Development and Verification of a Novel Three-Dimensional Aqueous Outflow Model for High-Throughput Drug Screening.

Area of Science:

• Ophthalmology
• Biotechnology
• Cell Biology

Background:

• Ophthalmic surgery involving bleb formation can lead to scarring, impaired fluid drainage, and increased intraocular pressure.
• Inflammatory cytokines in aqueous humor contribute to subconjunctival tissue fibrosis and surgical failure.

Purpose of the Study:

• To introduce a high-throughput screening platform for anti-fibrotic compounds.
• To evaluate verteporfin's efficacy in modulating the transforming growth factor beta (TGFβ)-SMAD2/3 pathway to prevent post-operative outflow reduction.

Main Methods:

• A microfluidic system with Human Tenon's capsule fibroblasts (HTCFs) in a 3D collagen matrix modeled aqueous humor drainage.
• Transforming growth factor beta 1 (TGFβ1) was used to induce fibrotic responses, and afferent pressure was monitored.
• Verteporfin was co-administered to assess its ability to counteract TGFβ1-induced pressure increases and cellular changes.

Main Results:

• TGFβ1 induced increased afferent pressure and myofibroblast transformation in HTCFs, characterized by cytoskeletal alterations.
• Verteporfin effectively mitigated these TGFβ1-induced functional and cellular changes.
• Immunofluorescence confirmed the reduction in cytoskeletal alterations following verteporfin treatment.

Conclusions:

• Modulating the TGFβ-SMAD2/3 pathway is a viable strategy to address post-operative fibrosis and improve filtration bleb outflow.
• Verteporfin demonstrates potential as a therapeutic agent to enhance wound healing and surgical success in ophthalmic procedures.
• The developed high-throughput screening platform serves as a valuable pre-clinical tool for identifying novel anti-fibrotic compounds.