Hypoxic conditions on the cornea exacerbate the binding and internalization of P. aeruginosa due to increased levels of CFTR expression and also induce basal NF-kappaB activation. Both of these responses probably exacerbate the effects of P. aeruginosa infection by allowing lower infectious doses of bacteria to induce disease and promote destructive inflammatory responses.
Objectives
This study used a rat contact lens (CL) model to test if high vs. low Dk lens wear caused changes in: 1) conjunctival Langerhans cell (LC) number or location; 2) Bcl-2 expression; and 3) infection risk.
Methods
Female, Lewis rats wore a high or low Dk CL continuously for 2 weeks. Afterward, corneas were harvested and processed for ADPase activity to identify Langerhans cells (LC), for immunostaining and for real time RT-PCR. CL wearing rats also were challenged with Pseudomonas aeruginosa by placing a bacterial-soaked CL on the eye followed by topical delivery of bacteria. After 48 hours, slit lamp examination and real time RT-PCR were used to evaluate the corneal response.
Results
Conjunctival LC were significantly increased after low vs. high Dk CL wear (p<0.0001). In contrast, conjunctival LC in non-lens wearing rats was not significantly different from the high Dk lens wearing group. Bcl-2 mRNA levels were significantly decreased in low vs. high Dk Cl wearing rats, while Bax, FasL, caspase 3 and caspase 9 levels were unchanged. Immunostaining for Bcl-2 showed fewer positively stained epithelial cells in the low vs. high Dk lens wearing group. After bacterial challenge, 30% of low vs. none of the high Dk CL wearing corneas became infected and showed increased mRNA levels for several pro-inflammatory cytokines/chemokines, inducible nitric oxide synthase (iNOS) and matrix metalloproteinase (MMP)-9.
Conclusion
Low vs. high Dk and/or no CL wear led to an increased number of conjunctival LC, decreased Bcl-2 levels, and increased the risk of bacterial infection.
Only Clear Care/AOSEPT Plus and BLUE Vision/BLUE SEPT showed high levels of antimicrobial activity against the cyst form of A. castellani. Oxysept 1 Step showed mild activity against the cysts and easyvision one step+ and OPTI-FREE Express with Aldox showed virtually no antiacanthamoeba activity against the cyst form after 6 hours of exposure.
Determination of a D value for specific test organisms is a component of the efficacy evaluation of new contact lens disinfecting solutions. This parameter is commonly defined as the time required for the number of surviving microorganisms to decrease 1 logarithmic unit. The assumption made in establishing a D value is that the rate of kill exhibits first-order kinetics under the specified conditions. Such exponential kill rates are seen with thermal contact lens disinfection systems. A comparison of the death rate kinetics for a variety of chemical contact lens disinfecting solutions was undertaken to ascertain the suitability of D-value determination for these chemical disinfectants. The active agents of these different solutions included hydrogen peroxide, thimerosal, chlorhexidine, tris(2-hydroxyethyl)tallow ammonium chloride, thimerosal, polyaminopropyl biguanide, and polyquaternium-1. The solutions were challenged with 106 CFU of either Pseudomonas aeruginosa, Serratia marcescens, or Staphylococcus hominis per ml, and survival rate was determined. This study clearly demonstrates the nonlinear nature of the inactivation curves for most contact lens chemical disinfecting solutions for the challenge organisms. D-value determination is, therefore, an inappropriate method of reporting the biocidal activity of these solutions.
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