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Establishing and functional testing of a canine corneal construct
Anke Werner1, Michael Braun, Stephan Reichl
1Institute of Pharmacology, Toxicology and Pharmacy, University of Veterinary Medicine Hannover, Hannover, Germany. anke.werner@tiho-hannover.de
Veterinary Ophthalmology
|December 3, 2008
Summary
This study established a novel three-dimensional canine corneal construct for in vitro drug testing. The model accurately mimics the in vivo cornea, enabling studies on drug effects on corneal cells.
Area of Science:
- Ophthalmology
- Biotechnology
- Drug Discovery
Background:
- Developing reliable in vitro models for ocular drug testing is crucial.
- Current models may not fully replicate the complex cellular environment of the canine cornea.
Purpose of the Study:
- To establish a protocol for creating a three-dimensional canine corneal construct for in vitro drug effect studies.
- To compare primary canine corneal cells with a rabbit corneal epithelial (RCE) cell line within this model.
Main Methods:
- A 5-week step-by-step assembly process in membrane inserts, including air-liquid interface culture for epithelial differentiation.
- Histological analysis, stimulation with lipopolysaccharides (LPS) and sodium dodecyl sulfate (SDS), and dexamethasone treatment to assess prostaglandin E(2) (PGE(2)) production.
- Immunohistochemistry to detect glucocorticoid receptor (GR) expression.
Main Results:
- A vital corneal construct protocol was successfully established, showing morphological similarity to the in vivo canine cornea.
- Glucocorticoid receptor (GR) protein was detected in all construct cell types.
- Corneal constructs demonstrated significantly increased PGE(2) production upon LPS/SDS stimulation, which was reduced by dexamethasone.
Conclusions:
- The developed corneal construct serves as a valuable in vitro system for testing drug effects.
- This model incorporates all three major corneal cell types, offering a more comprehensive cornea-like system for research.
