羊膜培养兔角膜缘上皮细胞及自体移植--角膜缘干细胞缺损的治疗

目的　探讨以人羊膜为载体培养兔角膜缘上皮细胞及其自体移植治疗全角膜缘干细胞缺损。方法　在8只兔右眼用正庚醇脱上皮和角膜缘环切的方法构建全角膜缘干细胞缺损模型2月。其中6只兔为实验组,活体取左眼角膜缘浅层小块,置羊膜上常规和气_液培养42天后进行自体移植治疗右眼角膜缘干细胞缺损;2只兔为对照组,直接用解冻无细胞人羊膜移植治疗右眼角膜缘干细胞缺损。进行细胞和术眼活体观察、组织学观察和电镜观察。结果　角膜缘上皮细胞在羊膜上生长良好,形成复层,细胞间的联结结构存在,细胞与羊膜组织粘附牢固。实验组移植手术后角膜迅速上皮化,恢复角膜表面光滑和透明,组织学观察和电镜观察呈现生理角膜上皮层的结构特点。但眼睑闭合不全可导致手术失败。对照组术后出现角膜缘干细胞缺损导致的角膜病变。结论　以羊膜为载体培养角膜缘上皮细胞后自体移植可有效地治疗角膜缘干细胞缺损导致的角膜病变。

Autologous transplantation of cultivated limbal corneal epithelium on amniotic membrane: An experimental treatment on total limbal deficiency in rabbits

Objective To investigate the outcome of using human amniotic membrane as substrates for cultivating rabbit limbal corneal epithelial cells and autologous transplanting them onto rabbits with total limbal deficiency. Methods An model of total limbal deficiency had been built in right eyes of 8 rabbits by n_heptanol debridement of the entire corneal epithelium followed by surgical removal of limbal rim for 2 months,6 of these rabbits were as experimental group. Small biopsy specimens from the limbal region of left eyes were taken and cultured on acellular amniotic membrane with air_liquid interface treatment for 44 days. Then autologous transplantation for total limbal deficiency was undertaken. Other 2 rabbits were as control group and received grafts of acellular amniotic membrane alone.Cultured cell, clinical, histological and ultrastructure examination were performed. Results The cultivated limbal corneal epithelial cells on the amniotic membrane grew well, formed stratified layers and cellular joins, and well attached to the underlying amniotic membrane. Corneas of experimental group rapidly epithelialized and clear ocular surface was reconstructed in postoperative period of transplantation. Histology and electron microscopy revealed the structural characteristics of normal corneal epithelium. However, lagophthalmos would induce clinical failure. Control group still had lesion of cornea resulted from total limbal deficiency. Conclusion Autologous transplantation of cultivated limbal corneal epithelium on amniotic membrane for corneal lesion from total limbal deficiency is feasible and effective.