Propionibacterium acnes biofilm is present in intervertebral discs of patients undergoing microdiscectomy

PLoS One. 2017 Apr 3;12(4):e0174518. doi: 10.1371/journal.pone.0174518. eCollection 2017.

Abstract

Background: In previous studies, Propionibacterium acnes was cultured from intervertebral disc tissue of ~25% of patients undergoing microdiscectomy, suggesting a possible link between chronic bacterial infection and disc degeneration. However, given the prominence of P. acnes as a skin commensal, such analyses often struggled to exclude the alternate possibility that these organisms represent perioperative microbiologic contamination. This investigation seeks to validate P. acnes prevalence in resected disc cultures, while providing microscopic evidence of P. acnes biofilm in the intervertebral discs.

Methods: Specimens from 368 patients undergoing microdiscectomy for disc herniation were divided into several fragments, one being homogenized, subjected to quantitative anaerobic culture, and assessed for bacterial growth, and a second fragment frozen for additional analyses. Colonies were identified by MALDI-TOF mass spectrometry and P. acnes phylotyping was conducted by multiplex PCR. For a sub-set of specimens, bacteria localization within the disc was assessed by microscopy using confocal laser scanning and FISH.

Results: Bacteria were cultured from 162 discs (44%), including 119 cases (32.3%) with P. acnes. In 89 cases, P. acnes was cultured exclusively; in 30 cases, it was isolated in combination with other bacteria (primarily coagulase-negative Staphylococcus spp.) Among positive specimens, the median P. acnes bacterial burden was 350 CFU/g (12 - ~20,000 CFU/g). Thirty-eight P. acnes isolates were subjected to molecular sub-typing, identifying 4 of 6 defined phylogroups: IA1, IB, IC, and II. Eight culture-positive specimens were evaluated by fluorescence microscopy and revealed P. acnes in situ. Notably, these bacteria demonstrated a biofilm distribution within the disc matrix. P. acnes bacteria were more prevalent in males than females (39% vs. 23%, p = 0.0013).

Conclusions: This study confirms that P. acnes is prevalent in herniated disc tissue. Moreover, it provides the first visual evidence of P. acnes biofilms within such specimens, consistent with infection rather than microbiologic contamination.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Biofilms / growth & development*
  • Diskectomy
  • Female
  • Gram-Positive Bacterial Infections / complications
  • Gram-Positive Bacterial Infections / microbiology
  • Humans
  • Intervertebral Disc / microbiology*
  • Intervertebral Disc / surgery
  • Intervertebral Disc Degeneration / etiology
  • Intervertebral Disc Degeneration / microbiology
  • Intervertebral Disc Displacement / etiology
  • Intervertebral Disc Displacement / microbiology*
  • Intervertebral Disc Displacement / surgery
  • Male
  • Middle Aged
  • Phenotype
  • Propionibacterium acnes / isolation & purification*
  • Propionibacterium acnes / pathogenicity
  • Propionibacterium acnes / physiology*
  • Young Adult