Introduction: The present study describes simulation of corneal epithelial injury and its regeneration using an in-vitro model of immortalized human corneal epithelial cells (HCE-T) growing as monolayer cultures.
Material and methods: The epithelial model was damaged using defined strengths by mechanical injury or partial damage using chemical detergents (SDS and acidified medium) and subsequently the epithelium was further cultivated using serum-containing and serum-free medium supplemented with varying concentrations of calcium pantothenat. After mechanical injury wound healing was evaluated using a photomicroscope over a period of up to 48 h whereas after chemical injury a cell viability assay was used to detect the course of ATP levels in the cell layers as an indicator for the metabolic activity.
Results: Depending on the kind of injury pantothenat showed a regeneration enhancing effect in the concentration range from 0.001% to 0.01%. However, a concentration of 0.1% pantothenat appeared to be regeneration inhibiting. The combination of pantothenat and serum was more beneficial for wound healing than pantothenat alone, whereas serum partly levelled the effect of pantothenat.
Conclusion: The described model allowed simulation of corneal epithelial injury and its regeneration, whereby the influence of the serum content and the kind of injury could be determined.