Hyperosmolarity induces notochordal cell differentiation with aquaporin3 upregulation and reduced N-cadherin expression

J Orthop Res. 2018 Feb;36(2):788-798. doi: 10.1002/jor.23715. Epub 2017 Sep 20.


The nucleus pulposus (NP) of intervertebral discs (IVD) undergoes dramatic changes with aging including loss of its gelatinous structure and large, vacuolated notochordal cells (NCs) in favor of a matrix-rich structure populated by small NP cells (sNPCs). NP maturation also involves a loading-pattern shift from pressurization to matrix deformations, and these events are thought to predispose to degeneration. Little is known of the triggering events and cellular alterations involved with NP maturation, which remains a fundamental open spinal mechanobiology question. A mouse IVD organ culture model was used to test the hypotheses that hyperosmotic overloading will induce NP maturation with transition of NCs to sNPCs while also increasing matrix accumulation and altering osmoregulatory and mechanotransductive proteins. Results indicated that static hyperosmolarity, as might occur during growth, caused maturation of NCs to sNPCs and involved a cellular differentiation process since known NC markers (cytokeratin-8, -19, and sonic hedgehog) persisted without increased cell apoptosis. Osmosensitive channels Aquaporin 3 (Aqp3) and transient receptor potential vanilloid-4 (TRPV4) expression were both modified with altered osmolarity, but increased Aqp3 with hyperosmolarity was associated with NC to sNPC differentiation. NC to sNPC differentiation was accompanied by a shift in cellular mechanotransduction proteins with decreased N-cadherin adhesions and increased Connexin 43 connexons. We conclude that hyperosmotic overloading can promote NC differentiation into sNPCs. This study identified osmolarity as a triggering mechanism for notochordal cell differentiation with associated shifts in osmoregulatory and mechanotransductive proteins that are likely to play important roles in intervertebral disc aging. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:788-798, 2018.

Keywords: intervertebral disc; mechanobiology; notochordal cell; nucleus pulposus; osmolarity.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Apoptosis
  • Aquaporin 3 / physiology*
  • Cadherins / metabolism*
  • Cell Differentiation*
  • Connexin 43 / metabolism
  • Extracellular Matrix / metabolism
  • Female
  • Humans
  • Mechanotransduction, Cellular
  • Mice, Inbred C57BL
  • Notochord / cytology
  • Nucleus Pulposus / cytology
  • Nucleus Pulposus / physiology*
  • Organ Culture Techniques
  • Osmotic Pressure*
  • TRPV Cation Channels / metabolism


  • Aqp3 protein, mouse
  • Cadherins
  • Cdh2 protein, mouse
  • Connexin 43
  • GJA1 protein, mouse
  • TRPV Cation Channels
  • Trpv4 protein, mouse
  • Aquaporin 3