RESUMEN
Proinflammatory cytokine such as interleukin (IL)-1ß causes inflammation of articular cartilage. In this current study, we explored the chondroprotective effects of long noncoding RNA (lncRNA) MALAT-1 on cell proliferation, apoptosis, and matrix metabolism in IL-1ß-induced inflammation in articular chondrocytes. Articular chondrocytes from knee joints of normal rats were isolated and cultured, followed by identification through observation of toluidine blue and COL II immunocytochemical stainings. The proliferation of chondrocytes at passage 2 was detected by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The inflammatory chondrocytes induced by 10 ng/mL IL-1ß were observed and identified by toluidine blue and COL II immunocytochemical stainings. pcDNA 3.1 and pcDNA-MALAT-1 were transfected in the chondrocytes. Ultrastructure of chondrocytes was observed by using a transmission electron microscope. The MTT assay was carried out to evaluate chondrocyte viability. Hoechst 33258 staining and flow cytometry were adopted to assess chondrocyte apoptosis. The chondrocytes at passage 2 with the biological characteristics of chondrocytes were used for subsequent experiments. In IL-1ß-treated chondrocytes, the growth rate of chondrocytes slowed down, the cells became narrow and long, the vacuoles were seen in the cells, and the morphology of the chondrocytes was irregular. The toluidine blue staining and the immunohistochemical staining of COL II became weaker. In response to IL-1ß induction, articular chondrocytes showed reduced MALAT-1 expression; moreover, obvious cartilage injury was observed with decreased chondrocyte viability and Col II expression and elevated chondrocyte apoptosis, MMP-13 expression, and p-JNK expression. With the treatment of pcDNA-MALAT-1, the cartilage injury was alleviated with increased chondrocyte viability and type II collagen (Col II) expression and reduced chondrocyte apoptosis, MMP-13 expression and p-JNK expression. Taken together these results, lncRNA MALAT-1 blocked the activation of the JNK signaling pathway; thereby, IL-1ß-induced inflammation in articular chondrocytes was reduced with enhanced chondrocyte proliferation and suppressed chondrocyte apoptosis and extracellular matrix degradation.
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Condrocitos/efectos de los fármacos , Interleucina-1beta/farmacología , MAP Quinasa Quinasa 4/genética , Sistema de Señalización de MAP Quinasas/genética , ARN Largo no Codificante/genética , Animales , Animales Recién Nacidos , Apoptosis/efectos de los fármacos , Apoptosis/genética , Cartílago Articular/citología , Cartílago Articular/metabolismo , Proliferación Celular/efectos de los fármacos , Condrocitos/metabolismo , Condrocitos/patología , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Regulación de la Expresión Génica , Inflamación , MAP Quinasa Quinasa 4/metabolismo , Metaloproteinasa 13 de la Matriz/genética , Metaloproteinasa 13 de la Matriz/metabolismo , Modelos Biológicos , Plásmidos/química , Plásmidos/metabolismo , Cultivo Primario de Células , ARN Largo no Codificante/agonistas , ARN Largo no Codificante/metabolismo , Ratas , TransfecciónRESUMEN
Our study assessed the effect of bone marrow mesenchymal stem cells (BMSCs) expressing inducible hepatocyte growth factor (HGF) on the recovery of femoral head necrosis (FHN). BMSCs were isolated by density gradient centrifugation. A recombinant AdTRE-HGF was constructed as the response plasmid and Adeno-X Tet-on as the regulator vector. The regulator and the response vectors were coinfected into BMSCs and induced at 0, 200, 500, 1000, and 1200 ng/mL doxycycline (Dox). After 3 days, the concentration of HGF was determined using enzyme-linked immunosorbent assay. Forty rabbits were selected to establish the FHN model and divided into 4 experimental groups. After the rabbits were killed by ketamine overdose, the restoration of FHN was assessed. The distribution of HGF-positive cells was observed by immunohistochemical method. Enzyme-linked immunosorbent assay results showed that 1000 ng/mL Dox induced the highest HGF expression level, even higher than the 1200 ng/mL Dox induction. The highest osteonecrosis incidence and empty lacunae percentage were found in group A compared with all the other groups (all P < 0.05). Furthermore, dramatically lower osteonecrosis incidence and empty lacunae percentage were found in group C compared with those of groups B and D (all P < 0.05). A significantly higher level of HGF protein was detected in group C compared with the other groups (all P < 0.05). Our study successfully developed the AdTRE-HGF, a recombinant adenovirus carrying HGF gene, for high expression of HGF in BMSCs. Importantly, introduction of BMSCs expressing HGF successfully produced the desired therapeutic effect in reversing FHN, in a Dox-dependent manner.
