Long-Term Phenotypic Study after Allogeneic Cultivated Corneal Limbal Epithelial Transplantation for Severe Ocular Surface Diseases
Long-Term Phenotypic Study after Allogeneic Cultivated Corneal Limbal Epithelial Transplantation for Severe Ocular Surface Diseases
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DOI：
10.1016/j.ophtha.2010.04.003
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发表时间：
2010-12-01
期刊：
影响因子：
13.7
通讯作者：
Kinoshita, Shigeru
中科院分区：
文献类型：
--
作者：
Nakamura, Takahiro;Sotozono, Chie;Kinoshita, Shigeru
目的：确定同种异体培养角膜缘上皮移植（CLET）后手术切除的移植物的长期上皮来源。设计：干预性病例报告。参与者：我们研究了2例角膜干细胞完全破坏的患者的2只眼; 1例Stevens-Johnson综合征患者的1只眼和1例持续性化学损伤。方法：将同种异体培养的角膜缘上皮片在人羊膜（AM）上移植到眼表。回植（1只眼，42个月后）或穿透性角膜移植术（1眼，75个月后）在初次移植手术后进行进一步的视力康复。主要结果测量：切除的移植物进行临床评估和光镜和透射电镜（TEM）检查和免疫组化分析。在临床结膜移植物中，TEM和免疫组化分析仅揭示了原始培养的角膜上皮细胞持续存在的小区域。邻近的结膜上皮细胞明显侵入大部分角膜表面（角蛋白3/12（-），Muc 5ac（+））。在临床角膜移植中，移植的同种异体培养角膜上皮细胞明显存活了很长时间（角蛋白3/12（+），Muc 5ac（-））;没有炎性细胞浸润，也没有AM底物溶解。我们推测同种异体CLET后移植物混浊的过程是移植培养角膜上皮细胞丢失的原因，随后结膜细胞侵入角膜表面。本研究结果证实，在临床评价的角膜移植物中，移植的培养角膜上皮细胞确实在角膜表面存活了很长时间，并保持了眼表完整性，即使移植的细胞是同种异体的。
Purpose: To determine the long-term epithelial lineage of origin of surgically removed grafts after allogeneic cultivated corneal limbal epithelial transplantation (CLET).Design: Interventional case reports.Participants: We studied 2 eyes from 2 patients with total corneal stem cell destruction; 1 eye was from a patient with Stevens-Johnson syndrome and 1 eye had sustained chemical injury.Methods: Allogeneic cultivated corneal limbal epithelial sheets on human amniotic membrane (AM) were transplanted onto the ocular surface. Regrafting (1 eye, 42 months later) or penetrating keratoplasty (1 eye, 75 months later) were performed after the initial transplantation procedure for further visual rehabilitation.Main Outcome Measures: The excised grafts were subjected to clinical evaluation and to light-and transmission electron microscopy (TEM) examination and to immunohistochemical analysis.Results: In clinically conjunctival grafts, TEM and immunohistochemical analysis disclosed only small areas where the original cultivated corneal epithelial cells persisted. Neighboring conjunctival epithelial cells had apparently invaded a large portion of the corneal surface (keratin 3/12(-), Muc5ac(+)). In clinically corneal grafts, transplanted allogeneic cultivated corneal epithelial cells clearly survived for a long period of time (keratin 3/12(+), Muc5ac(-)); there was no infiltration by inflammatory cells, nor was there dissolution of the AM substrate.Conclusions: We theorize that the process of graft opacification after allogeneic CLET is responsible for the loss of transplanted cultivated corneal epithelial cells and that this is followed by conjunctival cell invasion onto the corneal surface. The results of this study confirmed that in the clinically evaluated corneal graft, transplanted cultivated corneal epithelial cells indeed survived for a long period of time on the corneal surface and maintained ocular surface integrity, even though the transplanted cells were allogeneic.