A Porcine Anterior Segment Perfusion and Transduction Model With Direct Visualization of the Trabecular Meshwork

Area of Science:

• Ophthalmology
• Biomedical Engineering
• Genetics

Background:

• Developing reliable models for anterior segment research is crucial for understanding ocular diseases.
• Existing models using human donor eyes can be expensive and difficult to obtain.
• A need exists for a consistent, affordable, and high-quality model for trabecular meshwork (TM) studies.

Purpose of the Study:

• To establish a consistent, affordable, high-quality porcine anterior segment perfusion and transduction model.
• To enable direct visualization of the trabecular meshwork (TM).
• To serve as a viable alternative for TM bioengineering strategies.

Main Methods:

• Porcine anterior eye segments were cultured within 2 hours of death.
• A specialized petri dish allowed direct TM visualization.
• Gene delivery was achieved using feline immunodeficiency virus (FIV) vector GINSIN at varying concentrations (10^8 and 10^7 TU).
• Intraocular pressure (IOP) and gene expression were monitored for 14 days.

Main Results:

• The porcine model demonstrated a low failure rate (12%) and consistent anterior segment cultures.
• Gravity perfusion resulted in lower flow rates (3.7 μL/min) compared to fixed pump perfusion (3 μL/min).
• Higher vector concentration (GINSIN_8) caused a transient IOP increase, while GINSIN_7 did not.
• Trabecular meshwork architecture was preserved, and gene expression was successfully achieved.

Conclusions:

• The porcine anterior segment perfusion model is a high-quality, repeatable, and inexpensive alternative to human donor eyes.
• This model facilitates direct TM observation and TM bioengineering development.
• The transduction effects observed in the porcine model align with those seen in human, feline, and rodent models.