Culturing Bovine Nucleus Pulposus Explants by Balancing Medium Osmolarity

2011 ◽  
Vol 17 (11) ◽  
pp. 1089-1096 ◽  
Author(s):  
Bart van Dijk ◽  
Esther Potier ◽  
Keita Ito
Keyword(s):  
Nucleus Pulposus ◽  
Medium Osmolarity

Transplantation of Activated Nucleus Pulposus Cells after Cryopreservation: Efficacy Study in Canine Disc Degeneration Model

2014 ◽  
Vol 4 (1_suppl) ◽  
pp. s-0034-1376573-s-0034-1376573
Author(s):  
T. Nukaga ◽  
D. Sakai ◽  
A. Hiyama ◽  
T. Ishii ◽  
T. Nakai ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Disc Degeneration ◽  
Efficacy Study ◽  
Nucleus Pulposus Cells

Sirtuin 1 Maitains Survival via PKB Signaling in Degenerative Human Disc Nucleus Pulposus Cells*

2012 ◽  
Vol 39 (6) ◽  
pp. 563-573
Author(s):  
Da-Wu WANG ◽  
Zhen-Ming HU ◽  
Jie HAO ◽  
Bin HE ◽  
Qiang GAN ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Sirtuin 1 ◽  
Nucleus Pulposus Cells

ATP potently modulates anion channel-mediated excitatory amino acid release from cultured astrocytes

2002 ◽  
Vol 283 (2) ◽  
pp. C569-C578 ◽  
Author(s):  
Alexander A. Mongin ◽  
Harold K. Kimelberg
Keyword(s):  
Amino Acid ◽  
Excitatory Amino Acid ◽  
Atp Release ◽  
Anion Channel ◽  
P2y Receptors ◽  
Cell Swelling ◽  
Channel Blockers ◽  
Medium Osmolarity ◽  
Subsequent Activation

Volume-dependent ATP release and subsequent activation of purinergic P2Y receptors have been implicated as an autocrine mechanism triggering activation of volume-regulated anion channels (VRACs) in hepatoma cells. In the brain ATP is released by both neurons and astrocytes and participates in intercellular communication. We explored whether ATP triggers or modulates the release of excitatory amino acid (EAAs) via VRACs in astrocytes in primary culture. Under basal conditions exogenous ATP (10 μM) activated a small EAA release in 70–80% of the cultures tested. In both moderately (5% reduction of medium osmolarity) and substantially (35% reduction of medium osmolarity) swollen astrocytes, exogenous ATP greatly potentiated EAA release. The effects of ATP were mimicked by P2Y agonists and eliminated by P2Y antagonists or the ATP scavenger apyrase. In contrast, the same pharmacological maneuvers did not inhibit volume-dependent EAA release in the absence of exogenous ATP, ruling out a requirement of autocrine ATP release for VRAC activation. The ATP effect in nonswollen and moderately swollen cells was eliminated by a 5–10% increase in medium osmolarity or by anion channel blockers but was insensitive to tetanus toxin pretreatment, further supporting VRAC involvement. Our data suggest that in astrocytes ATP does not trigger EAA release itself but acts synergistically with cell swelling. Moderate cell swelling and ATP may serve as two cooperative signals in bidirectional neuron-astrocyte communication in vivo.

scholarly journals The distinct roles of myosin IIA and IIB under compression stress in nucleus pulposus cells

2021 ◽  
Vol 54 (2) ◽  
Author(s):  
Wencan Ke ◽  
Bingjin Wang ◽  
Wenbin Hua ◽  
Yu Song ◽  
Saideng Lu ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Compression Stress ◽  
Nucleus Pulposus Cells ◽  
Myosin Iia

scholarly journals Activation of nuclear factor-kappa B by TNF promotes nucleus pulposus mineralization through inhibition of ANKH and ENPP1