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Doxiciclina/farmacología , Necrosis de la Cabeza Femoral/terapia , Factor de Crecimiento de Hepatocito/biosíntesis , Factor de Crecimiento de Hepatocito/uso terapéutico , Células Madre Mesenquimatosas/metabolismo , Animales , Relación Dosis-Respuesta a Droga , Doxiciclina/administración & dosificación , Ensayo de Inmunoadsorción Enzimática , Vectores Genéticos , Factor de Crecimiento de Hepatocito/administración & dosificación , Masculino , ConejosRESUMEN
INTRODUCTION: Nontraumatic posterior atlantooccipital dislocation has only been rarely reported. In the current study, the authors reported an extremely rare case of nontraumatic posterior atlantooccipital dislocation associated with atlantoaxial instability. MATERIALS AND METHODS: A 47-year-old female was referred with a history of neck pain for 5 years. The patient had no history of trauma. The axial rotation of range of motion of the cervical spine was severely restricted. Posterior atlantooccipital dislocation with atlantoaxial instability was confirmed through conventional radiography, computed tomography and magnetic resonance imaging. We performed realignment of the dislocation and posterior occipitocervical (C0-C2) fusion. After the surgery, the patient's symptoms improved significantly and she manifested neurological improvement. CONCLUSION: To our knowledge, this lesion has not been reported previously. Anomalies of upper cervical spine may have induced this instability.
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Articulación Atlantoaxoidea/cirugía , Articulación Atlantooccipital/cirugía , Luxaciones Articulares/etiología , Inestabilidad de la Articulación/complicaciones , Vértebras Cervicales/cirugía , Femenino , Humanos , Luxaciones Articulares/diagnóstico , Luxaciones Articulares/cirugía , Inestabilidad de la Articulación/diagnóstico , Inestabilidad de la Articulación/cirugía , Imagen por Resonancia Magnética , Persona de Mediana Edad , Rango del Movimiento Articular , Fusión Vertebral/métodos , Tomografía Computarizada por Rayos XRESUMEN
The fast development of minimally invasive spine surgery in recent years is based on the advance of endoscopic microsurgery techniques, computer science and medical imaging, as well as the growing concerning of medical humanities. The concept of minimally invasive and precise targeting therapy has been penetrating into various areas of surgery, and minimal tissue damage and fewer complications are the new directions of minimally invasive spine surgery. In this article we review some advances in precise spinal surgery including percutaneous lumbar discectomy, microendoscopic discectomy, computer-assisted orthopedic surgery and robot surgery.
