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Multilayered Fabrication Assembly Technique to Engineer a Corneal Stromal Equivalent
Julia Fernández-Pérez1,2, Peter W Madden1,2, Mark Ahearne1,2
1Department of Mechanical, Manufacturing and Biomedical Engineering, School of Engineering, Trinity College Dublin, University of Dublin, Dublin, Ireland.
Bio-Protocol
|April 15, 2021
Summary
This study presents a novel method for creating corneal tissue engineering constructs. By layering decellularized porcine corneas with cell-laden hydrogels, high cellularity is achieved rapidly for potential transplantation applications.
Area of Science:
- Ophthalmology
- Biomaterials Science
- Regenerative Medicine
Background:
- Donor shortage for human corneal transplantation necessitates tissue engineering solutions.
- Current synthetic corneal substitutes fail to support normal human cell phenotype.
- Decellularized animal corneas offer topographical and biochemical cues but have slow cell infiltration.
Purpose of the Study:
- To develop a rapid method for creating corneal constructs with high cellularity.
- To utilize tissue-derived materials for improved corneal regeneration.
- To overcome limitations of existing decellularized corneal scaffolds.
Main Methods:
- Decellularized porcine corneal sheets were prepared via cryosectioning, detergent/nuclease treatment, and air drying.
- Corneal stromal cells were encapsulated in collagen type I hydrogel.
- Alternating layers of decellularized corneal sheets and cell-laden hydrogel were stacked to form constructs.
Main Results:
- The developed protocol enables rapid production of corneal constructs.
- High cellularity is achieved throughout the entire construct.
- The method utilizes readily available, tissue-derived materials.
Conclusions:
- This layered approach offers a promising strategy for corneal tissue engineering.
- The method ensures uniform and high cell density, crucial for corneal function.
- This technique addresses the limitations of previous decellularized corneal scaffolds.
