Explorative Study of Modulatory Effects of Notochordal Cell‐Derived Extracellular Vesicles on the IL‐1β‐Induced Catabolic Cascade in Nucleus Pulposus Cell Pellets and Explants
ABSTRACT Background Cell‐free regenerative strategies, such as notochordal cell (NC)‐derived extracellular vesicles (EVs), are an attractive alternative in developing new therapies for intervertebral disc (IVD) degeneration. NC‐EVs have been reported to elicit matrix anabolic effects on nucleus pulp...
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Wiley
2025-03-01
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| Online Access: | https://doi.org/10.1002/jsp2.70043 |
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| author | J. C. vanMaanen F. C. Bach J. W. Snuggs K. Ito M. H. M. Wauben C. L. Le Maitre M. A. Tryfonidou |
| author_facet | J. C. vanMaanen F. C. Bach J. W. Snuggs K. Ito M. H. M. Wauben C. L. Le Maitre M. A. Tryfonidou |
| author_sort | J. C. vanMaanen |
| collection | DOAJ |
| description | ABSTRACT Background Cell‐free regenerative strategies, such as notochordal cell (NC)‐derived extracellular vesicles (EVs), are an attractive alternative in developing new therapies for intervertebral disc (IVD) degeneration. NC‐EVs have been reported to elicit matrix anabolic effects on nucleus pulposus cells from degenerated IVDs cultured under basal conditions. However, the degenerative process is exacerbated by pro‐inflammatory cytokines contributing to the vicious degenerative cycle. Therefore, this study explores whether NC‐EVs modulate interleukin (IL)‐1β‐mediated pro‐inflammatory responses in the degenerating disc. Methods This study utilized two IL‐1β induced pro‐catabolic culture models; a dog 3D nucleus pulposus (NP) cell pellet culture and a human patient‐derived, ex vivo NP tissue culture system. Porcine NC‐EVs were generated from NC‐conditioned medium by differential centrifugation followed by size exclusion chromatography. Donor matched EV‐depleted media were generated by overnight ultracentrifugation, whereafter the EV‐depleted NCCM supernatant was subjected to size exclusion chromatography. To investigate whether observed effects were EV‐associated, NC‐EVs conditions were compared to EV‐depleted controls in the absence and presence of IL‐1β. Results The size and concentration of NC‐EVs were quantified by nanoparticle tracking analysis, which showed minimal donor variation and confirmed depletion of EVs in the EV‐depleted media. In the IL‐1β‐induced catabolic cascade, the NC‐EVs did not elicit anabolic effects at the matrix level nor did they rescue the pro‐catabolic phenotype within dog pellets. Modification of the CCL2 secretion seemed to be context dependent in the human explants: where EVs treatment stimulated CCL2 secretion but in the presence of IL‐1β this effect was counteracted. Secretion of IL‐6 and C‐X‐C motif chemokine ligand 1 was significantly decreased in NC‐EV + IL‐1β vs. control+IL‐1β but not compared to EV‐depleted human explant controls. Altogether, this data provides evidence for a protective modulatory role of NC‐EVs. Considering the homeostatic function EVs exert, inherently encompassing subtle biologic modifications, the current study may have lacked sufficient power to demonstrate statistical significance in a sample set with evident donor variation. Conclusions NC‐EVs may modulate the production of specific cytokines and chemokines in human degenerate explants when the key pro‐inflammatory cytokine IL‐1β is present. Implementation of the technical EV‐depleted controls in further studies is essential to robustly demonstrate that these effects are EV‐mediated and not associated with other secreted factors co‐isolated during EV‐isolation. |
| format | Article |
| id | doaj-art-850c4e4e98264efd8978644b03bc01aa |
| institution | Kabale University |
| issn | 2572-1143 |
| language | English |
| publishDate | 2025-03-01 |
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| spelling | doaj-art-850c4e4e98264efd8978644b03bc01aa2025-08-20T03:48:45ZengWileyJOR Spine2572-11432025-03-0181n/an/a10.1002/jsp2.70043Explorative Study of Modulatory Effects of Notochordal Cell‐Derived Extracellular Vesicles on the IL‐1β‐Induced Catabolic Cascade in Nucleus Pulposus Cell Pellets and ExplantsJ. C. vanMaanen0F. C. Bach1J. W. Snuggs2K. Ito3M. H. M. Wauben4C. L. Le Maitre5M. A. Tryfonidou6Department of Clinical Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht NetherlandsDepartment of Clinical Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht NetherlandsDivision of Clinical Medicine, Faculty of Health University of Sheffield Sheffield UKOrthopedic Biomechanics, Department of Biomedical Engineering Eindhoven University of Technology Eindhoven NetherlandsDepartment of