Fuchs endothelial corneal dystrophy FECD is a progressive corneal disease that impacts the structure and stiffness of the Descemets membrane DM, a protein layer that serves as a matrix for the corneal endothelial cells CECs. These structural alterations of the DM contribute to the loss of the CECs resulting in corneal edema and blindness. Oxidative stress and TGF-b pathways have been implicated in endothelial cell loss and endothelial to mesenchymal transition of CECs in FECD. Ascorbic acid AA is found at high concentrations in FECD and its impacts on CECs survival has been investigated. However, TGF-b and AA effects on the composition and rigidity of the CECs matrix remain unknown. In this study, we investigated the effect of AA, TGF-b1 and TGF-b3 on the deposition, ultrastructure, stiffness, and composition of the extracellular matrix ECM secreted by primary bovine corneal endothelial cells BCECs. Immunofluorescence and electron microscopy post-decellularization demonstrated a robust deposition and distinct structure of ECM in response to treatments. AFM measurements showed that the modulus of the matrix in BCECs treated with TGF-b1 and TGF-b3 was significantly lower than the controls. There was no difference in the stiffness of the matrix between the AA-treated cell and controls. Gene Ontology analysis of the proteomics results revealed that AA modulates the oxidative stress pathway in the matrix while TGF-b induces the expression of matrix proteins collagen IV, laminin and lysyl oxidase homolog 1. Molecular pathways identified in this study demonstrate the differential role of soluble factors in pathogenesis of the FECD.
[doi:10.25345/C5J854]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: Moo ; Fuchs endothelial corneal dystrophy ; corneal disease ; cornea
Principal Investigators: (in alphabetical order) |
Vijay Krishna Raghunathan, Department of Basic Sciences, College of Optometry, d Department of Biomedical Engineering, Cullen College of Engineering, University of Houston, USA |
Submitting User: | brettsp1 |
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Owner | Reanalyses | |
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