Tissue-Engineered Endothelial Keratoplasty with Controlled Cell Density: Toward Super TEEKs

Area of Science:

• Ophthalmology
• Biomaterials Science
• Regenerative Medicine

Background:

• Endothelial keratoplasty is a leading treatment for corneal endothelial disorders, but donor tissue shortages necessitate alternatives.
• Tissue-engineered endothelial keratoplasty (TEEK) aims to replicate native endothelial grafts using cultured cells on a scaffold.
• Current TEEK development focuses on mimicking natural grafts and addressing donor tissue limitations.

Purpose of the Study:

• To demonstrate proof of concept for controlling endothelial cell density (ECD) in tissue-engineered endothelial keratoplasty (TEEK).
• To evaluate the quality and viability of endothelial cells cultured on a novel bioengineered scaffold.
• To establish a foundation for creating "super TEEKs" with supra-physiological cell densities.

Main Methods:

• Utilized femtosecond-laser-cut lens capsule discs as biocompatible scaffolds for endothelial cell culture.
• Varied seeding densities to control the final endothelial cell density (ECD) in 12 TEEK samples.
• Characterized cell morphology and differentiation using anti-NCAM staining and the CellPose AI algorithm.
• Assessed cell viability at 28 days using Hoechst 33342 and Calcein-AM staining.

Main Results:

• Achieved controlled endothelial cell density by adjusting seeding density per mm².
• Median cell viability reached 98% at 28 days of culture.
• The highest-density group achieved a median viable ECD of 3.245 cells/mm², exceeding physiological levels.

Conclusions:

• Demonstrated the feasibility of controlling ECD in TEEK through precise seeding density.
• The developed TEEK exhibits high cell viability and quality, suitable for transplantation.
• This approach enables the bioengineering of "super TEEKs" with enhanced therapeutic potential, addressing donor tissue scarcity.