Slow Interstitial Fluid Flow Activates TGF-β Signaling and Drives Fibrotic Responses in Human Tenon Fibroblasts.

Abstract

BACKGROUND:

Fibrosis limits the success of filtering glaucoma surgery. We employed 2D and 3D in vitro models to assess the effects of fluid flow on human tenon fibroblasts (HTF).

METHODS:

HTF were exposed to continuous or pulsatile fluid flow for 48 or 72 h, at rates expected at the transscleral outflow site after filtering surgery. In the 2D model, the F-actin cytoskeleton and fibronectin 1 (FN1) were visualized by confocal immunofluorescence microscopy. In the 3D model, mRNA and whole cell lysates were extracted to analyze the expression of fibrosis-associated genes by qPCR and Western blot. The effects of a small-molecule inhibitor of the TGF-β receptor ALK5 were studied.

RESULTS:

Slow, continuous fluid flow induced fibrotic responses in the 2D and 3D models. It elicited changes in cell shape, the F-actin cytoskeleton, the deposition of FN1 and activated the intracellular TGF-β signaling pathway to induce expression of fibrosis-related genes, such as CTGF, FN1 and COL1A1. ALK5-inhibition reduced this effect. Intermittent fluid flow also induced fibrotic changes, which decreased with increasing pause duration.

CONCLUSIONS:

Slow interstitial fluid flow is sufficient to induce fibrosis, could underlie the intractable nature of fibrosis following filtering glaucoma surgery and might be a target for antifibrotic therapy.

• Authors: Cornelius Jakob Wiedenmann, Charlotte Gottwald, Kosovare Zeqiri, Janne Frömmichen, Emma Bungert, Moritz Gläser, Jeanne Ströble, Robert Lohmüller, Thomas Reinhard, Jan Lübke, Günther Schlunck
• Journal: Cells (Cells) Vol. 12 Issue 17 (09 04 2023) ISSN: 2073-4409 [Electronic] Switzerland
• PMID: 37681937 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Actins

Topics

• Humans
• Extracellular Fluid
• Actin Cytoskeleton
• Cytoskeleton
• Actins
• Glaucoma