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TGF-β-Mediated Modulation of Cell-Cell Interactions in Postconfluent Maturing Corneal Endothelial Cells
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DOI：10.1167/iovs.63.11.3.png)
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Abstract
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PURPOSE. Transforming growth factor-beta (TGF-beta) is known to influence many cell functions. In the corneal endothelium, TGF-beta 1 exerts contextual effects, promoting endothelial-mesenchymal transition in proliferating cells and enhancing barrier integrity in early confluent maturing cells. Herein, we studied how TGF-beta isoforms participate in the formation of corneal endothelial intercellular junctions. METHODS. Corneal endothelial cells (CECs) were cultured using a two-phase media approach. When CECs reached confluence, the proliferation medium was replaced with maturation medium, which was supplemented or not with TGF-beta isoforms. The cell morphology (circularity index), intercellular junction protein expression, transendothelial electrical resistance (TEER), and permeability of 7-day postconfluent CECs were assessed. Gene transcription and signaling pathways that were activated following maturation in the presence of TGF-beta 2 were also studied. The beneficial effect of TGF-beta 2 on CEC maturation was evaluated using ex vivo corneas mounted on a corneal bioreactor. RESULTS. The results showed increases in circularity index, membrane localization of junction-related proteins, and TEER when TGF-beta isoforms were individually added during the maturation phase, and TGF-beta 2 was the most effective isoform. Gene profiling revealed an increase in extracellular matrix-related gene expression. In ex vivo cell adhesion experiments, CECs that were matured in the presence of TGF-beta 2 had a higher circularity index and cell density and exhibited cell membrane-localized junction-related protein expression at earlier time points. CONCLUSIONS. These results suggest that TGF-beta 2 can strengthen cell-cell and cell-substrate adhesion, which accelerates barrier integrity establishment and thus enhances CEC functionality.
Keywords:
corneal endothelial cells
TGF-beta
intercellular junctions
extracellular matrix
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