组织工程人角膜内皮在维持新西兰兔角膜透明中的效果研究

为了评价组织工程人角膜内皮(TE-HCE)维持动物角膜透明的效果,利用来自非转染人角膜内皮(HCE)细胞系的核型正常单克隆细胞株为种子细胞、以去上皮层修饰羊膜为载体支架体外重建的TE-HCE,对新西兰兔进行了穿透性角膜内皮移植试验,利用肉眼观察了移植兔眼的水肿和免疫排斥情况,利用裂隙灯显微镜检测了移植兔角膜的透明情况,并利用荧光显微镜检测了移植兔角膜内皮细胞的CM-DiI标记。对撕除后板层新西兰兔眼进行TE-HCE穿透性角膜内皮移植实验的观察和检测结果显示,移植后的兔眼角膜没有出现明显的水肿和排斥等不良反应,使移植兔角膜维持透明的时间长达100d,且角膜内皮移植区的细胞均为带有CM-DiI标记、来自TE-HCE的种子细胞。由此可见,体外重建TE-HCE移植后能使新西兰兔角膜长期保持透明,有望作为HCE的替代物用于角膜内皮失代偿和角膜内皮盲的临床移植和治疗。

Studies on effects of tissue-engineered human corneal endothelia on maintaining corneal transparency of New Zealand white rabbits

To evaluate corneal transparency maintaining effects of tissue-engineered human corneal endothelia (TE-HCEs), the TE-HCEs were in vitro reconstructed with seeder cells of a monoclonal cell strain with a normal karyotype, screened out from an untransfected HCE cell line, and scaffold carriers of modified denuded amniotic membrane. Penetrating keratoplasty with TE-HCEs was performed in New Zealand white rabbits whose corneal posterior lamellars had been ripped off. The status of corneal edema and immune rejection of the transplanted eyes was observed and monitored by the naked eye, the corneal transparency of transplanted eyes was examined by a slit-lamp biomicroscope, and the CM-DiI label of cells was validated by a fluorescent microscope. The observation and examination results of transplanted rabbit eyes showed no negative responses such as corneal edema and immune rejection, and the corneas of transplanted eyes maintained transparency for as long as 100 days. Also, the cells in the transplanted area of the corneal endothelium, with a CM-DiI label, were all the seeder cells from reconstructed TE-HCEs. In conclusion, in vitro reconstructed TE-HCEs can maintain long term corneal transparency of the transplanted eyes of New Zealand White Rabbits be used  promisingly as HCE equivalents for clinical therapy of corneal endothelium uncompensation and primary corneal endotheliopathy.