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PLoS By Category | Recent PLoS Articles
Infectious Diseases - Surgery

Monitoring Bacterial Burden, Inflammation and Bone Damage Longitudinally Using Optical and µCT Imaging in an Orthopaedic Implant Infection in Mice
Published: Wednesday, October 17, 2012
Author: Jared A. Niska et al.

by Jared A. Niska, Jeffrey A. Meganck, Jonathan R. Pribaz, Jonathan H. Shahbazian, Ed Lim, Ning Zhang, Brad W. Rice, Ali Akin, Romela Irene Ramos, Nicholas M. Bernthal, Kevin P. Francis, Lloyd S. Miller

Background

Recent advances in non-invasive optical, radiographic and µCT imaging provide an opportunity to monitor biological processes longitudinally in an anatomical context. One particularly relevant application for combining these modalities is to study orthopaedic implant infections. These infections are characterized by the formation of persistent bacterial biofilms on the implanted materials, causing inflammation, periprosthetic osteolysis, osteomyelitis, and bone damage, resulting in implant loosening and failure.

Methodology/Principal Findings

An orthopaedic implant infection model was used in which a titanium Kirshner-wire was surgically placed in femurs of LysEGFP mice, which possess EGFP-fluorescent neutrophils, and a bioluminescent S. aureus strain (Xen29; 1×103 CFUs) was inoculated in the knee joint before closure. In vivo bioluminescent, fluorescent, X-ray and µCT imaging were performed on various postoperative days. The bacterial bioluminescent signals of the S. aureus-infected mice peaked on day 19, before decreasing to a basal level of light, which remained measurable for the entire 48 day experiment. Neutrophil EGFP-fluorescent signals of the S. aureus-infected mice were statistically greater than uninfected mice on days 2 and 5, but afterwards the signals for both groups approached background levels of detection. To visualize the three-dimensional location of the bacterial infection and neutrophil infiltration, a diffuse optical tomography reconstruction algorithm was used to co-register the bioluminescent and fluorescent signals with µCT images. To quantify the anatomical bone changes on the µCT images, the outer bone volume of the distal femurs were measured using a semi-automated contour based segmentation process. The outer bone volume increased through day 48, indicating that bone damage continued during the implant infection.

Conclusions/Significance

Bioluminescent and fluorescent optical imaging was combined with X-ray and µCT imaging to provide noninvasive and longitudinal measurements of the dynamic changes in bacterial burden, neutrophil recruitment and bone damage in a mouse orthopaedic implant infection model.

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