RESUMEN
SUMMARY: Intervertebral disc degeneration (IVDD) is induced by nucleus pulposus (NP) dysfunction as a result of massive loss of NP cells. It has been reported that the acidic microenvironment of the intervertebral disc (IVD) can induce NP cell pyroptosis, and that up-regulation of periostin (POSTN) expression has a negative effect on NP cell survival. However, the relationship between the acidic environment, POSTN expression level and NP cell pyroptosis is unclear. Therefore, the aim of this study was to explore the relationship between acidic environment and POSTN expression level in NP cells, as well as the effect of POSTN in acidic environment on NP cell pyroptosis. NP cells were obtained from the lumbar vertebrae of Sprague Dawley (SD) male rats. These cells were divided into normal and acidic groups according to whether they were exposed to 6 mM lactic acid solution. And NP cells in the acidic group were additionally divided into three groups: (1) Blank group: no transfection; (2) NC group: cells transfected with empty vector plasmid; (3) sh-POSTN group: cells transfected with sh-POSTN plasmid to knock down the expression level of POSTN. Quantitative real-time PCR (qRT-PCR) and western blot was performed to assess the expression of POSTN at the mRNAand protein levels. CCK8 was used to evaluate cell survival. Western blot, in addition, was performed to examine acid-sensing ion channels (ASIC)-related proteins. And pyroptosis was detected by ELISA and western blot. The expression level of POSTN was significantly increased in NP cells in acidic environment. Knockdown of POSTN expression promoted the survival of NP cells in acidic environment and reduced the protein levels of ASIC3 and ASIC1a in NP cells. Moreover, knockdown of POSTN expression decreased the pyroptosis proportion of NP cells and the levels of pro-inflammatory cytokines interleukin (IL)-1β and IL-18. The levels of pyroptosis-related proteins NLRP3, ASC, cleaved-Caspase-1, and cleaved-GSDMD were also affected by the decreased POSTN expression. The extracellular acidic environment created by lactic acid solution activated NLRP3 inflammatory vesicle-induced caspase-1 to get involved in NP cell pyroptosis by up-regulating POSTN expression.
La degeneración del disco intervertebral (DDIV) es inducida por una disfunción del núcleo pulposo (NP) como resultado de una pérdida masiva de células NP. Se ha informado que el microambiente ácido del disco intervertebral (DIV) puede inducir la piroptosis de las células NP y que la regulación positiva de la expresión de periostina (POSTN) tiene un efecto negativo en la supervivencia de las células NP. Sin embargo, la relación entre el ambiente ácido, el nivel de expresión de POSTN y la piroptosis de las células NP es poco clara. Por lo tanto, el objetivo de este estudio fue explorar la relación entre el ambiente ácido y el nivel de expresión de POSTN en células NP, así como el efecto de POSTN en ambiente ácido sobre la piroptosis de las células NP. Las células NP se obtuvieron de las vertebras lumbares de ratas macho Sprague Dawley (SD). Estas células se dividieron en grupos normales y ácidos según se expusieron a una solución de ácido láctico 6 mM. Las células NP en el grupo ácido se dividieron adicionalmente en tres grupos: (1) Grupo en blanco: sin transfección; (2) grupo NC: células transfectadas con plásmido vector vacío; (3) grupo sh-POSTN: células transfectadas con plásmido sh-POSTN para reducir el nivel de expresión de POSTN. Se realizó una PCR cuantitativa en tiempo real (qRT-PCR) y una transferencia Western para evaluar la expresión de POSTN en los niveles de ARNm y proteína. Se utilizó CCK8 para evaluar la supervivencia celular. Además, se realizó una transferencia Western para examinar las proteínas relacionadas con los canales iónicos sensibles al ácido (ASIC). La piroptosis se detectó mediante ELISA y Western blot. El nivel de expresión de POSTN aumentó significativamente en células NP en ambiente ácido. La eliminación de la expresión de POSTN promovió la supervivencia de las células NP en un ambiente ácido y redujo los niveles de proteína de ASIC3 y ASIC1a en las células NP. Además, la eliminación de la expresión de POSTN disminuyó la proporción de piroptosis de las células NP y los niveles de citocinas proinflamatorias interleucina (IL) - 1β e IL-18. Los niveles de proteínas relacionadas con la piroptosis NLRP3, ASC, Caspasa-1 escindida y GSDMD escindida también se vieron afectados por la disminución de la expresión de POSTN. El ambiente ácido extracelular creado por la solución de ácido láctico activó la caspasa-1 inducida por vesículas inflamatorias NLRP3 para involucrarse en la piroptosis de las células NP mediante la regulación positiva de la expresión de POSTN.
