| Objective To investigate the efficacy of ozone therapy(Ozone) on brain edema in an experimental rat model of traumatic brain injury(TBI) and then to compare its effects with hyperbaric oxygen(HBO) therapy in this entity.Methods One hundred and twenty male S-D rats were randomly divided into four groups: Sham(n=30), TBI(n=30), TBI +HBO(n=30) and TBI +Ozone(n=30), the last three groups with successful modeling were randomly divided into 5 subgroups(each group n=6) for analysis on days 1, 3, 5 and 7 and one group for histopathologic analysis, respectively. TBI rats were induced to mild fluid percussion injury. HBO was administered twice daily at a 2.3-atm pressure for 90 minutes. Ozone therapy was set as daily intraperitoneal injections of 0.7 mg/kg ozone/oxygen gas mixture. HBO and Ozone were continued for 7 days after the induction of TBI. Brain tissues and blood sample were harvested for biochemical analysis at corresponding time points(days 1, 3, 5 and 7). Neural functional status of rats was assessed by modified neurological severity score(m NSS). Malondialdehyde(MDA) levels, superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) activities were determined in brain homogenates. The brain water content was administrated by the means of wet-dry weight. The group for histopathological analysis were performing MRI via animal machine, especially in T2 WI for obversing edema at 6h, 1d, 3d, 5d and 7d after the corresponding treatments. Rats in this group were killed at the eighth day,and their brain tissues were obtained for histopathologic analyses by hematoxylin-eosin staining.Results The m NSS scores in the HBO, Ozone and TBI groups were significantly higher than those in Sham group. The m NSS scores in the HBO and Ozone groups were lower than those in the TBI group on days 3, 4, 5, 6 and 7, and then those in Ozone group were lower than TBI on days 5, 6 and 7. The three groups at 6h, 1d and2 d scores had no significant difference(p > 0.05). Compared with the other threegroups at the same time, the SOD content of sham group was significantly higher(p <0.05); TBI, HBO and Ozone groups had no significant difference in 1d and no statistical significance(p < 0.05); HBO and Ozone groups of SOD content were significantly higher than those in TBI group in 3d, 5d and 7d(p < 0.05), and its no difference between The first two groups(p > 0.05); The change of GSH-Px content groups of rats had the trend consistent with the SOD. Compared with the other three groups at the same time, the MDA content of Sham group was significantly lower; There was no significant different among TBI, HBO and Ozone groups at the1 d and 3d, and no statistical significance(p < 0.05); The content of MDA in HBO and Ozone groups at the 5d and 7d was significantly lower than those in the TBI group(p < 0.05), and no difference between(p > 0.05). Except of the 7d, the brain water content of the sham group was significantly lower compared with the other three groups at the same time. There was no significant difference among TBI, HBO and Ozone group at the 1d and 3d, and no statistical significance(p < 0.05). This level of HBO group and Ozone group in the 5d was significantly lower than the TBI group(p < 0.05), and its no difference between(p > 0.05). In the 7d, Ozone brain water content was lower than the HBO group(p < 0.05), and then there was no significant difference between Ozone group and Sham group(p > 0.05). The result of the T2 WI showed that the hyperbaric oxygen and ozone had therapeutic effect on acute phase of severe head trauma, promoted the absorption of hematoma,contributing to the improvement in intracranial conditions, but only to observe from the nuclear magnetic there had no obvious difference. The edema volume comparison shows that it was significantly lower(p < 0.05) in sham group than those in the other three groups at the same time. TBI group, HBO group and Ozone group had no significant difference at the same time and no statistical significance(p > 0.05).Compared to Sham group, both brain water content and brain edema volume in T2 WI were increased at day 1, reaching a maximum at day 3, and then gradually decreased in the others. HE staining did not show any structural changes in Sham group.Necrosis and apoptotic neurocytes were found mostly in TBI group. Necrosis and normal neurocytes were found in HBO and Ozone group, but less of necrosis was in the last.1. The m NSS scores in the HBO, Ozone and TBI groups were significantly higher than