RESUMEN
The role of oxidative stress in ligamentum flavum (LF) hypertrophy has not been elucidated. We hypothesize that oxidative stress induces inflammatory responses and the subsequent fibrotic processes in LF, via activation of the Akt and MAPK pathways. Specimens of LFs were collected during surgeries for lumbar disc herniation (LDH) or lumbar spinal stenosis (LSS). Part of the LF specimens underwent analyses for ROS, fibrotic markers, and inflammatory mediators, with the remainder minced for cell cultures. The cell cultures were treated with H2O2, after which the cells were lysed and analyzed via western blotting. The specimens of the LSS patients showed increased infiltration of inflammatory cells and were stained positively for MMP-3, MMP-9, vimentin, and fibronectin. The LF of the LSS patients had increased oxidative stress and inflammation compared to that of the LDH patients. In vitro analyses demonstrated that oxidative stress rapidly activated the Akt and MAPK pathways. Inflammatory mediators, iNOS and NF-κB, and fibrotic markers, including TGF-ß, ß-catenin, α-SMA and vimentin, were significantly upregulated after induction of oxidative stress. Oxidative stress activated the intrinsic apoptotic pathway. These findings revealed that oxidative stress is one of the etiological factors of LF hypertrophy, which might provide new insights into treatment approaches.
Asunto(s)
Apoptosis , Mediadores de Inflamación/metabolismo , Desplazamiento del Disco Intervertebral/enzimología , Ligamento Amarillo/enzimología , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Estrés Oxidativo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Estenosis Espinal/enzimología , Adulto , Factores de Edad , Anciano , Apoptosis/efectos de los fármacos , Proteínas Reguladoras de la Apoptosis/metabolismo , Células Cultivadas , Femenino , Fibrosis , Humanos , Peróxido de Hidrógeno/toxicidad , Hipertrofia , Desplazamiento del Disco Intervertebral/patología , Ligamento Amarillo/efectos de los fármacos , Ligamento Amarillo/patología , Masculino , Persona de Mediana Edad , Estrés Oxidativo/efectos de los fármacos , Transducción de Señal , Estenosis Espinal/patologíaRESUMEN
BACKGROUND: This is an immunohistologic study of gene expression between patients and controls.This study aims to evaluate expression of the catalase gene in hypertrophied ligamentum flavum (LF) specimens obtained from patients with lumbar spinal canal stenosis (LSCS).LSCS is one of the most common spinal disorders. It is well known that LF hypertrophy plays an important role in the onset of LSCS. Although degenerative changes, aging, and mechanical stress are all thought to contribute to hypertrophy and fibrosis of the LF, the precise pathogenesis of LF hypertrophy remains unknown. Previous genetic studies have tried to determine the mechanism of LF hypertrophy. However, the association between catalase gene expression and LF hypertrophy has not yet been explored. METHODS: LF specimens were surgically obtained from 30 patients with spinal stenosis (LSCS group) and from 30 controls with lumbar disc herniation (LDH group). LF thickness was measured at the thickest point using calipers to an accuracy of 0.01âmm during surgical intervention. The extent of LF elastin degradation and fibrosis were graded (grades 0-4) by hematoxylin and eosin staining and Masson trichrome staining, respectively. The resulting LF measurements, histologic data, and immunohistologic results were then compared between the 2 groups. RESULTS: The average LF thickness was significantly higher in the LSCS group than in the LDH group (5.99 and 2.95âmm, respectively, Pâ=â.004). Elastin degradation and fibrosis of the LF were significantly more severe in spinal stenosis samples than in the disc herniation samples (3.04â±â0.50 vs 0.79â±â0.60, Pâ=â.007; 3.01â±â0.47 vs 0.66â±â0.42, Pâ=â.009, respectively). Significantly lower expression of catalase was observed in the perivascular area of LF samples obtained from patients with LSCS compared with controls (61.80â±â31.10 vs 152.80â±â41.13, respectively, Pâ=â.009). CONCLUSION: Our findings suggest that decreased expression of catalase is associated with LF hypertrophy in patients with LSCS.