Asunto(s)
Animales , Masculino , Ratas , Ácidos/química , Moléculas de Adhesión Celular/metabolismo , Degeneración del Disco Intervertebral , Núcleo Pulposo/fisiopatología , Ensayo de Inmunoadsorción Enzimática , Moléculas de Adhesión Celular/genética , Supervivencia Celular , Western Blotting , Ratas Sprague-Dawley , Ambiente , Reacción en Cadena en Tiempo Real de la Polimerasa , Núcleo Pulposo/citología , Proteína con Dominio Pirina 3 de la Familia NLRRESUMEN
Fibrocartilaginous embolism (FCEM) and acute, non-compressive nucleus pulposus extrusion (ANNPE) are non-compressive myelopathies that are difficult to differentiate. The definitive diagnosis is obtained only with histology, but the presumptive diagnosis is made through clinical signs and imaging tests. The aim of this study is to report the imaging tests performed for the diagnosis of a neurological clinical case and discuss the best diagnostic method. After attending the patient, complementary tests were requested. Radiography results showed no change. The computed tomography diagnostic impression indicated distal protrusion between C6-C7, T11-T12, T13-L1 followed by mild spinal cord compression defined by the presence of a ventral hyperattenuating region. Magnetic resonance (RMI), showed a slight T2W hypersignal, well delimited in the gray matter, lateralized to the right, over the cranial third of C7. Concluding that the magnetic resonance is the method that brought more information for the diagnosis, in which the others were not described medullary alterations pertinent to FCEM and ANNPE. With their fair prognosis, the absence of histological diagnosis of these diseases may be a limiting factor in this study and, in relation to the RMI alterations being very similar between FCEM and ANNPE it is not possible to diagnose fully accurately.
A embolia fibrocartilaginosa (EFC) e a extrusão aguda não compressiva do núcleo pulposo (EANCNP) são mielopatias não compressivas de difícil diferenciação. O diagnóstico definitivo é obtido apenas com a histologia, mas o diagnóstico presuntivo é feito por meio de sinais clínicos e exames de imagem. O objetivo deste trabalho é relatar os exames de imagem realizados para o diagnóstico de um caso clínico neurológico e discutir o melhor método diagnóstico. Após o atendimento do paciente, foram solicitados exames complementares. Os resultados da radiografia não mostraram nenhuma alteração. A impressão diagnóstica da tomografia computadorizada indicou protrusão distal entre C6-C7, T11-T12, T13-L1, seguida de leve compressão medular definida pela presença de região hiperatenuante ventral. À ressonância magnética (RM), apresentava discreto hipersinal em T2W, bem delimitado na substância cinzenta, lateralizado à direita, sobre o terço cranial de C7. Concluiu-se que a ressonância magnética é o método que mais trouxe informações para o diagnóstico, os demais métodos não foram descritos alterações medulares pertinentes à EFC e à EANCNP. Com seu prognóstico favorável, a ausência de diagnóstico histológico dessas doenças pode ser um fator limitante neste estudo. Em relação às alterações do RM serem muito semelhantes entre EFC e EANCNP, não é possível diagnosticar com total precisão.
Asunto(s)
Animales , Perros , Enfermedades de la Médula Espinal/veterinaria , Espectroscopía de Resonancia Magnética , Tomografía Computarizada por Rayos X/veterinaria , Enfermedades de los Perros , Embolia/veterinaria , Núcleo Pulposo/anomalíasRESUMEN
Ozone therapy has been used to treat disc herniation for more than four decades. There are several papers describing results and mechanism of action. However, it is very important to define the characteristics of extruded disc herniation. Although ozone therapy showed excellent results in the majority of spinal diseases, it is not yet fully accepted within the medical community. Perhaps it is partly due to the fact that, sometimes, indications are not appropriately made. The objective of our work is to explain the mechanisms of action of ozone therapy on the extruded disc herniation. Indeed, these mechanisms are quite different from those exerted by ozone on the protruded disc herniation and on the degenerative disc disease because the inflammatory response is very different between the various cases. Extruded disc herniation occurs when the nucleus squeezes through a weakness or tear in the annulus. Host immune system considers the nucleus material to be a foreign invader, which triggers an immune response and inflammation. We think ozone therapy modulates this immune response, activating macrophages, which produce phagocytosis of extruded nucleus pulposus. Ozone would also facilitate the passage from the M1 to M2 phase of macrophages, going from an inflammatory phase to a reparative phase. Further studies are needed to verify the switch of macrophages.
