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用于眼表重建的体外扩增干细胞同种异体移植物的结果及DNA分析

Outcomes and DNA analysis of ex vivo expanded stem cell allograft for ocular surface reconstruction.

作者信息

Daya Sheraz M, Watson Adam, Sharpe Justin R, Giledi Osama, Rowe Andrea, Martin Robin, James S Elizabeth

机构信息

Corneo-Plastic Unit, Queen Victoria Hospital, East Grinstead, West Sussex, United Kingdom.

出版信息

Ophthalmology. 2005 Mar;112(3):470-7. doi: 10.1016/j.ophtha.2004.09.023.

Abstract

PURPOSE

To investigate the outcome of a new technique of ex vivo expanded stem cell allograft for limbal stem cell deficiency (LSCD), and to characterize the ocular surface genotype after surgery.

DESIGN

Retrospective noncomparative case series.

PARTICIPANTS

Ten eyes of 10 patients with profound LSCD arising from ectodermal dysplasia (3 eyes), Stevens-Johnson syndrome (3 eyes), chemical injury (2 eyes), thermal injury (1 eye), and rosacea blepharoconjunctivitis (1 eye).

INTERVENTION

Allogeneic corneal limbal stem cells were cultured on plastic and transplanted to the recipient eye after removal of conjunctival pannus. Amniotic membrane was applied in a bandage capacity. The procedure was combined with other reconstructive surgery in 2 cases. Nine patients received systemic cyclosporin A immunosuppression, and the DNA genotype was investigated with surface impression cytology.

MAIN OUTCOME MEASURES

Parameters of LSCD, including vascularization, conjunctivalization, inflammation, epithelial defect, photophobia, and pain.

RESULTS

The mean follow-up period was 28 months (range, 12-50). Seven of 10 eyes (70%) had improved parameters of LSCD at final follow-up and were considered successes. Four (40%) had improved visual acuity, including 3 having had further procedures for visual rehabilitation. Three patients failed to improve-1 with a thermal burn and lid deformity, 1 with Stevens-Johnson syndrome and severe dry eye, and 1 with ectodermal dysplasia who developed an epithelial defect at 26 months. DNA analysis of the first 7 cases showed no ex vivo donor stem cell DNA present beyond 9 months.

CONCLUSIONS

Ex vivo expanded stem cell allograft is a useful technique for restoring the ocular surface in profound LSCD. The absence of donor DNA beyond 9 months suggests that ongoing immunosuppression may be unnecessary and raises questions regarding the origin of the host corneal epithelium.

摘要

目的

研究一种用于治疗角膜缘干细胞缺乏症(LSCD)的体外扩增干细胞同种异体移植新技术的效果,并对手术后的眼表基因型进行特征分析。

设计

回顾性非对照病例系列。

研究对象

10例患有严重LSCD的患者的10只眼,病因包括外胚层发育不良(3只眼)、史蒂文斯-约翰逊综合征(3只眼)、化学伤(2只眼)、热烧伤(1只眼)和酒渣鼻性睑结膜炎(1只眼)。

干预措施

将同种异体角膜缘干细胞在塑料培养皿上培养,在切除结膜血管翳后移植到受体眼。应用羊膜作为绷带。该手术在2例患者中与其他重建手术联合进行。9例患者接受全身性环孢素A免疫抑制治疗,并通过表面印记细胞学研究DNA基因型。

主要观察指标

LSCD的参数,包括血管化、结膜化、炎症、上皮缺损、畏光和疼痛。

结果

平均随访期为28个月(范围12 - 50个月)。10只眼中有7只(70%)在最终随访时LSCD参数有所改善,被视为成功。4只眼(40%)视力有所提高,其中3只眼接受了进一步的视力康复手术。3例患者未改善,1例因热烧伤和眼睑畸形,1例因史蒂文斯-约翰逊综合征和严重干眼,1例因外胚层发育不良在26个月时出现上皮缺损。对前7例患者的DNA分析显示,9个月后未检测到体外供体干细胞DNA。

结论

体外扩增干细胞同种异体移植是恢复严重LSCD眼表的一种有用技术。9个月后未检测到供体DNA表明持续免疫抑制可能不必要,并引发了关于宿主角膜上皮来源的问题。

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