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Susceptibility

In Vivo Gentamicin Susceptibility Test for Prevention of Bacterial Biofilms in Bone Tissue and on Implants

Louise Kruse Jensen, Thomas Bjarnsholt, Kasper N. Kragh, Bent Aalbæk, Nicole Lind Henriksen, Sophie Amalie Blirup, Karen Pankoke, Andreas Petersen, Henrik Elvang Jensen
Louise Kruse Jensen
aDepartment of Veterinary and Animal Sciences, University of Copenhagen, Copenhagen, Denmark
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Thomas Bjarnsholt
bCosterton Biofilm Center, Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark
cDepartment of Clinical Microbiology, Copenhagen University Hospital, Copenhagen, Denmark
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Kasper N. Kragh
bCosterton Biofilm Center, Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark
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Bent Aalbæk
aDepartment of Veterinary and Animal Sciences, University of Copenhagen, Copenhagen, Denmark
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Nicole Lind Henriksen
aDepartment of Veterinary and Animal Sciences, University of Copenhagen, Copenhagen, Denmark
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Sophie Amalie Blirup
aDepartment of Veterinary and Animal Sciences, University of Copenhagen, Copenhagen, Denmark
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Karen Pankoke
aDepartment of Veterinary and Animal Sciences, University of Copenhagen, Copenhagen, Denmark
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Andreas Petersen
dStatens Serum Institut, Copenhagen, Denmark
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Henrik Elvang Jensen
aDepartment of Veterinary and Animal Sciences, University of Copenhagen, Copenhagen, Denmark
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DOI: 10.1128/AAC.01889-18
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ABSTRACT

The objective of this study was to set up an in vivo gentamicin susceptibility test for biofilm prevention in bone tissue and on implants. Twenty-five pigs were allocated to six groups. Pigs in group A (n = 6) were inoculated with saline. Pigs in groups B (n = 6), C (n = 3), D (n = 3), E (n = 3), and F (n = 4) were inoculated with 10 μl saline containing 104 CFU of Staphylococcus aureus. Different concentrations based on the MIC of gentamicin for the specific strain were added to the 10-μl inoculum for groups C (160× MIC), D (1,600× MIC), E (16,000× MIC), and F (160,000× MIC). The inocula were injected into a predrilled tibial implant cavity, followed by insertion of a steel implant (2 by 15 mm). The pigs were euthanized after 5 days. In vitro, all the doses used were found to be bactericidal after up to 6 h. All implant cavities of pigs inoculated with bacteria and bacteria plus 160× MIC or 1,600× MIC of gentamicin were positive for S. aureus. In animals in each of groups E (16,000× MIC) and F (160,000× MIC), 2/3 and 1/4 of the implant cavities were S. aureus positive, respectively. By grouping groups C and D (<10,000× MIC) and groups E and F (>10,000× MIC), a significant decrease in the number of implant-attached bacteria was seen only between the high-MIC-value group and group B. Histologically, it was demonstrated that 1,600×, 16,000×, and 160,000× MIC resulted in a peri-implant tissue reaction comparable to that in saline-inoculated animals. In vivo, the antimicrobial tolerance of the inoculated planktonic bacteria was increased by in vivo-specific factors of acute inflammation. This resulted in bacterial aggregation and biofilm formation, which further increased the gentamicin tolerance. Thus, susceptibility patterns in vitro might not reflect the actual in vivo susceptibility locally within a developing infectious area.

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In Vivo Gentamicin Susceptibility Test for Prevention of Bacterial Biofilms in Bone Tissue and on Implants
Louise Kruse Jensen, Thomas Bjarnsholt, Kasper N. Kragh, Bent Aalbæk, Nicole Lind Henriksen, Sophie Amalie Blirup, Karen Pankoke, Andreas Petersen, Henrik Elvang Jensen
Antimicrobial Agents and Chemotherapy Jan 2019, 63 (2) e01889-18; DOI: 10.1128/AAC.01889-18

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In Vivo Gentamicin Susceptibility Test for Prevention of Bacterial Biofilms in Bone Tissue and on Implants
Louise Kruse Jensen, Thomas Bjarnsholt, Kasper N. Kragh, Bent Aalbæk, Nicole Lind Henriksen, Sophie Amalie Blirup, Karen Pankoke, Andreas Petersen, Henrik Elvang Jensen
Antimicrobial Agents and Chemotherapy Jan 2019, 63 (2) e01889-18; DOI: 10.1128/AAC.01889-18
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KEYWORDS

aminoglycosides
animal models
biofilms
susceptibility testing

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