Fas ligand plays an important role for the production of pro-inflammatory cytokines in intervertebral disc nucleus pulposus cells

2012 ◽  
Vol 31 (4) ◽  
pp. 608-615 ◽  
Author(s):  
Junya Yamamoto ◽  
Koichiro Maeno ◽  
Toru Takada ◽  
Kenichiro Kakutani ◽  
Takashi Yurube ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Inflammatory Cytokines ◽  
Fas Ligand ◽  
Nucleus Pulposus Cells ◽  
Pro Inflammatory Cytokines

Inhibition of the Extracellular Signal-regulated Kinase Pathway Restores the Discogenic Phenotype of Inflammatory Intervertebral Disc Cells

2021 ◽  
Author(s):  
Adel Tekari ◽  
Alessandro Marazza ◽  
Benjamin Gantenbein
Keyword(s):  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Inflammatory Cytokines ◽  
Gelatin Zymography ◽  
Erk Pathway ◽  
Nucleus Pulposus Cells ◽  
Necrosis Factor Alpha ◽  
Extracellular Signal Regulated Kinase ◽  
Extracellular Signal ◽  
Pro Inflammatory Cytokines

Abstract Background: Intervertebral disc (IVD) degeneration is a spinal disease caused by trauma and/or repetitive mechanical overloading of the spine which triggers inflammatory response pathways. Long-term disc inflammation may lead to development of spinal pseudoarthrosis. The aim of the present study was to elucidate the role of the extracellular signal-regulated kinase (ERK) pathway in inflammation-induced IVD cells. Methods: Inflammatory human nucleus pulposus cells (NPC) were stimulated using tumor necrosis factor alpha (TNFα) and the ERK pathway was blocked using a selective molecule-based inhibitor U0126. Gene expression of catabolic and anabolic events, pro-inflammatory, and NPC markers were investigated. The enzymatic activity of matrix metalloproteinases (MMP)2/9 were determined by gelatin zymography. The cytoxicity of U0126 concentrations on NPC was quantified using resazurin assay, and the specificity of U0126 on ERK1/2 signaling was determined.Results: The pro-inflammatory cytokines like MMP3/13 and interleukin 6 in nucleus pulposus (NP) inflammatory conditions were down-regulated by U0126 and a trend towards an increase of the NP-specific collagen type 2, aggrecan and keratin 19 was observed suggesting a recovery of the NP phenotype. U0126 does not seem to have effect on prostaglandin production, aggrecanases and some anabolic genes. We confirmed that U0126 selectively blocks the ERK phosphorylation and U0126 affects the cells metabolic activity only for high concentrations. Conclusions: Inhibition of ERK signaling down-regulates important metalloproteinase, pro-inflammatory cytokines, and up-regulates NP markers in order to restore the discogenic phenotype of inflammatory NPC.

Tissue Engineering of the Intervertebral Disc With Cultured Nucleus Pulposus Cells Using Atelocollagen Scaffold and Growth Factors

Spine ◽  
2012 ◽  
Vol 37 (6) ◽  
pp. 452-458 ◽  
Author(s):  
Kwang-Il Lee ◽  
Seong-Hwan Moon ◽  
Hyang Kim ◽  
Un-Hye Kwon ◽  
Ho-Joong Kim ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Growth Factors ◽  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Nucleus Pulposus Cells

scholarly journals Synergistic Effects of Acidic pH and Pro-Inflammatory Cytokines IL-1β and TNF-α for Cell-Based Intervertebral Disc Regeneration

2020 ◽  
Vol 10 (24) ◽  
pp. 9009
Author(s):  
Chiara Borrelli ◽  
Conor T. Buckley
Keyword(s):  
Intervertebral Disc ◽  
Cell Viability ◽  
Inflammatory Cytokines ◽  
Cell Function ◽  
Synergistic Effects ◽  
Dominant Factor ◽  
Disc Regeneration ◽  
Tnf Α ◽  
Intervertebral Disc Regeneration ◽  
Pro Inflammatory Cytokines

The intervertebral disc (IVD) relies mainly on diffusion through the cartilaginous endplates (CEP) to regulate the nutrient and metabolites exchange, thus creating a challenging microenvironment. Degeneration of the IVD is associated with intradiscal acidification and elevated levels of pro-inflammatory cytokines. However, the synergistic impact of these microenvironmental factors for cell-based therapies remains to be elucidated. The aim of this study was to investigate the effects of low pH and physiological levels of interleukin-1ß (IL-1β) and tumour necrosis factor-α (TNF-α) on nasal chondrocytes (NCs) and subsequently compare their matrix forming capacity to nucleus pulposus (NP) cells in acidic and inflamed culture conditions. NCs and NP cells were cultured in low glucose and low oxygen at different pH conditions (pH 7.1, 6.8 and 6.5) and supplemented with physiological levels of IL-1β and TNF-α. Results showed that acidosis played a pivotal role in influencing cell viability and matrix accumulation, while inflammatory cytokine supplementation had a minor impact. This study demonstrates that intradiscal pH is a dominant factor in determining cell viability and subsequent cell function when compared to physiologically relevant inflammatory conditions. Moreover, we found that NCs allowed for improved cell viability and more effective NP-like matrix synthesis compared to NP cells, and therefore may represent an alternative and appropriate cell choice for disc regeneration.

scholarly journals Ligustrazine Prevents Intervertebral Disc Degeneration via Suppression of Aberrant TGFβ Activation in Nucleus Pulposus Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-9
Author(s):  
Shufen Liu ◽  
Yuhao Cheng ◽  
Yuqi Tan ◽  
Jingcheng Dong ◽  
Qin Bian
Keyword(s):  
Growth Factor ◽  
Mouse Model ◽  
Intervertebral Disc ◽  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Intervertebral Disc Degeneration ◽  
Ex Vivo ◽  
Tissue Growth ◽  
Nucleus Pulposus Cells

Objectives. Aberrant transforming growth factor β (TGFβ) activation is detrimental to both nucleus pulposus (NP) cells and cartilage endplates (CEPs), which can lead to intervertebral disc degeneration (IDD). Ligustrazine (LIG) reduces the expression of inflammatory factors and TGFβ1 in hypertrophic CEP to prevent IDD. In this study, we investigate the effects of LIG on NP cells and the TGFβ signaling. Design. LIG was injected to the lumbar spinal instability (LSI) mouse model. The effect of LIG was evaluated by intervertebral disc (IVD) score in the LSI mouse model. The expression of activated TGFβ was examined using immunostaining with pSmad2/3 antibody. The upright posture (UP) rat model was also treated and evaluated in the same manner to assess the effect of LIG. In ex vivo study, IVDs from four-week old mice were isolated and treated with 10−5, 10−6, and 10−7 M of LIG. We used western blot to detect activated TGFβ expression. TGFβ-treated human nucleus pulposus cells (HNPCs) were cotreated with optimized dose of LIG in vitro. Immunofluorescence staining was performed to determine pSmad2/3, connective tissue growth factor (CCN2), and aggrecan (ACAN) expression levels. Results. IVD score and the percentage of pSmad2/3+ NP cells were low in LIG-treated LSI mice in comparison with LSI mice, but close to the levels in the Sham group. Similarly, LIG reduced the overexpression of TGFβ1 in NP cells. The inhibitory effect of LIG was dose dependent. A dose of 10−5 M LIG not only strongly attenuated Smad2/3 phosphorylation in TGFβ-treated IVD ex vivo but also suppressed pSmad2/3, CCN2, and ACAN expression in TGFβ-treated NP cells in vitro. Conclusions. LIG prevents IDD via suppression of TGFβ overactivation in NP cells.

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