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Vértebras Lumbares/cirugía , Procedimientos Quirúrgicos Mínimamente Invasivos , Endoscopía , Humanos , Microcirugia , Procedimientos Quirúrgicos RobotizadosRESUMEN
OBJECTIVE: To investigate the mechanism of chlorogenic acid (CGA) on H2O2-induced apoptosis in the rat nucleus pulposus cells (NPCs). METHODS: NPCs were isolated from SD rats and cultured in vitro. Cultured cells (P3) were randomly divided into normal control group, H2O2 group, CGA + H2O2 group, CGA group and LY294002 pretreatment group. The apoptosis and ROS production of rNPCs was detected by flow cytometry. The expressions of p-Akt, BCL-2 and Akt were analyzed by Western blot. RESULTS: Compared with normal control group, in the H2O2 group, the production of ROS and the apoptosis rate significantly increased in rNPCs; CGA treatment inhibited ROS production and cell apoptosis, while increased the expression of p-Akt and BCL-2; LY294002, a PI3Kinse inhibitor, not only decreased the expression of p-Akt and BCL-2, but also obviously increased ROS production and cell apoptosis. CONCLUSION: Chlorogenic acid can protect NPCs against apoptosis by oxidative stress through decreasing reactive oxygen species production and increasing anti-apoptotic protein BCL-2 expression in NPCs by activation of PI3K-Akt signaling pathways.
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Apoptosis/efectos de los fármacos , Ácido Clorogénico/farmacología , Disco Intervertebral/efectos de los fármacos , Fosfatidilinositol 3-Quinasas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Animales , Antioxidantes/farmacología , Western Blotting , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Femenino , Citometría de Flujo , Regulación de la Expresión Génica/efectos de los fármacos , Peróxido de Hidrógeno/toxicidad , Disco Intervertebral/citología , Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/prevención & control , Estrés Oxidativo/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Ratas , Transducción de Señal/efectos de los fármacosRESUMEN
This study aimed to reveal the specific role of early growth response protein 1 (EGR1) and nuclear receptor 4A3 (NR4A3) in nucleus pulposus cells (NPCs) and the related molecular mechanism and to identify a new strategy for treating intervertebral disc degeneration (IVDD). Bioinformatics analysis was used to explore and predict IVDD-related differentially expressed genes, and chromatin immunoprecipitation sequencing (ChIP-seq) revealed NR4A3 as the EGR1 target gene. An in vitro NPC model induced by tributyl hydrogen peroxide (TBHP) and a rat model induced by fibrous ring acupuncture were established. Western blotting, quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemical staining, immunofluorescence staining, and flow cytometry were used to detect the effects of EGR1 and NR4A3 knockdown and overexpression on NPC apoptosis and the expression of extracellular matrix (ECM) anabolism-related proteins. Interactions between EGR1 and NR4A3 were analyzed via ChIP-qPCR and dual luciferase assays. EGR1 and NR4A3 expression levels were significantly higher in severely degenerated discs (SDD) than in mildly degenerated discs (MDD), indicating that these genes are important risk factors in IVDD progression. ChIP-seq and RNA-seq revealed NR4A3 as a direct downstream target of EGR1, and this finding was verified by ChIP-qPCR and dual luciferase reporter experiments. Remarkably, the rescue experiments showed that EGR1 promotes TBHP-induced NPC apoptosis and impairs ECM anabolism, dependent on elevated NR4A3 expression. In summary, the EGR1-NR4A3 axis mediates the progression of NPC apoptosis and ECM impairment and is a potential therapeutic target in IVDD.
