Abstract
Objective
The aim of this study was to evaluate the GSH effect on functional and histological recovery after experimental spinal cord injury in rats.
Methods
Forty Wistar rats were subjected to spinal cord injury through the Multicenter Animal Spinal Cord Injury Study (MASCIS) Impactor system. The rats were sorted and divided into four groups, as follows: Group 1 ‒ Laminectomy and spinal cord injury; Group 2 ‒ Laminectomy, spinal cord injury and Saline Solution (SS) 0.9%; Group 3 ‒ Laminectomy, spinal cord injury, and GSH; and Group 4 ‒ lLaminectomy without spinal cord injury. GSH and SS were administered intraperitoneally. Groups 1 and 4 received no intervention.
Results
The rats were evaluated for locomotor function recovery at seven different times by the Basso, Beattie, and Bresnahan (BBB) scale on days 2, 7, 14, 21, 28, 35, and 42 after the spinal cord injury. On day 42, the rats were sacrificed to analyze the histological findings of the injured spinal cord. In the group submitted to GSH, our experimental study revealed better functional scores on the BBB scale, horizontal ladder scale, and cranial and caudal axon count. The differences found were statistically significant in BBB scores and axonal count analysis.
Conclusion
This study demonstrated that using glutathione in experimental spinal trauma can lead to better functional recovery and improved axonal regeneration rate in Wistar rats submitted to experimental spinal cord injury.
Keywords
Glutathione; Spinal cord injuries; Central nervous system/injuries; Rats; Antioxidants/Drug effects
Highlights
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Glutathione is important in protecting secondary spinal cord injury from oxidative stress.
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As the secondary injury progresses, depletion in its reduced form occurs.
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Using glutathione in traumatic spinal cord injury could help control the neurological injury.
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In an experimental setting, the use of glutathione provided better functional scores after traumatic spinal cord injury.
Highlights
-
Glutathione is important in protecting secondary spinal cord injury from oxidative stress.
-
As the secondary injury progresses, depletion in its reduced form occurs.
-
Using glutathione in traumatic spinal cord injury could help control the neurological injury.
-
In an experimental setting, the use of glutathione provided better functional scores after traumatic spinal cord injury.
Introduction
Spinal cord injury occurs by two mechanisms; primary injury is caused by direct trauma to the spinal cord, and secondary injury is a consequence of primary injury, resulting in a sequence of self-destructive biochemical events that can last for hours or days leading to dysfunction and cell death.11 Ducker TB, Salcman M, Perot PL Jr, Ballantine D. Experimental spinal cord trauma, I: Correlation of blood flow, tissue oxygen and neurologic status in the dog. Surg Neurol. 1978;10(1):60-3.,22 Ray SK, Matzelle DD, Wilford GG, Hogan EL, Banik NL. Cell death in spinal cord injury (SCI) requires de novo protein synthesis. Calpain inhibitor E-64-d provides neuroprotection in SCI lesion and penumbra. Ann N Y Acad Sci. 2001;939:436-49. These events are characterized by a pattern of progressive activation of several pathophysiological mechanisms, including microvascular perfusion changes, inflammation, the release of oxygen free radicals and lipid peroxidation of the cell membrane, deregulation of ion cell flow control, and cell death by apoptosis.11 Ducker TB, Salcman M, Perot PL Jr, Ballantine D. Experimental spinal cord trauma, I: Correlation of blood flow, tissue oxygen and neurologic status in the dog. Surg Neurol. 1978;10(1):60-3.
