GEORG SCHMORL PRIZE OF THE GERMAN SPINE SOCIETY (DWG) 2018: combined inflammatory and mechanical stress weakens the annulus fibrosus: evidences from a loaded bovine AF organ culture

  • Taryn Saggese
  • Graciosa Q. Teixeira
  • Kelly Wade
  • Lydia Moll
  • Anita Ignatius
  • Hans-Joachim Wilke
  • Raquel M. Goncalves
  • Cornelia Neidlinger-WilkeEmail author
Original Article



The pathomechanism of annulus fibrosus (AF) failure is still unknown. We hypothesise that mechanical overload and an inflammatory microenvironment contribute to AF structural weakening. Therefore, the objective of this study was to investigate the influence of these factors on the AF, particularly the translamellar bridging network (TLBN) which connects the AF lamellae.


A bovine AF organ culture (AF-OC) model of standardised AF rings was used to study the individual and combined effects of cyclic tensile strain (CTS) and IL-1β (1 ng/mL) culture medium supplementation. AF-OCs were analysed for PGE2 production (ELISA) and deposition of IL-6, COX-2, fibrillin, and MMP3 in the tissue (immunohistochemistry, IHC). The mechanical strength of the TLBN was evaluated using a peel test to measure the strength required to separate an AF segment along a lamellar bound.


The combination of CTS + IL-1β led to a significant increase in PGE2 production compared to Control (p < 0.01). IHC evaluations showed that the CTS + IL-1β group exhibited higher production of COX-2 and MMP3 within the TLBN regions compared to the adjacent lamellae and a significant increase in IL-6 ratio compared to Control (p < 0.05). A significant decrease in the annular peel strength was observed in the CTS + IL1β group compared to Control (p < 0.05).


Our findings suggest that CTS and IL-1β act synergistically to increase pro-inflammatory and catabolic molecules within the AF, particularly the TLBN, leading to a weakening of the tissue. This standardised model enables the investigation of AF/TLBN structure–function relationship and is a platform to test AF-focused therapeutics.

Graphical abstract

These slides can be retrieved under Electronic Supplementary Material.


Annulus fibrosus Organ culture Inflammation Mechanical loading Disc herniation 



The authors of the present study wish to thank the local certified abattoir Fleischmarkt Donautal for kindly providing to us the bovine tails. We wish to thank Mrs. Iris Baum, Mrs. Patrizia Horny, Mrs. Marion Tomo, Mrs. Anna Weigl, Mrs. Anastasia Raiber, Mr. Zhiyao Yong and Mr. Alexander Vogel for the excellent support. We also acknowledge the German Spine Foundation (Deutsche Wirbelsäulenstitung), the German Academic Exchange Service (DAAD), the Conselho de Reitores das Universidades Portuguesas, the Ulm University (L.SBN.0157) and the Alexander von Humboldt Foundation for their financial support. The funding agencies did not have any involvement in the study design, data collection/analysis/interpretation, manuscript preparation or in the decision to submit the manuscript for publication.

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.

Ethical approval

This study did not involve animal experiments or any studies with human participants performed by any of the authors. The bovine tails were obtained from cattle which were slaughtered for alimentary purposes.

Supplementary material

586_2019_5901_MOESM1_ESM.pptx (450 kb)
Supplementary material 1 (PPTX 450 kb)
586_2019_5901_MOESM2_ESM.docx (660 kb)
Supplementary material 2 (DOCX 660 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of Orthopaedic Research and Biomechanics, Trauma Research CentreUlm UniversityUlmGermany
  2. 2.Instituto de Investigação e Inovação em Saúde (i3S)Universidade do PortoPortoPortugal
  3. 3.Instituto de Engenharia Biomédica (INEB)Universidade do PortoPortoPortugal
  4. 4.Instituto de Ciências Biomédicas Abel Salazar (ICBAS)Universidade do PortoPortoPortugal

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