ESCRS - FP28.13 - Protective Effect Of Ovds With Various Biochemical Constituents Against Hydrogen Peroxide-Induced Oxidative Damage To Corneal Endothelial Cells

Protective Effect Of Ovds With Various Biochemical Constituents Against Hydrogen Peroxide-Induced Oxidative Damage To Corneal Endothelial Cells

Published 2025 - 43rd Congress of the ESCRS

Reference: FP28.13 | Type: Free paper | DOI: 10.82333/qs2n-y014

Authors: Hussien Mahmoud Ismail 1 , Sherif Nabil Ragaei 2 , Mahmoud Ismail* 3

1Cornea,Nour El Hayah Eye Hospital,Cairo,Egypt, 2Ophthalmology,University of Lausanne, Jules-Gonin Eye Hospital,Lausanne,Switzerland, 3Ophthalmology ,Al Azhar University,Cairo,Egypt

Purpose

The role of biochemical constituents of ophthalmic viscosurgical devices (OVDs) in protecting corneal endothelial cells (CECs) from oxidative damage is not well understood. Hydrogen peroxide (H2O2) has been shown to represent reactive oxygen species (ROS) generated during phacoemulsification. We hypothesize that OVDs with different constituents reduce oxidative damage to CECs to varying extents using an in vitro cell culture H2O2 assay. The 3 test OVDs consisted of 1) chondroitin sulfate and hyaluronic acid (CS-HA), 2) sorbitol and hyaluronic acid (sorbitol-HA) and 3) HA alone, with Balanced Salt Solution (BSS) as the CEC damage positive control.

Setting

Experimental bench-top study

Methods

Confluent rabbit CECs were cultured in 96-well plates and exposed to a 40% OVD, 60% media mixture with 6mM H2O2 for 2.5h, then assessed for CEC damage using a lactate dehydrogenase (LDH) assay. 200µL of the mixtures containing CS-HA (N=36), sorbitol-HA (N=18), HA alone (N=18), or BSS control (N=36) were added to each well, and incubated for 2.5h with 5% CO2. 100µL of supernatant was removed from each well and transferred to a plate with 100µL of BSS. 100µL of the LDH assay reagent was added to 100µL of the mixture and incubated at room temperature for 35min. The magnitude of Absorbance, proportional to CEC damage and measured at 500nm in a plate reader, was normalized to the control group. A 1-Way ANOVA was conducted to compare %CEC damage.

Results

Mean, SD and 95% CIs were calculated for each study group. Mean %CEC damage was significantly different among all groups (p<0.001). The lowest %CEC damage was found with CS-HA (n=36, 24.7 ± 2.5; Mean ± 95% CI) followed by Sorbitol-HA (n=18, 36.2 ± 4.8; Mean ± 95% CI) then HA (n=18, 51.8 ± 7.3; Mean ± 95% CI) normalized to BSS (n=36, 100.0 ± 3.5; Mean ± 95% CI). CS-HA had significantly less CEC damage when compared to Sorbitol-HA and HA alone (p<0.001).

Conclusions

OVD biochemical composition influences CEC oxidative damage. OVDs that contain CS-HA provided the least oxidative damage, suggesting greater CEC protection when compared to OVDs containing sorbitol-HA and HA alone. All OVDs provided better protection than BSS alone.