CCN family member 2/connective tissue growth factor (CCN2/CTGF) is regulated by Wnt–β-catenin signaling in nucleus pulposus cells

Abstract Background The aims of this study were to investigate the gene expression of CCN family members in rat intervertebral disc (IVD) cells and to examine whether Wnt–β-catenin signaling regulates the expression of CCN family 2 (CCN2)/connective tissue growth factor (CTGF) in rat nucleus pulposus (NP) cells. Methods The gene expression of CCN family members were assessed in rat IVD cells using real-time reverse transcription polymerase chain reaction (RT-PCR). The expression pattern of CCN2 was also assessed in rat IVD cells using western blot and immunohistochemical analyses. Gain-of-function and loss-of-function experiments were performed to identify the mechanisms by which Wnt–β-catenin signaling influences the activity of the CCN2 promoter. To further determine if the mitogen-activated protein kinase (MAPK) pathway is required for the Wnt–β-catenin signaling-induced regulation of CCN2 expression in the NP cells, CCN2 expression was analyzed by reporter assay, RT-PCR and western blot analysis. Results CCN2 messenger RNA (mRNA) and protein were expressed in rat IVDs. Expression of CCN2 was significantly higher than for mRNA of other CCN family members in both rat NP and annulus fibrosus (AF) cells. The relative activity of the CCN2 promoter decreased 24 h after treatment with 6-bromoindirubin-3′-oxime (1.0 μM) (0.773 (95% 0.735, 0.812) P = 0.0077) in NP cells. In addition, treatment with the WT–β-catenin vector (500 ng) significantly decreased CCN2 promoter activity (0.688 (95% 0.535, 0.842) P = 0.0063), whereas β-catenin small interfering RNA (500 ng) significantly increased CCN2 promoter activity (1.775 (95% 1.435, 2.115) P < 0.001). Activation of Wnt–β-catenin signaling decreased the expression of CCN2 mRNA and protein by NP cells. Regulation of CCN2 by Wnt–β-catenin signaling involved the MAPK pathway in rat NP cells. Conclusions This study shows that Wnt–β-catenin signaling regulates the expression of CCN2 through the MAPK pathway in NP cells. Understanding the balance between Wnt–β-catenin signaling and CCN2 is necessary for developing therapeutic alternatives for the treatment of IVD degeneration

CCN family member 2/connective tissue growth factor (CCN2/CTGF) is regulated by Wnt–β-catenin signaling in nucleus pulposus cells

Abstract Background The aims of this study were to investigate the gene expression of CCN family members in rat intervertebral disc (IVD) cells and to examine whether Wnt–β-catenin signaling regulates the expression of CCN family 2 (CCN2)/connective tissue growth factor (CTGF) in rat nucleus pulposus (NP) cells. Methods The gene expression of CCN family members were assessed in rat IVD cells using real-time reverse transcription polymerase chain reaction (RT-PCR). The expression pattern of CCN2 was also assessed in rat IVD cells using western blot and immunohistochemical analyses. Gain-of-function and loss-of-function experiments were performed to identify the mechanisms by which Wnt–β-catenin signaling influences the activity of the CCN2 promoter. To further determine if the mitogen-activated protein kinase (MAPK) pathway is required for the Wnt–β-catenin signaling-induced regulation of CCN2 expression in the NP cells, CCN2 expression was analyzed by reporter assay, RT-PCR and western blot analysis. Results CCN2 messenger RNA (mRNA) and protein were expressed in rat IVDs. Expression of CCN2 was significantly higher than for mRNA of other CCN family members in both rat NP and annulus fibrosus (AF) cells. The relative activity of the CCN2 promoter decreased 24 h after treatment with 6-bromoindirubin-3′-oxime (1.0 μM) (0.773 (95% 0.735, 0.812) P = 0.0077) in NP cells. In addition, treatment with the WT–β-catenin vector (500 ng) significantly decreased CCN2 promoter activity (0.688 (95% 0.535, 0.842) P = 0.0063), whereas β-catenin small interfering RNA (500 ng) significantly increased CCN2 promoter activity (1.775 (95% 1.435, 2.115) P < 0.001). Activation of Wnt–β-catenin signaling decreased the expression of CCN2 mRNA and protein by NP cells. Regulation of CCN2 by Wnt–β-catenin signaling involved the MAPK pathway in rat NP cells. Conclusions This study shows that Wnt–β-catenin signaling regulates the expression of CCN2 through the MAPK pathway in NP cells. Understanding the balance between Wnt–β-catenin signaling and CCN2 is necessary for developing therapeutic alternatives for the treatment of IVD degeneration

Prevalence of diffuse idiopathic skeletal hyperostosis (DISH) assessed with whole-spine computed tomography in 1479 subjects

Abstract Background Computed tomography (CT) analyses have reported that the prevalence of diffuse idiopathic skeletal hyperostosis (DISH) in Japan is 8.7–27.1%. However, these data were obtained using chest-abdominal CT, and no evaluations of sagittal, coronal, and axial images using whole-spine CT have been reported. The aim of this study was to investigate the prevalence and characteristic of DISH by whole spinal CT. Methods Participants were patients who had experienced trauma who had undergone whole-spine CT scanning based on the initial clinical practice guidelines for trauma in our institute from April 2015 to February 2018. The subjects were > 20 years old and 1479 were included in the analysis. The presence and distribution of DISH and clinical parameters such as age and sex were reviewed retrospectively according to the location of DISH. Results The overall prevalence of DISH was 19.5% (n = 289). Subjects with DISH were older than those without. DISH was located in the thoracic spine in 65.1% and thoracolumbar spine in 24.2% of patients. More than 80% of ligamentous ossifications associated with DISH occurred at T8 (n = 255, 88%), T9 (n = 262, 91%), and T10 (n = 247, 85%). Most of the ossification occurred to the right anterior of the vertebral body, and there were few ossifications in the areas in contact with the artery and vein. Conclusions The prevalence of DISH based on whole-spine CT was 19.5%. Ossification was noted more often at T8, T9, and T10, and to the right anterior of the vertebral body. It is for the first time report that we have studied the location of ossification in detail using the axial images of whole spine CT. We hope this study will enhance the understanding of the characteristics of DISH

Comparative Study of Cage Subsidence in Single-Level Lateral Lumbar Interbody Fusion

We investigated the incidence and clinical features of cage subsidence after single-level lateral lumbar interbody fusion (LLIF). We studied a retrospective cohort of 59 patients (34 males, 25 females; mean age, 68.9 years) who received single-level LLIF. Patients were classified into subsidence and no-subsidence groups. Cage subsidence was defined as any violation of either endplate, classified using radiographs and computed tomography (CT) images. After one year, we compared patient characteristics, surgical parameters, radiological findings, pain scores, and fusion status. We also compared the Hounsfield unit (HU) endplate value obtained on CT preoperatively. Twenty patients (33.9%) had radiographic evidence of interbody cage subsidence. There were significant differences between the subsidence and no-subsidence groups in sex, cage height, fusion rate, and average HU value of both endplates (p &lt; 0.05). There were no significant differences in age, height, weight, or body mass index. Moreover, there were no significant differences in global alignment and Numerical Rating Scale change in low back pain, leg pain, and numbness. Despite suggestions that patients with lower HU values might develop cage subsidence, our results showed that cage subsidence after single-level LLIF was not associated with low back pain, leg pain, or numbness one year post-operation