Abstract
Purpose: :
To examine the effects of Sodium Lauryl Sulphate (SLS) on irritancy within an artificial stem cell based limbal corneal construct in comparison with explanted bovine eye to show the construct is a viable alternative to in vivo testing.
Methods: :
Compressed Collagen gels were seeded with low passage bovine keratocytes and epithelial cells to create a 3D artificial cornea1. These were treated with up to 1% SLS, a known ocular irritant to induce a response similar to that seen with in vivo studies. Histology and the MTT assay were used on both the artificial cornea and fresh bovine eye to allow comparison of the two systems for signs of irritancy over the course of 30 minutes.
Results: :
The artificial cornea showed similar results in the MTT assay, histological examination, PCR analysis and Immunohistochemistry to bovine cornea in a manner that supports previously published data2. We showed a concentration dependent reduction in cell viability and organization that was concordant with data obtained from bovine cornea. We show that there is comparable IC50 results for both explants and artificial constructs across all measured SLS concentrations backed up with MTT data and histological examination of disruption to the epithelial and stromal layers.
Conclusions: :
We have demonstrated that our artificial cornea has similar properties to native tissue in response to irritant factors. This paves the way for further research into the effects of other irritants and allows us to do so in a reproducible and defined way without the need for in vivo testing usually associated with such testing.
Keywords: cornea: basic science • cornea: epithelium • cornea: stroma and keratocytes