Journal
JOURNAL OF ORTHOPAEDIC RESEARCH
Volume 33, Issue 9, Pages 1311-1319Publisher
WILEY
DOI: 10.1002/jor.22907
Keywords
Staphylococcus aureus; biofilm; implant; osteomyelitis; scanning electron microscopy
Categories
Funding
- AOTrauma
- National Institutes of Health [P30AR061307, P50AR054041, S10RR026542, R43AI85844, T32AR53459]
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While it is well known that Staphylococcus aureus establishes chronic implant-associated osteomyelitis by generating and persisting in biofilm, research to elucidate pathogen, and host specific factors controlling this process has been limited due to the absence of a quantitative in vivo model. To address this, we developed a murine tibia implant model with ex vivo region of interest (ROI) imaging analysis by scanning electron microscopy (SEM). Implants were coated with Staphylococcus aureus strains (SH1000, UAMS-1, USA300LAC) with distinct in vitro biofilm phenotypes, were used to infect C57BL/6 or Balb/c mice. In contrast to their in vitro biofilm phenotype, results from all bacteria strains in vivo were similar, and demonstrated that biofilm on the implant is established within the first day, followed by a robust proliferation phase peaking on Day 3 in Balb/c mice, and persisting until Day 7 in C57BL/6 mice, as detected by SEM and bioluminescent imaging. Biofilm formation peaked at Day 14, covering similar to 40% of the ROI coincident with massive agr-dependent bacterial emigration, as evidenced by large numbers of empty lacunae with few residual bacteria, which were largely culture negative (80%) and PCR positive (87.5%), supporting the clinical relevance of this implant model. (C) 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.
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