Filev Filip, Oezcan Ceprail, Feuerstacke Jana, Linke Stephan J, Wulff Birgit, Hellwinkel Olaf J C
Department of Ophthalmology, University Medical Center Hamburg-Eppendorf, Martitistr. 52, 20246, Hamburg, Germany.
Department of Forensic Medicine, University Medical Center Hamburg-Eppendorf, Martitistr. 52, 20246, Hamburg, Germany.
Cell Tissue Bank. 2017 Mar;18(1):91-98. doi: 10.1007/s10561-016-9596-z. Epub 2016 Dec 7.
Dextran is added to corneal culture medium for at least 8 h prior to transplantation to ensure that the cornea is osmotically dehydrated. It is presumed that dextran has a certain toxic effect on corneal endothelium but the degree and the kinetics of this effect have not been quantified so far. We consider that such data regarding the toxicity of dextran on the corneal endothelium could have an impact on scheduling and logistics of corneal preparation in eye banking. In retrospective statistic analyses, we compared the progress of corneal endothelium (endothelium cell loss per day) of 1334 organ-cultured corneal explants in media with and without dextran. Also, the influence of donor-age, sex and cause of death on the observed dextran-mediated effect on endothelial cell counts was studied. Corneas cultured in dextran-free medium showed a mean endothelium cell count decrease of 0.7% per day. Dextran supplementation led to a mean endothelium cell loss of 2.01% per day; this reflects an increase by the factor of 2.9. The toxic impact of dextran was found to be time dependent; while the prevailing part of the effect was observed within the first 24 h after dextran-addition. Donor age, sex and cause of death did not seem to have an influence on the dextran-mediated toxicity. Based on these findings, we could design an algorithm which approximately describes the kinetics of dextran-toxicity. We reproduced the previously reported toxic effect of dextran on the corneal endothelium in vitro. Additionally, this is the first work that provides an algorithmic instrument for the semi-quantitative calculation of the putative endothelium cell count decrease in dextran containing medium for a given incubation time and could thus influence the time management and planning of corneal transplantations.
在移植前至少8小时将右旋糖酐添加到角膜培养基中,以确保角膜发生渗透性脱水。据推测,右旋糖酐对角膜内皮有一定毒性作用,但目前这种作用的程度和动力学尚未量化。我们认为,关于右旋糖酐对角膜内皮毒性的此类数据可能会影响眼库中角膜制备的时间安排和物流。在回顾性统计分析中,我们比较了1334个器官培养角膜外植体在含和不含右旋糖酐的培养基中角膜内皮的进展情况(每天内皮细胞损失)。此外,还研究了供体年龄、性别和死亡原因对观察到的右旋糖酐介导的内皮细胞计数影响。在无右旋糖酐培养基中培养的角膜显示内皮细胞计数平均每天下降0.7%。添加右旋糖酐导致内皮细胞平均每天损失2.01%;这反映出增加了2.9倍。发现右旋糖酐的毒性影响是时间依赖性的;而这种影响的主要部分在添加右旋糖酐后的最初24小时内观察到。供体年龄、性别和死亡原因似乎对右旋糖酐介导的毒性没有影响。基于这些发现,我们可以设计一种算法,大致描述右旋糖酐毒性的动力学。我们在体外重现了先前报道的右旋糖酐对角膜内皮的毒性作用。此外,这是第一项提供算法工具的工作,用于在给定孵育时间下对含右旋糖酐培养基中假定的内皮细胞计数减少进行半定量计算,从而可能影响角膜移植的时间管理和规划。