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Original Investigation |

Inhibition of Otopathogenic Biofilms by Organoselenium-Coated Tympanostomy Tubes

James C. Wang, BS1; Phat L. Tran, PhD2; Rob Hanes, PhD3; Joehassin Cordero, MD4; John Marchbanks, MD4; Ted W. Reid, PhD2; Jane A. Colmer-Hamood, PhD5; Abdul N. Hamood, PhD5
[+] Author Affiliations
1School of Medicine, Texas Tech University Health Sciences Center, Lubbock
2Department of Ophthalmology and Visual Sciences, Texas Tech University Health Sciences Center, Lubbock
3Selenium Ltd, Austin, Texas
4Division of Otolaryngology–Head and Neck Surgery, Department of Surgery, Texas Tech University Health Sciences Center, Lubbock
5Department of Immunology and Molecular Microbiology, Texas Tech University Health Sciences Center, Lubbock
JAMA Otolaryngol Head Neck Surg. 2013;139(10):1009-1016. doi:10.1001/jamaoto.2013.4690.
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Importance  Tube occlusion and post–tympanostomy tube otorrhea (PTTO) are 2 major sequelae of tympanostomy tube placement. Plugging negates the function of the tympanostomy tubes and, along with chronic PTTO, can be financially burdensome owing to repeated surgical procedures and additional treatments.

Objective  To investigate the effectiveness of an organoselenium (OSe) coating on Donaldson tympanostomy tubes in inhibiting biofilm formation on the tympanostomy tubes.

Design  In vitro microbiologic study; all experiments were performed in a Texas Tech University Health Sciences Center basic sciences laboratory.

Interventions  Inhibition of biofilm formation was investigated by incubating OSe-coated vs uncoated (control) tympanostomy tubes in a nutrient broth containing either Staphylococcus aureus (Sa) expressing green fluorescent protein (GFP), nontypeable Haemophilus influenzae (NTHi) expressing GFP, or Moraxella catarrhalis (Mc) for 48 hours at 37°C. All biofilms were quantified via colony-forming unit (CFU) assays. The Sa and NTHi biofilms were visualized using confocal laser-scanning microscopy (CLSM) and analyzed using the COMSTAT program.

Main Outcomes and Measures  The CFU assays, CLSM, and COMSTAT analysis revealed that compared with uncoated control tympanostomy tubes, OSe-coated tympanostomy tubes are able to inhibit Sa, NTHi, and Mc biofilm formation.

Results  The Sa and NTHi developed thick mature biofilms containing considerable biomass on uncoated tympanostomy tubes as determined by CLSM and COMSTAT analysis, while the OSe coating on the tympanostomy tubes drastically inhibited biofilm formation by Sa and NTHi. Quantitative CFU analysis revealed that this reduction in biofilm formation was significant, 6 logs for Sa (P < .001) and 4 logs for NTHi (P = .02). OSe coating also inhibited biofilm formation by Mc with a 4.5-log reduction (P < .001).

Conclusions and Relevance  The OSe coating is a potential long-lasting agent to prevent biofilm development on tympanostomy tubes by otopathogens.

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Figures

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Figure 1.
Staphylococcus aureus AH133 That Has Formed a Biofilm on the Outer and Inner Surfaces of Uncoated Tympanostomy Tubes

Biofilms were developed as described in the text, and the outer (A) and inner (B) surfaces of the tympanostomy tubes were examined by confocal laser-scanning microscopy. C and D, Three-dimensional reconstruction of A and B, respectively (NIS-Elements software, version 2.2; Nikon Instruments).

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Figure 2.
Organoselenium-Coated Tympanostomy Tubes That Inhibited Development of Biofilm by Otopathogens

Staphylococcus aureus–AH133 (A), nontypeable Haemophilus influenzae–86-028NP (B), and Moraxella catarrhalis–43617 (C) were allowed to develop 48-hour biofilms, and the number of micro-organisms (colony-forming units per tympanostomy tube) was determined. Values represent the mean of at least 3 independent experiments ± standard error of the mean. CFU indicates colony-forming units; OSe, organoselenium.

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Figure 3.
Organoselenium-Coated Tympanostomy Tubes That Inhibited Development of Staphylococcus aureus–AH133 and Nontypeable Haemophilus influenzae–86-028NP Biofilm

Representative 48-hour biofilms formed on outer (A) and inner (B) surfaces of uncoated and organoselenium (OSe)-coated tympanostomy tubes with 3-dimensional reconstructions below. Numbers indicate corresponding location of structures visualized on tympanostomy tubes with location on images of Donaldson-type tympanostomy tubes (used with permission of Medtronic Inc).

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Figure 4.
Organoselenium-Coated Tympanostomy Tubes Stored for 9 Months That Inhibited Biofilm Development by Staphylococcus aureus–AH133

Biofilms were developed for 48 hours, and the number of colony-forming units (CFUs) per tympanostomy tube present in the biofilm was determined. Values represent the mean of at least 3 independent experiments ± standard error of the mean. OSe indicates organoselenium.

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Figure 5.
Organoselenium-Coated Tympanostomy Tubes That Inhibited Planktonic Growth of Staphylococcus aureus–AH133 in Biofilm Medium

Biofilms were developed, and planktonic cells were harvested from the medium and quantified by colony-forming unit (CFU) assay. Values represent the mean of at least 3 independent experiments ± standard error of the mean. OSe indicates organoselenium.

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