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Int. J. Mol. Sci. 2016, 17(10), 1640; doi:10.3390/ijms17101640

An Inflammatory Nucleus Pulposus Tissue Culture Model to Test Molecular Regenerative Therapies: Validation with Epigallocatechin 3-Gallate

1
Institute for Biomechanics, ETH Zurich, Hoenggerbergring 64, CH-8093 Zurich, Switzerland
2
Health Department, ZHAW—Zurich University of Applied Sciences, Technikumstrasse 71, CH-8401 Winterthur, Switzerland
3
Department of Biomedical Engineering, Eindhoven University of Technology, Postbus 513, 5600 MB Eindhoven, The Netherlands
4
Competence Center for Applied Biotechnology and Molecular Medicine, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland
*
Author to whom correspondence should be addressed.
Academic Editor: Paula Andrade
Received: 23 July 2016 / Revised: 15 September 2016 / Accepted: 19 September 2016 / Published: 27 September 2016
(This article belongs to the Special Issue Natural Anti-Inflammatory Agents)
View Full-Text   |   Download PDF [6323 KB, uploaded 27 September 2016]   |  

Abstract

Organ cultures are practical tools to investigate regenerative strategies for the intervertebral disc. However, most existing organ culture systems induce severe tissue degradation with only limited representation of the in vivo processes. The objective of this study was to develop a space- and cost-efficient tissue culture model, which represents degenerative processes of the nucleus pulposus (NP). Intact bovine NPs were cultured in a previously developed system using Dyneema jackets. Degenerative changes in the NP tissue were induced either by the direct injection of chondroitinase ABC (1–20 U/mL) or by the diffusion of interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) (both 100 ng/mL) from the culture media. Extracellular matrix composition (collagens, proteoglycans, water, and DNA) and the expression of inflammatory and catabolic genes were analyzed. The anti-inflammatory and anti-catabolic compound epigallocatechin 3-gallate (EGCG, 10 µM) was employed to assess the relevance of the degenerative NP model. Although a single injection of chondroitinase ABC reduced the proteoglycan content in the NPs, it did not activate cellular responses. On the other hand, IL-1β and TNF-α significantly increased the mRNA expression of inflammatory mediators IL-6, IL-8, inducible nitric oxide synthase (iNOS), prostaglandin-endoperoxide synthase 2 (PTGS2) and matrix metalloproteinases (MMP1, MMP3, and MMP13). The cytokine-induced gene expression in the NPs was ameliorated with EGCG. This study provides a proof of concept that inflammatory NP cultures, with appropriate containment, can be useful for the discovery and evaluation of molecular therapeutic strategies against early degenerative disc disease. View Full-Text
Keywords: nucleus pulposus; organ culture model; degenerative disc disease; IL-1β; TNF-α; chondroitinase ABC; epigallocatechin 3-gallate; inflammation; extracellular matrix nucleus pulposus; organ culture model; degenerative disc disease; IL-1β; TNF-α; chondroitinase ABC; epigallocatechin 3-gallate; inflammation; extracellular matrix
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Krupkova, O.; Hlavna, M.; Amir Tahmasseb, J.; Zvick, J.; Kunz, D.; Ito, K.; Ferguson, S.J.; Wuertz-Kozak, K. An Inflammatory Nucleus Pulposus Tissue Culture Model to Test Molecular Regenerative Therapies: Validation with Epigallocatechin 3-Gallate. Int. J. Mol. Sci. 2016, 17, 1640.

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