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Acta Neurochirurgica

, Volume 156, Issue 12, pp 2365–2378 | Cite as

Shock wave trauma leads to inflammatory response and morphological activation in macrophage cell lines, but does not induce iNOS or NO synthesis

  • Mattias Günther
  • Stefan Plantman
  • Caroline Gahm
  • Anders Sondén
  • Mårten Risling
  • Tiit Mathiesen
Experimental research - Brain Injury

Abstract

Background

Experimental CNS trauma results in post-traumatic inflammation for which microglia and macrophages are vital. Experimental brain contusion entails iNOS synthesis and formation of free radicals, NO and peroxynitrite. Shock wave trauma can be used as a model of high-energy trauma in cell culture. It is known that shock wave trauma causes sub-lytic injury and inflammatory activation in endothelial cells. Mechanical disruption of red blood cells can induce iNOS synthesis in experimental systems. However, it is not known whether trauma can induce activation and iNOS synthesis in inflammatory cell lines with microglial or macrophage lineage. We studied the response and activation in two macrophage cell lines and the consequence for iNOS and NO formation after shock wave trauma.

Methods

Two macrophage cell lines from rat (NR8383) and mouse (RAW264.7) were exposed to shock wave trauma by the Flyer Plate method. The cellular response was investigated by Affymetrix gene arrays. Cell survival and morphological activation was monitored for 24 h in a Cell-IQ live cell imaging system. iNOS induction and NO synthesis were analyzed by Western blot, in cell Western IR-immunofluorescence, and Griess nitrite assay.

Results

Morphological signs of activation were detected in both macrophage cell lines. The activation of RAW264.7 was statistically significant (p < 0.05), but activation of NR8383 did not pass the threshold of statistical significance alpha (p > 0.05). The growth rate of idle cells was unaffected and growth arrest was not seen. Trauma did not result in iNOS synthesis or NO induction. Gene array analyses showed high enrichment for inflammatory response, G-protein coupled signaling, detection of stimulus and chemotaxis. Shock wave trauma combined with low LPS stimulation instead led to high enrichment in apoptosis, IL-8 signaling, mitosis and DNA-related activities. LPS/IFN-ɣ stimulation caused iNOS and NO induction and morphological activation in both cell lines.

Conclusions

Shock wave trauma by the Flyer Plate method caused an inflammatory response and morphological signs of activation in two macrophage cell lines, while iNOS induction appeared to require humoral signaling by LPS/IFN-ɣ. Our findings indicated that direct energy transfer by trauma can activate macrophages directly without humoral mediators, which comprises a novel activation mechanism of macrophages.

Keywords

Shock wave trauma Flyer Plate iNOS Nitric oxide Macrophage activation NR8383 RAW264.7 Post-traumatic inflammation 

Notes

Acknowledgments

The authors would like to thank Elisabeth Malm for excellent technical assistance. We would also like to thank the core facility at Novum, BEA, Bioinformatics and Expression Analysis, which is supported by the board of research at the Karolinska Institute and the research committee at the Karolinska Hospital. This study was funded by ALF Stockholms Läns Landsting and The Swedish Defense Agency.

Conflict of interest

All authors certify that they have NO affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript.

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Copyright information

© Springer-Verlag Wien 2014

Authors and Affiliations

  • Mattias Günther
    • 1
    • 3
  • Stefan Plantman
    • 1
  • Caroline Gahm
    • 3
  • Anders Sondén
    • 2
  • Mårten Risling
    • 1
  • Tiit Mathiesen
    • 3
  1. 1.Department of Neuroscience, Section of Experimental TraumatologyKarolinska InstitutetStockholmSweden
  2. 2.Department of SurgerySöder HospitalStockholmSweden
  3. 3.Department of Clinical Neuroscience, Section of NeurosurgeryKarolinska InstitutetStockholmSweden

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