2 Ray SK, Matzelle DD, Wilford GG, Hogan EL, Banik NL. Cell death in spinal cord injury (SCI) requires de novo protein synthesis. Calpain inhibitor E-64-d provides neuroprotection in SCI lesion and penumbra. Ann N Y Acad Sci. 2001;939:436-49.-33 Beattie MS, Farooqui AA, Bresnahan JC. Review of current evidence for apoptosis after spinal cord injury. J Neurotrauma. 2000;17(10):915-25. In the acute phase, necrosis occurs in the central gray matter, mainly in the first hour after injury, followed by edema and hemorrhage in the following hours. This process results in ischemia caused by reduced blood flow to the affected spinal cord segment. This reduction in blood flow may be caused by a change in the vertebral canal, significant hemorrhage and edema of the spinal cord, or a decrease in systemic blood pressure. Ischemia creates a chain of biochemical reactions resulting in cell death. Then, inflammatory cells migrate to the injured site, and glial proliferation occurs simultaneously. The formation of scar tissue and cysts occurs in the chronic phase, in the period of one to four weeks, due to the proliferation of astrocytes and hypertrophy, forming a glial scar.44 Ding WX, Shen HM, Ong CN. Critical role of reactive oxygen species and mitochondrial permeability transition in microcystin-induced rapid apoptosis in rat hepatocytes. Hepatology. 2000;32(3):547-55.,55 Barros Filho TEP, Taricco MA, Oliveira RP, Greve JMA, Santos LCR, Napoli MMM. Estudo epidemiológico dos pacientes com traumatismo da coluna vertebral e déficit neurológico, internados no Instituto de Ortopedia e Traumatologia do Hospital das Clínicas da Faculdade de Medicina da USP. Rev Hosp Clin Fac Med Sao Paulo. 1990;45(3):123-126.
Over the years, the attempt to obtain more effective treatment for spinal cord injury was based on four basic approaches: surgical, physical, biological, and pharmacological.66 Bracken MB, Shepard MJ, Holford TR, Leo-Summers L, Aldrich EF, Fazl M, et al. Methylprednisolone or tirilazad mesylate administration after acute spinal cord injury: 1-year follow up. Results of the third National Acute Spinal Cord Injury randomized controlled trial. J Neurosurg. 1998;89(5):699-706. In this context, research by pharmacological approach to control secondary injuries has gained prominence in the last decade.77 Foret A, Quertainmont R, Botman O, Bouhy D, Amabili P, Brook G, et al. Stem cells in the adult rat spinal cord: plasticity after injury and treadmill training exercise. J Neurochem. 2010;112(3):762-72. Pharmacological therapy has a promising role in preventing secondary injury. Neurons and glial cells from the central nervous system are particularly prone to oxidative and electrophilic stress due to many factors, including a high content of polyunsaturated fatty acids, a high rate of oxidative metabolic activity, intense production of reactive oxygen metabolites, and relatively low antioxidant capacity. Thus, increased oxidative stress is considered a marker of spinal cord injury.33 Beattie MS, Farooqui AA, Bresnahan JC. Review of current evidence for apoptosis after spinal cord injury. J Neurotrauma. 2000;17(10):915-25.
Glutathione (GSH) is a molecule produced by almost all cells of the body, with a fundamental role in maintaining the redox state of the cell and protecting it from oxidative stress. GSH has low molecular weight and still acts as a cosubstrate for several other antioxidant enzymes, such as GSH peroxidase and GSH transferase. The main cells of the nervous system, neurons, have relatively low levels of GSH compared to other organs, such as the liver, which makes them especially vulnerable to oxidative stress. On the other hand, GSH is present in high concentrations in astrocytes, glial cells that still provide substrates for synthesizing GSH in neurons. Neuros incubation with glial cells for 24 hours increased levels of GSH in the neurons. The main substrates for GSH synthesis are cysteine, glutamate, and glycine. In addition, GSH can also be an important neuromodulator of the central nervous system.88 Logan MP, Parker S, Shi R. Glutathione and ascorbic acid enhance recovery of Guinea pig spinal cord white matter following ischemia and acrolein exposure. Pathobiology. 2005;72(4):171-8.
The aim of this study is to evaluate, in a standardized model, the nerve regeneration and motor functional recovery of Wistar rats with surgically induced paraplegia in response to GSH.
Material and methods
Group 1 - Laminectomy + Spinal Cord Injury without intervention - rats submitted to laminectomy and spinal cord injury without any intervention for control;
Group 2 - Laminectomy + Contused Spinal Cord Injury + intraperitoneal administration of 0.9% Saline Solution (SS) - rats submitted to spinal cord injury and 0.9% Saline Solution (SS) for comparison with the intervention group - control group/placebo;
Group 3 - Laminectomy + Contused Spinal Cord Injury + intraperitoneal administration of GSH (8 mg/kg) - rats submitted to spinal cord injury and intraperitoneal administration of GSH;
Group 4 - Laminectomy only, without suffering from spinal cord injury and without any intervention.
