Induced pluripotent stem cell-based therapy for spinal regeneration

EU H2020
A Vernengo, Z Li, S Grad, M Alini

Background

Notochordal cells (NCs) are a promising candidate for cell-based therapies for intervertebral disc (IVD) degeneration. A multitude of in vitro studies in 2D monolayers and 3D hydrogels indicate beneficial molecular effects of NCs on resident IVD cells, like increased protection from apoptosis and stimulation of GAG production. In this work, we evaluated the therapeutic effects of NC-based therapy in an ex vivo bovine organ culture model of IVD degeneration. In this model, degeneration was induced enzymatically with injection of the enzyme collagenase. In previous studies, we showed that collagenase II induces significant losses in disc height, formation of a void at the site of injection, and degenerative shifts in gene expression by 7 days of dynamically loaded culture. Here, it was hypothesized that the injection of porcine-derived notochordal cells after 7 days of collagenase digestion would have regenerative effects on the bovine tissue by enhancing GAG content, upregulating anabolic gene expression, and mitigating IVD height loss.

 

 

Results

Gross observations of collagenase-digested specimens at day 21 of culture revealed greater structural integrity for the collagenase-digested IVDs which received notochordal cells (+NC) compared to the untreated group (-NC, Figure A). The glycosaminoglycan (GAG) content in the inner annulus fibrosus (iAF) at day 21 was higher for the +NC group compared to the –NC group (p=0.04, Figure B). Mean loss of viability in the iAF and nucleus pulposus (NP region) was significantly greater at day 21 for the -NC group (67.6 ± 31.1%) versus +NC group (34.19 ± 19.9%, p=0.009, Figure C). 

 

A) Macromorphological properties of collagenase-degenerated bovine IVDs with or without notochordal cell treatment (+NC or -NC, respectively) compared to day 0 intact specimen. B) Mean GAG content of collagenase-digested bovine IVDs with or without notochordal cell treatment. C) Average percent change in cell viability over 21 days with or without notochordal cell treatment.
  • Publication

    Lee NN, Salzer E, Bach FC, Bonilla AF, Cook JL, Gazit Z, Grad S, Ito K, Smith LJ, Vernengo A, Wilke HJ, Engiles JB, Tryfonidou MA. A comprehensive tool box for large animal studies of intervertebral disc degeneration. JOR Spine. 2021 Jun 14;4(2):e1162. DOI: 10.1002/jsp2.1162. eCollection 2021 Jun. 

    Basatvat S, Bach FC, Barcellona MN, Binch AL, Buckley CT, Bueno B, Cahine NO, Creemers LB, Dudli S, Fearing B, Ferguson SJ, Gansau J, Gantenbein B, Gawri R, Glaeser JD, Grad S, Guerrero J, Haglund L, Hernandez PA, Hoyland JA, Huang C, Iatridis JC, Illien-Junger S, Jing L, Kraus P, Laagland LT, Lang G, Leung V, Li Z, Lufkin T, van Maanen JC, McDonnell EE, Panebiancho CJ, Presciutti SM, Rao S, Richardson SM, Romerein S, Schmitz TC, Schol J, Setton L, Sheyn D, Snuggs JW, Sun Y, Tan X, Tryfonidou MA, Vo N, Wang D, Williams B, Williams R, Yoon ST, Le Maitre CL. 
    Harmonization and standardization of nucleus pulposus cell extraction and culture methods. JOR Spine. 2023;epub Jan 10. https://www.doi.org/10.1002/jsp2.1238

    Salzer E, Schmitz TC, Mouser VH, Vernengo A, Gantenbein B, Jansen JU, Neidlinger-Wilke C, Wilke HJ, Grad S, Le Maitre CL, Tryfonidou MA, Ito K. Ex vivo intervertebral disc cultures: degeneration-induction methods and their implications for clinical translation. Eur Cell Mater. 2023;45:88-112
     

    Vernengo A, Bumann H, Kluser N, Soubrier A, Šećerović A, Gewiess J, Jansen JU, Neidlinger-Wilke C, Wilke H-J, Grad S. Chemonucleolysis combined with dynamic loading for inducing degeneration in bovine caudal intervertebral discs. Front Bioeng Biotechnol. 2023;11:1178938. https://doi.org/10.3389/fbioe.2023.1178938 

     
  • Presentation
    Vernengo A, Bumann H, Kluser N, Soubrier A, Secerovic A, Zuncheddu D, Williams R, Snuggs J, Janai R, Sammon C, Jansen JU, Neidlinger-Wilke C, Wilke HJ, Le Maitre C, Grad S. Injectable hydrogel-encapsulated porcine notochordal cells implanted into an ex vivo model of intervertebral disc degeneration. 2023 ORS (poster)
     
  • Partner

    Tryfonidou M (Prof), University of Utrecht, Netherlands

    Creemers L (PhD), University Medical Centre Utrecht, Netherlands

    Ito K (Prof), Technical University of Eindhoven, Netherlands

    Guicheux J (Prof), University of Nantes, France

    Pandit A (Prof), National University of Galway, Ireland

    Wilke H-J (Prof), University of Ulm, Germany

    Gantenbein B (Prof), University of Bern, Switzerland

    Jorgensen C (Prof), Institute National de la Sante, France

    Templin M, Naturwissenschaftliches und Medizinisches Institut, Germany

    Le Maitre C (Prof), Sheffield Hallam University, United Kingdom

    Vadala G, University Campus Biomedico, Rom, Italy

    De Boer M, Ntrans Technologies, Netherlands

    Noel D, University of Montpellier, France

    Isasi R, University of Miami, US

    Kienle A, Spineserv Gmbh, Germany

    Chan D, The University of Hong Kong, Hong Kong

    Buljovcic Z, Pharmalex Gmbh, Germany

    Lether I, National Reumafonds, Netherlands

     

     

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