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.
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).
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/jsp2.1238Salzer 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
PresentationVernengo 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)
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