Area “A”: Cranial region (proximal) to the injury;
Area “B”: Central region with spinal cord injury;
Area “C”: Caudal region (distal) to the injury.
The segments of the spinal cord (“A”, “B”, and “C”) underwent two types of histological analysis: tissue injury and axonal regeneration.1111 Letaif OB. Avaliação do efeito do estrógeno na lesão medular experimental em ratos [dissertação]. São Paulo: Faculdade de Medicina da Universidade de São Paulo; 2014. For the qualitative analysis, the B-area slides were evaluated for necrosis, hemorrhage, hyperemia, degeneration of the nervous substance, and cellular infiltrate. A score ranging from 0 to 3 (absent, discrete, moderate, and intense) was attributed to the findings in each section of the bone marrow histologically studied.1212 Grossman SD, Rosenberg LJ, Wrathall JR. Temporal-spatial pattern of acute neuronal and glial loss after spinal cord contusion. Exp Neurol. 2001;168(2):273-82.
For the quantitative analysis of the proximal (“A”) and distal (“C”) areas, two fields of each segment were selected and randomly chosen by the pathologist. The photos were analyzed using Sigma Scan Pro 5.0 software for counting axon fibers. Only neurons with a diameter ≥ 15 μm were considered for counting.1313 Bertelli JA, dos Santos AR, Taleb M, Calixto JB, Mira JC, Ghizoni MF. Long interpositional nerve graft consistently induces incomplete motor and sensory recovery in the rat. An experimental model to test nerve repair. J Neurosci Methods. 2004;134(1):75-80. The number of regenerated axons in the distal segment and the number of proximal axons were applied to the following formula1212 Grossman SD, Rosenberg LJ, Wrathall JR. Temporal-spatial pattern of acute neuronal and glial loss after spinal cord contusion. Exp Neurol. 2001;168(2):273-82.
13 Bertelli JA, dos Santos AR, Taleb M, Calixto JB, Mira JC, Ghizoni MF. Long interpositional nerve graft consistently induces incomplete motor and sensory recovery in the rat. An experimental model to test nerve repair. J Neurosci Methods. 2004;134(1):75-80.-1414 Grossman SD, Rosenberg LJ, Wrathall JR. Relationship of altered glutamate receptor subunit mRNA expression to acute cell loss after spinal cord contusion. Exp Neurol. 2001;168(2):283-289. to calculate an axonal Regeneration Index (RI):
For statistical analysis, the nonparametric Kruskal-Wallis test was used to compare more than two groups, the Mann-Whitney test was used in the paired comparison between two groups, and one-way analysis of variance (ANOVA) was used for the analysis of BBB, cell count, and histological analysis. To analyze the follow-up of each group, the ANOVA test for repeated measures was used. The type I error with p ≤ 0.05 was accepted.
Results
In the BBB evaluation, from day 7 after spinal cord injury, a statistically significant difference was observed in locomotor function in all four groups (p < 0.001) and the comparison of improvement of motor function (p < 0.001). In addition, statistically significant differences were observed from day 14 (p ≤ 0.05) among the groups. In the weekly evolution among the groups, no statistical difference was found in the first week among Group 1 (laminectomy with spinal cord injury), Group 2 (laminectomy with spinal cord injury and SS), and Group 3 (laminectomy with spinal cord injury and GSH). However, there was a statistical difference between the SS and GSH groups in the second week, which was not maintained in the third and fourth weeks. From the fifth week, there was a statistically significant difference between the GSH and SS groups, with better results in the GSH group.
The horizontal ladder evaluation showed a statistical difference between the SS and GSH groups from the second week, which did not remain in the third and fourth weeks. From the fifth week, there was a statistically significant difference between the GSH and SS groups, with better results in the GSH group.
Tissue evaluation of the spinal cord injury area (B-area) showed degeneration, hemorrhage, hyperemia, cellular infiltrate, and necrosis. The variables were described as absent, discrete, moderate, and intense. Group 4 without spinal cord injury did not present areas of degeneration, hemorrhage, hyperemia, cellular infiltrate, or necrosis, as expected. No statistically significant difference among the groups submitted to spinal cord injury (p > 0.05).
