Mastropasqua Leonardo, Calienno Roberta, Lanzini Manuela, Nubile Mario, Colabelli-Gisoldi Rossella Annamaria, De Carlo Luca, Pocobelli Augusto
Department of Medicine and Ageing Sciences, Ophthalmology Clinic. University "G. d' Annunzio" of Chieti-Pescara, Italy.
Azienda Ospedaliera San Giovanni-Addolorata, Rome, Italy.
Mol Vis. 2016 Jul 10;22:748-60. eCollection 2016.
To correlate a biomicroscopic evaluation, an in vivo confocal microscopy examination, and impression cytologic findings of the corneal center and sclerocorneal limbus after cultured limbal stem cell transplantation and to test the effectiveness of in vivo confocal microscopy as a diagnostic procedure in ocular surface cell therapy reconstructive surgery.
Six eyes of six patients affected by limbal stem cell deficiency after chemical burns underwent ex vivo expanded limbal stem cell transplantation (two eyes) and ex vivo expanded limbal stem cell transplantation with subsequent penetrating keratoplasty (four eyes) to restore corneal transparency. One year after surgery, all patients underwent a biomicroscopic evaluation, central cornea impression cytology to detect cytokeratin 12 (CK12) positivity, and in vivo confocal microscopy of the central cornea and the sclerocorneal limbus to investigate the epithelial cellular morphology, limbal architecture, and corneal inflammation level.
Impression cytology analysis showed CK12 positivity in five of six cases, in concordance with the biomicroscopic evaluation. Confocal microscopy pointed out irregular limbal architecture with the absence of the palisades of Vogt in all cases; the central epithelial morphology presented clear corneal characteristics in three cases and irregular morphology in the remaining three.
After successful ex vivo expanded limbal stem cell transplantation, in the presence of a complete anatomic architecture subversion, documented by support of in vivo confocal microscopy, the sclerocorneal limbus seemed to maintain its primary function. In vivo confocal microscopy confirmed the procedure was a non-invasive, efficacious diagnostic ocular surface procedure in the case of cell therapy reconstructive surgery.
将培养的角膜缘干细胞移植后角膜中央和角膜缘的生物显微镜评估、活体共聚焦显微镜检查及印迹细胞学检查结果进行关联，并测试活体共聚焦显微镜作为眼表细胞治疗重建手术诊断方法的有效性。
6例化学伤后角膜缘干细胞缺乏患者的6只眼接受了体外扩增角膜缘干细胞移植（2只眼）及体外扩增角膜缘干细胞移植联合穿透性角膜移植术（4只眼），以恢复角膜透明度。术后1年，所有患者均接受生物显微镜评估、中央角膜印迹细胞学检查以检测细胞角蛋白12（CK12）阳性情况，以及中央角膜和角膜缘的活体共聚焦显微镜检查，以研究上皮细胞形态、角膜缘结构及角膜炎症水平。
印迹细胞学分析显示6例中有5例CK12阳性，与生物显微镜评估结果一致。共聚焦显微镜检查指出所有病例的角膜缘结构均不规则，无沃格特栅栏；3例中央上皮形态呈现清晰的角膜特征，其余3例形态不规则。
成功进行体外扩增角膜缘干细胞移植后，在活体共聚焦显微镜证实存在完整解剖结构破坏的情况下，角膜缘似乎仍维持其主要功能。活体共聚焦显微镜证实该方法在细胞治疗重建手术中是一种非侵入性、有效的眼表诊断方法。