Asunto(s)
Catalasa/metabolismo , Ligamento Amarillo/enzimología , Ligamento Amarillo/patología , Estenosis Espinal/enzimología , Estenosis Espinal/patología , Adulto , Anciano , Elastina/metabolismo , Femenino , Fibrosis/enzimología , Fibrosis/patología , Expresión Génica , Humanos , Hipertrofia/enzimología , Hipertrofia/patología , Desplazamiento del Disco Intervertebral/enzimología , Desplazamiento del Disco Intervertebral/patología , Desplazamiento del Disco Intervertebral/cirugía , Ligamento Amarillo/cirugía , Vértebras Lumbares , Masculino , Persona de Mediana Edad , Tamaño de los Órganos , Estudios Retrospectivos , Estenosis Espinal/cirugíaRESUMEN
STUDY DESIGN: Immunohistological study. OBJECTIVE: To elucidate the role of matrix metalloproteinases (MMPs), hypoxia-inducible factor-1α (HIF), and vascular endothelial growth factor (VEGF) in the hypertrophied ligamentum flavum (LF) obtained from patients with lumbar spinal stenosis (LSS). SUMMARY OF BACKGROUND DATA: The most common spinal disorder in the elderly is LSS, which results in part from LF hypertrophy. Although prior histologic and immunochemical studies have been performed in this area, the pathophysiology of loss of elasticity and hypertrophy is not completely understood. METHODS: LF samples of 38 patients with LSS were harvested during spinal decompression. Twelve LF samples obtained from patients with disk herniation and no visible degeneration on preoperative magnetic resonance imaging were obtained as controls. Samples were dehydrated and paraffin embedded. For immunohistochemical determination of VEGF, HIF, and MMPs 1, 3, and 9 expression, slices were stained with VEGF, HIF, and MMP antibody dilution. Neovessel density and number of elastic fibers were counted after Masson-Goldner staining. LF hypertrophy and cross-sectional area (CSA) were measured on T1-weighted magnetic resonance imaging. RESULTS: MMPs 1, 3, 9 and VEGF expression were significantly increased in the hypertrophy group (P<0.05). HIF expression was negative in both groups. Vessel density was increased in the hypertrophy group, although this was not statistically significant. The number of elastic fibres was significantly higher in the control group. In the hypertrophy group, LF thickness was significantly increased, whereas CSA was significantly decreased. There was a statistical correlation between LF thickness, CSA, MMP, and VEGF expression in the hypertrophy group (P<0.05). CONCLUSIONS: LF hypertrophy is accompanied by increased MMPs 1, 3, 9 and VEGF expression. Neovessel density is increased in hypertrophied LF. HIF is not expressed in hypertrophied LF.
Asunto(s)
Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Ligamento Amarillo/enzimología , Ligamento Amarillo/patología , Metaloproteinasas de la Matriz/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Demografía , Femenino , Humanos , Hipertrofia/patología , Desplazamiento del Disco Intervertebral , Ligamento Amarillo/diagnóstico por imagen , Vértebras Lumbares/enzimología , Vértebras Lumbares/patología , Imagen por Resonancia Magnética , Masculino , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 3 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Persona de Mediana Edad , Radiografía , Estenosis Espinal/enzimología , Estenosis Espinal/patología , Adulto JovenRESUMEN
STUDY DESIGN: Human ligamentum flavum (LF) was examined for the activity level of matrix metalloproteinase-3 (MMP-3) in degenerative spondylolithesis (DS) patients using immunohistochemistry, Western blot, reverse transcriptase-polymerase chain reaction (RT-PCR), and quantitative real-time PCR. OBJECTIVE: To investigate the hypothesis that the activity of MMP-3 is elevated in LF of DS patients, which might contribute to DS pathogenesis. SUMMARY OF BACKGROUND DATA: MMP-3 is a proteinase produced by connective tissue cells and is responsible for the degradation and modification of extracellular matrix molecules. MMP-3 activity has been established in articular cartilage, synovial membrane, and intervertebral discs, but not in the LF. METHODS: The experimental group consisted of 18 patients with DS and the control group consisted of 18 patients with spinal stenosis (SS) without any instabilities. MMP-3 expression was measured with in situ using immunohistochemistry and both for mRNA and protein levels. RESULTS: The MMP-3 positive cell ratio in the LF observed in DS patients was substantially higher than in SS patients (P = 0.030). In Western blot, the average optical density (OD) of MMP-3 was higher in LF of DS than of SS (P = 0.028). There was greater MMP-3 expression in DS patients as quantified by RT-PCR (P = 0.004). CONCLUSION: Our study shows that MMP-3 expression in the LF of DS patients was significantly higher than in SS patients. Increased MMP-3 expression may be associated with the degenerative changes of LF in DS patients comprising one of the mechanisms of pathogenesis in DS.