those in Sham group.The m NSS scores in the HBO and Ozone groups were lower than those in the TBI group in 3d, 4d, 5d, 6d and 7d and then those in Ozone group were lower than TBI on day 5, 6 and 7. The three groups at 6h, 1d and 2d in score had no significant difference(p > 0.05).2. Compared with the other three groups at the same time,the SOD content of sham group was significantly higher(p < 0.05); TBI group, HBO group and Ozone group had no significant difference in 1 d and no statistical significance(p < 0.05); HBO and Ozone groups of SOD content were significantly higher than those in TBI group in 3d, 5d and 7d(p < 0.05), and its no difference between The first two groups(p >0.05); The change of GSH-Px content groups of rats had the trend consistent with the SOD. Compared with the other three groups at the same time, the MDA content of Sham group was significantly lower; There was no significant different among TBI,HBO and Ozone group at the 1d and 3d and no statistical significance(p < 0.05); The content of MDA in HBO and Ozone groups at the 5d and 7d was significantly lower than those in the TBI group(p < 0.05), and its no difference between(p > 0.05).3. Except of the 7d, the brain water content of the sham group was significantly lower compared with the other three groups at the same time. There was no significant difference among TBI, HBO and Ozone group at the 1d and 3d, and no statistical significance(p < 0.05). This level of HBO group and Ozone group in the 5d was significantly lower than the TBI group(p < 0.05), and its no difference between them(p > 0.05). In the 7d, Ozone brain water content was lower than the HBO group(p <0.05), and then there was no significant difference between Ozone and Sham groups(p > 0.05).4. The result of the MRI T2 WI showed that the hyperbaric oxygen and ozone had therapeutic effect on acute phase of severe head trauma, promoted the absorption of hematoma, contributing to the improvement in intracranial conditions, but only to observe from the nuclear magnetic there had no obvious difference. The edema volume comparison shows that it was significantly lower(p < 0.05) in sham group than those in the other three groups at the same time. TBI group, HBO group and Ozone group had no significant difference at the same time and no statisticalsignificance(p > 0.05). Compared to Sham group, both brain water content and brain edema volume in T2 WI were increased at day 1, reaching a maximum at day 3, and then gradually decreased in the others.5. HE staining did not show any structural changes in Sham group. Necrosis and apoptotic neurocytes were found mostly in TBI group. Necrosis and normal neurocytes were found in HBO and Ozone group, but less of necrosis was in the last.In the Sham and Ozone group, histopathologic scores were lower than in the HBO group, but it was not statistically significant.Conclusion These findings suggest that Ozone and HBO might improve traumatic cerebral edema because of both oxidative stress and inflammation better than HBO, and a greater benefit was received for Ozone comparing with HBO.1. TBI can cause the body a series of pathophysiological changes: including neurobehavioral, imaging, histopathology and the corresponding change of cell and molecular level.2. Nerve function and activities was significantly damaged after TBI. The accurate and objective MRI results can show the change of the craniocerebral trauma, and then brain tissue morphology significantly changed at the same time, The cortex and hippocampus nerve fine structure was severely damaged. Moreover, the oxidative stress increased in the body and antioxidant enzyme system was damaged. The large numbers of free radicals were produced and inflammatory cytokines released,resulting in late onset neuron degeneration, causing nerve cell necrosis and apoptosis.3. By a certain time of hyperbaric oxygen and Ozone treatment after TBI, relative to the Sham group, the m NSS and MDA content decreased, the SOD and GSH-Px activity increased. It is confirmed that the situation of intracranial hematoma improved on T2 WI, the brain water content was significantly lower. Morever, HE dying showed that nerve cells had got improved in the form, and then it showed a certain treatment from the aspects of morphology.4. In aspect of improving the m NSS, decreasing the brain water content and reducing the histopathological injury, etc. Ozone is superior to hyperbaric oxygen. |
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