April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Neutrophil-enhanced Pseudomonas aeruginosa Biofilms on Silicone Hydrogel Contact Lenses
Author Affiliations & Notes
  • Geoffrey W. Burnham
    Ophthalmology, Univ of Texas Southwestern Med Ctr, Dallas, Texas
  • H D. Cavanagh
    Ophthalmology, Univ Texas Southwestern Med Ctr, Dallas, Texas
  • Danielle M. Robertson
    Ophthalmology, Univ Texas Southwestern Med Ctr, Dallas, Texas
  • Footnotes
    Commercial Relationships  Geoffrey W. Burnham, None; H. D. Cavanagh, None; Danielle M. Robertson, None
  • Footnotes
    Support  NIH R01 EY018219 (DMR), EY020799, OneSight Research Foundation, Dallas, TX (DMR), & a Career Development Award (DMR) & an unrestricted grant from Research to Prevent Blindness, Inc., New York, NY
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 6506. doi:
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    • Get Citation

      Geoffrey W. Burnham, H D. Cavanagh, Danielle M. Robertson; Neutrophil-enhanced Pseudomonas aeruginosa Biofilms on Silicone Hydrogel Contact Lenses. Invest. Ophthalmol. Vis. Sci. 2011;52(14):6506.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : In animal models, extended contact lens wear with concurrent bacterial challenge results in significant neutrophil accumulation. Recently we have reported that Pseudomonas aeruginosa (PA) biofilms formed on hydrogel lenses are significantly increased in the presence of neutrophils.The purpose of this study is to evaluate neutrophil-enhanced PA biofilm formation on silicone hydrogel contact lenses.

Methods: : A fully invasive corneal isolate that has been stably conjugated to GFP, strain PA6487, was used in all experiments. Neutrophils were isolated from human blood using a percoll gradient separation and activated by brief exposure to phorbol 12-myristate 13-acetate. Neutrophils were utilized at a concentration of 8x106cells/well. Unworn lotrafilcon A lenses were incubated in 1mL of bacterial suspension at a concentration of 1x108CFU/mL for 2 hours. After being washed in PBS, lenses were incubated overnight in RPMI with 2% Heat inactivated platelet poor plasma either with or without neutrophils. The lenses were incubated in their respective solutions for 24 hours at 37OC. Biofilm formation was evaluated using laser scanning confocal microscopy and colony forming unit (CFU) analysis.

Results: : Primary attachment of PA was confirmed at 2 hours by confocal microscopy and CFU analysis. No bacteria were harvested from the non-bacteria exposed control lens. After a 24 hour incubation in the presence of neutrophils, confocal microscopy demonstrated increased density and an alteration of the architecture of the biofilm. CFU analysis confirmed an increase in viable PA in the neutrophil-enhanced biofilm (p=0.002).

Conclusions: : These data demonstrate that intense corneal inflammation associated with PA contamination results in enhanced biofilm formation on silicone hydrogel contact lenses. Further studies are required to determine the efficacy of currently available cleaning regimens against neutrophil-enhanced biofilms on contact lens surfaces.

Keywords: contact lens • pseudomonas • inflammation 
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