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DOS Kongressen 2017 ·
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Bacterial biofilms: A possible mechanism for chronic
infection in patients with lumbar disc herniation – A
prospective proof-of-concept study using fluores-
cence in-situ hybridization.
Søren Ohrt-Nissen, Blaine Fritz, Jonas Walbom, Kasper Kragh, Thomas Bjarn-
sholt, Benny Dahl , Claus Manniche
Department of Orthopaedic Surgery, Spine Unit, Rigshospitalet; Department
of Immunology and Microbiology, University of Copenhagen, Faculty of Health
Sciences; Department of Orthopaedic Surgery, Spine Unit, Rigshospitalet; De-
partment of Immunology and Microbiology, University of Copenhagen, Faculty
of Health Sciences; Department of Clinical Microbiology, Rigshospitalet; Spine
surgery, Texas children’s hospital; Spine Centre of Southern Denmark, Institute
of Regional Health Service, University of Southern Denmark
Background:
A relationship has been suggested between lumbar interverte-
bral disc herniation (LDH) and chronic bacterial infection frequently involving
P. acnes, which is known to cause chronic infection through the formation of
biofilm.
Purpose / Aim of Study:
To assess whether a disc infection involving biofilm
formation is present in patients with LDH.
Materials and Methods:
Patients with LDH undergoing primary discectomy
were prospectively included. Patients operated for spinal fractures or deformi-
ties were included as controls. Bacterial 16S rDNA was purified and amplified by
real-time polymerase chain reaction (PCR). Sanger sequencing was performed
on PCR positive samples. Formalin- fixed paraffin embedded tissue sections
were stained using fluorescence in situ hybridization with peptide nucleic acid
probes (one P. acnes specific probe and one universal bacterial probe). To visual-
ize bacterial aggregates, tissue sections were examined for the first 50 included
patients by confocal laser scanning microscopy (CLSM).
Findings / Results:
A total of 51 LDH patients and 14 controls were included.
Bacterial DNA was detected by PCR in 16/51 samples in the LDH group and
7/14 controls (p=0.215). Sequencing identified bacteria in 9/16 and 6/7 PCR
positive samples in the LDH and control groups, respectively. CLSM demon-
strated tissue-embedded bacterial aggregates with host inflammatory cells in
7/44 LDH patients and no controls. Only one sample positive for aggregates by
CLSM was positive for bacterial 16S rDNA by PCR.
Conclusions:
CLSM demonstrated bacterial aggregates and inflammatory cells
in 16% of LDH patients, suggesting chronic bacterial infection. Discordance be-
tween molecular and microscopic analyses highlights the importance of a dual-
approach diagnostic strategy to discriminate infection versus contamination.
No conflicts of interest reported
139.