International Center for Ocular Physiopathology (ICOP), The Veneto Eye Bank Foundation, Zelarino, Venice, Italy.
Br J Ophthalmol. 2014 Aug;98(8):1120-5. doi: 10.1136/bjophthalmol-2013-304466. Epub 2014 May 30.
To describe a novel submerged hydro-separation (SubHyS) technique followed by anterior corneal dissection to prepare a Descemet endothelial graft (DEG) for Descemet's membrane endothelial keratoplasty from human donor corneas.
30 human donor corneas were immersed in liquid (organ culture (OC) storage medium). Using a 25-gauge needle, approximately 0.3 mL of OC was injected (SubHyS) in the posterior stroma to create a liquid bubble. The bubbled cornea was mounted onto a modified artificial chamber with the epithelial side facing the air. The endothelium was protected with a viscoelastic solution. The anterior cornea was excised with a Barron radial vacuum trephine and the residual peripheral stroma was removed manually using micro-scissors. The DEG was dismounted and washed. The endothelial cell density (ECD) and mortality of the prepared DEG was determined. All the DEGs were preserved in deturgescent medium for 7 days using a cornea claw which was fixed to the sclera. ECD and mortality were checked post preservation.
Complete detachment of Descemet's membrane and stroma was achieved in all 30 cases. Bubble burst was observed in five cases (excluded from the study) due to overfilling of the liquid. The average diameter of the excised DEG was 10.96 mm. The average endothelial cell loss post preservation was 27.69%. Histological analysis confirmed elimination of the residual stroma (n=13).
The DEGs can be preserved in a deturgescent medium for up to 7 days. The procedure provides a standardised, pre-validated (quality assured), pre-separated, no-touch, ready-to-use tissue and also reduces the preparation time. Further, the tissues can be trephined as per the surgeon's convenience and can either be rolled or a contact lens could be used for final delivery of the DEG using a surgical glide.
描述一种新型的浸没式水分离（SubHyS）技术，随后进行前角膜解剖，从人供体角膜中制备用于 Descemet 膜内皮角膜移植的 Descemet 内皮移植物（DEG）。
将 30 个人供体角膜浸入液体（器官培养（OC）储存介质）中。使用 25 号针头，在后基质中注射约 0.3ml 的 OC，以产生液体气泡。将起泡的角膜安装到带有改良人工室的装置上，使上皮面朝向空气。用粘性溶液保护内皮。用 Barron 径向真空环钻切除前角膜，并使用微型剪刀手动切除残留的周边基质。卸下 DEG 并清洗。确定制备的 DEG 的内皮细胞密度（ECD）和死亡率。所有的 DEG 都在角膜爪的作用下，用 Deturgescent 介质保存在角膜上，持续 7 天。角膜爪固定在巩膜上。保存后检查 ECD 和死亡率。
在所有 30 例中均成功实现了 Descemet 膜和基质的完全分离。由于液体填充过多，有 5 例（排除在研究之外）观察到气泡破裂。切除的 DEG 的平均直径为 10.96mm。保存后的平均内皮细胞丢失率为 27.69%。组织学分析证实消除了残留的基质（n=13）。
DEG 可以在 Deturgescent 介质中保存长达 7 天。该程序提供了标准化、预先验证（质量保证）、预先分离、无接触、即用型组织，同时还缩短了准备时间。此外，根据外科医生的方便，组织可以进行环钻切割，然后使用手术滑液将 DEG 滚动或使用隐形眼镜进行最终输送。
