PEER-REVIEWED PUBLICATION

2023

TGF-β Isoforms Affect the Planar and Subepithelial Fibrogenesis of Human Conjunctival Fibroblasts in Different Manners

Watanabe M, Tsugeno Y, et al.

Biomedicines

Sapporo Medical University

RESEARCH SUMMARY

This study investigated how three isoforms of transforming growth factor beta (TGF-β1, -β2, and -β3) distinctly influence planar and subepithelial fibrosis mechanisms in human conjunctival fibroblasts (HconF). Using both two-dimensional monolayers and three-dimensional spheroids, the authors demonstrated isoform-specific differences in extracellular matrix (ECM) gene expression, cell proliferation, and mechanical stiffness. TGF-β1 and TGF-β2 increased epithelial resistance and stiffness, whereas TGF-β3 uniquely reduced monolayer TEER while simultaneously inducing spheroid compaction and stiffening. qPCR confirmed differential regulation of collagen I, IV, VI, fibronectin, and α-SMA. These results reveal that TGF-β isoforms distinctly control conjunctival fibrogenesis and may inform anti-scarring strategies for ocular surgery and glaucoma treatment.

A CellScale MicroSquisher was used to quantify the mechanical stiffness of live 3D human conjunctival fibroblast spheroids following treatment with TGF-β-1, TGF-β-2, or TGF-β-3. Individual spheroids were compressed to half of their diameter while measuring ultra-low force and displacement, enabling calculation of spheroid stiffness as a biomechanical indicator of subepithelial fibrogenesis. These CellScale measurements were critical for revealing isoform-dependent mechanical remodeling of conjunctival microtissues and for demonstrating that changes in spheroid stiffness occur independently of cellular metabolic activity. The MicroSquisher data provided a direct mechanical link between biochemical signaling and fibrotic tissue remodeling in 3D ocular models.

AUTHORS

Megumi Watanabe, Yuri Tsugeno, Tatsuya Sato, Araya Umetsu, Nami Nishikiori, Masato Furuhashi, Hiroshi Ohguro.

PUBLICATION DETAILS

JOURNAL

Biomedicines

YEAR

2023

INSTITUTIONS

Sapporo Medical University

COUNTRIES

Japan

INSTRUMENT USED

MicroSquisher

TESTING METHODS

Compression TestingMicro-Mechanical TestingUltra Low Force Testing

RESEARCH APPLICATIONS

Fibrosis & Tissue RemodelingMechanotransductionMicrotissue and Spheroid MechanicsOphthalmic Biomechanics & Corneal Tissue Engineering

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