Regulation of corneal stromal cell behavior by modulating curvature using a hydraulically-controlled organ chip array

Area of Science:

• Ophthalmology
• Biomechanical Engineering
• Cell Biology

Background:

• Corneal curvature abnormalities are key drivers of ectatic diseases.
• The mechanobiological effects of these geometric stresses on corneal stromal cells are not well understood.

Purpose of the Study:

• To investigate the cellular and molecular responses of corneal keratocytes, fibroblasts, and myofibroblasts to disease-relevant geometric stresses.
• To elucidate the role of mechanotransduction in the pathogenesis of corneal ectatic diseases.

Main Methods:

• Development of a hydraulically controlled curvature array chip to mimic corneal geometries (33-56D).
• Analysis of cellular responses including proliferation, phenotypic transformation, alignment, and contractility.
• RNA sequencing to identify molecular pathways affected by geometric stress.

Main Results:

• Geometric stress induced significant proliferative enhancement and fibrotic activation in keratocytes (downregulation of ALDH3A1, upregulation of α-SMA).
• Fibroblasts exhibited curvature-dependent orthogonal alignment, recapitulating corneal lamellar organization.
• Myofibroblasts showed enhanced contractile responses, and RNA sequencing revealed activation of TGF-β/SMAD signaling, ECM-receptor interactions, and inflammatory pathways.
• Cell-type-specific extracellular matrix remodeling was observed, with keratocytes losing homeostatic control, fibroblasts depositing matrix, and myofibroblasts degrading it.

Conclusions:

• Curvature-induced mechanotransduction is a fundamental driver of corneal ectatic disease progression.
• Geometric stress actively determines corneal pathology, rather than being a passive consequence.