Nanofibrous PCL-Based Human Trabecular Meshwork for Aqueous Humor Outflow Studies

Area of Science:

• Biomedical Engineering
• Ophthalmology
• Cell Biology

Background:

• Primary open-angle glaucoma involves optic nerve degeneration, with high intraocular pressure (IOP) as a key risk factor.
• The human trabecular meshwork (HTM), particularly the juxtacanalicular tissue (JCT), regulates aqueous humor (AH) outflow and IOP.
• Current in vitro JCT models are limited, hindering research into AH outflow and HTM degeneration.

Purpose of the Study:

• To develop a functional in vitro JCT model using a polycaprolactone (PCL) nanofibrous scaffold.
• To investigate the morphology, cell behavior, and drug responsiveness within the engineered JCT model.
• To provide a platform for studying HTM cell biology and screening drugs affecting trabecular outflow.

Main Methods:

• Fabrication of a PCL nanofibrous scaffold for JCT recapitulation.
• Seeding human trabecular meshwork cells onto the scaffold.
• Characterization using SEM, qPCR, and immunochemistry for HTM markers.
• Construction of a pressure-sensitive perfusion system to assess outflow facility.
• Testing drug responsiveness with dexamethasone and netarsudil.

Main Results:

• The PCL scaffold supported HTM cell morphology and function, mimicking the JCT.
• Engineered JCT exhibited characteristic HTM cell markers (myocilin, fibronectin, collagen IV) and genetic expression.
• The model demonstrated functional outflow characteristics and responsiveness to IOP-lowering drugs.
• Successful recapitulation of HTM cell biology and drug effects in vitro.

Conclusions:

• A novel, functional in vitro JCT model was successfully developed using PCL nanofibrous scaffolds.
• This model effectively mimics key aspects of the human JCT, including cell morphology, marker expression, and drug response.
• The developed model serves as a valuable tool for understanding HTM cell biology, AH outflow regulation, and for pharmaceutical drug screening in glaucoma research.