚度,A组在术后1,2,4,8wk各取1例植片制备光镜标本进行组织病理学检查。对照组均取术后2wk植片做光镜检查。结果:A组共11例植片术后3d水肿渐消退,植片渐透明,并在1wk内保持完全透明。4例在术后1～2wk因排斥反应而混浊,其中3例于混浊后4wk逐渐恢复透明,1例持续混浊。6例在术后12～24wk植片持续透明,光镜观察显示,移植的内皮细胞在前房内成连续单层排列。B组植片持续水肿混浊,但较C组轻,未见免疫排斥反应发生;2wk植片光镜观察羊膜与基质贴附较好无皱折。C植片持续高度水肿混浊,在长达24wk的观察中未发现恢复透明现象。2wk植片光镜观察完全没有后弹力层及内皮细胞层,基质高度水肿,结构疏松紊乱。结论:以人羊膜为载体移植的人脐静脉内皮细胞在前房环境内存活24wk,并可行使角膜内皮细胞的屏障及液泵功能。与角膜基质贴附紧密,不发生免疫排斥反应。

Experiment of endothelial cell replacement with the cultured human umbilicus vascular endothelium in cats

AIM:To evaluate the feasibility of cultured human umbilicus vascular endothelium replacing the function of cat’s native endothelium. METHODS:The third passaged cultured human umbilicus vascular endothelium were planted on the processed human amniotic membrane.After they confluented to a monolayer,the endothelium and human amniotic membrane were transplanted to the cat’s cornea to replace its native endothelium and Descement’s membrane(group A).Meanwhile,transplanting amniotic membrane solely(group B) and depriving of its native endothelium and Descement’s membrane but nothing was transplanted(group C) as control groups. After operation,the operated eyes were observed by slitlamp every day and taken photos also.At different time point,the operated corneas were examinated by light microscopy. RESULTS:In 12 cases of the group A,11 cases kept clear about one week post operation.But after two weeks,4 grafts showed the signs of rejection,thus resulting to the failure of the grafts.Among these failure grafts,3 cases spontaneously cleared after 4 weeks.Six cases kept clear all the time point post operation.The light and electric microscope showed that the transplanted human umbilicus vascular endothelium can carry out the barrier and pump function as well as the native corneal endothelium.4 cases of the group B showed no sign of rejection.In 4 cases of the group C,their cornea became edema step by step. CONCLUSION:The transplanted human umbilicus vascular endothelium can adapt to the anterior chamber environment and show pump and barrier function.The amniotic membrane can attached to the stoma tightly and show no signs of immunoreactions.