Asunto(s)
Ligamento Amarillo/enzimología , Metaloproteinasa 3 de la Matriz/metabolismo , Estenosis Espinal/enzimología , Espondilolistesis/enzimología , Anciano , Western Blotting , Femenino , Humanos , Inmunohistoquímica , Ligamento Amarillo/metabolismo , Vértebras Lumbares/enzimología , Vértebras Lumbares/metabolismo , Vértebras Lumbares/patología , Metaloproteinasa 3 de la Matriz/genética , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Estenosis Espinal/genética , Espondilolistesis/genéticaRESUMEN
BACKGROUND: To assess the effect of 3 different surgical approaches on paraspinal muscle atrophy in patients undergoing lumbar back surgery, we compared their pre- and postoperative CT scans and their serum Hb, CRP, and CPK levels. METHODS: The study population consisted of 71 patients who had undergone lumbar back surgery with microscopic posterior decompression without fusion. We examined the effect on paraspinal muscle atrophy of 3 different approaches to the spinal canal. Group 1 (n = 19) underwent unilateral paraspinal dissection from the spinous process with cutting of the spinous process. In group 2 (n = 24), we used modified bilateral decompression via hemilaminectomy, and group 3 (n = 28) was treated by modified bilateral decompression via spinous process splitting. We measured the levels of CPK, Hb, and CRP preoperatively and on the first postoperative day, and compared the preoperative volume of the paraspinal muscle with the volume measured 1 year after the operation. RESULTS: Age, sex, operative time, and CRP and Hb levels were not statistically different among the 3 groups. The postoperative elevation of CPK was significantly lower in groups 2 and 3 than in group 1. Group 3 manifested a significantly lower degree of atrophic changes of the paraspinal muscle than groups 1 and 2. CONCLUSIONS: We found that among the 3 approaches evaluated, modified bilateral decompression via spinous process splitting is less invasive, facilitates preservation of the paraspinal muscle, and is a useful approach to posterior spinal elements resulting in decreased muscle damage.
Asunto(s)
Descompresión Quirúrgica , Disección/métodos , Atrofia Muscular Espinal/prevención & control , Canal Medular , Estenosis Espinal/cirugía , Adulto , Anciano , Anciano de 80 o más Años , Creatina Quinasa/sangre , Disección/efectos adversos , Femenino , Humanos , Laminectomía , Vértebras Lumbares , Masculino , Persona de Mediana Edad , Atrofia Muscular Espinal/etiología , Estudios Retrospectivos , Estenosis Espinal/enzimologíaRESUMEN
STUDY DESIGN: Immunohistochemical and behavioral study using a rat cauda equina compression model. OBJECTIVE: To investigate, after cauda equina compression by spinal canal stenosis (SCS), Rho activation in the spinal cord and cauda equina, and the effect of intrathecal administration of a Rho kinase inhibitor on hypoalgesia and motor dysfunction. SUMMARY OF BACKGROUND DATA: Compression of the cauda equina caused by SCS is a common clinical disorder associated with sensory disturbance and intermittent claudication. Cauda equina compression is thought to reduce blood flow and result in nerve degeneration caused by various cytokines. Rho, a member of the small GTPases, is a signal transmitter. It promotes Wallerian degeneration, decreases blood flow in the spinal cord and brain, and increases expression of several cytokines. Currently, Rho kinase inhibitor is used clinically to treat progressive nerve damage due to cerebrovascular disorders. However, its effect for SCS has not been evaluated. METHODS: Forty-two 6-week-old male Sprague-Dawley rats (200-250 g) were used. For the SCS model (n = 27), a small piece of silicon was placed under the lamina of the fourth lumbar vertebra. In the sham-operated group, laminectomies were performed at L5 only (n = 15). We examined mechanical sensitivity and motor function using von Frey hairs and a treadmill, and immunohistochemically localized Rho in the spinal ventral neurons, axons, and Schwann cells in the cauda equina. We also examined the effects of intrathecally administered Rho kinase inhibitor for hypoalgesia or motor dysfunction caused by SCS. RESULTS: We observed motor dysfunction and hypoalgesia and activated Rho-immunoreactive cells in spinal ventral neuroreported to induce neurite and axonal outgrowth in the spinal cord and brain after nervous system injury. In addition, 1 report showed that Rho kinase was involved in Wallerian degeneration that was rescued by Rho kinase inhibitor. Furthermore, it is thought that Rho is involved in TNF-alpha and interleukin (IL) production in the central nervous system, and the production was inhibited by administering Rho kinase inhibitor in the central nervous system. Regardns, axons, and Schwann cells in the cauda equina. Intrathecal administration of Rho kinase inhibitor improved mechanical hypoalgesia and motor dysfunction caused by SCS. CONCLUSION: Activated Rho may play an important role in nerve damage in the cauda equina in SCS. Rho kinase inhibitor may be a useful tool in determining the pathomechanism of cauda equina syndrome caused by SCS.