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一种用于角膜移植物低温保存的新型储存解决方案:一项实验研究。

A new storage solution for the hypothermic preservation of corneal grafts: an experimental study.

机构信息

Department of Forensic Medicine, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.

Massachusetts Eye and Ear Infirmary, Boston, MA, 02114, USA.

出版信息

Cell Tissue Bank. 2020 Sep;21(3):507-521. doi: 10.1007/s10561-020-09838-z. Epub 2020 May 25.

Abstract

In this experimental study we used for the first time Tiprotec as a solution for corneal preservation and cold storage. We compared the resultant endothelial cell morphology and viability with this obtained after preservation of the ex-vivo corneas with both usual standard techniques: conventional cold storage (using Eusol-C) and organ culture. This prospective, in vitro, 3-armed parallel study was performed with the use of 90 porcine corneas (examined for their endothelial quality and transparency) randomly selected for preservation in three storage methods (each 30 corneas): organ culture, standard cold storage (Eusol-C) and experimental cold storage (Tiprotec) Endothelium cell quantity and quality as well as corneal opacification were assessed. The degree of endothelial transparency was significantly reduced over time with all preservation media, without any significant difference among the three groups at any point of time. A reduction in endothelial cell density was also observed with all three preservation media after 30 days of storage without statistically significant differences between groups. The number of hexagonal and pentagonal endothelium cells was significantly reduced overtime in all media with significantly more hexagonal and pentagonal in the organ culture group compared to the cold storage groups. We could show that the cryopreservation medium Tiprotec, used until now for the preservation of vascular grafts, was of similar quality compared to the medium Eusol-C for the hypothermic storage of corneal tissue for an extended period of time up to 30 days. In comparison to organic culture with culture medium KII, both Tiprotec and Eusol-C were found less effective in preserving endothelial cell quality, as assessed by the morphometric analysis, and viability, as assessed by the degree of vacuolization at least up to the 30th day of storage. However, both, Tiprotec- and Eusol-C-preserved corneas demonstrated a certain capacity to recover after their submission in organ culture.

摘要

在这项实验研究中,我们首次将 Tiprotec 用作角膜保存和冷藏的溶液。我们将保存后的角膜内皮细胞形态和活力与使用常规标准技术(传统冷藏(使用 Eusol-C)和器官培养)保存的结果进行了比较。这项前瞻性、体外、三臂平行研究使用了 90 只猪角膜(检查其内皮质量和透明度),随机选择了三种保存方法(每组 30 只角膜):器官培养、标准冷藏(Eusol-C)和实验冷藏(Tiprotec)。评估了内皮细胞数量和质量以及角膜混浊度。随着时间的推移,所有保存介质的内皮透明度均显著降低,在任何时间点三组之间均无显着差异。在用所有三种保存介质保存 30 天后,还观察到内皮细胞密度降低,各组之间无统计学差异。在用所有三种保存介质保存 30 天后,六边形和五边形内皮细胞的数量也随着时间的推移而显着减少,与冷藏组相比,器官培养组中的六边形和五边形内皮细胞数量明显更多。我们证明,直到现在用于保存血管移植物的冷冻保存介质 Tiprotec 与 Eusol-C 等用于长时间(长达 30 天)低温保存角膜组织的介质相比,质量相当。与用 KII 培养基进行的有机培养相比,Tiprotec 和 Eusol-C 在通过形态计量分析评估内皮细胞质量和通过空泡化程度评估活力方面都不太有效,至少在 30 天的储存期内。然而,Tiprotec 和 Eusol-C 保存的角膜在提交器官培养后都显示出一定的恢复能力。

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