Using In Vivo Confocal Microscopy to Detect the Wound Healing Process after DSAEK and NDSAEK

• Main Authors: Jen-Pin Sun, 孫仁彬
• Other Authors: 陳偉勵
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/90594393568786635348

碩士 === 國立臺灣大學 === 臨床醫學研究所 === 101 === 【Background and Study Purpose】 Although traditional penetrating keratoplasty (PK) was a widely accepted procedure, descemet''s stripping automated endothelial keratoplasty (DSAEK) becomes popular and has replaced PK in the treatment of endothelial disorders. DSAEK is unique for the smaller wound size, lesser astigmatism, lesser chance of intraoperative expulsive hemorrhage, faster/better visual outcome and faster wound healing. However, there still exist many unknown factors for a better surgery. Most surgeons removed Descemet’s membrane and endothelial layer during DSAEK surgery follower by insertion of posterior corneal lamella. However, still some surgeons keep Descemet’s membrane and endothelial layer before the attachment of the posterior corneal lamella (n-DSAEK). They claim that the remained Descemet’s membrane/ endothelial layer between donor and recipient corneas will not inference corneal clarity and visual prognosis. The purpose of this project is to use New Zealand White Rabbit as the experimental model to find out the best surgical condition for DSAEK. 【Material and Method】 We will use femtosecond laser machine to create donor corneal posterior lamellas. The posterior corneal lamellas will be transplanted to rabbit corneas with or without stripping of Descemet’s membrane/corneal endothelial layer. After operation, the rabbit eyes will receive in vivo confocal microscopic examination every two weeks for totally 3 months. The Descemet’s membrane/corneal endothelial layer between donor and recipient corneas will be examined by morphology and Z-profile. At post-operative 3 months, the rabbit eyes will be obtained, and examined by anterior segment OCT to evaluate the attachment and healing of the transplanted grafts. In addition, the tissue section will be examined by H&E staining, immunohistochemical staining and transmission electron microscopy. 【Results】 The interface recipient endothelial cell density in nDSAEK gradually decreased in the follow-up period. The interface haziness of DSAEK and nDSAEK diminished gradually, and nDSAEK has greater interface opacity than DSAEK at all time points examined. There was no significant difference of total corneal thickness between DSAEK and nDSAEK at all time points. Tissue section using H&E staining showed preserved interface recipient Descemet’s membrane and decreased endothelial cell density three months after nDSAEK. Transmission electron microscopy revealed morphologically changed interface recipient endothelial cell and some collagen fibers similar as in graft stroma in recipient endothelial cell in nDSAEK three months after operation. The recipient endothelial cell didn’t express markers of pump function and epithelial cell by immunohistochemical staining. However some recipient endothelial cell went apoptotic pathway. 【Conclusion】 NDSAEK has similar surgical results as DSAEK in all parameters measured in this study. However, the interface haziness was greater than DSAEK, and recipient endothelial cell at the interface underwent morphological change and showed some phagocytic activity during the whole observational process. The endothelial cell density decreased due to cell apoptosis