Abstract
Introduction
Resolution of intervertebral disc (IVD) degeneration-associated inflammation is a pre-requisite for tissue regeneration. Here, a pro-inflammatory disc organ culture model from bovine origin was established,1 and a therapy based on the intra-discal delivery of an anti-inflammatory drug (diclofenac).
Material and Methods
Bovine caudal IVD punches were needle-punctured and additionally stimulated with lipopolysaccharide (10 µg/mL) or interleukin-1β (IL-1β, 10–100 ng/mL) for 48 hours. An intradiscal therapeutic approach was tested based on a non-steroidal anti-inflammatory drug, diclofenac (Df), alone or incorporated into nanoparticles, previously developed.2
Results
IL-1β-treated IVD organ cultures showed a statistically significant up-regulation of pro-inflammatory markers (IL-6, IL-8, PGE2) and metalloproteases (MMP1, MMP3) expression while ECM proteins (Collagen II, Aggrecan) were significantly down-regulated. The injection of the anti-inflammatory drug was able to reduce the levels of pro-inflammatory cytokines and MMPs and surprisingly increased ECM protein levels.
Conclusion
These results point the intradiscal application of anti-inflammatory drugs as promising therapeutics for disc degeneration. Moreover, the organ culture model established could be used to address cellular/molecular mechanisms that regulate inflammation and IVD degeneration, and moreover to test novel therapeutic drugs, thus reducing the number of animals in in vivo experimentation.
Acknowledgments
This work was supported by European funds from FP7-EU-project GENODISC (HEALTH-F2–2008–201626), by German funds from the German Spine Foundation (Deutsche Wirbelsäulenstitung), by the Luso-German Integrated Actions Program (DAAD/CRUP, ref. A20/12), by FEDER funds through the Programa Operacional Factores de Competitividade – COMPETE, by Portuguese funds through QREN (project NORTE-07–0124-FEDER-000005) co-financed by North Portugal Regional Operational (ON.2-O Novo Norte). Graciosa Q. Teixeira and Catarina L. Pereira also acknowledge FCT for their PhD grants (SFRH/BD/88429/2012 and SFRH/BD/85779/2012, respectively).
Teixeira GQ, et al. Tissue Eng 2015; (Part C). Accepted for publication
Gonçalves RM, et al. J Mater Sci Mater Med 2015