The mean axonal RI demonstrated a statistical difference among the groups added together. Group 3 presented a higher mean RI than groups 1 and 2 and lower than Group 4. The group submitted to intervention with GSH showed statistically significant improvement concerning the other groups submitted to spinal cord injury favorable to intervention with GSH (p < 0.05), and, as expected, statistically significant difference to the group submitted to laminectomy without spinal cord injury in favor of the latter.
Discussion
There are no medications approved that are capable of reversing neural tissue damage.1515 Yamamoto N, Yamamoto S, Inagaki F, Kawaichi M, Fukamizu A, Kishi N, et al. Role of Deltex-1 as a transcriptional regulator downstream of the Notch receptor. J Biol Chem. 2001;276(48):45031-40. Research has sought different strategies to improve functional recovery after spinal cord trauma, focusing on reducing secondary damage and stimulating tissue regeneration. Physical means are used, such as hypothermia, oxygen therapy and exercises, substances that promote the improvement of the inflammatory environment and control of free radicals, and therapies with different cell types to promote neuronal recovery, using other animal models.1616 Bresnahan JC, Beattie MS, Todd FD 3rd, Noyes DH. A behavioral and anatomical analysis of spinal cord injury produced by a feedback-controlled impaction device. Exp Neurol. 1987;95(3):548-70.
17 Noble LJ, Wrathall JR. Correlative analyses of lesion development and functional status after graded spinal cord contusive injuries in the rat. Exp Neurol. 1989;103(1):34-40.
18 Marcon RM, Barros Filho TEP, Oliveira RP, Cristante AF, Taricco MA, Colares G, et al. Experimental study on the action of methylprednisolone on wistar rats before spinal cord injury. Acta Ortop Bras. 2010;18(1):26-30.
19 Souza FI, Cristante AF, Marcon RM, Ferreira R, Dos Santos GB, de Barros Filho TEP, et al. Transdermal monosialoganglioside with laser in the treatment of spinal cord lesion in rats. Acta Ortop Bras. 2013;21(2):87-91.
20 Cristante AF, Barros Filho TE, Marcon RM, Letaif OB, Rocha ID. Therapeutic approaches for spinal cord injury. Clinics. 2012;67(10):1219-1224.
21 Cristante AF, Damasceno ML, Barros Filho TE, de Oliveira RP, Marcon RM, da Rocha ID. Evaluation of the effects of hyperbaric oxygen therapy for spinal cord lesion in correlation with the moment of intervention. Spinal Cord. 2012;50(7):502-6.
22 Marcon RM, Cristante AF, de Barros Filho TE, de Oliveira RP, dos Santos GB. Potentializing the effects of GM1 by hyperbaric oxygen therapy in acute experimental spinal cord lesion in rats. Spinal Cord. 2010;48(11):808-13.
23 Guimarães PE, Fridman C, Gregório SP, Kalil EM, Cristante AF, Teixeira WGJ, et al. DNA polymorphisms as tools for spinal cord injury research. Spinal Cord. 2009;47(2):171-5.
24 Guimarães PE, Fridman C, Gregório SP, Kalil EM, de Barros Filho TEP, Cristante AF, et al. WITHDRAWN: Association between polymorphisms in genes of inflammatory response and axonal repair with spinal cord injury. Spinal Cord. 2007. doi: 10.1038/sj.sc.3102122. Online ahead of print.
https://doi.org/10.1038/sj.sc.3102122....
-2525 Narazaki DK, Barros Filho TE, Oliveira CR, Cristante AF, Iutaka AS, Marcon RM, et al. Spinal cord regeneration: the action of neurotrophin-3 in spinal cord injury in rats. Clinics. 2006;61(5):453-60.
Our study used the standardized model to investigate the effects of GSH, an antioxidant, in the experimental spinal cord injury.2626 Woodward JS, Freeman LW. Ischemia of the spinal cord; an experimental study. J Neurosurg. 1956;13(1):63-72.
27 Ducker TB, Kindt GW, Kempf LG. Pathological findings in acute experimental spinal cord trauma. J Neurosurg. 1971;35(6):700-8.
28 Kim S-J, Ko W-K, Han G-H, Lee D, Lee Y, Sheen S-H, et al. Chirality-dependent anti-inflammatory effect of glutathione after spinal cord injury in an animal model Pharmaceuticals2021;14(8):792.
