体外培养角膜内皮细胞移植的实验研究

Experimental study on transplantation of cultured corneal endothelial cells in vitro

目的 探讨体外培养角膜内皮细胞移植的手术方法及移植后在活体的生理功能。方法 植片 :以直径 7 5mm、厚 10 0 μm、冷冻脱水保存的猪角膜后弹力膜 (Descemet′smembrane ,DM ) /基质为载体 ,原代接种兔角膜内皮细胞 ,体外培养 7～ 10d ,用于移植。受眼 :新西兰兔 3 0只 ( 3 0眼 ) ,实验组 2 0眼 ,对照组 10眼 ,全角膜范围去除术眼内皮细胞 ,5周后施行手术 ;做直径 9mm、3 / 4角膜厚的带蒂前层角膜瓣并掀置一侧 ,再用直径 7 2 5mm的环钻钻去中央后层角膜 ,形成植床 ;将植片置于植床 ,8-0可吸收线间断缝合 ,10 -0尼龙线间断缝合前层角膜瓣 ,术后观察 1～ 12个月。结果 术后角膜持续透明 6个月 14眼 ( 14 / 18、 77% ) ,12个月 8眼 ( 8/ 14、 5 7% ) ;术后 6个月内皮细胞密度 ( 193 4± 3 0 7)个 /mm2 ,角膜厚度平均为 ( 3 92± 3 9) μm。 结论 以异种DM /基质为载体培养的兔角膜内皮细胞 ,行同种异体移植后 ,能够在活体上成活 ,并具有正常内皮细胞的生物学功能 ,维持角膜透明 ,深板层角膜内皮移植术是体外培养内皮细胞移植的一种有效术式。

Objective To evaluate the efficacy for transplantation of cultured corneal endothelial cells in vitro.Methods Donor button:using a cell carrier which were isolated from porcine eye corneal posterior lameller,7.5mm in diameter,100μm in thickness,the endothelial cells were removed and preserved in glycerine.Rabbits corneal endothelial cells were seeded onto DM surface of carrier and cultured in vitro for 7～10 days,then be used for transplantation.Recipient bed:recipient eyes from 30 New Zealand white rabbits (30eyes),experimental group 20 eyes,control group 10 eyes.A centered,hinged flap 9 mm in diameter,and three-fourths in thickness of cornea was maded.The central posterior lamellar were excised using a 7.25mm trephine.The donor button was placed in the recipient bed and sutured using 8-0 absorbable suture,the flap was sutured using 10-0 nylon suture.Postoperative observation for 1～12 months.Results 14 eyes (14/18,77%)cornea maintained clear for 6 months affer surgery,8 eyes (8/14,57%) cornea maintained clear for 12 months.Endothelial cells density averaged (1934±307) cells/mm 2,corneal thickness averaged (392±39)μm at 6 months after surgery.Conclusions cultured corneal endothelial cells in vitro can be transplantation and have normal endothelial cell function in the recipient and maintained cornea clear.