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Apoptosis , Proteína 1 de la Respuesta de Crecimiento Precoz , Degeneración del Disco Intervertebral , Núcleo Pulposo , Estrés Oxidativo , Receptores de Hormona Tiroidea , Adulto , Animales , Femenino , Humanos , Masculino , Persona de Mediana Edad , Ratas , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Proteína 1 de la Respuesta de Crecimiento Precoz/metabolismo , Proteína 1 de la Respuesta de Crecimiento Precoz/genética , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/genética , Degeneración del Disco Intervertebral/patología , Proteínas del Tejido Nervioso , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patología , Ratas Sprague-Dawley , Receptores de Esteroides/metabolismo , Receptores de Esteroides/genética , Receptores de Hormona Tiroidea/metabolismo , Receptores de Hormona Tiroidea/genética , Regulación hacia ArribaRESUMEN
Bone marrow-derived mesenchymal stem cells (BM-MSCs) transplantation is widely adopted for the curing of osteonecrosis of femoral head (ONFH) in recent years. Furthermore, it is known that introducing hepatocyte growth factor (HGF) into BM-MSCs will greatly improve the therapeutic effect of stem-cell therapy owing to the great angiogenic and anti-fibrotic capabilities of HGF. However, continuing overexpression of HGF in vivo may cause sarcomas, such as Kaposi's sarcoma. Aiming at enhancing the therapeutic effect and preventing the side effects of HGF-modified stem-cell transplantation on ONFH, we sought to construct a gene regulation system to control HGF expression in BM-MSCs rigorously and accurately. We selected baculovirus as the gene vector and introduced pTet-on advanced system into that. Finally, a virus vector vAc(rtTA2s-Ptight-HGF) was successfully built and delivered into BM-MSCs to regulate the accurate expression of HGF. As shown in the results, different levels of HGF expression were verified by ELISA and Western blot with different induction doses of doxycycline (DOX). There was a dose-response relationship between them, and the optimum dose of DOX to induce HGF expression in BM-MSCs in vitro was 1 µg/mL. We conclude that it is feasible to regulate HGF expression in BM-MSCs by baculovirus-mediated one-off transduction.
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Baculoviridae/genética , Factor de Crecimiento de Hepatocito/metabolismo , Células Madre Mesenquimatosas/metabolismo , Animales , Células de la Médula Ósea/citología , Proliferación Celular , Células Cultivadas , Doxiciclina/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Vectores Genéticos/genética , Vectores Genéticos/metabolismo , Factor de Crecimiento de Hepatocito/genética , Masculino , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/patología , ConejosRESUMEN
OBJECTIVE: To improve the treatment effect of patients with L5 S1 lumber disc herniation (LDH) with a narrow interlaminar window, we proposed an alternative approach to percutaneous endoscopic interlaminar discectomy (PEID) via the laminoplasty technique. METHODS: Fifteen L5 S1 LDH patients (7 men and 8 women; age range, 22 to 56 years; median age, 34 years; 9 left, 6 right) were enrolled in the present study retrospectively. The interlaminar windows of all patients were narrow (the transverse diameter of the L5 S1 interlaminar window is equal to or less than that of L4-5 ). Percutaneous laminoplasty and endoscopic interlaminar discectomy surgery were undergone by all patients from July 2018 to July 2019. All operations were completed under local anesthesia. The target laminoplasty area was the safety zone, use of which avoids both transverse and exit nerve roots. Under fluoroscopic guidance or clear endoscopic visualization, the trephines were used to enlarge the interlaminar window, which allowed the working cannula to enter the spinal canal but avoid nerve roots and the dural sac. The preoperative/postoperative visual analogue scale (VAS) scores and Oswestry disability index (ODI) were statistically analyzed. The modified MacNab criterion was used to assess the clinical effects. The radiological outcomes were evaluated by MRI and CT. SPSS 19.0 software was used for the statistical evaluation. RESULTS: The operative time ranged from 70 to 120 min, with a median time of 92 min, and the fluoroscopy times ranged from 8 to 12, with a median of 9.7 times. The body mass index (BMI) of patients ranged from 18.10 to 26.06, with a median of 22.04. All patients were followed up in the outpatient department for at least 12 months after surgery. At the last follow up, the average VAS-Back score of the study patients was reduced from 5.33 ± 2.09 to 2.00 ± 1.20 (P < 0.001) and the average VAS-Leg score was reduced from 7.53 ± 1.69 to 1.47 ± 0.92 (P < 0.001). The average ODI scores improved from 47.87 ± 11.41 to 12.93 ± 3.24 (P < 0.01). According to the modified MacNab criteria, 11 cases achieved excellent results and 4 cases achieved good results. All of the operations were successful. There wertr no nerve root injuries, dural tears, or other complications. CONCLUSION: The laminoplasty approach for PEID provides a safe and useful alternative for the treatment of L5-S1 LDH patients with a narrow interlaminar window.
