title: Soluble factors from the notochordal-rich intervertebral disc inhibit endothelial cell invasion and vessel formation in the presence and absence of pro-inflammatory cytokines.
abstract: BACKGROUND
  Chronic low back pain can be associated with the pathological ingrowth of blood vessels and nerves into intervertebral discs (IVDs). The notochord patterns the IVD during development and is a source of anti-angiogenic soluble factors such as Noggin and Chondroitin sulfate (CS). These factors may form the basis for a new minimally invasive strategy to target angiogenesis in the IVD.
  
  
  OBJECTIVE
  To examine the anti-angiogenic potential of soluble factors from notochordal cells (NCs) and candidates Noggin and CS under healthy culture conditions and in the presence of pro-inflammatory mediators.
  
  
  METHODS
  NC conditioned media (NCCM) was generated from porcine NC-rich nucleus pulposus tissue. To assess the effects of NCCM, CS and Noggin on angiogenesis, cell invasion and tubular formation assays were performed using human umbilical vein endothelial cells (HUVECs) ± tumor necrosis factor alpha (TNFα [10 ng/ml]). vascular endothelial growth factor (VEGF)-A, MMP-7, interleukin-6 (IL-6) and IL-8 mRNA levels were assessed using qRT-PCR.
  
  
  RESULTS
  NCCM (10 & 100%), CS (10 and 100 μg) and Noggin (10 and 100 ng) significantly decreased cell invasion of HUVECs with and without TNFα. NCCM 10% and Noggin 10 ng inhibited tubular formation with and without TNFα and CS 100 μg inhibited tubules in Basal conditions whereas CS 10 μg inhibited tubules with TNFα. NCCM significantly decreased VEGF-A, MMP-7 and IL-6 mRNA levels in HUVECs with and without TNFα. CS and Noggin had no effects on gene expression.
  
  
  CONCLUSIONS
  We provide the first evidence that soluble factors from NCs can inhibit angiogenesis by suppressing VEGF signaling. Notochordal-derived ligands are a promising minimally invasive strategy targeting neurovascular ingrowth and pain in the degenerated IVD.
authors.affiliation: Leni and Peter W. May Department of Orthopedics, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. Electronic address: Marisa.cornejo@mountsinai.org.
authors.forename: M C
authors.initials: MC
authors.lastname: Cornejo
authors.affiliation: Leni and Peter W. May Department of Orthopedics, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. Electronic address: Samuel.cho@mountsinai.org.
authors.forename: S K
authors.initials: SK
authors.lastname: Cho
authors.affiliation: Cardiovascular Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. Electronic address: Chiara.giannarelli@mssm.edu.
authors.forename: C
authors.initials: C
authors.lastname: Giannarelli
authors.affiliation: Leni and Peter W. May Department of Orthopedics, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. Electronic address: James.iatridis@mssm.edu.
authors.forename: J C
authors.initials: JC
authors.lastname: Iatridis
authors.affiliation: Leni and Peter W. May Department of Orthopedics, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA. Electronic address: Devina.purmessur@mssm.edu.
authors.forename: D
authors.initials: D
authors.lastname: Purmessur
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D002352:Carrier Proteins / Q000494:pharmacology*; D002809:Chondroitin Sulfates / Q000494:pharmacology*; D016207:Cytokines / Q000235:genetics* / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D015850:Interleukin-6 / Q000235:genetics; D016209:Interleukin-8 / Q000187:drug effects / Q000235:genetics; D007403:Intervertebral Disc / Q000196:embryology / Q000378:metabolism*; D020783:Matrix Metalloproteinase 7 / Q000187:drug effects / Q000235:genetics; D009389:Neovascularization, Pathologic / Q000378:metabolism*; D009672:Notochord / Q000196:embryology / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism*; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000187:drug effects / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000187:drug effects / Q000235:genetics; D000099176:Noggin Protein
chemical_list: D020533:Angiogenesis Inhibitors; D002352:Carrier Proteins; D002809:Chondroitin Sulfates; D016207:Cytokines; D015850:Interleukin-6; D016209:Interleukin-8; D020783:Matrix Metalloproteinase 7; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; D000099176:Noggin Protein; C508606:CXCL8 protein, human; C508600:IL6 protein, human; C467484:VEGFA protein, human; C060932:MMP7 protein, human
keywords: Angiogenesis; Back pain; Intervertebral disc; Notochordal cells; TNFα