Extracellular matrix nitration alters growth factor release and activates bioactive complement in human retinal pigment epithelial cells.

PloS One
Mark A FieldsL V Del Priore

Abstract

We have shown previously that non-enzymatic nitration (NEN) of the extracellular matrix (ECM), which serves as a model of Bruch's membrane (BM) aging, has a profound effect on the behavior of the overlying retinal pigment epithelial (RPE) cells, including altered phagocytic ability, reduced cell adhesion, and inhibition of proliferation. We know that transplanted RPE monolayers will encounter a hostile sub-RPE environment, including age-related alterations in BM that may compromise cell function and survival. Here we use our previous NEN model of BM aging to determine the effects of NEN of the ECM on growth factor release and complement activation in RPE cells. Human induced-pluripotent stem cells (iPSCs) were differentiated into RPE cells, and confirmed by immunohistochemistry, confocal microscopy, and polymerase chain reaction. IPSC-derived RPE cells were plated onto RPE-derived ECM under untreated or nitrite-modified conditions. Cells were cultured for 7 days and barrier function measured by transepithelial resistance (TER). Vascular endothelial growth factor (VEGF), pigment epithelium-derived factor (PEDF), and complement component C3a were measured using enzyme-linked immunosorbent assay (ELISA). On average nitrite-modifie...Continue Reading

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Citations

Oct 14, 2017·Arteriosclerosis, Thrombosis, and Vascular Biology·Yang LinMervin C Yoder
Sep 14, 2019·Current Opinion in Cell Biology·Paulo S Caceres, Enrique Rodriguez-Boulan
Jan 29, 2021·International Journal of Molecular Sciences·Arunbalaji PugazhendhiBalamurali Ambati
Feb 22, 2021·Progress in Retinal and Eye Research·Sarah de JongAnneke I den Hollander
Mar 23, 2021·Cellular and Molecular Life Sciences : CMLS·Angela ArmentoSimon J Clark
Sep 21, 2020·Progress in Retinal and Eye Research·Sayon Roy, Dongjoon Kim
Aug 8, 2021·International Journal of Molecular Sciences·Blanca Chinchilla, Rosario Fernandez-Godino

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Methods Mentioned

BETA
biopsies
ELISA

Software Mentioned

GraphPad
Prism

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