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应用角膜活性保存仪建立单纯疱疹病毒Ⅰ型树突状和地图状角膜溃疡的体外模型。

Ex vivo model of herpes simplex virus type I dendritic and geographic keratitis using a corneal active storage machine.

机构信息

Corneal Graft Biology, Engineering and Imaging Laboratory, Health Innovation Campus, Faculty of Medicine, Jean Monnet University, Saint-Etienne, France.

Department of Ophthalmology, University Hospital, Saint-Etienne, France.

出版信息

PLoS One. 2020 Jul 22;15(7):e0236183. doi: 10.1371/journal.pone.0236183. eCollection 2020.

DOI:10.1371/journal.pone.0236183
PMID:32697805
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7375596/
Abstract

BACKGROUND

Herpetic keratitis (HK) models using whole human corneas are essential for studying virus-host relationships, because of high species specificity and the role of interactions between corneal cell populations that cell culture cannot reproduce. Nevertheless, the two current corneal storage methods (hypothermia and organ culture (OC)) do not preserve corneas in good physiological condition, as they are characterized by epithelial abrasion, stromal oedema, and excessive endothelial mortality.

METHODS

To rehabilitate human corneas intended for scientific use, we used an active storage machine (ASM) that restores two physiological parameters that are essential for corneal homeostasis: intraocular pressure and storage medium renewal (21mmHg and 2.6 μL/min, respectively). ASM storage regenerates a normal multilayer epithelium in 2 weeks. We infected six pairs of corneas unsuitable for graft by inoculating the epithelium with herpes simplex virus type 1 (HSV-1), and compared each ASM-stored cornea with the other cornea stored in the same medium using the conventional OC method.

RESULTS

Only corneas in the ASM developed a dendritic (n = 3) or geographic (n = 2) epithelial ulcer reproducing typical HSV-1-induced clinical lesions. Corneas in OC showed only extensive desquamations. None of the uninfected controls showed epithelial damage. Histology, immunohistochemistry, transmission electron microscopy and polymerase chain reaction on corneal tissue confirmed infection in all cases (excluding negative controls).

CONCLUSIONS

The ASM provides an innovative ex vivo model of HK in whole human cornea that reproduces typical epithelial lesions.

摘要

背景

使用完整的人眼角膜的单纯疱疹性角膜炎（HK）模型对于研究病毒-宿主关系至关重要，因为物种特异性高，角膜细胞群体之间的相互作用是细胞培养无法复制的。然而，目前的两种角膜储存方法（低温保存和器官培养（OC））并不能使角膜保持良好的生理状态，因为它们的特点是上皮磨损、基质水肿和内皮细胞死亡率过高。

方法

为了恢复用于科学研究的人眼角膜，我们使用了一种主动储存机（ASM），它恢复了角膜内稳态所必需的两个生理参数：眼内压和储存介质更新（分别为 21mmHg 和 2.6μL/min）。ASM 储存可在 2 周内再生正常的多层上皮。我们通过在角膜上皮接种单纯疱疹病毒 1 型（HSV-1）感染了六对不适合移植的角膜，然后将每对 ASM 储存的角膜与使用传统 OC 方法储存的另一对角膜进行比较。

结果

只有在 ASM 中储存的角膜才会出现树枝状（n=3）或地图状（n=2）的上皮溃疡，再现了典型的 HSV-1 诱导的临床病变。OC 中的角膜仅表现为广泛的脱屑。未感染的对照组角膜均未出现上皮损伤。角膜组织的组织学、免疫组织化学、透射电子显微镜和聚合酶链反应证实了所有病例（排除阴性对照组）的感染。

结论

ASM 提供了一种新颖的全人眼角膜 HK 体外模型，可再现典型的上皮病变。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/a87456dff947/pone.0236183.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/ef1cf428b53f/pone.0236183.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/12f2b3b82b8e/pone.0236183.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/01723d16a9a8/pone.0236183.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/e21f57315de0/pone.0236183.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/8f542571b5c6/pone.0236183.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/01fe61dc042a/pone.0236183.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/a87456dff947/pone.0236183.g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/ef1cf428b53f/pone.0236183.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/12f2b3b82b8e/pone.0236183.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/01723d16a9a8/pone.0236183.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/e21f57315de0/pone.0236183.g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/8f542571b5c6/pone.0236183.g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/01fe61dc042a/pone.0236183.g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/37b6/7375596/a87456dff947/pone.0236183.g007.jpg

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