人脐静脉血管内皮细胞在体移植替代猴角膜内皮细胞的形态学分析

目的　使用去除后弹力层的恒河猴自体角膜为细胞载体，将培养的人脐静脉血管内皮细胞（ｈｕｍａｎｕｍｂｉｌｉｃａｌｖｅｉｎｅｎ-ｄｏｔｈｅｌｉａｌｃｅｌｌｓ，ＨＵＶＥＣ）种植到角膜内表面，观察ＨＵＶＥＣ替代恒河猴角膜内皮细胞的功能情况以及ＨＵＶＥＣ在恒河猴眼内生长的情况。方法　取恒河猴６只随机分为３组:实验组（３只）、实验对照组（２只）、空白对照组（１只）。实验组:用离心沉淀法将培养的ＨＵＶＥＣ移植到去除后弹力层的恒河猴自体角膜内表面，之后将自体角膜缝回植床；实验对照组:将撕除部分后弹力层的术眼角膜植片原位缝回植床；空白对照组:取下术眼角膜植片不做任何处理原位缝回植床。术后观察各组角膜植片透明情况；实验组及实验对照组于术后３０ｄ及６０ｄ、空白对照组于术后６０ｄ行术眼摘除，标本做病理切片、ＣＤ３４免疫组化及扫描电镜，观察房角结构及ＨＵＶＥＣ在角膜植片内表面形态分布。结果　实验组角膜植片维持了一定的厚度和透明性，而实验对照组角膜植片发生严重大泡性改变。病理切片示实验组角膜内表面可见一层细胞生长，ＣＤ３４染色阳性，提示为血管内皮细胞；实验对照组角膜内表面未见任何细胞生长；空白对照组角膜植片保留完整后弹力层及内皮细胞层。扫描电镜示实验组有ＨＵＶＥＣ单层在角膜内表面生长但有大量白细胞聚集及少量细胞碎片嵌顿于小梁网；实验对照组角膜内表面残留胶原纤维样物质，无细胞生长；空白对照组见完整六边形角膜内皮细胞层。结论　ＨＵＶＥＣ能够在撕除后弹力层的恒河猴角膜内表面生长并发挥一定的屏障作用，维持角膜的厚度和透明性，但会产生较重的排斥反应。

Microstructure observation of human vascular endothelial cells transplantation in autograft to replace monkey corneal endothelial cells

Objective Using the Descemet’ s membrane-removed thesus autologous corneal as the cell carrier and transplant the cultured human umbilical vein endothelial cells ( HUVEC ) to the inner surface of the comea.to observe the function of HUVEC after alternative thesus monkey corneal endothelial cell and the growth of the HUVEC in the thesus monkey eye corneal surface. Methods The experimented monkeys were divided int0 3 groups : experimental group ( 3 cases) , experimental control group ( 2 cases) and blank control group ( I case) . In experimental group , cultured HUVEC were transplanted onto thesus monkey posterior corneal surface without corneal endothelium ;In experimental control group ,the detached corneal Descemet membrane of thesus monkey fixed in suit without cultured HUVEC transplantation;ln blank control group , a simple penetrativity corneal transplantation without destroy the descemet membrane performed. The transparent of implants in each group was observed after corneal transplantation. Histopathology of cornea grafts, immunohistochemistry and scanning electron microscope were performed at 30 days and 60 days postoperatively in order to observe the appearance of HUVEC onto the monkey posterior corneal surface and the structure of anterior chamber angle. Results The corneal implants of experimental group kept transparency and thickness rn some extent while the experimental control group showed severity corneal edema and bullae. Histopathology of corneal grafts showed that a cells layer with CD34 staining positive reaction growth on posterior corneal surface of corneal button in the experimental group ,which indicated that the layer of cells were HUVEC.no cell and material like membrane could be seen on that of the experimental control group ;A layer of corneal endothelial cells and Descemet ’ s membrane were visible on that of the blank control group. Scanning electron microscope showed that a layer of HUVEC were growing on the posterior surface of cornea button with plenty of leucocytes and small amounts of cell debris could be found to block up the trabecula meshwork, collagen fibers remained on the posterior surface of cornea button of the experimental control group with no cell grown; The complete corneal endothelial cells layer with six-fringe shape could be found in the blank control group. Conclusion HUVEC can survive and enduce the barrier function at the posterior surface of thesus monkey cornea without corneal endothelial cells, and keep the cornea button transparency and normal thickness in some extent .but severe rejection will happen.