Asunto(s)
Inflamación/tratamiento farmacológico , Desplazamiento del Disco Intervertebral/tratamiento farmacológico , Núcleo Pulposo/patología , Ozono/uso terapéutico , Humanos , Factores Inmunológicos/uso terapéutico , Inflamación/complicaciones , Inflamación/inmunología , Desplazamiento del Disco Intervertebral/complicaciones , Desplazamiento del Disco Intervertebral/inmunología , Dolor de la Región Lumbar/etiología , Núcleo Pulposo/efectos de los fármacos , Núcleo Pulposo/inmunología , Ozono/farmacologíaRESUMEN
Lumbar disc herniation is a common disease characterized by the degeneration of intervertebral discs (IVDs), accompanied by imbalance of metabolic and inflammatory homeostasis. Current studies establish that IVD degeneration is induced by increased apoptosis of nucleus pulposus (NP) cells. However, the underlying mechanisms of NP cell survival/apoptosis are not well elucidated. Here, we reveal a novel mechanism by which mTORC1 signaling controls NP cell survival through regulating metabolic homeostasis. We demonstrated that hyperactivated mTORC1 activity induced by inflammatory cytokines engenders the apoptosis of NP cells, whereas pharmacological inhibition of mTORC1 activity promotes NP cell survival. Using an integrative approach spanning metabolomics and biochemical approaches, we showed that mTORC1 activation enhanced glucose metabolism and lactic acid production, and therefore caused NP cell apoptosis. Our study identified mTORC1 in NP cells as a novel target for IVD degeneration, and provided potential strategies for clinical intervention of lumbar disc herniation.
Asunto(s)
Degeneración del Disco Intervertebral , Núcleo Pulposo , Apoptosis , Humanos , Inflamación/tratamiento farmacológico , Degeneración del Disco Intervertebral/tratamiento farmacológico , Diana Mecanicista del Complejo 1 de la RapamicinaRESUMEN
Lumbar disc herniation is a common disease characterized by the degeneration of intervertebral discs (IVDs), accompanied by imbalance of metabolic and inflammatory homeostasis. Current studies establish that IVD degeneration is induced by increased apoptosis of nucleus pulposus (NP) cells. However, the underlying mechanisms of NP cell survival/apoptosis are not well elucidated. Here, we reveal a novel mechanism by which mTORC1 signaling controls NP cell survival through regulating metabolic homeostasis. We demonstrated that hyperactivated mTORC1 activity induced by inflammatory cytokines engenders the apoptosis of NP cells, whereas pharmacological inhibition of mTORC1 activity promotes NP cell survival. Using an integrative approach spanning metabolomics and biochemical approaches, we showed that mTORC1 activation enhanced glucose metabolism and lactic acid production, and therefore caused NP cell apoptosis. Our study identified mTORC1 in NP cells as a novel target for IVD degeneration, and provided potential strategies for clinical intervention of lumbar disc herniation.
Asunto(s)
Humanos , Degeneración del Disco Intervertebral/tratamiento farmacológico , Núcleo Pulposo , Apoptosis , Diana Mecanicista del Complejo 1 de la Rapamicina , Inflamación/tratamiento farmacológicoRESUMEN
PURPOSE: Chordoma is a rare tumor of the skeletal system that is characterized by a high recurrence rate and treatment resistance. Given the common finding of immune dysregulation in chordoma, immunotherapy has emerged as potential treatment option. As an important immune checkpoint regulator, we evaluated cytotoxic T-lymphocyte antigen-4 (CTLA-4) expression and its prognostic significance for patients with chordoma of the spine. METHODS: CTLA-4 expression was analyzed immunohistochemically in 32 chordoma tissues and 14 nucleus pulposus tissues to examine the specificity of CTLA-4 expression in chordoma. Univariate log-rank analysis was used to evaluate the association of CTLA-4 expression in tumor cells and tumor-infiltrating lymphocytes (TILs) with survival. Cox multivariate analysis was used to identify independent factors of survival. RESULTS: Positive CTLA-4 expression was observed in all of the TILs and tumor cell cytoplasm, and partial in the membrane or in both the membrane and nucleus, with a markedly higher positivity rate than that observed in normal nucleus tissues. Higher CTLA-4 expression in the tumor but not in TILs was significantly associated with shorter continuous disease-free survival (CDFS) and overall survival (OS). CTLA-4 expression in tumor cells and TILs were independent predictors for CDFS, whereas only tumor cell expression was a significant predictor of OS. Furthermore, the combination of CTLA-4 expression in the tumor and TILs had higher prognostic value. CONCLUSIONS: Targeting CTLA-4 may be a potential novel therapeutic strategy for chordoma patients.
Asunto(s)
Antígeno CTLA-4/metabolismo , Cordoma/metabolismo , Linfocitos Infiltrantes de Tumor/metabolismo , Neoplasias de la Columna Vertebral/metabolismo , Adolescente , Adulto , Anciano , Niño , Cordoma/inmunología , Cordoma/mortalidad , Cordoma/terapia , Supervivencia sin Enfermedad , Femenino , Humanos , Inmunoterapia , Masculino , Persona de Mediana Edad , Núcleo Pulposo/metabolismo , Pronóstico , Modelos de Riesgos Proporcionales , Estudios Retrospectivos , Neoplasias de la Columna Vertebral/inmunología , Neoplasias de la Columna Vertebral/mortalidad , Neoplasias de la Columna Vertebral/terapia , Adulto JovenRESUMEN
RESUMEN La vértebra limbus es una rara condición poco descrita en la literatura médica. Se caracteriza por una herniación marginal intracorporal del núcleo pulposo que resulta en la separación de un fragmento óseo de forma triangular. Usualmente se presenta en niños o adolescentes, principalmente en la columna lumbar, y ocasiona dolor que, en la mayoría de los casos, mejora con el tratamiento con antiinflamatorios.
