The Time Course of Hydroxyl Radical Formation following Spinal Cord Injury: The Possible Role of the Iron-Catalyzed Haber-Weiss Reaction

This study explores whether the hydroxyl radical (^•OH)—one of the most destructive reactive oxygen species—plays a role in secondary spinal cord injury (SCI). First, we measured the time course of ^•OH formation in rat spinal tissue after impact SCI by administering salicylate as a trapping agent into the intrathecal space of the cord and measuring the hydroxylation products of salicylate, 2,3- and 2,5-dihydroxybenzoic acid (2,3- and 2,5-DHBA) by HPLC. The 2,3-DHBA concentration was significantly higher in injured spinal tissue than in sham controls at 5 min, 1 and 3 h, but not at 5 h post-injury. Second, we generated ^•OH by administering H₂O₂ and FeCl₂/EDTA (Fenton's reagents) at the concentrations produced by SCI into the gray matter of the cord for 4 h and found that it induced significant cell loss at 24 h post-^•OH exposure. Mn (III) tetrakis (4-benzoic acid) porphyrin(MnTBAP)—a broad spectrum reactive species scavenger—significantly reduced ^•OH-induced cell death. Finally, we generated superoxide and administered FeCl₃/EDTA in the intrathecal space of the cord at the concentration produced by SCI and measured extracellular ^•OH formation in the gray matter of the cord by microdialysis sampling. We found that the levels of ^•OH significantly increased compared to the pre-administration level, indicating that ^•OH can be produced in vivo by the iron-catalyzed Haber-Weiss reaction. All together, we demonstrated that ^•OH is an endogenous secondary damaging agent following SCI and the metal-catalyzed Haber-Weiss reaction may contribute to early ^•OH formation after SCI.