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Antibodies directed against integration host factor mediate biofilm clearance from nasopore

Authors

  • Kathleyn A. Brandstetter BA,

    1. Department of Otolaryngology–Head and Neck Surgery, The Ohio State University, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, U.S.A.
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  • Joseph A. Jurcisek BS,

    1. Center for Microbial Pathogenesis, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, U.S.A.
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  • Steven D. Goodman PhD,

    1. Center for Microbial Pathogenesis, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, U.S.A.
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  • Lauren O. Bakaletz PhD,

    1. Department of Otolaryngology–Head and Neck Surgery, The Ohio State University, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, U.S.A.
    2. Center for Microbial Pathogenesis, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, U.S.A.
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  • Subinoy Das MD

    Corresponding author
    1. Department of Otolaryngology–Head and Neck Surgery, The Ohio State University, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, U.S.A.
    2. Center for Microbial Pathogenesis, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio, U.S.A.
    • Send correspondence to Subinoy Das, MD, 915 Olentangy River Road, Suite 4000, Columbus, OH 43212. E-mail: Subinoy.Das@osumc.edu

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  • Editor's Note: This Manuscript was accepted for publication on April 5, 2013.

  • This manuscript was presented at the 2013 Combined Sections Meeting, The Triological Society, in Scottsdale, Arizona, January 26, 2013.

  • Financial Disclosure: This work was supported, in part, from a grant from the NIH/NIDCD 5R01-DC011818 (Bakaletz). The authors have no other funding, financial relationships, or conflicts of interest to disclose.

Abstract

Objectives/Hypothesis

Intranasal resorbable packing, such as Nasopore, is commonly used during sinus surgery despite a paucity of evidence that demonstrates clinical benefit. We theorized that Nasopore supports bacterial growth and biofilm formation. The DNABII family of bacterial nucleic acid binding proteins stabilizes the extracellular polymeric substance of the biofilm, thus protecting bacteria from host defenses and traditional antibiotics. We tested the hypothesis that use of anti-IHF antibodies in conjunction with antibiotics would enhance biofilm eradication from Nasopore.

Study Design

In vitro experiments.

Methods

Nontypeable Haemophilus influenzae (NTHI) biofilms were grown on Nasopore. Following 24-hour incubation, biofilms were incubated for an additional 16 hours with either medium alone, naïve rabbit serum, rabbit anti-IHF serum, amoxicillin/clavulanate, or anti-IHF serum + amoxicillin/clavulanate. Computer statistics (COMSTAT) analysis was performed on images of biofilms obtained via confocal microscopy.

Results

NTHI readily formed a biofilm on Nasopore. Treatment with amoxicillin/clavulanate alone mediated an increase in biomass by 92% to 6.63 μ23 compared to incubation in sterile medium alone (3.46 μ23). Treatment with anti-IHF alone reduced the biomass by 77% to 1.29 μ23 compared to incubation with naïve rabbit serum (5.53 μ23). Anti-IHF + amoxicillin/clavulanate reduced biomass by 88% to 0.66 μ23 (P <0.02) compared to incubation with naïve rabbit serum.

Conclusion

Antibiotics alone were ineffective in eradicating NTHI biofilms that had formed on Nasopore in vitro. Anti-IHF antibodies plus amoxicillin/clavulanate therapy synergistically reduced biofilm biomass by 88%. These data support clinical studies for the use of anti-IHF combined with antibiotics to reduce biofilm formation on intranasal packing.

Level of Evidence

N/A. Laryngoscope, 123:2626–2632, 2013

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