29 Stewart AN, Glaser EP, Mott CA, Bailey WM, Sullivan PG, Patel SP, et al. Advanced age and neurotrauma diminish glutathione and impair antioxidant defense after spinal cord injury. J Neurotrauma. 2022;39(15-16):1075-89.-3030 Wyss PO, Richter JK, Zweers P, Brust AK, Funk C, Zoelch N, et al. Glutathione in the pons is associated with clinical status improvements in subacute spinal cord injury. Invest Radiol. 2023;58(2):131-8. GSH plays an important role in controlling reactive oxygen species that arise after spinal cord injury and are responsible for the propagation of tissue injury. Different studies demonstrate the GSH in its reduced form when there is injury by oxygen free radicals in different cell types, being even more important in neuronal cells. This substance is inserted in the secondary injury cycle to stop the propagation of cell damage, thus having a neuroprotective effect. Some studies demonstrate this effect in several cell types.2828 Kim S-J, Ko W-K, Han G-H, Lee D, Lee Y, Sheen S-H, et al. Chirality-dependent anti-inflammatory effect of glutathione after spinal cord injury in an animal model Pharmaceuticals2021;14(8):792.
29 Stewart AN, Glaser EP, Mott CA, Bailey WM, Sullivan PG, Patel SP, et al. Advanced age and neurotrauma diminish glutathione and impair antioxidant defense after spinal cord injury. J Neurotrauma. 2022;39(15-16):1075-89.-3030 Wyss PO, Richter JK, Zweers P, Brust AK, Funk C, Zoelch N, et al. Glutathione in the pons is associated with clinical status improvements in subacute spinal cord injury. Invest Radiol. 2023;58(2):131-8.
The difference in the results of the several functional scores evaluated, presenting a statistically significant improvement in the BBB scale and improvement without a statistically significant difference in the horizontal ladder test, may be due to the difference in the scores, with the BBB scale being more detailed to its evaluation. However, both tests are validated in the literature for motor evaluation in experimental studies with rats.
The histological analysis regarding the degeneration, hemorrhage, hyperemia, cellular infiltrate, and necrosis was performed quantitatively to enable statistical analysis. However, this generates some degree of subjectivity for the scores evaluated for each variable. In this analysis, there was no statistically significant difference. On the other hand, the evaluation of neuronal count through the axonal RI was higher in the group submitted to intervention with GSH compared to the other groups submitted to spinal cord injury.
The secondary cellular events that follow the initial traumatic mechanical injury contribute to the propagation of tissue damage associated with cell damage caused by the release of oxygen free radicals and the reduction of mechanisms and substances responsible for maintaining cellular homeostasis, highlighting the role of GSH in its reduced form.2929 Stewart AN, Glaser EP, Mott CA, Bailey WM, Sullivan PG, Patel SP, et al. Advanced age and neurotrauma diminish glutathione and impair antioxidant defense after spinal cord injury. J Neurotrauma. 2022;39(15-16):1075-89.,3030 Wyss PO, Richter JK, Zweers P, Brust AK, Funk C, Zoelch N, et al. Glutathione in the pons is associated with clinical status improvements in subacute spinal cord injury. Invest Radiol. 2023;58(2):131-8. Furthermore, associated with oxidative stress and inflammatory reaction, the harmful effect of vascular and ischemic disorders and homeostasis in this propagation of secondary damage is added, which can also be addressed with different therapeutic strategies, making the approach to spinal cord trauma increasingly multifactorial.3131 Hall ED, Wolf DL, Braughler JM. Effects of a single large dose of methylprednisolone sodium succinate on experimental posttraumatic spinal cord ischemia: dose-response and time-action analysis. J Neurosurg. 1984;61(1):124-30.
32 Hall ED. The neuroprotective pharmacology of methylprednisolone. J Neurosurg. 1992;76(1):13-22.-3333 Bracken MB, Shepard MJ, Hellenbrand KG, Collins WF, Leo LS, Freeman DF, et al. Methylprednisolone and neurological function 1 year after spinal cord injury: results of the National Acute Spinal Cord Injury Study. J Neurosurg. 1985;63(5):704-13.
Conclusion
This study demonstrated that using glutathione in the experimental spinal cord trauma scenario leads to better statistically significant functional recovery through the BBB score and improvement of the axonal regeneration index in Wistar rats submitted to experimental spinal cord injury.
Data availability
The authors confirm that the data supporting the findings of this study are available within the article. Furthermore, the data sets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.
