不同移植手术治疗角膜缘干细胞缺损的实验研究  

An experimental study treatment of total limbal deficiency in rabbits by autograft or allograft with cultured limbal corneal epithelium.

作　　者：庆惠玲[1] 王丽娅[1] 赵承彦[1] 杜连心[1] 

机构地区：[1]河南省眼科研究所,河南郑州450003

出　　处：《眼外伤职业眼病杂志》2007年第4期245-248,共4页Journal of Injuries and Occupational Diseases of the Eye with Ophthalmic Surgeries

基　　金：河南省科技攻关基金资助项目(0524410035)

摘　　要：目的探讨以人羊膜为载体培养的角膜缘干细胞,自体及异体移植治疗全角膜缘干细胞缺损。方法制作兔眼角膜缘干细胞完全缺损3个月的模型。实验动物随机分为自体移植组和异体移植组,前者取对侧眼角膜缘组织,后者取异体兔眼角膜缘组织,均以去除上皮细胞的羊膜基底膜为载体,培养12d后行角膜缘干细胞羊膜移植术。术后观察3个月,以角膜上皮染色、角膜浑浊和新生血管3项指标进行临床疗效评定,通过病理检查评估术后角膜上皮修复情况,印迹细胞学检查移植前后角膜上皮的细胞表型。结果体外培养的兔角膜缘干细胞可在羊膜上粘附生长并增生,体外培养12d可形成复层。自体移植组和部分异体移植组术后角膜上皮逐渐愈合,透明度提高,基质细胞浸润减轻,新生血管减退或消失。印迹细胞学检查显示:移植前角膜上皮细胞PAS阳性,而移植后转为阴性;组织病理学显示:移植前角膜上皮大部分缺损,移植后呈现角膜上皮结构。部分异体移植组术后出现了免疫排斥反应。结论兔自体角膜缘干细胞羊膜移植术可重建眼表;免疫排斥反应仍是异体角膜缘干细胞羊膜移植术失败的主要原因。Objective To investigate the outcome of using human amniotic membrane as substrates for cultivating rabbit limbal corneal epithelial cells , and autologous or allogenous transplanting them onto rabbits with total limbal deficiency. Methods As model of total limbal deficiency had. been built in right eyes of 15 rabbits by surgical removal of limbal rim for 3 months. They were randomly divided into 2 groups of 10： autologous group （5） and allogenous group （5）. small biopsy specimens from the limbal region of 10 left eyes were taken and cultured on acellular amniotic membrane . When it was cultured about 12 days, 5 of the right eyes then received grafts of amniotic membrane containing autologous cultured epithelial cells ; 5 of the right eyes then received grafts of amniotic membrane containing allogenouscuhured epithelial cells. To observe for 3 months after transplantation, the restoration of corneal epithelium was evaluated by pathology , and the cells phenotype of corneal epithelium was examined by impression cytology. Results Limbal epithelial cells can be cultured and proliferated on the amniotic membrane, stratified layers of epithelium were formed after 12 days. Corneas of autologous （5） or allogenous （3） transplanting gradually epithelialized and progressive decrease of vascularity and stromal infiltration in the limbal and peripheral zone. The examination of impression cytology shew that corneal epithelium before transplantation： PAS （ ＋ ）, corneal epithelium after transplantation： PAS （ - ）. The histopathology shew that corneal epithelium before transplantation was deficience mostly, corneal epithelium after transplantation was restored. However corneas of allogenous （2） transplanting had happened immunology rejection. Conclusions Autologous transplantation of cultivated limbal corneal epithelium on amniotic membrane can reconstruct corneal epithelium. However, immunology rejection is main reason of limbal epithelial allogenous transplantation.

关 键 词：角膜缘干细胞 羊膜 自体移植 异体移植 角膜缘上皮缺损 

分 类 号：R779.65[医药卫生—眼科]