纤维蛋白胶体外重建兔角膜上皮的实验研究

目的利用组织工程技术体外重建角膜上皮组织,为重建角膜表面提供良好的移植材料和方法。方法以等量凝血酶和纤维蛋白原作用生成的纤维蛋白胶为载体(实验组),种植兔角膜缘组织块,体外培养重建角膜上皮层,每天记录细胞生长覆盖面积,从生长特性、光镜特征、超微结构特点以及免疫细胞化学方面进行观察检测。结果角膜缘上皮细胞在纤维蛋白胶上黏附生长增殖,体外培养7d左右即形成单层上皮细胞,2周左右形成的复层上皮细胞与纤维蛋白胶构成角膜上皮组织经检测与生理状态下的角膜上皮组织相近。实验组与对照组(置盖玻片为载体)比较细胞生长覆盖面积第4天至第7天差异有显著性(P<0.05)。结论以纤维蛋白胶为载体,体外培养的角膜上皮组织片可作为眼表重建角膜上皮移植良好的来源。

Experimental Study on Reconstruction of Rabbit Cornea Epithelium in Vitro Using Fibrin Sealant

Objective To investigatte reconstruct corneal epithelial tissue in vitro by tissue engineer technique to provide corneal epithelial transplantation materials and methods for corneal surface reconstruction. Methods Rabbit corneal limbal epithelial primary cells were seeded on fibrin glue which were produced by equivalence thrombin and fibrinogen and the composite corneal epithelial were cultured in vitro and the compound epithelial tissue were examined by light and electron microscopy, immunocytochemical behavior. Percentage coverage of cell growth was recorded every day. Results Limbal epithelial cells were cultured and proliferated on fibrin glue, single layer cells were formed after 7 days, 3~4 layers epithelial were formed after 10~14 days. The compound epithelial tissue was composed of fibrin glue and stratified epithelial cells, which express epithelial cytokeratin(ck1). The cultured corneal epithelium was quite similar to the normal epithelial layers in vivo. The percentage coverage of cornea epithelial cells in experimental group was obviously improved from the fourth to seventh day.Conclusion The corneal epithelia cultured on the fibrin glue proves to be potential grafts for ocular transplantation.