Abstract
Purpose: :
To investigate the efficacy of redox fluorometry in evaluating the state of cultured human corneal endothelial cells and to evaluate the protective effects of balanced salt solution (BSS) and BSS plus against ultrasonic energy in vitro.
Methods: :
Human corneal endothelial cells (HCECs) from fresh banked human donor tissue not suitable for transplantation were harvested and cultured. A phacoemulsification probe with a 30° round, 1.1mm TurboSonicsÿ ABSTM Tip (Alcon Surgical, Fort Worth, Texas) was introduced into culture dishes filled with BSS and BSS plus. Cellular autofluorescence images were obtained using a Zeiss inverted microscope (Thornwood, NY, Axiovert 200M). The redox fluorometric ratio, which can be related to oxidative stress, was calculated as the net value of fluorescence from the DAPI channel divided by net value of fluorescence from the FITC channel after subtraction of background.
Results: :
HCECs exposed to increasing phacoemulsification times and increasing ultrasonic energy levels displayed dose dependent decreases in measured redox ratios. BSS and BSS plus caused a significant difference of redox ratios compared to the PBS control. (p<0.001)
Conclusions: :
Redox fluorometry microscopy is a useful tool to evaluate human corneal endothelial damage induced by the oxidative stress of phacoemulsification.
Keywords: cornea: endothelium • image processing • oxidation/oxidative or free radical damage