Recreating a human trabecular meshwork outflow system on microfabricated porous structures

Area of Science:

• Biomedical Engineering
• Ophthalmology
• Cell Biology

Background:

• Glaucoma, a leading cause of irreversible blindness, is linked to elevated intraocular pressure (IOP).
• The human trabecular meshwork (HTM) regulates IOP by controlling aqueous humor outflow.
• A lack of effective in vitro HTM models hinders glaucoma research and therapeutic development.

Purpose of the Study:

• To engineer a functional in vitro human trabecular meshwork (HTM) model.
• To investigate HTM cell behavior and outflow physiology.
• To facilitate the discovery of novel anti-glaucoma therapeutics.

Main Methods:

• Development of an in vitro HTM model using micropatterned, porous SU-8 scaffolds.
• Optimization of scaffold properties and cell culture conditions for HTM cell growth.
• Characterization of the bioengineered HTM using F-actin staining and immunocytochemistry.
• Construction of a perfusion chamber with integrated pressure sensing for outflow facility assessment.
• Evaluation of drug responsiveness using latrunculin B, an IOP-lowering agent.

Main Results:

• The engineered HTM model successfully recapitulated functional HTM morphology and cellular characteristics.
• The model expressed key HTM markers, including α-smooth muscle actin, myocilin, and αß-crystallin.
• Outflow facility was successfully measured, and the model demonstrated responsiveness to latrunculin B.
• The developed system allows for the study of HTM cell biology and drug screening.

Conclusions:

• A robust in vitro HTM model system has been successfully developed.
• This model system is suitable for understanding HTM cell biology and screening anti-glaucoma agents.
• The model accelerates the discovery of therapeutics targeting trabecular outflow facility for glaucoma treatment.