Abstract

P 189

Influence of dexpanthenol on regeneration of human corneal epithelial monolayers

Hahne M., Reichl S.
Technische Universität Carolo-Wilhelmina zu Braunschweig, Institut für Pharmazeutische Technologie, Braunschweig

Objective: The study describes the evaluation of an in-vitro model of human corneal epithelial cells (HCE-T) growing as monolayers to define regenerative and wound healing enhancing effects. On the basis of two different methods the influence of dexpanthenol on the regeneration of HCE-T cells after mechanical injury as well as partial damage using a chemical detergent (sodiumdodecylsulfate-SDS) was analysed.
Methods: SV40 immortalized, in calcium-pantothenate-free medium cultivated HCE-T cells were used. Cells were cultivated confluent and subsequently a 1200 µm wide wound was scratched using a cell scraper. Afterwards it was further cultivated with medium, that contains increasing concentrations of dexpanthenol up to 0,1%. Wound healing was observed using a microscope over a period of 24 h and regenerated epithelial area was evaluated. Furthermore the regeneration of the cells was analysed by using a cell viability assay (CellTiter-Glo), that detects the ATP-level of the cells as indication for the metabolic activity. Confluent monolayers of HCE-T cells in 96 well-plates were damaged with medium containing 0,013% SDS. Afterwards the cells were further incubated with medium, that contains increasing concentrations of dexpanthenol as well. The increase of the ATP-level during the regeneration phase was analysed and used to evaluate the regeneration.
Results: Surprisingly both methods showed the best results of regeneration and wound healing in the absence of dexpanthenol. The regeneration decelerated with increasing dexpanthenol-concentration significantly. In the medium without dexpanthenol 60% of the wound area was closed within 24 h, whereas in medium containing 0,1% only 33% was regenerated. The viability of the SDS-treated cell layer (ATP-level) decreased with increasing dexpanthenol-concentration as well. Concentrations of 0,5% or higher showed even cytotoxic character.
Conclusions: In summary dexpanthenol seems to have an inhibiting effect on the regeneration of corneal epithelial cells cultivated as monolayers. Further experiments including multilayered epithelial cultures as well as organotypical cornea constructs will show the effect of dexpanthenol on more complex structures.

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