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. Design 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.

IL‐1β‐regulating angiogenic factors expression in perforated temporomandibular disk cells via NF‐κB pathway

“…In this study, VEGF and Ang‐1 was up‐regulated in perforated disk cells like other cartilage angiogenesis, including intervertebral disk and TMJ cartilage . In TMJ disk, the invaded vessels are formed from pre‐existing ones which are mainly at disk periphery, such as retrodiscal tissues.…”

Molecular Therapy for Degenerative Disc Disease: Clues from Secretome Analysis of the Notochordal Cell-Rich Nucleus Pulposus

“…Similarly, other studies have demonstrated increased proteoglycan synthesis in NP cells treated with notochordal cell derived conditioned medium (NCCM) in vitro 22. A recent study also demonstrated the anti-angiogeneic effects of NCCM in vitro 23. These studies provide compelling evidence that NCs secrete proteins, which may confer anabolic characteristics, maintain a homeostatic microenvironment in NP and resists DDD.…”

Meniscus, articular cartilage and nucleus pulposus: a comparative review of cartilage-like tissues in anatomy, development and function

“…Fas ligand, a type II transmembrane protein of the tumor necrosis factor family, expressed by normal NP cells, could cause apoptosis in vascular endothelial cells and subsequently inhibit blood vessel infiltration (Sun et al ., 2013). Additionally, Nog and chondroitin sulfate released from notochordal cells inhibited angiogenesis by suppressing vascular endothelial growth factor signaling (Cornejo et al ., 2015). Nerves in the discs, either accompanying the vessels or occurring independently, are branches of the sinuvertebral nerve or the gray rami communicantes (Johnson et al , 2001; Raj, 2008).…”