Farias Roberta J M, Sousa Luciene B, Lima Filho Acácio A S, Lourenço Andréa C S, Tanakai Márcia H, Freymuller Edna
Federal University of São Paulo, São Paulo, Brazil.
Cornea. 2008 Aug;27(7):791-4. doi: 10.1097/ICO.0b013e31816ed54c.
Cornea storage for longer periods is still a challenge for corneal surgeons. The purpose of this study was to find a method to lyophilize corneas for anterior lamellar transplant and to evaluate them by light and transmission electronic microscopy.
Corneal flaps were created by using a microkeratome. Corneas were lyophilized with a cryoprotectant (2.3 mol sacarousis for 40 minutes) and without a cryoprotectant in a lyophilization machine (Modulyon D). The corneas were rehydrated with distilled water, balanced saline solution (BSS), and phosphate-buffered saline, after which they were evaluated by microscopy. A cornea that did not undergo lyophilization served as a control.
Lyophilization without a cryoprotectant did not preserve the corneal structure. This finding was also observed when lyophilizing and rehydrating the corneas with distilled water or phosphate-buffered saline. We found that lyophilizing corneas and rehydrating them with 11 mL of BSS for 30 minutes preserved the general corneal structure, the parallelism of the collagen fibers, the Bowman layer, and the epithelial basement membrane for 15 and 30 days and for as long as 1 year or more.
Lyophilization with sacarousis and rehydration with BSS may be a good method for anterior lamellar transplantation.
对于角膜外科医生而言，长时间保存角膜仍是一项挑战。本研究的目的是找到一种用于前板层移植的角膜冻干方法，并通过光学显微镜和透射电子显微镜对其进行评估。
使用微型角膜刀制作角膜瓣。将角膜在冻干机（Modulyon D）中使用冷冻保护剂（2.3摩尔蔗糖，40分钟）和不使用冷冻保护剂进行冻干。用蒸馏水、平衡盐溶液（BSS）和磷酸盐缓冲盐水对角膜进行复水，之后通过显微镜进行评估。未进行冻干的角膜作为对照。
不使用冷冻保护剂进行冻干不能保留角膜结构。在用蒸馏水或磷酸盐缓冲盐水对角膜进行冻干和复水时也观察到了这一现象。我们发现，用11毫升BSS对冻干角膜进行30分钟的复水可在15天和30天以及长达1年或更长时间内保留角膜的总体结构、胶原纤维的平行排列、Bowman层和上皮基底膜。
用蔗糖进行冻干并用BSS进行复水可能是前板层移植的一种良好方法。
