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Related Experiment Video

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An “All-laser” Endothelial Transplant
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Tissue-Engineered Endothelial Keratoplasty with Controlled Cell Density: Toward Super TEEKs.

Inès Aouimeur1, Louise Coulomb1, Sofiane Fraine1

  • 1Laboratory for Biology, Engineering, and Imaging for Ophthalmology, BiiO, Faculty of Medicine, Health & Innovation Campus, Jean Monnet University, Saint-Etienne, France.

Tissue Engineering. Part A
|September 23, 2025
PubMed
Summary

Tissue-engineered endothelial keratoplasty (TEEK) offers a promising alternative to donor corneas. Researchers demonstrated control over endothelial cell density in TEEK, achieving high cell viability and paving the way for advanced corneal treatments.

Keywords:
corneal bioengineeringcorneal endotheliumhuman anterior lens capsulehuman corneal endothelial cells (hCECs)viable tissue-engineered endothelial keratoplasty (TEEK)

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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.