Ethics approval
This study was approved by the Ethics internal review boards of the Biosciences Institute under the number 956/2018.
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FundingNo funding was received for this project.
References
-
1Ducker TB, Salcman M, Perot PL Jr, Ballantine D. Experimental spinal cord trauma, I: Correlation of blood flow, tissue oxygen and neurologic status in the dog. Surg Neurol. 1978;10(1):60-3.
-
2Ray SK, Matzelle DD, Wilford GG, Hogan EL, Banik NL. Cell death in spinal cord injury (SCI) requires de novo protein synthesis. Calpain inhibitor E-64-d provides neuroprotection in SCI lesion and penumbra. Ann N Y Acad Sci. 2001;939:436-49.
-
3Beattie MS, Farooqui AA, Bresnahan JC. Review of current evidence for apoptosis after spinal cord injury. J Neurotrauma. 2000;17(10):915-25.
-
4Ding WX, Shen HM, Ong CN. Critical role of reactive oxygen species and mitochondrial permeability transition in microcystin-induced rapid apoptosis in rat hepatocytes. Hepatology. 2000;32(3):547-55.
-
5Barros Filho TEP, Taricco MA, Oliveira RP, Greve JMA, Santos LCR, Napoli MMM. Estudo epidemiológico dos pacientes com traumatismo da coluna vertebral e déficit neurológico, internados no Instituto de Ortopedia e Traumatologia do Hospital das Clínicas da Faculdade de Medicina da USP. Rev Hosp Clin Fac Med Sao Paulo. 1990;45(3):123-126.
-
6Bracken MB, Shepard MJ, Holford TR, Leo-Summers L, Aldrich EF, Fazl M, et al. Methylprednisolone or tirilazad mesylate administration after acute spinal cord injury: 1-year follow up. Results of the third National Acute Spinal Cord Injury randomized controlled trial. J Neurosurg. 1998;89(5):699-706.
-
7Foret A, Quertainmont R, Botman O, Bouhy D, Amabili P, Brook G, et al. Stem cells in the adult rat spinal cord: plasticity after injury and treadmill training exercise. J Neurochem. 2010;112(3):762-72.
-
8Logan MP, Parker S, Shi R. Glutathione and ascorbic acid enhance recovery of Guinea pig spinal cord white matter following ischemia and acrolein exposure. Pathobiology. 2005;72(4):171-8.
-
9Rodrigues NR. Padronização da lesão de medula espinal em ratos Wistar [tese]. São Paulo: Faculdade de Medicina da Universidade de São Paulo; 1999.
-
10Santos GB, Cristante AF, Marcon RM, Souza FI, Barros Filho TEP, Damasceno ML. Spinal cord injury experimental model and motion evaluation protocol in wistar rats. Acta Ortop Bras. 2011;19(2):87-91.
-
11Letaif OB. Avaliação do efeito do estrógeno na lesão medular experimental em ratos [dissertação]. São Paulo: Faculdade de Medicina da Universidade de São Paulo; 2014.
-
12Grossman SD, Rosenberg LJ, Wrathall JR. Temporal-spatial pattern of acute neuronal and glial loss after spinal cord contusion. Exp Neurol. 2001;168(2):273-82.
-
13Bertelli JA, dos Santos AR, Taleb M, Calixto JB, Mira JC, Ghizoni MF. Long interpositional nerve graft consistently induces incomplete motor and sensory recovery in the rat. An experimental model to test nerve repair. J Neurosci Methods. 2004;134(1):75-80.
-
14Grossman SD, Rosenberg LJ, Wrathall JR. Relationship of altered glutamate receptor subunit mRNA expression to acute cell loss after spinal cord contusion. Exp Neurol. 2001;168(2):283-289.
-
15Yamamoto N, Yamamoto S, Inagaki F, Kawaichi M, Fukamizu A, Kishi N, et al. Role of Deltex-1 as a transcriptional regulator downstream of the Notch receptor. J Biol Chem. 2001;276(48):45031-40.
-
16Bresnahan JC, Beattie MS, Todd FD 3rd, Noyes DH. A behavioral and anatomical analysis of spinal cord injury produced by a feedback-controlled impaction device. Exp Neurol. 1987;95(3):548-70.