A B S T R A C T The limbus vertebra is a rare condition poorly described in the medical literature. Marginal intrabody herniation of the nucleus pulposus resulting in the separation of a triangular bone fragment. It usually occurs in children or adolescents mainly in the lumbar spine and causes pain that in most cases improves with treatment with anti-inflammatories.
Asunto(s)
Humanos , Niño , Adolescente , Adulto , Columna Vertebral , Terapéutica , Dolor de la Región Lumbar , Huesos , Núcleo Pulposo , AntiinflamatoriosRESUMEN
Many compounds of ginsenosides show anti-inflammatory properties. However, their anti-inflammatory effects in intervertebral chondrocytes in the presence of inflammatory factors have never been shown. Increased levels of pro-inflammatory cytokines are generally associated with the degradation and death of chondrocytes; therefore, finding an effective and nontoxic substance that attenuates the inflammation is worthwhile. In this study, chondrocytes were isolated from the nucleus pulposus tissues, and the cells were treated with ginsenoside compounds and IL-1ß, alone and in combination. Cell viability and death rate were assessed by CCK-8 and flow cytometry methods, respectively. PCR, western blot, and immunoprecipitation assays were performed to determine the mRNA and protein expression, and the interactions between proteins, respectively. Monomeric component of ginsenoside Rd had no toxicity at the tested range of concentrations. Furthermore, Rd suppressed the inflammatory response of chondrocytes to interleukin (IL)-1ß by suppressing the increase in IL-1ß, tumor necrosis factor (TNF)-α, IL-6, COX-2, and inducible nitric oxide synthase (iNOS) expression, and retarding IL-1ß-induced degradation of chondrocytes by improving cell proliferation characteristics and expression of aggrecan and COL2A1. These protective effects of Rd were associated with ubiquitination of IL-1 receptor accessory protein (IL1RAP), blocking the stimulation of IL-1ß to NF-κB. Bioinformatics analysis showed that NEDD4, CBL, CBLB, CBLC, and ITCH most likely target IL1RAP. Rd increased intracellular ITCH level and the amount of ITCH attaching to IL1RAP. Thus, IL1RAP ubiquitination promoted by Rd is likely to occur by up-regulation of ITCH. In summary, Rd inhibited IL-1ß-induced inflammation and degradation of intervertebral disc chondrocytes by increasing IL1RAP ubiquitination.
Asunto(s)
Condrocitos/efectos de los fármacos , Ginsenósidos/farmacología , Proteína Accesoria del Receptor de Interleucina-1/metabolismo , Interleucina-1beta/efectos de los fármacos , Degeneración del Disco Intervertebral/metabolismo , Adulto , Anciano , Agrecanos/metabolismo , Supervivencia Celular/efectos de los fármacos , Condrocitos/citología , Condrocitos/metabolismo , Ciclooxigenasa 2/metabolismo , Dinoprostona/metabolismo , Femenino , Ginsenósidos/metabolismo , Humanos , Inflamación/metabolismo , Interleucina-1beta/metabolismo , Dolor de la Región Lumbar/metabolismo , Masculino , Persona de Mediana Edad , Óxido Nítrico Sintasa/metabolismo , Núcleo Pulposo/citología , Núcleo Pulposo/efectos de los fármacos , Núcleo Pulposo/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , UbiquitinaciónRESUMEN
Many compounds of ginsenosides show anti-inflammatory properties. However, their anti-inflammatory effects in intervertebral chondrocytes in the presence of inflammatory factors have never been shown. Increased levels of pro-inflammatory cytokines are generally associated with the degradation and death of chondrocytes; therefore, finding an effective and nontoxic substance that attenuates the inflammation is worthwhile. In this study, chondrocytes were isolated from the nucleus pulposus tissues, and the cells were treated with ginsenoside compounds and IL-1β, alone and in combination. Cell viability and death rate were assessed by CCK-8 and flow cytometry methods, respectively. PCR, western blot, and immunoprecipitation assays were performed to determine the mRNA and protein expression, and the interactions between proteins, respectively. Monomeric component of ginsenoside Rd had no toxicity at the tested range of concentrations. Furthermore, Rd suppressed the inflammatory response of chondrocytes to interleukin (IL)-1β by suppressing the increase in IL-1β, tumor necrosis factor (TNF)-α, IL-6, COX-2, and inducible nitric oxide synthase (iNOS) expression, and retarding IL-1β-induced degradation of chondrocytes by improving cell proliferation characteristics and expression of aggrecan and COL2A1. These protective effects of Rd were associated with ubiquitination of IL-1 receptor accessory protein (IL1RAP), blocking the stimulation of IL-1β to NF-κB. Bioinformatics analysis showed that NEDD4, CBL, CBLB, CBLC, and ITCH most likely target IL1RAP. Rd increased intracellular ITCH level and the amount of ITCH attaching to IL1RAP. Thus, IL1RAP ubiquitination promoted by Rd is likely to occur by up-regulation of ITCH. In summary, Rd inhibited IL-1β-induced inflammation and degradation of intervertebral disc chondrocytes by increasing IL1RAP ubiquitination.
