Departments of Ophthalmology.
Immunology; and.
Cornea. 2021 Jul 1;40(7):842-850. doi: 10.1097/ICO.0000000000002498.
To analyze the phenotype of the corneal epithelium in patients with long-term follow-up who underwent autologous cultivated oral mucosal epithelial transplantation (COMET) using in vivo confocal microscopy (IVCM) and impression cytology with immunofluorescence staining (ICIF).
Thirteen eyes from patients with severe limbal stem cell deficiency, who underwent COMET at least 48 months before, were recruited in this noncomparative cohort study. After eye examination, IVCM and ICIF were performed. Clinical manifestations of the cornea were evaluated and compared with epithelial findings detected by IVCM and ICIF [cytokeratin (CK) 3, CK7, and CK12]. Two corneal buttons derived from patients receiving the corneal transplantation post-COMET were sent for immunohistochemistry (CK3, CK6, CK7, CK12, paired box gene 6, p63, zonula occludens-1, and integrin β -1).
The mean age of patients was 51.2 ± 20.6 years, and the mean follow-up time since COMET was 78.7 ± 16.3 months. Six of 13 eyes showed clinically successful COMET. In these eyes, IVCM demonstrated predominant cornea-like epithelium and ICIF reported positivity for CK3 and CK12, confirming the presence of oral mucosal and corneal epithelium. Meanwhile, 7 eyes showed total conjunctivalization, corresponding with substantial conjunctival epithelium detected by IVCM and positivity for conjunctival (CK7) and oral mucosal epithelial (CK3) markers detected by ICIF. The immunohistochemistry of corneal buttons stained positive for oral mucosal, corneal epithelial, and stem cell markers (CK3, CK12, and p63).
In long-term follow-up of COMET, epithelium of successful patients demonstrated cornea-like phenotype, whereas failed cases revealed mainly conjunctival phenotype. However, there were evidences that oral mucosal epithelial cells remained across the cornea in both successful and failed COMET as detected by IVCM and ICIF.
通过活体共聚焦显微镜（IVCM）和免疫荧光染色印迹细胞学（ICIF）分析接受自体培养口腔黏膜上皮移植（COMET）后长期随访患者的角膜上皮表型。
本非对照队列研究招募了 13 名曾接受 COMET 治疗的严重角膜缘干细胞缺陷患者的 13 只眼，这些患者在接受 COMET 治疗至少 48 个月后接受了此项研究。眼部检查后进行 IVCM 和 ICIF。评估角膜的临床表现，并将其与 IVCM 和 ICIF 检测到的上皮发现进行比较[细胞角蛋白（CK）3、CK7 和 CK12]。从接受 COMET 后角膜移植的患者中切取两个角膜纽扣，用于免疫组织化学（CK3、CK6、CK7、CK12、配对盒基因 6、p63、闭合蛋白-1 和整合素β-1）。
患者的平均年龄为 51.2±20.6 岁，自 COMET 以来的平均随访时间为 78.7±16.3 个月。13 只眼中有 6 只眼的 COMET 临床结果成功。在这些眼中，IVCM 显示出主要的角膜样上皮，ICIF 报告 CK3 和 CK12 阳性，证实存在口腔黏膜和角膜上皮。同时，7 只眼表现为完全结膜化，与 IVCM 检测到的大量结膜上皮和 ICIF 检测到的结膜（CK7）和口腔黏膜上皮（CK3）标志物阳性相对应。角膜纽扣的免疫组织化学染色阳性，表明存在口腔黏膜、角膜上皮和干细胞标志物（CK3、CK12 和 p63）。
在 COMET 的长期随访中，成功患者的上皮表现出角膜样表型，而失败患者则主要表现出结膜表型。然而，通过 IVCM 和 ICIF 检测，在成功和失败的 COMET 中均有证据表明口腔黏膜上皮细胞仍存在于角膜中。
