兔自体骨髓间充质干细胞移植对角膜缘干细胞损伤的初步疗效研究

目的探讨兔自体骨髓间充质干细胞(BMSCs)移植对角膜缘干细胞(LSCs)损伤的初步疗效。方法 36只大白兔分为三组:BMSCs组、LSCs组与羊膜对照组,每组12只。BMSCs组采用附着有BMSCs的羊膜治疗角膜缘干细胞损伤,LSCs组将附着有角膜内皮细胞的羊膜治疗角膜缘干细胞损伤,羊膜对照组则仅将羊膜用缝合线固定于浅层巩膜上治疗角膜缘干细胞损伤。于术前、术后1周、术后2周、3周和4周对眼表的形态进行评分;并于术前、术后1周、4周、8周利用聚焦显微镜观察细胞密度和表面积。结果在移植手术实施之前三组兔的眼表形态综合评分之间的差异无统计学意义(P0.05);手术之后4周BMSCs组和LSCs组兔的眼表形态评分得以显著改善,差异有统计学意义(P0.05);手术之后4周羊膜对照组兔的眼表形态评分与手术前相比,差异无统计学意义(P0.05);手术之后4周BMSCs组和LSCs组之间的眼表形态评分之间差异无统计学意义(P0.05)。与羊膜对照组相比,其它两组手术4周之后的眼表形态评分改善更显著,差异有统计学意义(P0.05)。经过共焦显微镜观察,BMSCs组兔移植1周之后的细胞形态为圆形,而非初始的梭边形,细胞较小,密度较高,而随着时间推移,个数逐渐减少,体积变大。随着时间的推移BMSCs组与LSCs组兔移植后的平均细胞密度出现下降。手术前BMSCs组与LSCs组细胞表面积差异无统计学意义(P0.05);手术后1周、4周BMSCs组的细胞表面积显著低于LSCs组,差异有统计学意义(P0.05),手术之后8周和12周时BMSCs组与LSCs组细胞表面积差异无统计学意义(P0.05)。结论与角膜缘干细胞相似,兔自体骨髓间充质干细胞移植治疗角膜缘干细胞可以重建受损的眼表,减轻水肿,抑制血管生成。

A preliminary study on the efficacy of autologous transplantation of mesenchymal stem cells from bone marrow for treatment of corneal limbus stem cell injury

Objective To explore the efficacy of transplantation in rabbits with autologous bone marrow mesenchymal stem cells（ BMSCs） for treatment of injury of limbal stem cells（ LSCs）. Methods A total of 36 rabbits were divided into three groups：BMSCs group,LSCs group and amniotic membrane control group,12 rabbits in each group. Rabbits in BMSCs group were applied with BMSCs attached to amniotic membrane for treatment of corneal limbal stem cell damage,rabbits in LSCs group were applied with corneal endothelial cells attached to amniotic membrane for treatment of corneal limbal stem cell injury,and rabbits in amniotic membrane control group were only applied with amniotic membrane fixed on superficial sclera for treatment of corneal limbal stem cell injury with suture. Morphological scores on surface of eyes were recorded before opera-tion,1 week,2 weeks,3 weeks and 4 weeks after surgery,and ocular surface area and cell density were observed before operation,and 1 week, 4 weeks,8 weeks after surgery by using confocal microscopy. Results There was no statistically significant difference in ocular surface shape comprehensive scores between 3 groups of rabbits in results before implementation of transplantation operation（ P 〉0. 05）;rabbit ocular surface morphology scoring could be obviously improved in BMSCs group and LSCs group at 4 weeks after operation,there was statistically significant difference（ P 〈0. 05）;there was no significant difference in rabbit scores of control group with operation at 4 weeks after the amnion eye pheno-typic state compared with pre operation（ P 〉0. 05）;the difference in ocular surface morphology scores at 4 weeks after operation between BMSCs group and LSCs group had no statistical significance（ P 〉0. 05）. In comparison with amniotic membrane control group,the ocular surface mor-phology scores in other 2 groups had been improved more significantly,there was statistically significant difference,there was statistically signifi-cant difference（ P 〈0. 05）at 4