The impact of inoculation parameters on the pathogenesis of contact lens-related infectious keratitis.
OBJECTIVE
Contact lens wear predisposes to Pseudomonas aeruginosa keratitis, but the mechanisms involved remain unclear. An in vivo model was used to study lens inoculation conditions enabling disease.


METHODS
Custom-made hydrogel contact lenses were fitted to rats after incubation in P. aeruginosa approximately 10(11) cfu/mL (3 hours) or approximately 10(3) cfu/mL (24 hours). Another group was inadvertently inoculated with a suction pen previously used with high inocula, but rinsed in ethanol and stored dry (6 months). Some corneas were tissue paper-blotted to cause fluorescein staining before lens fitting. Contralateral eyes were untreated. Twenty-four hours after disease detection, lenses were transferred to naive rats or examined by confocal microscopy before homogenization to quantify viable bacteria. After lens removal, corneas were washed to collect nonadherent bacteria and were analyzed by immunohistochemistry.


RESULTS
All eyes challenged with unworn contaminated lenses developed keratitis after approximately 7 to 10 days. Disease delay and severity were unaffected by inoculum parameters or tissue blotting but occurred sooner with lenses transferred from infected eyes ( approximately 2 days). Worn lenses and corneal washes contained infecting bacteria. Posterior, not anterior, lens surfaces harbored P. aeruginosa biofilms that penetrated the lens matrix. Diseased corneas showed an infiltration of phagocytes and T-lymphocytes.


CONCLUSIONS
P. aeruginosa induces keratitis in this lens-wearing model after a single inoculation. Delayed disease onset was interesting considering the greater keratitis risk during extended wear. Infection did not require the disruption of corneal barrier function before lens wear and occurred without exposure to lens care solutions. The data suggest that keratitis involves biofilm formation or other bacterial adaptations in vivo.
School of Optometry, University of California, Berkeley, California 94720-2020, USA.
Connie
C
Tam
James J
JJ
Mun
David J
DJ
Evans
Suzanne M J
SM
Fleiszig
D000818:Animals; D001422:Bacterial Adhesion; D001431:Bacteriological Techniques; D018441:Biofilms; D003263:Contact Lenses, Hydrophilic / Q000382:microbiology*; D003320:Corneal Ulcer / Q000378:metabolism*; D004195:Disease Models, Animal; D015818:Eye Infections, Bacterial / Q000382:microbiology*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D018613:Microscopy, Confocal; D011552:Pseudomonas Infections / Q000382:microbiology*; D011550:Pseudomonas aeruginosa / Q000502:physiology*; D051381:Rats; D011917:Rats, Inbred Lew
D020136:Hydrogel, Polyethylene Glycol Dimethacrylate