title: Quantifying the natural history of biofilm formation in vivo during the establishment of chronic implant-associated Staphylococcus aureus osteomyelitis in mice to identify critical pathogen and host factors.
abstract: 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 ∼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.
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York.
authors.forename: Kohei
authors.initials: K
authors.lastname: Nishitani
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York.
authors.forename: Werasak
authors.initials: W
authors.lastname: Sutipornpalangkul
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York.|Pathology and Laboratory Medicine, University of Rochester Medical Center, Rochester, New York.
authors.forename: Karen L
authors.initials: KL
authors.lastname: de Mesy Bentley
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York.
authors.forename: John J
authors.initials: JJ
authors.lastname: Varrone
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York.
authors.forename: Sheila N
authors.initials: SN
authors.lastname: Bello-Irizarry
authors.affiliation: Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
authors.forename: Hiromu
authors.initials: H
authors.lastname: Ito
authors.affiliation: Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
authors.forename: Shuichi
authors.initials: S
authors.lastname: Matsuda
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York.|Department of Orthopedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York.
authors.forename: Stephen L
authors.initials: SL
authors.lastname: Kates
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York.
authors.forename: John L
authors.initials: JL
authors.lastname: Daiss
authors.affiliation: Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, New York.|Department of Orthopedics and Rehabilitation, University of Rochester Medical Center, Rochester, New York.
authors.forename: Edward M
authors.initials: EM
authors.lastname: Schwarz
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D018441:Biofilms; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D004195:Disease Models, Animal; D005260:Female; D008422:Materials Testing; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008855:Microscopy, Electron, Scanning; D010019:Osteomyelitis / Q000382:microbiology*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D019736:Prostheses and Implants / Q000382:microbiology*; D016459:Prosthesis-Related Infections / Q000382:microbiology*; D013045:Species Specificity; D013203:Staphylococcal Infections / Q000517:prevention & control*; D013234:Stem Cells; D013977:Tibia / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D020099:Coated Materials, Biocompatible
keywords: Staphylococcus aureus; biofilm; implant; osteomyelitis; scanning electron microscopy
references.citation: JAMA. 2012 Sep 26;308(12):1227-36
references.pmid: 23011713
references.citation: J Arthroplasty. 2012 Sep;27(8 Suppl):61-5.e1
references.pmid: 22554729
references.citation: J Microbiol Methods. 2012 Aug;90(2):115-8
references.pmid: 22542521
references.citation: J Orthop Res. 2012 Jul;30(7):1176-80
references.pmid: 22228044
references.citation: PLoS Pathog. 2012;8(4):e1002626
references.pmid: 22496652
references.citation: Infect Immun. 2012 Feb;80(2):733-41
references.pmid: 22104103
references.citation: Proc Natl Acad Sci U S A. 2011 Dec 13;108(50):20172-7
references.pmid: 22123973
references.citation: Infect Immun. 2011 Dec;79(12):5010-8
references.pmid: 21947772
references.citation: J Bacteriol. 2010 Nov;192(21):5663-73
references.pmid: 20817770
references.citation: J Orthop Res. 2010 Jan;28(1):55-61
references.pmid: 19610092
references.citation: N Engl J Med. 2009 Aug 20;361(8):787-94
references.pmid: 19692690
references.citation: FEMS Immunol Med Microbiol. 2008 Jan;52(1):13-22
references.pmid: 18081847
references.citation: Appl Environ Microbiol. 2008 Jan;74(2):470-6
references.pmid: 18039822
references.citation: J Orthop Res. 2008 Jan;26(1):96-105
references.pmid: 17676625
references.citation: N Engl J Med. 2007 Aug 16;357(7):654-63
references.pmid: 17699815
references.citation: J Orthop Sci. 2007 Mar;12(2):178-84
references.pmid: 17393274
references.citation: Am J Infect Control. 2005 Sep;33(7):385-91
references.pmid: 16153484
references.citation: Infect Immun. 1995 Sep;63(9):3373-80
references.pmid: 7642265
references.citation: Annu Rev Microbiol. 1994;48:585-617
references.pmid: 7826020
references.citation: J Bacteriol. 2004 Jul;186(14):4665-84
references.pmid: 15231800
references.citation: J Orthop Res. 2014 Oct;32(10):1389-96
references.pmid: 24992290
references.citation: Antimicrob Agents Chemother. 2014 Aug;58(8):4353-61
references.pmid: 24841260
references.citation: Infect Immun. 2014 May;82(5):2125-34
references.pmid: 24614654
references.citation: Nat Rev Microbiol. 2014 Jan;12(1):49-62
references.pmid: 24336184
references.citation: BMC Infect Dis. 2013;13:47
references.pmid: 23356488
references.citation: Antimicrob Agents Chemother. 2013 Feb;57(2):855-63
references.pmid: 23208713
references.citation: Antimicrob Agents Chemother. 2012 Nov;56(11):5923-37
references.pmid: 22964248