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用于角膜保存的重组人血清白蛋白。

Recombinant human serum albumin for corneal preservation.

作者信息

Parekh Mohit, Elbadawy Hossein, Salvalaio Gianni, Amoureux Marie-Claude, Di Iorio Enzo, Fortier Denis, Ponzin Diego, Ferrari Stefano, Ruzza Alessandro

机构信息

International Center for Ocular Physiopathology (ICOP), The Veneto Eye Bank Foundation, Venice, Italy.

Department of Molecular Medicine, School of Biomedicine, University of Padova, Padova, Italy.

出版信息

Acta Ophthalmol. 2018 Feb;96(1):e79-e86. doi: 10.1111/aos.13498. Epub 2017 Jun 21.

DOI:10.1111/aos.13498
PMID:28636255
Abstract

PURPOSE

To study the performance of a completely synthetic organ culture (OC) preservation system containing recombinant human serum albumin (rHSA) for preservation of human donor corneas.

METHODS

Twenty-four paired donor corneas were randomly collected, and one cornea from each donor was preserved in synthetic (experimental) and serum-based media (control). The tissues were assessed at day 0; after 6 days of preservation at room temperature (RT) in Cornea Trans and Cornea Prep II ; after 28 days at 31°C in Cornea Syn [with rHSA] and Cornea Max [with foetal calf serum (FCS)] and; 4-day post deswelling in Cornea Trans and Cornea Jet . Thickness was determined with optical coherence tomography (OCT) and transparency with a validated, custom device. Morphology, endothelial cell density (ECD) and mortality were observed after treating the tissues with Trypan blue and sucrose. Glucose uptake by the cells was analysed. Data were compared using non-parametric paired Wilcoxon tests with p < 0.05 deemed significant. Histology using periodic acid-schiff (PAS), expressions of p63, CK12, αSMA and ZO-1 were analysed, and cell apoptosis postpreservation was studied.

RESULTS

Corneas stored in synthetic media showed a higher and statistically significant value as compared to serum-based media in terms of viable endothelial cell density (VECD), mortality, morphology and glucose uptake postpreservation. Histology showed presence of all the layers, all the markers were expressed, and no apoptosis was observed in either media.

CONCLUSION

The new synthetic preservation system containing rHSA (and other confidential constituents) showed better preservation effects than traditional media containing serum.

摘要

目的

研究一种含有重组人血清白蛋白(rHSA)的全合成器官培养(OC)保存系统对人供体角膜的保存性能。

方法

随机收集24对供体角膜,每个供体的一只角膜保存在合成(实验)培养基和基于血清的培养基(对照)中。在第0天对组织进行评估;在角膜转运液和角膜预处理液II中于室温(RT)保存6天后;在含rHSA的角膜合成液和含胎牛血清(FCS)的角膜最大保存液中于31°C保存28天后;以及在角膜转运液和角膜喷射液中复水4天后进行评估。用光学相干断层扫描(OCT)测定厚度,用经过验证的定制设备测定透明度。在用台盼蓝和蔗糖处理组织后观察形态、内皮细胞密度(ECD)和死亡率。分析细胞对葡萄糖的摄取。使用非参数配对Wilcoxon检验比较数据,p < 0.05被认为具有统计学意义。使用过碘酸希夫(PAS)进行组织学分析,分析p63、CK12、αSMA和ZO-1 的表达,并研究保存后的细胞凋亡情况。

结果

与基于血清的培养基相比,保存在合成培养基中的角膜在保存后的存活内皮细胞密度(VECD)、死亡率、形态和葡萄糖摄取方面显示出更高且具有统计学意义的值。组织学显示所有层均存在,所有标志物均有表达,且两种培养基中均未观察到细胞凋亡。

结论

含有rHSA(和其他保密成分)的新型合成保存系统比含血清的传统培养基显示出更好的保存效果。

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