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用于角膜组织工程的新型纳米纤维支架

[New nanofibrous scaffold for corneal tissue engineering].

作者信息

Salehi S, Grünert A K, Bahners T, Gutmann J S, Steuhl K P, Czugala M, Singer B B, Fuchsluger T A

机构信息

Deutsches Textilforschungszentrum Nord-West gGmbH, Universität Duisburg-Essen, Krefeld.

Augenklinik, Universitätsklinikum Düsseldorf, Heinrich-Heine-Universität, Düsseldorf.

出版信息

Klin Monbl Augenheilkd. 2014 Jun;231(6):626-30. doi: 10.1055/s-0034-1368533. Epub 2014 Jun 18.

Abstract

BACKGROUND

An estimated 10 million people suffer worldwide from vision loss caused by corneal damage. For the worst cases, the only available treatment is transplantation with human donor corneal tissue. However, in numerous countries there is a considerable shortage of corneal tissue of good quality, leading to various efforts to develop tissue substitutes. The present study aims to introduce a nanofibrous scaffold of poly(glycerol sebacate) PGS as a biodegradable implant, for the corneal tissue engineering.

MATERIALS AND METHODS

Nanofibrous scaffolds were produced from PGS and poly(ε-caprolactone) (PCL) by a modified electro-spinning process. The biocompatibility of the material was tested in vitro by colorimetric MTT assay on days 3, 5, and 7 to test the cell viability of human corneal endothelium cells (HCEC). To examine a potential immunological reaction of the scaffolds, samples were exposed to mononuclear cells derived from peripheral blood (PBMCs). After an incubation period of 3 days, supernatants were assayed for apoptotic assessment and immunogenic potentials by annexin V FITC//propidium iodide and flow-cytometric analysis.

RESULTS

We could successfully demonstrate that cultivation of HCECs on PGS/PCL scaffolds was possible. Compared to day 3, cell density determined by microplate absorbance was significantly higher after 7 days of cultivation (p < 0.0001). According to the MTT data, none of the samples showed toxicity. Apoptotic assessments by FACS analysis showed that no composition stimulated apoptosis or activated PBMCs occurred. All the compositions were inert for native as well as activated T/B/NK cells and monocytes. It can be concluded that leukocytes and their activity was not affected by the scaffolds.

CONCLUSION

A tissue-like scaffold mimicking the human stroma could be developed. The results indicate that PGS/PCL scaffolds could be considered as ideal candidates for corneal tissue engineering as they are biocompatible in contact to corneal endothelial cells and blood cells.

摘要

背景

据估计,全球有1000万人因角膜损伤而视力丧失。对于最严重的病例,唯一可用的治疗方法是用人供体角膜组织进行移植。然而,在许多国家,优质角膜组织严重短缺,促使人们做出各种努力来开发组织替代物。本研究旨在引入一种聚癸二酸甘油酯(PGS)纳米纤维支架作为可生物降解植入物,用于角膜组织工程。

材料与方法

通过改良的静电纺丝工艺,由PGS和聚己内酯(PCL)制备纳米纤维支架。通过比色法MTT试验在第3、5和7天对材料的生物相容性进行体外测试,以检测人角膜内皮细胞(HCEC)的细胞活力。为了检查支架的潜在免疫反应,将样品暴露于外周血来源的单核细胞(PBMC)。孵育3天后,通过膜联蛋白V FITC/碘化丙啶和流式细胞术分析检测上清液的凋亡评估和免疫原性潜力。

结果

我们成功证明了在PGS/PCL支架上培养HCEC是可行的。与第3天相比,培养7天后通过微孔板吸光度测定的细胞密度显著更高(p < 0.0001)。根据MTT数据,所有样品均未显示毒性。通过FACS分析进行的凋亡评估表明,没有任何组合物刺激凋亡或激活PBMC。所有组合物对天然以及活化后的T/B/NK细胞和单核细胞均呈惰性。可以得出结论,白细胞及其活性不受支架影响。

结论

可以开发出一种模仿人基质的组织样支架。结果表明,PGS/PCL支架可被视为角膜组织工程的理想候选材料,因为它们与角膜内皮细胞和血细胞接触时具有生物相容性。

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