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Agata K. Krzyzanowska ◽  
Robert J. Frawley ◽  
Sheela Damle ◽  
Tony Chen ◽  
Miguel Otero ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Inflammatory Cytokines ◽  
Intervertebral Disc Degeneration ◽  
Inflammatory Cytokine ◽  
Mineral Content ◽  
Nuclear Factor Kappa B ◽  
Nuclear Factor ◽  
Matrix Mineralization ◽  
Phenotypic Changes ◽  
Cellular Alterations

AbstractSpontaneous mineralization of the nucleus pulposus (NP) has been observed in cases of intervertebral disc degeneration (IDD). Inflammatory cytokines have been implicated in mineralization of multiple tissues through their modulation of expression of factors that enable or inhibit mineralization, including TNAP, ANKH or ENPP1. This study examines the underlying factors leading to NP mineralization, focusing on the contribution of the inflammatory cytokine, TNF, to this pathologic event. We show that human and bovine primary NP cells express high levels of ANKH and ENPP1, and low or undetectable levels of TNAP. Bovine NPs transduced to express TNAP were capable of matrix mineralization, which was further enhanced by ANKH knockdown. TNF treatment or overexpression promoted a greater increase in mineralization of TNAP-expressing cells by downregulating the expression of ANKH and ENPP1 via NF-κB activation. The increased mineralization was accompanied by phenotypic changes that resemble chondrocyte hypertrophy, including increased RUNX2 and COL10A1 mRNA; mirroring the cellular alterations typical of samples from IDD patients. Disc organ explants injected with TNAP/TNF- or TNAP/shANKH-overexpressing cells showed increased mineral content inside the NP. Together, our results confirm interactions between TNF and downstream regulators of matrix mineralization in NP cells, providing evidence to suggest their participation in NP calcification during IDD.

scholarly journals Towards Tissue-Specific Stem Cell Therapy for the Intervertebral Disc: PPARδ Agonist Increases the Yield of Human Nucleus Pulposus Progenitor Cells in Expansion

Surgeries ◽  
2021 ◽  
Vol 2 (1) ◽  
pp. 92-104
Author(s):  
Xingshuo Zhang ◽  
Julien Guerrero ◽  
Andreas S. Croft ◽  
Katharina A.C. Oswald ◽  
Christoph E. Albers ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Progenitor Cells ◽  
Nucleus Pulposus ◽  
Nucleus Pulposus Cell ◽  
Treated Group ◽  
Peroxisome Proliferator ◽  
Peroxisome Proliferator Activated Receptor ◽  
Hematopoietic Stem ◽  
Heterogeneous Nucleus ◽  
Red Dye

(1) Background: Low back pain (LBP) is often associated with intervertebral disc degeneration (IVDD). Autochthonous progenitor cells isolated from the center, i.e., the nucleus pulposus, of the IVD (so-called nucleus pulposus progenitor cells (NPPCs)) could be a future cell source for therapy. The NPPCs were also identified to be positive for the angiopoietin-1 receptor (Tie2). Similar to hematopoietic stem cells, Tie2 might be involved in peroxisome proliferator-activated receptor delta (PPARδ) agonist-induced self-renewal regulation. The purpose of this study was to investigate whether a PPARδ agonist (GW501516) increases the Tie2+ NPPCs’ yield within the heterogeneous nucleus pulposus cell (NPC) population. (2) Methods: Primary NPCs were treated with 10 µM of GW501516 for eight days. Mitochondrial mass was determined by microscopy, using mitotracker red dye, and the relative gene expression was quantified by qPCR, using extracellular matrix and mitophagy-related genes. (3) The NPC’s group treated with the PPARδ agonist showed a significant increase of the Tie2+ NPCs yield from ~7% in passage 1 to ~50% in passage two, compared to the NPCs vehicle-treated group. Furthermore, no significant differences were found among treatment and control, using qPCR and mitotracker deep red. (4) Conclusion: PPARδ agonist could help to increase the Tie2+ NPCs yield during NPC expansion.

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