J Luo1, N Li, J P Robinson, R Shi. 1. Center for Paralysis Research, Department of Basic Medical Sciences, School of Veterinary Medicine, Purdue University, West Lafayette, Indiana 47907, USA.
Abstract
STUDY DESIGN: In vitro studies using isolated guinea pig spinal cord. OBJECTIVES: To develop an alternative model using isolated guinea pig spinal cord, which can be used to screen antioxidants for in vivo SCI treatment. SETTING: Department of Basic Medical Sciences, Purdue University, West Lafayette, Indiana, USA. METHODS: The compression injury was induced by a constant-displacement of 5-s compression of spinal cord using a modified forceps possessing a spacer. Reactive oxygen species (ROS) were evaluated using three distinct methods: fluorescence microscopy, lipid peroxidation assay, and flow cytometry. RESULTS: The injury-mediated ROS increases are comparable with other in vivo studies and consistent with our previous observation using a similar injury model and measured with electrophysiological and anatomical technique. Further, ascorbic acid, hypothermia, or the combination of both significantly suppressed superoxide and lipid peroxidation. The combination treatment was the most effective when compared with ascorbic acid or hypothermia alone. CONCLUSION: This in vitro model has the advantage of replicating some of the in vivo conditions while gaining the ability to control the experimental conditions. This in vitro model is suitable to study the mechanisms of ROS generation and degradation and can also be used to critically evaluate the effective suppressor of ROS in the contents of spinal cord traumatic injury.
STUDY DESIGN: In vitro studies using isolated guinea pig spinal cord. OBJECTIVES: To develop an alternative model using isolated guinea pig spinal cord, which can be used to screen antioxidants for in vivo SCI treatment. SETTING: Department of Basic Medical Sciences, Purdue University, West Lafayette, Indiana, USA. METHODS: The compression injury was induced by a constant-displacement of 5-s compression of spinal cord using a modified forceps possessing a spacer. Reactive oxygen species (ROS) were evaluated using three distinct methods: fluorescence microscopy, lipid peroxidation assay, and flow cytometry. RESULTS: The injury-mediated ROS increases are comparable with other in vivo studies and consistent with our previous observation using a similar injury model and measured with electrophysiological and anatomical technique. Further, ascorbic acid, hypothermia, or the combination of both significantly suppressed superoxide and lipid peroxidation. The combination treatment was the most effective when compared with ascorbic acid or hypothermia alone. CONCLUSION: This in vitro model has the advantage of replicating some of the in vivo conditions while gaining the ability to control the experimental conditions. This in vitro model is suitable to study the mechanisms of ROS generation and degradation and can also be used to critically evaluate the effective suppressor of ROS in the contents of spinal cord traumatic injury.
Authors: Martin Tribus; Johannes Galehr; Patrick Trojer; Gerald Brosch; Peter Loidl; Florentine Marx; Hubertus Haas; Stefan Graessle Journal: Eukaryot Cell Date: 2005-10