New observations on bicarbonate-pH effects on thickness changes of rabbit corneas under silicone oil in vitro

Specular microscopy was used to measure thickness changes of stroma-endothelium preparations from rabbit corneas in vitro. The preparations were first bathed on both sides for 90 min with different bicarbonate-Ringer solutions (2 to 50 mM bicarbonate) maintained in equilibrium with 5% CO2-air (pH 6.2 to 7.9). During this equilibration, the stroma attained a stable thickness that was inversely related to the hydrostatic pressure (20 to 100 cm H2O) applied to the endothelial surface. After equilibration and at 20 cm H2O pressure, covering the anterior stromal surface with silicone oil (Dow Corning 200 dielectric fluid, 20 cs viscosity) resulted in stromal thinning. The rate of this deturgescence increased (from 5 to 75 micron/h) as the equilibration solution bicarbonate level increased from 2 to 30 mM. The net size of the thickness change was also related to the equilibration bicarbonate level. However, indirect studies on the cornea using phenol red indicator and pH electrode measurements of solutions revealed that the stromal bathing solutions became more alkaline under the silicone oil layer. CO2 is soluble in silicone oil. Implications of these CO2 and pH effects on mechanisms of corneal deturgescence are discussed.