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使用人去细胞角膜板层和培养的人角膜内皮细胞构建的生物工程移植物治疗兔角膜内皮损伤模型。

Treatment of corneal endothelial damage in a rabbit model with a bioengineered graft using human decellularized corneal lamina and cultured human corneal endothelium.

机构信息

Ophthalmology Department, Hospital Ramón y Cajal, Madrid, Spain.

Cell Engineering Laboratory, La Paz Hospital Research Institute, iDIPAZ, Madrid, Spain.

出版信息

PLoS One. 2019 Nov 21;14(11):e0225480. doi: 10.1371/journal.pone.0225480. eCollection 2019.

Abstract

OBJECTIVE

We aimed to investigate the functionality of human decellularized stromal laminas seeded with cultured human corneal endothelial cells as a tissue engineered endothelial graft (TEEK) construct to perform endothelial keratoplasty in an animal model of corneal endothelial damage.

METHODS

Engineered corneal endothelial grafts were constructed by seeding cultured human corneal endothelial cell (hCEC) suspensions onto decellularized human corneal stromal laminas with various coatings. The functionality and survival of these grafts with cultured hCECs was examined in a rabbit model of corneal endothelial damage after central descemetorhexis. Rabbits received laminas with and without hCECs (TEEK and control group, respectively).

RESULTS

hCEC seeding over fibronectin-coated laminas provided an optimal and consistent endothelial cell count density and polygonal shape on the decellularized laminas, showing active pump fuction. Surgery was performed uneventfully as standard Descemet stripping automated endothelial keratoplasty (DSAEK). Corneal transparency gradually recovered in the TEEK group, whereas haze and edema persisted for up to 4 weeks in the controls. Histologic examination showed endothelial cells of human origin covering the posterior surface of the graft in the TEEK group.

CONCLUSIONS

Grafting of decellularized stroma carriers re-surfaced with human corneal endothelial cells ex vivo can be a readily translatable method to improve visual quality in corneal endothelial diseases.

摘要

目的

我们旨在研究经培养的人角膜内皮细胞接种的人去细胞化基质层的功能,作为组织工程化内皮移植物(TEEK)构建体,在角膜内皮损伤的动物模型中进行内皮角膜移植。

方法

通过将培养的人角膜内皮细胞(hCEC)悬浮液接种到具有各种涂层的去细胞化人角膜基质层上来构建工程化角膜内皮移植物。在中央去内皮化的角膜内皮损伤兔模型中检查这些带有和不带有 hCEC 的移植物(TEEK 和对照组)的功能和培养 hCEC 的存活率。

结果

纤连蛋白涂层的基质层上的 hCEC 接种提供了最佳且一致的去细胞化层上的内皮细胞计数密度和多边形形状,显示出活跃的泵功能。手术按照标准的 Descemet 剥离自动角膜内皮移植术(DSAEK)进行。在 TEEK 组中,角膜透明度逐渐恢复,而在对照组中,混浊和水肿持续长达 4 周。组织学检查显示,人源内皮细胞覆盖了 TEEK 组移植物的后表面。

结论

体外重新表面化带有人类角膜内皮细胞的去细胞化基质载体的移植可以是一种易于转化的方法,以改善角膜内皮疾病的视觉质量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/15e8/6871783/f52929ef489a/pone.0225480.g001.jpg

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