A technique to harvest Descemet's membrane with viable endothelial cells for selective transplantation.
A technique to harvest Descemet's membrane with viable endothelial cells for selective transplantation.
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DOI：
10.1016/j.ajo.2004.09.068
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发表时间：
2005-02
影响因子：
4.2
通讯作者：
Teresa S. Ignacio;Thao T B Nguyen;M. Sarayba;P. Sweet;O. Piovanetti;R. Chuck;A. Behrens
中科院分区：
文献类型：
--
作者：
Teresa S. Ignacio;Thao T B Nguyen;M. Sarayba;P. Sweet;O. Piovanetti;R. Chuck;A. Behrens
目的描述一种利用人工前房获取角膜后弹力膜（DM）和角膜内皮细胞的手术技术，用于角膜内皮细胞移植.方法将7只供体眼的角膜巩膜扣状体内皮面朝上安装在人工前房上.保持内皮侧与其通常的厚度相同，用9.0 mm环钻在Schwalbe线内的后表面上进行手动环钻，深度刚好超过DM。该腔室充满空气，使供体角膜的内皮侧呈现凸形构型。使用钝的睫状体分离刮刀将DM沿着其内皮与后基质分离。滴用台盼蓝0.3%和茜素红S 0.2%（n = 6）。在光学显微镜下检查染色的DM并拍照以计算内皮细胞损伤的百分比。在未染色的角膜上进行组织学检查。在所有7只眼睛中，成功地从后基质中去除了携带内皮的DM。尽管DM看起来非常易碎，但所有样本均被完全取出，且未破裂。活体染色显示平均内皮细胞损失为8.46%（标准差（SD）6.9）。直接光学显微镜显示的保存内皮细胞的morphology.CONCLUSIONSThis技术似乎是一个安全和简单的方法来收获DM内皮细胞移植。进一步的研究正在进行中，以确定通过小角膜切口插入所获得的具有内皮细胞的健康DM的最佳方法。
PURPOSETo describe a surgical technique using an artificial anterior chamber to facilitate harvest of Descemet’s membrane (DM) and endothelium for corneal endothelial cell transplantation.DESIGNLaboratory investigation.METHODSCorneoscleral buttons of seven human donor eyes were mounted endothelial side up on an artificial anterior chamber. Keeping the endothelial side with its usual concavity, a manual trephination was made on the posterior surface with a 9.0-mm trephine, inside the Schwalbe line and just past the DM in depth. The chamber was filled with air, causing the endothelial side of the donor cornea to assume a convex configuration. The DM along with its endothelium was separated from the posterior stroma using a blunt cyclodialysis spatula. Drops of trypan blue 0.3% and alizarin red S 0.2% (n = 6) were applied. The stained DMs were examined under a light microscope and photographed to calculate the percentage of endothelial cell damage. Histology was done on the unstained cornea.RESULTSThe DM carrying endothelium was successfully removed from the posterior stroma in all seven eyes. Although the DM appears to be very friable, all samples were removed in toto without rupture. Vital staining showed a mean endothelial cell loss of 8.46% (standard deviation (SD) 6.9). Direct light microscopy demonstrated the preservation of endothelial cell morphology.CONCLUSIONSThis technique appears to be a safe and straightforward method to harvest DM for endothelial cell transplantation. Further studies are underway to determine the optimal method of insertion of the obtained healthy DM with endothelial cells through small corneal incisions.