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Imaging Bacteria and Biofilms on Hardware and Periprosthetic Tissue in Orthopedic Infections

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Microbial Biofilms

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1147))

Abstract

Infection is a major complication of total joint arthroplasty (TJA) surgery, and even though it is now as low as 1 % in some hospitals, the increasing number of primary surgeries translates to tens of thousands of revisions due to prosthetic joint infection (PJI). In many cases the only solution is revision surgery in which the hardware is removed. This process is extremely long and painful for patients and is a considerable financial burden for the health-care system. A significant proportion of the difficulties in diagnosis and treatment of PJI are associated with biofilm formation where bacteria attach to the surface of the prosthesis and periprosthetic tissue and build a 3-D biofilm community encased in an extracellular polymeric slime (EPS) matrix. Bacteria in biofilms have a low metabolic rate which is thought to be a major contributor to their recalcitrance to antibiotic treatment. The diagnosis of biofilm infections is difficult due to the fact that bacteria in biofilms are not readily cultured with standard clinical microbiology techniques. To identify and visualize in situ biofilm bacteria in orthopedic samples, we have developed protocols for the collection of samples in the operating room, for molecular fluorescent staining with 16S rRNA fluorescence in situ hybridization (FISH), and for imaging of samples using confocal laser scanning microscopy (CLSM). Direct imaging is the only method which can definitively identify biofilms on implants and complements both culture and culture-independent diagnostic methods.

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Acknowledgments

We thank S. Conti, MD, G. Altman MD, D. Altman MD, and N. Sotereanos, MD, Orthopedic Department, Allegheny General Hospital, Pittsburgh, PA, for providing the samples; S. Kathju, MD, PhD, University of Pittsburgh School of Medicine, Pittsburgh, PA and from the Center for Genomic Sciences, Allegheny-Singer Research Institute, Pittsburgh, PA; G.D. Ehrlich, PhD, C.J. Post, MD, PhD, and J.W. Costerton, PhD, for protocol development and provision of resources; and Mary O’Toole for her help in the preparation of the manuscript.

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Authors and Affiliations

  1. Center for Genomic Sciences, Allegheny-Singer Research Institute, Pittsburgh, USA

    Laura Nistico & Paul Stoodley

  2. National Center for Advanced Tribology, University of Southampton, Southampton, UK

    Luanne Hall-Stoodley & Paul Stoodley

  3. Center for Microbial Interface Biology and Department of Orthopeadics, The Ohio State University, 716 Biomedical Research Tower, 460 W 12th Avenue, Columbus, OH, 43210, USA

    Luanne Hall-Stoodley & Paul Stoodley

Authors
  1. Laura Nistico
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  2. Luanne Hall-Stoodley
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  3. Paul Stoodley
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Corresponding author

Correspondence to Paul Stoodley .

Editor information

Editors and Affiliations

  1. Fondazione Santa Lucia IRCCS Microbial Biofilm Laboratory, Rome, Italy

    Gianfranco Donelli

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Nistico, L., Hall-Stoodley, L., Stoodley, P. (2014). Imaging Bacteria and Biofilms on Hardware and Periprosthetic Tissue in Orthopedic Infections. In: Donelli, G. (eds) Microbial Biofilms. Methods in Molecular Biology, vol 1147. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0467-9_8

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  • DOI: https://doi.org/10.1007/978-1-4939-0467-9_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0466-2

  • Online ISBN: 978-1-4939-0467-9

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