Asunto(s)
Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Anciano , Condrocitos/efectos de los fármacos , Ginsenósidos/farmacología , Interleucina-1beta/efectos de los fármacos , Proteína Accesoria del Receptor de Interleucina-1/metabolismo , Degeneración del Disco Intervertebral/metabolismo , Dinoprostona/metabolismo , Supervivencia Celular/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismo , Dolor de la Región Lumbar/metabolismo , Óxido Nítrico Sintasa/metabolismo , Condrocitos/citología , Condrocitos/metabolismo , Ginsenósidos/metabolismo , Ciclooxigenasa 2/metabolismo , Agrecanos/metabolismo , Interleucina-1beta/metabolismo , Ubiquitinación , Núcleo Pulposo/citología , Núcleo Pulposo/efectos de los fármacos , Núcleo Pulposo/metabolismo , Inflamación/metabolismoRESUMEN
STUDY DESIGN: ADAMTS5-deficient and wild type (WT) mice were chronically exposed to tobacco smoke to investigate effects on intervertebral disc degeneration (IDD). OBJECTIVE: The aim of this study was to demonstrate a role for ADAMTS5 in mediating tobacco smoking-induced IDD. SUMMARY OF BACKGROUND DATA: We previously demonstrated that chronic tobacco smoking causes IDD in mice because, in part, of proteolytic destruction of disc aggrecan. However, it was unknown which matrix proteinase(s) drive these detrimental effects. METHODS: Three-month-old WT (C57BL/6) and ADAMTS5 mice were chronically exposed to tobacco smoke (four cigarettes/day, 5âday/week for 6 months). ADAMTS-mediated cleavage of disc aggrecan was analyzed by Western blot. Disc total glycosaminoglycan (GAG) content was assessed by dimethyl methylene blue assay and safranin O/fast green histology. Vertebral osteoporosity was measured by microcomputed tomography. Human nucleus pulposus (hNP) cell cultures were also exposed directly to tobacco smoke extract (TSE), a condensate containing the water-soluble compounds inhaled by smokers, to measure ADAMTS5 expression and ADAMTS-mediated cleavage of aggrecan. Activation of nuclear factor (NF)-κB, a family of transcription factors essential for modulating the cellular response to stress, was measured by immunofluorescence assay. RESULTS: Genetic depletion of ADAMTS5 prevented vertebral bone loss, substantially reduced loss of disc GAG content, and completely obviated ADAMTS-mediated proteolysis of disc aggrecan within its interglobular domain (IGD) in mice following exposure to tobacco smoke. hNP cell cultures exposed to TSE also resulted in upregulation of ADAMTS5 protein expression and a concomitant increase in ADAMTS-mediated cleavage within aggrecan IGD. Activation of NF-κB, known to be required for ADAMTS5 gene expression, was observed in both TSE-treated hNP cell cultures and disc tissue of tobacco smoke-exposed mice. CONCLUSION: The findings demonstrate that ADAMTS5 is the primary aggrecanase mediating smoking-induced disc aggrecanolysis and IDD. Mouse models of chronic tobacco smoking are important and useful for probing the mechanisms of disc aggrecan catabolism and IDD. LEVEL OF EVIDENCE: N/A.
