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Fate of endothelial cells after intrastromal implantation of Descemet's membrane-endothelial cell tissue.
Cell and Tissue Banking ( IF 1.5 ) Pub Date : 2020-06-01 , DOI: 10.1007/s10561-020-09840-5
Pietro Rosetta 1 , Alessandro Ruzza 2 , Mohit Parekh 2, 3 , Paola Gallon 2 , Riccardo Vinciguerra 1 , Stefano Ferrari 2 , Harminder S Dua 4 , Diego Ponzin 2 , Paolo Vinciguerra 5, 6
Affiliation  

In non-Descemet Stripping Automated Endothelial Keratoplasty (nDSAEK), the host DM and endothelium are not removed surgically before the introduction of the posterior lamellar graft; the result is that the patient has both the healthy donor endothelium and the diseased or residual host endothelium. Conversely, DSAEK tissues, that are inserted with inverted polarity (upside down), do not survive and the graft fails. While the mechanism of endothelial cell transplantation is clear, the fate of the endothelial cells retained between two stromal interfaces and their physiological role, if any, is not well understood. The aim of our study was therefore to evaluate the viability of a healthy endothelial-Descemet’s membrane (EDM) graft after the insertion into a stromal pocket of a recipient donor cornea. Research corneas (n = 52) were divided into three groups: Group A, where an EDM (obtained from another cornea) with good endothelium was inserted in a stromal pocket endothelium side down; Group B, consisting of control corneas with a stromal pocket but without EDM insertion; and Group C, pre-stripped membranes resting on their stroma (not in a stromal pocket). The tissues were preserved in tissue culture medium for 21 days at 31 °C. Parameters including viability of endothelial cells, expression of tight junctions (ZO-1) and thickness were evaluated. After 21 days, all the membranes inserted within the stromal pocket of Group A survived, although an average endothelial cell loss of 30.1% (± 18.10) and a mortality of 10.2% (± 22.86) were recorded. Qualitative analysis using triple staining with Hoechst, ethidium homodimer and calcein AM confirmed the mortality. ZO-1 was expressed where the cells were present, showing good integrity of tight junctions. Group C showed an average endothelial cell loss of 1.9% (± 3.38), a mortality of 0.02% (± 0.07) and a higher expression of ZO-1. An EDM graft with endothelium facing downwards can survive in a stromal pocket for at least 3 weeks, with an overall cell mortality of 30%. Further studies are needed to evaluate the possible outcomes of the insertion of a healthy intrastromal EDMs with reverse polarity and in edematous corneas.

中文翻译:

Descemet的膜内皮细胞组织基质内植入后内皮细胞的命运。

在非Descemet剥离自动角膜内皮移植术(nDSAEK)中,在引入后板状移植物之前不通过手术去除宿主DM和内皮。结果是患者同时拥有健康的供体内皮和患病或残留的宿主内皮。相反,以相反极性插入的DSAEK组织(倒置)无法存活,并且移植物会失败。尽管内皮细胞移植的机制尚不清楚,但内皮细胞在两个基质界面之间保留的命运以及它们的生理作用(如果有的话)尚不清楚。因此,我们的研究目的是评估将健康的内皮-德耳曼膜(EDM)移植物插入受体供体角膜的基质袋后的活力。研究性角膜(n = 52)分为三组:A组,将具有良好内皮细胞的EDM(从另一只角膜获得)插入到基质袋内皮细胞的下方。B组,由具有角膜袋但无EDM插入的对照角膜组成;C组,预剥离的膜搁在它们的基质上(不在基质袋中)。将组织在31℃下在组织培养基中保存21天。评价包括内皮细胞存活力,紧密连接的表达(ZO-1)和厚度的参数。21天后,尽管记录的平均内皮细胞损失为30.1%(±18.10),死亡率为10.2%(±22.86),但插入A组基质袋的所有膜均得以存活。使用Hoechst进行三重染色的定性分析,乙二胺均二聚体和钙黄绿素AM证实了死亡率。ZO-1在存在细胞的地方表达,显示紧密连接的良好完整性。C组显示平均内皮细胞损失为1.9%(±3.38),死亡率为0.02%(±0.07),ZO-1的表达更高。内皮面向下的EDM移植物可以在基质袋中存活至少3周,总细胞死亡率为30%。需要进一步的研究来评估插入健康的具有相反极性的水基质角膜内电火花的可能结果。内皮面向下的EDM移植物可以在基质袋中存活至少3周,总细胞死亡率为30%。需要进一步的研究来评估插入健康的具有相反极性的水基质角膜内电火花的可能结果。内皮朝下的EDM移植物可以在基质袋中存活至少3周,总细胞死亡率为30%。需要进一步的研究来评估插入健康的具有相反极性的水基质角膜内电火花的可能结果。
更新日期:2020-06-01
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