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Discectomía Percutánea/métodos , Endoscopía/métodos , Degeneración del Disco Intervertebral/cirugía , Desplazamiento del Disco Intervertebral/cirugía , Laminoplastia/métodos , Vértebras Lumbares/cirugía , Adulto , Evaluación de la Discapacidad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Dimensión del Dolor , Estudios Retrospectivos , Adulto JovenRESUMEN
BACKGROUNDS: Transforaminal percutaneous endoscopic discectomy (TF-PELD) and interlaminar percutaneous endoscopic discectomy (IL-PELD) are the most common alternative treatments of lumbar disc herniation. The aim of this study was to compare the operation time duration and X-ray exposure as well as outcomes of TF-PELD and IL-PELD as indicated by the published clinical evidences within randomized trials. METHODS: We included randomized, controlled studies reporting operation duration and X-ray exposure as well as clinical outcome evaluations, comparing TF-PELD to IL-PELD with a minimum of 10 patients per group. The included data measures were operation duration, X-ray exposure and postoperation evaluations. Data were synthesized and analyzed using ReviewManager version 5.3. Publication bias was evaluated via funnel plot. The Cochran Q test and the degree of inconsistency (I2) were used to assess heterogeneity. Lowly biased and heterogenous dichotomous data were calculated by odds ratio and continuous data were calculated by mean difference (MD) with 95% confidence intervals (CI). RESULTS: Thirteen studies published from January 1970 to March 2018, with a total of 770 lumbar disc herniation patients, including 361 cases of TF-PELD and 409 cases of IL-PELD, were finally included. Meta-analysis of data extracted from these studies revealed that the postoperation outcomes of both surgery methods did not differ significantly, but the surgery duration was significantly shorter in the IL-PELD group than in the TF-PELD group (MD 21.69; 95% CI 12.94-30.27; Pâ=â.00001), and the fluoroscopy times demanded in the IL-PELD group was significantly fewer than those in the TF-PELD group (MD 7.57; 95% CI 6.22-8.93; Pâ=â.00001). CONCLUSION: The main finding of the study is that IL-PELD approach can decrease radiation exposure as their demanded duration of operation and fluoroscopy times were significantly shorter and fewer in the IL-PELD group, which they achieve similar outcomes comparing to TF-PELD. The study is limited at a lack of samples with lumbar disc herniation levels out of L5/S1. The findings implicate selection of IL-PELD approach over TF-PELD at applicable circumstances for lower lumbar disc herniation. Physicians should consider this data when choosing between TF-PELD and IL-PELD.
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Discectomía Percutánea/métodos , Desplazamiento del Disco Intervertebral/cirugía , Vértebras Lumbares/cirugía , Fluoroscopía/estadística & datos numéricos , Humanos , Tempo Operativo , Complicaciones Posoperatorias/epidemiología , Ensayos Clínicos Controlados Aleatorios como Asunto , Estudios RetrospectivosRESUMEN
Accumulating evidences indicate the elevated expression of αB-Crystallin (Cryab) is implicated in tumorigenesis. However, the expression and biologic role of Cryab in osteosarcoma (OS) are still unknown. In this study, we showed that Cryab expression was elevated in OS tissues and cell lines, and down-regulation of Cryab in MG-63 and U-2OS cells led to a decline in the cells' aggressiveness, and reduced secretion of matrix metalloproteinase-9 (MMP-9) in vitro, and lower metastasis potential in vivo. Further study indicated that the Cryab expression was positively associated with the activity of ERK1/2 which is responsible for the cells' aggressiveness and MMP-9 secretion. Clinically, our data confirmed that the high level of Cryab was associated with shorten survival and tumor recurrence for the postoperative OS patients. Together, our results indicate that high level of Cryab is a new adverse outcomes marker for OS patients and may be used as a new therapeutic target.