Abstract
Purpose :
Corneal endothelial cells exhibit attenuated expression and distribution of function-related markers following cell expansion. Various strategies are investigated worldwide to counterbalance this effect. Here we investigate if mimicking the hydrostatic pressure of the anterior chamber can modulate corneal endothelial morphology and functionality.
Methods :
Cultivated corneal endothelial cells (P2-P3; n = 6 populations) were seeded on devitalized corneas (n = 10 pairs). Native corneas and devitalized corneas were respectively used as positive (n = 2 pairs) and negative controls (n = 3 pairs). Corneas were placed in artificial anterior chambers and subjected to a hydrostatic pressure between 0.3 and 0.4 psi during 4-5 days. Unpressured control corneas were maintained in cell culture dishes. Pictures of the corneas were taken following the experiment to assess stromal transparency. Morphology, corneal thickness and distribution of ZO-1, n-cadherin, b-catenin, Na+/K+ ATPase pump and HCO3- cotransporter was evaluated by electron microscopy, histological staining and immunofluorescences.
Results :
Pressure treated corneas were more transparent than the controls. Thickness was accordingly reduced by 38.4±5.5% for cultivated endothelium and 32.2±9.6% for native endothelium. Negative controls change in transparency and thickness were marginal. Pressure treated cells showed none or at most marginal difference in morphology and distribution of ZO-1, n-cadherin, b-catenin, Na+/K+ ATPase pump and HCO3- cotransporters and failed to recreate a phenotype similar to native corneas.
Conclusions :
Results show that in presence of the endothelium, pressure caused an amelioration of corneal transparency associated with stromal thinning but without a change in cell morphology or in the presence and distribution of function-associated markers. This suggests that anterior chamber pressure may have an endothelial functionality unrelated role in supporting corneal transparency.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.