器官培养角膜保存法保存兔角膜的观察研究

The experimental study of organ culture preservation of rabbit cornea and clinical application

目的 观察器官培养角膜保存法保存角膜活性的效果 ,探讨该方法临床应用的可行性。方法 兔角膜 2 8只 ,分为两组 ,实验组 16只眼 ,对照组 12只眼。实验组用 31℃器官培养液保存 ,对照组用K 液 4℃保存。两组中每 4只角膜分别保存 3、7、14d。实验组 4只角膜保存 14d后移入运输液中培养 3d。保存前测角膜厚度 ,做内皮细胞形态学检查和台盼蓝染色。保存后不同时间做相同检查和茜素红染色 ,并用透射电镜观察角膜内皮细胞超微结构。在取出角膜前从所有保存瓶抽取保存液样本做细菌和真菌的培养。临床应用器官培养保存 1周的角膜行穿透性角膜移植术 1例。结果 实验组和对照组角膜内皮细胞形态、活性率 1周后差异有显著性。 2组角膜厚度差异有显著性。透射电镜检查 ,实验组 7d和 14d角膜内皮细胞超微结构完整。对照组 7d后线粒体肿胀 ,14d可见细胞结构破坏。穿透性角膜移植术临床病例随访 2年 ,角膜移植片基本透明。结论 器官培养液保存角膜的效果确切 ,活性保存时间明显长于K 液保存。

Objective To observe the effects of corneal preservation by organ culture and to disscuss the feasibility of this method. Methods 28 rabbit corneas were divided into two groups,16 for experimental group and 12 for control group. In the experimental group, every 4 corneas were preserved in 31℃ organ culture media for 3,7,14 days respectively. 4 corneas were shifted to 31℃ organ culture media with 5% daxtran 500 and kept for 3 days after 14 days preservation in organ culture. In the control group every 4 corneas were preserved in 4 ℃ K-solution for 3, 7, 14 days respectively. Before preservation the thickness of corneas was measured and endothelium was evaluated for morphology and trypan blue staining. After presrvation, the same check items and staining of alizarin red were made.The sterility of media was checked in the same way with porcine cornea preservation media. Results There were signifcant differences in morphology of endothelium between the two groups after 7 days preservation. The rate of viable endothelium of experimental group is statistically higher than that of the control group at 7 and 14 days of preservation. There were statistical differences in corneal thickness between the two groups. The results of TEM showed that corneal endothelium maintained integrity at 7 and 14 days of preservation in the experimental group. In the control group, mitochondria swelled at 7 days, the ultrrastructure of endothelium was distroyed at 14 days of preservation. Conclusion The method of corneal preservation in organ culture is effective. The time used for maintaining viability of corneal endothelium with this method is longer than middle term preservation medium. According to the present condition of our hospital, it is possible to use this method to preserve corneas for clinical work after further studies.