Asunto(s)
Proteína ADAMTS5/deficiencia , Degeneración del Disco Intervertebral/metabolismo , Disco Intervertebral/metabolismo , Fumar Tabaco/efectos adversos , Fumar Tabaco/metabolismo , Proteína ADAMTS5/biosíntesis , Adulto , Animales , Células Cultivadas , Femenino , Humanos , Disco Intervertebral/patología , Degeneración del Disco Intervertebral/patología , Degeneración del Disco Intervertebral/prevención & control , Masculino , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , FN-kappa B/metabolismo , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patología , Fumar Tabaco/patologíaRESUMEN
A extrusão discal aguda e não compressiva é caracterizada pela extrusão de caráter agudo/hiperagudo e não compressivo do núcleo pulposo de um disco intervertebral não degenerado. Pode ser chamada de hérnia de disco de baixo volume e alta velocidade ou explosões discais e geralmente está associado a exercícios intensos ou episódios traumáticos. O núcleo pulposo é fortemente forçado através de uma pequena fissura no ânulo fibroso dorsal, provocando uma contusão espinhal. Este relato tem como objetivo apresentar um caso de provável extrusão aguda de núcleo pulposo não compressiva. Foi atendido um cão macho, três anos e seis meses de idade, maltês, pesando 4,1kg. Como queixa principal, o proprietário relatou dificuldade locomotora e dor à manipulação há um dia, sem histórico de trauma. Foi constatada paraparesia não ambulatória de início agudo com ausência de propriocepção e dor superficial em membros pélvicos e dor à palpação epaxial da coluna toracolombar. A ressonância magnética (RM) evidenciou extensa área de hipersinal em segmento toracolombar da medula espinhal, sem sinais de compressão medular e de atenuação da intensidade do núcleo pulposo do disco intervertebral L1-L2. Foi feito diagnóstico presuntivo de mielopatia focal não compressiva com edema medular de todo segmento toracolombar, característico de uma extrusão aguda de núcleo pulposo não compressiva. Foi prescrito tratamento com anti-inflamatório esteroidal, analgésico, repouso absoluto e protocolo de reabilitação com acupuntura e fisioterapia. Após sete dias de tratamento, o animal recuperou a sensibilidade dolorosa superficial em membros pélvicos e evoluiu para paraparesia ambulatória. Os resultados deste relato sugerem que a RM pode ser útil para fazer um diagnóstico presuntivo em cães com histórico e sinais clínicos compatíveis. Além disso, o tratamento conservativo em extrusões discais não compressivas é preconizado e o paciente pode apresentar boa recuperação.(AU)
Acute and non-compressive disc extrusion is characterized by the acute character of extrusion of the nucleus pulposus without real compression of the spine. It has been called low-volume and high speed disc herniation or disc explosions, and usually is associated with an intense exercise or traumatic episode. This report aims to present a case of an acute extrusion of nucleus pulposus with no compression of the spinal cord. A 3.5 year-old male dog of the Maltes breed, weighing 4.1kg was presented at the Veterinary Hospital with locomotion disorders and pain during manipulation with no history of trauma. At the physical and neurological examination, non-ambulatory paraparesis of acute onset with absence of proprioception and superficial pain in hind limbs was found, as well as pain on palpation of epaxial thoracolumbar spine. Magnetic resonance imaging (MRI) showed extensive hyper intense area in the thoracolumbar spinal cord, with no signs of spinal cord compression, and decreased intensity of the nucleus pulposus of the L1-L2 intervertebral disc. Additionally, a spinal cord edema in all thoracolumbar segments was seen that is characteristic of an acute extrusion of non-compressive nucleus pulposus. A presumptive diagnosis of non-compressive myelopathy was assumed. The dog was prescribed steroidal anti-inflammatory, analgesic, absolute rest and rehabilitation protocol, including acupuncture and physiotherapy. The patient recovered superficial pain in the pelvic limbs and evolved into ambulatory paraparesis after seven days. The results of this report suggested that MRI can be useful for making a presumptive diagnosis in dogs with a history of compatible clinical signs. Moreover, the conservative treatment in non-compressive disc extrusions can be feasible.(AU)
Asunto(s)
Animales , Perros , Disco Intervertebral/patología , Núcleo Pulposo/patología , Traumatismos de la Médula Espinal/veterinaria , Espectroscopía de Resonancia MagnéticaRESUMEN
A extrusão discal aguda e não compressiva é caracterizada pela extrusão de caráter agudo/hiperagudo e não compressivo do núcleo pulposo de um disco intervertebral não degenerado. Pode ser chamada de hérnia de disco de baixo volume e alta velocidade ou explosões discais e geralmente está associado a exercícios intensos ou episódios traumáticos. O núcleo pulposo é fortemente forçado através de uma pequena fissura no ânulo fibroso dorsal, provocando uma contusão espinhal. Este relato tem como objetivo apresentar um caso de provável extrusão aguda de núcleo pulposo não compressiva. Foi atendido um cão macho, três anos e seis meses de idade, maltês, pesando 4,1kg. Como queixa principal, o proprietário relatou dificuldade locomotora e dor à manipulação há um dia, sem histórico de trauma. Foi constatada paraparesia não ambulatória de início agudo com ausência de propriocepção e dor superficial em membros pélvicos e dor à palpação epaxial da coluna toracolombar. A ressonância magnética (RM) evidenciou extensa área de hipersinal em segmento toracolombar da medula espinhal, sem sinais de compressão medular e de atenuação da intensidade do núcleo pulposo do disco intervertebral L1-L2. Foi feito diagnóstico presuntivo de mielopatia focal não compressiva com edema medular de todo segmento toracolombar, característico de uma extrusão aguda de núcleo pulposo não compressiva. Foi prescrito tratamento com anti-inflamatório esteroidal, analgésico, repouso absoluto e protocolo de reabilitação com acupuntura e fisioterapia. Após sete dias de tratamento, o animal recuperou a sensibilidade dolorosa superficial em membros pélvicos e evoluiu para paraparesia ambulatória. Os resultados deste relato sugerem que a RM pode ser útil para fazer um diagnóstico presuntivo em cães com histórico e sinais clínicos compatíveis. Além disso, o tratamento conservativo em extrusões discais não compressivas é preconizado e o paciente pode apresentar boa recuperação.(AU)