Biomolecular Health Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht NetherlandsDivision of Clinical Medicine, Faculty of Health University of Sheffield Sheffield UKDepartment of Clinical Sciences, Faculty of Veterinary Medicine Utrecht University Utrecht NetherlandsABSTRACT Background Cell‐free regenerative strategies, such as notochordal cell (NC)‐derived extracellular vesicles (EVs), are an attractive alternative in developing new therapies for intervertebral disc (IVD) degeneration. NC‐EVs have been reported to elicit matrix anabolic effects on nucleus pulposus cells from degenerated IVDs cultured under basal conditions. However, the degenerative process is exacerbated by pro‐inflammatory cytokines contributing to the vicious degenerative cycle. Therefore, this study explores whether NC‐EVs modulate interleukin (IL)‐1β‐mediated pro‐inflammatory responses in the degenerating disc. Methods This study utilized two IL‐1β induced pro‐catabolic culture models; a dog 3D nucleus pulposus (NP) cell pellet culture and a human patient‐derived, ex vivo NP tissue culture system. Porcine NC‐EVs were generated from NC‐conditioned medium by differential centrifugation followed by size exclusion chromatography. Donor matched EV‐depleted media were generated by overnight ultracentrifugation, whereafter the EV‐depleted NCCM supernatant was subjected to size exclusion chromatography. To investigate whether observed effects were EV‐associated, NC‐EVs conditions were compared to EV‐depleted controls in the absence and presence of IL‐1β. Results The size and concentration of NC‐EVs were quantified by nanoparticle tracking analysis, which showed minimal donor variation and confirmed depletion of EVs in the EV‐depleted media. In the IL‐1β‐induced catabolic cascade, the NC‐EVs did not elicit anabolic effects at the matrix level nor did they rescue the pro‐catabolic phenotype within dog pellets. Modification of the CCL2 secretion seemed to be context dependent in the human explants: where EVs treatment stimulated CCL2 secretion but in the presence of IL‐1β this effect was counteracted. Secretion of IL‐6 and C‐X‐C motif chemokine ligand 1 was significantly decreased in NC‐EV + IL‐1β vs. control+IL‐1β but not compared to EV‐depleted human explant controls. Altogether, this data provides evidence for a protective modulatory role of NC‐EVs. Considering the homeostatic function EVs exert, inherently encompassing subtle biologic modifications, the current study may have lacked sufficient power to demonstrate statistical significance in a sample set with evident donor variation. Conclusions NC‐EVs may modulate the production of specific cytokines and chemokines in human degenerate explants when the key pro‐inflammatory cytokine IL‐1β is present. Implementation of the technical EV‐depleted controls in further studies is essential to robustly demonstrate that these effects are EV‐mediated and not associated with other secreted factors co‐isolated during EV‐isolation.https://doi.org/10.1002/jsp2.70043CCL2degenerationdoghuman explantintervertebral disc |
| spellingShingle | J. C. vanMaanen F. C. Bach J. W. Snuggs K. Ito M. H. M. Wauben C. L. Le Maitre M. A. Tryfonidou Explorative Study of Modulatory Effects of Notochordal Cell‐Derived Extracellular Vesicles on the IL‐1β‐Induced Catabolic Cascade in Nucleus Pulposus Cell Pellets and Explants JOR Spine CCL2 degeneration dog human explant intervertebral disc |
| title | Explorative Study of Modulatory Effects of Notochordal Cell‐Derived Extracellular Vesicles on the IL‐1β‐Induced Catabolic Cascade in Nucleus Pulposus Cell Pellets and Explants |
| title_full | Explorative Study of Modulatory Effects of Notochordal Cell‐Derived Extracellular Vesicles on the IL‐1β‐Induced Catabolic Cascade in Nucleus Pulposus Cell Pellets and Explants |
| title_fullStr | Explorative Study of Modulatory Effects of Notochordal Cell‐Derived Extracellular Vesicles on the IL‐1β‐Induced Catabolic Cascade in Nucleus Pulposus Cell Pellets and Explants |
| title_full_unstemmed | Explorative Study of Modulatory Effects of Notochordal Cell‐Derived Extracellular Vesicles on the IL‐1β‐Induced Catabolic Cascade in Nucleus Pulposus Cell Pellets and Explants |
| title_short | Explorative Study of Modulatory Effects of Notochordal Cell‐Derived Extracellular Vesicles on the IL‐1β‐Induced Catabolic Cascade in Nucleus Pulposus Cell Pellets and Explants |
| title_sort | explorative study of modulatory effects of notochordal cell derived extracellular vesicles on the il 1β induced catabolic cascade in nucleus pulposus cell pellets and explants |
| topic | CCL2 degeneration dog human explant intervertebral disc |
| url | https://doi.org/10.1002/jsp2.70043 |
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