Disruption of contact lens-associated Pseudomonas aeruginosa biofilms formed in the presence of neutrophils.
OBJECTIVE
To evaluate the capacity of neutrophils to enhance biofilm formation on contact lenses by an infectious Pseudomonas aeruginosa (PA) corneal isolate. Agents that target F-actin and DNA were tested as a therapeutic strategy for disrupting biofilms formed in the setting of neutrophils in vitro and for limiting the infectious bioburden in vivo.


METHODS
Biofilm formation by infectious PA strain 6294 was assessed in the presence of neutrophils on a static biofilm plate and on unworn etafilcon A soft contact lenses. A d-isomer of poly(aspartic acid) was used alone and with DNase to reduce biofilm formation on test contact lenses. The gentamicin survival assay was used to determine the effectiveness of the test compound in reducing subsequent intracellular bacterial load in the corneal epithelium in a contact lens infection model in the rabbit.


RESULTS
In a static reactor and on hydrogel lenses, PA biofilm density was enhanced 30-fold at 24 hours in the presence of neutrophils (P < 0.0001). The combination of DNase and anionic poly(aspartic acid) reduced the PA biofilms formed in the presence of activated neutrophils by 79.2% on hydrogel contact lenses (P < 0.001). An identical treatment resulted in a 41% reduction in internalized PA in the rabbit corneal epithelium after 24 hours (P = 0.03).


CONCLUSIONS
These results demonstrate that PA can exploit the presence of neutrophils to form biofilm on contact lenses within a short time. Incorporation of F-actin and DNA represent a mechanism for neutrophil-induced biofilm enhancement and are targets for available agents to disrupt pathogenic biofilms formed on contact lenses and as a treatment for established corneal infections.
Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9057, USA. danielle.robertson@utsouthwestern.edu
Danielle M
DM
Robertson
Quinn M
QM
Parks
Robert L
RL
Young
Jennifer
J
Kret
Katie R
KR
Poch
Kenneth C
KC
Malcolm
David P
DP
Nichols
Michelle
M
Nichols
Meifang
M
Zhu
H Dwight
HD
Cavanagh
Jerry A
JA
Nick
D000818:Animals; D001422:Bacterial Adhesion / Q000502:physiology; D058491:Bacterial Load; D018441:Biofilms / Q000187:drug effects*; D003263:Contact Lenses, Hydrophilic / Q000382:microbiology*; D003320:Corneal Ulcer / Q000382:microbiology / Q000517:prevention & control; D003851:Deoxyribonucleases / Q000494:pharmacology*; D004359:Drug Therapy, Combination; D019573:Epithelium, Corneal / Q000382:microbiology; D015818:Eye Infections, Bacterial / Q000382:microbiology / Q000517:prevention & control*; D006801:Humans; D008689:Methacrylates; D009504:Neutrophils / Q000502:physiology*; D010455:Peptides / Q000494:pharmacology*; D011552:Pseudomonas Infections / Q000382:microbiology / Q000517:prevention & control; D011550:Pseudomonas aeruginosa / Q000502:physiology*; D011817:Rabbits
D008689:Methacrylates; D010455:Peptides; C114176:etafilcon; C017645:polyaspartate; D003851:Deoxyribonucleases
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