Acute and non-compressive disc extrusion is characterized by the acute character of extrusion of the nucleus pulposus without real compression of the spine. It has been called low-volume and high speed disc herniation or disc explosions, and usually is associated with an intense exercise or traumatic episode. This report aims to present a case of an acute extrusion of nucleus pulposus with no compression of the spinal cord. A 3.5 year-old male dog of the Maltes breed, weighing 4.1kg was presented at the Veterinary Hospital with locomotion disorders and pain during manipulation with no history of trauma. At the physical and neurological examination, non-ambulatory paraparesis of acute onset with absence of proprioception and superficial pain in hind limbs was found, as well as pain on palpation of epaxial thoracolumbar spine. Magnetic resonance imaging (MRI) showed extensive hyper intense area in the thoracolumbar spinal cord, with no signs of spinal cord compression, and decreased intensity of the nucleus pulposus of the L1-L2 intervertebral disc. Additionally, a spinal cord edema in all thoracolumbar segments was seen that is characteristic of an acute extrusion of non-compressive nucleus pulposus. A presumptive diagnosis of non-compressive myelopathy was assumed. The dog was prescribed steroidal anti-inflammatory, analgesic, absolute rest and rehabilitation protocol, including acupuncture and physiotherapy. The patient recovered superficial pain in the pelvic limbs and evolved into ambulatory paraparesis after seven days. The results of this report suggested that MRI can be useful for making a presumptive diagnosis in dogs with a history of compatible clinical signs. Moreover, the conservative treatment in non-compressive disc extrusions can be feasible.(AU)
Asunto(s)
Animales , Perros , Disco Intervertebral/patología , Traumatismos de la Médula Espinal/veterinaria , Núcleo Pulposo/patología , Espectroscopía de Resonancia MagnéticaRESUMEN
RNA extraction from the nucleus pulposus of intervertebral discs has been extensively used in orthopedic studies. We compared two methods for extracting RNA from the nucleus pulposus: liquid nitrogen grinding and enzyme digestion. The RNA was detected by agarose gel electrophoresis, and the purity was evaluated by absorbance ratio using a spectrophotometer. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression was assayed by reverse transcription-polymerase chain reaction (RT-PCR). Thirty human lumbar intervertebral discs were used in this study. The liquid nitrogen-grinding method was used for RNA extraction from 15 samples, and the mean RNA concentration was 491.04 ± 44.16 ng/mL. The enzyme digestion method was used on 15 samples, and the mean RNA concentration was 898.42 ± 38.64 ng/mL. The statistical analysis revealed that there was a significant difference in concentration between the different methods. Apparent 28S, 18S, and 5S bands were detectable in RNA extracted using the enzyme digestion method, whereas no 28S or 18S bands were detected in RNA extracted using the liquid nitrogen-grinding method. The GAPDH band was visible, and no non-specific band was detected in the RT-PCR assay by the enzyme digestion method. Therefore, the enzyme digestion method is an efficient and easy method for RNA extraction from the nucleus pulposus of intervertebral discs for further intervertebral disc degeneration-related studies.
Asunto(s)
Degeneración del Disco Intervertebral/genética , Disco Intervertebral/metabolismo , Núcleo Pulposo/metabolismo , ARN/aislamiento & purificación , Gliceraldehído-3-Fosfato Deshidrogenasas/biosíntesis , Humanos , ARN/genéticaRESUMEN
This study aims to explore the effect of microRNA-21 (miR-21) on the proliferation of human degenerated nucleus pulposus (NP) by targeting programmed cell death 4 (PDCD4) tumor suppressor. NP tissues were collected from 20 intervertebral disc degeneration (IDD) patients, and from 5 patients with traumatic spine fracture. MiR-21 expressions were tested. NP cells from IDD patients were collected and divided into blank control group, negative control group (transfected with miR-21 negative sequences), miR-21 inhibitor group (transfected with miR-21 inhibitors), miR-21 mimics group (transfected with miR-21 mimics) and PDCD4 siRNA group (transfected with PDCD4 siRNAs). Cell growth was estimated by Cell Counting Kit-8; PDCD4, MMP-2,MMP-9 mRNA expressions were evaluated by qRT-PCR; PDCD4, c-Jun and p-c-Jun expressions were tested using western blot. In IDD patients, the expressions of miR-21 and PDCD4 mRNA were respectively elevated and decreased (both P<0.05). The miR-21 expressions were positively correlated with Pfirrmann grades, but negatively correlated with PDCD4 mRNA (both P<0.001). In miR-21 inhibitor group, cell growth, MMP-2 and MMP-9 mRNA expressions, and p-c-Jun protein expressions were significantly lower, while PDCD4 mRNA and protein expressions were higher than the other groups (all P<0.05). These expressions in the PDCD4 siRNA and miR-21 mimics groups was inverted compared to that in the miR-21 inhibitor group (all P<0.05). MiR-21 could promote the proliferation of human degenerated NP cells by targeting PDCD4, increasing phosphorylation of c-Jun protein, and activating AP-1-dependent transcription of MMPs, indicating that miR-21 may be a crucial biomarker in the pathogenesis of IDD.
