David Lockington1, Pankaj Agarwal2, David Young3, Muriel Caslake4, Kanna Ramaesh2. 1. Tennent Institute of Ophthalmology, Glasgow, United Kingdom. Electronic address: davidlockington@hotmail.com. 2. Tennent Institute of Ophthalmology, Glasgow, United Kingdom. 3. Department of Statistics and Modelling Science, University of Strathclyde, Glasgow, United Kingdom. 4. Institute of Cardiovascular and Medical Sciences, University of Glasgow, Glasgow, United Kingdom.
Abstract
OBJECTIVE: Amniotic membrane (AM) is used to manage various debilitated ocular surface conditions. The impact of oxidative stress and free radicals on the ocular surface is increasingly being recognized. Hyaluronic acid (HA) has anti-inflammatory properties and is abundantly present in AM. In this in vitro pilot study, we investigated the potential of AM for intrinsic free radical scavenging properties. METHODS: Strips of AM were incubated in sealed tubes with hydrogen peroxide (H2O2). After being sonicated, uptake of reactive oxygen species (ROS) was measured by the Amplex Red Hydrogen Peroxide/Peroxidase assay. For comparison, 1630 kDA HA was used. RESULTS: There was uptake of ROS by all AM samples, which decreased with increasing concentrations of H2O2. Mean ROS uptake for 5 different AMs at 1 hour was significantly greater for 50 μM (83%; SD 11.7, SEM 5.23) compared with 100 μM (67%; SD 20.48, SEM 9.16; p = 0.028; 95% CI 2.8-29.2). The HA comparison group showed similar uptake and trend. CONCLUSION: This pilot study demonstrates that AM is able to remove ROS from its environment. Demonstrating total antioxidant capacity in AM provides evidence for use as a free radical scavenger. The antioxidant properties of AM and the contribution from HA require more research.
OBJECTIVE: Amniotic membrane (AM) is used to manage various debilitated ocular surface conditions. The impact of oxidative stress and free radicals on the ocular surface is increasingly being recognized. Hyaluronic acid (HA) has anti-inflammatory properties and is abundantly present in AM. In this in vitro pilot study, we investigated the potential of AM for intrinsic free radical scavenging properties. METHODS: Strips of AM were incubated in sealed tubes with hydrogen peroxide (H2O2). After being sonicated, uptake of reactive oxygen species (ROS) was measured by the Amplex Red Hydrogen Peroxide/Peroxidase assay. For comparison, 1630 kDA HA was used. RESULTS: There was uptake of ROS by all AM samples, which decreased with increasing concentrations of H2O2. Mean ROS uptake for 5 different AMs at 1 hour was significantly greater for 50 μM (83%; SD 11.7, SEM 5.23) compared with 100 μM (67%; SD 20.48, SEM 9.16; p = 0.028; 95% CI 2.8-29.2). The HA comparison group showed similar uptake and trend. CONCLUSION: This pilot study demonstrates that AM is able to remove ROS from its environment. Demonstrating total antioxidant capacity in AM provides evidence for use as a free radical scavenger. The antioxidant properties of AM and the contribution from HA require more research.
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