-
17Noble LJ, Wrathall JR. Correlative analyses of lesion development and functional status after graded spinal cord contusive injuries in the rat. Exp Neurol. 1989;103(1):34-40.
-
18Marcon RM, Barros Filho TEP, Oliveira RP, Cristante AF, Taricco MA, Colares G, et al. Experimental study on the action of methylprednisolone on wistar rats before spinal cord injury. Acta Ortop Bras. 2010;18(1):26-30.
-
19Souza FI, Cristante AF, Marcon RM, Ferreira R, Dos Santos GB, de Barros Filho TEP, et al. Transdermal monosialoganglioside with laser in the treatment of spinal cord lesion in rats. Acta Ortop Bras. 2013;21(2):87-91.
-
20Cristante AF, Barros Filho TE, Marcon RM, Letaif OB, Rocha ID. Therapeutic approaches for spinal cord injury. Clinics. 2012;67(10):1219-1224.
-
21Cristante AF, Damasceno ML, Barros Filho TE, de Oliveira RP, Marcon RM, da Rocha ID. Evaluation of the effects of hyperbaric oxygen therapy for spinal cord lesion in correlation with the moment of intervention. Spinal Cord. 2012;50(7):502-6.
-
22Marcon RM, Cristante AF, de Barros Filho TE, de Oliveira RP, dos Santos GB. Potentializing the effects of GM1 by hyperbaric oxygen therapy in acute experimental spinal cord lesion in rats. Spinal Cord. 2010;48(11):808-13.
-
23Guimarães PE, Fridman C, Gregório SP, Kalil EM, Cristante AF, Teixeira WGJ, et al. DNA polymorphisms as tools for spinal cord injury research. Spinal Cord. 2009;47(2):171-5.
-
24Guimarães PE, Fridman C, Gregório SP, Kalil EM, de Barros Filho TEP, Cristante AF, et al. WITHDRAWN: Association between polymorphisms in genes of inflammatory response and axonal repair with spinal cord injury. Spinal Cord. 2007. doi: 10.1038/sj.sc.3102122. Online ahead of print.
» https://doi.org/10.1038/sj.sc.3102122. -
25Narazaki DK, Barros Filho TE, Oliveira CR, Cristante AF, Iutaka AS, Marcon RM, et al. Spinal cord regeneration: the action of neurotrophin-3 in spinal cord injury in rats. Clinics. 2006;61(5):453-60.
-
26Woodward JS, Freeman LW. Ischemia of the spinal cord; an experimental study. J Neurosurg. 1956;13(1):63-72.
-
27Ducker TB, Kindt GW, Kempf LG. Pathological findings in acute experimental spinal cord trauma. J Neurosurg. 1971;35(6):700-8.
-
28Kim S-J, Ko W-K, Han G-H, Lee D, Lee Y, Sheen S-H, et al. Chirality-dependent anti-inflammatory effect of glutathione after spinal cord injury in an animal model Pharmaceuticals2021;14(8):792.
-
29Stewart AN, Glaser EP, Mott CA, Bailey WM, Sullivan PG, Patel SP, et al. Advanced age and neurotrauma diminish glutathione and impair antioxidant defense after spinal cord injury. J Neurotrauma. 2022;39(15-16):1075-89.
-
30Wyss PO, Richter JK, Zweers P, Brust AK, Funk C, Zoelch N, et al. Glutathione in the pons is associated with clinical status improvements in subacute spinal cord injury. Invest Radiol. 2023;58(2):131-8.
-
31Hall ED, Wolf DL, Braughler JM. Effects of a single large dose of methylprednisolone sodium succinate on experimental posttraumatic spinal cord ischemia: dose-response and time-action analysis. J Neurosurg. 1984;61(1):124-30.
-
32Hall ED. The neuroprotective pharmacology of methylprednisolone. J Neurosurg. 1992;76(1):13-22.
-
33Bracken MB, Shepard MJ, Hellenbrand KG, Collins WF, Leo LS, Freeman DF, et al. Methylprednisolone and neurological function 1 year after spinal cord injury: results of the National Acute Spinal Cord Injury Study. J Neurosurg. 1985;63(5):704-13.
Publication Dates
-
Publication in this collection
24 June 2024 -
Date of issue
2024
History
-
Received
6 July 2023 -
Accepted
29 Mar 2024