Asunto(s)
Proteínas Reguladoras de la Apoptosis/metabolismo , Proliferación Celular/fisiología , MicroARNs/metabolismo , Núcleo Pulposo/metabolismo , Proteínas de Unión al ARN/metabolismo , Adulto , Anciano , Proteínas Reguladoras de la Apoptosis/análisis , Proteínas Reguladoras de la Apoptosis/genética , Western Blotting , Recuento de Células , Células Cultivadas , Femenino , Expresión Génica , Humanos , Degeneración del Disco Intervertebral/metabolismo , Luciferasas , Masculino , Metaloproteinasa 2 de la Matriz/análisis , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/análisis , Metaloproteinasa 9 de la Matriz/genética , MicroARNs/análisis , MicroARNs/genética , Persona de Mediana Edad , Núcleo Pulposo/citología , ARN Mensajero/análisis , Proteínas de Unión al ARN/análisis , Proteínas de Unión al ARN/genética , Valores de Referencia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Fracturas de la Columna Vertebral/metabolismo , Factores de TiempoRESUMEN
STUDY DESIGN: In vitro biomechanical laboratory study. OBJECTIVES: The purpose of this study is to evaluate a mechanical treatment to create a degenerative motion segment and the ability of nucleus augmentation to restore biomechanics. SUMMARY OF BACKGROUND: In cases with an intact annulus fibrosus, the replacement or augmentation of the nucleus pulposus alone may provide a less invasive option to restore normal biomechanics and disk height when compared with spinal fusion or total disk replacement. Laboratory testing allows these changes to be fully characterized. However, without preexisting pathology, nucleus augmentation therapies are difficult to evaluate in vitro. METHODS: The present study evaluated pure moment bending and compressive biomechanics in 3 states (n=6): (1) intact, (2) after creep loading and nucleus disruption to induce degenerative biomechanical changes, and (3) after nucleus augmentation through an injectable polymer (DiscCell). RESULTS: Neutral zone and ROM were increased in all modes of bending after the degenerative treatment. The most sensitive mode of bending was lateral bending, with intact ROM (20.0±2.9 degrees) increased to 22.3±2.6 degrees after degenerative treatment and reduced to 18.4±1.6 degrees after injection of the polymer. All bending ROM and NZ changes induced by the degenerative treatment were reversed by nucleus augmentation. CONCLUSIONS: This material was shown to be effective at altering motion segment biomechanics and restoring disk height during time zero tests. This technique may provide a model to examine the time zero performance of a nucleus augmentation device/material.
Asunto(s)
Núcleo Pulposo/fisiología , Análisis de Varianza , Animales , Anillo Fibroso/fisiología , Fenómenos Biomecánicos , Modelos Animales de Enfermedad , Imagenología Tridimensional , Técnicas In Vitro , Fijadores Internos , Degeneración del Disco Intervertebral/etiología , Presión , Rango del Movimiento Articular/fisiología , Ovinos , Fusión Vertebral , Estrés Mecánico , Tomógrafos Computarizados por Rayos X , Tomografía Computarizada por Rayos XRESUMEN
This study aims to explore the effect of microRNA-21 (miR-21) on the proliferation of human degenerated nucleus pulposus (NP) by targeting programmed cell death 4 (PDCD4) tumor suppressor. NP tissues were collected from 20 intervertebral disc degeneration (IDD) patients, and from 5 patients with traumatic spine fracture. MiR-21 expressions were tested. NP cells from IDD patients were collected and divided into blank control group, negative control group (transfected with miR-21 negative sequences), miR-21 inhibitor group (transfected with miR-21 inhibitors), miR-21 mimics group (transfected with miR-21 mimics) and PDCD4 siRNA group (transfected with PDCD4 siRNAs). Cell growth was estimated by Cell Counting Kit-8; PDCD4, MMP-2,MMP-9 mRNA expressions were evaluated by qRT-PCR; PDCD4, c-Jun and p-c-Jun expressions were tested using western blot. In IDD patients, the expressions of miR-21 and PDCD4 mRNA were respectively elevated and decreased (both P<0.05). The miR-21 expressions were positively correlated with Pfirrmann grades, but negatively correlated with PDCD4 mRNA (both P<0.001). In miR-21 inhibitor group, cell growth, MMP-2 and MMP-9 mRNA expressions, and p-c-Jun protein expressions were significantly lower, while PDCD4 mRNA and protein expressions were higher than the other groups (all P<0.05). These expressions in the PDCD4 siRNA and miR-21 mimics groups was inverted compared to that in the miR-21 inhibitor group (all P<0.05). MiR-21 could promote the proliferation of human degenerated NP cells by targeting PDCD4, increasing phosphorylation of c-Jun protein, and activating AP-1-dependent transcription of MMPs, indicating that miR-21 may be a crucial biomarker in the pathogenesis of IDD.