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Miechia A. Esco, Linda D. Hazlett, Michelle Kurpakus-Wheater; Pseudomonas aeruginosa Binds to Extracellular Matrix Deposited by Human Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2002;43(12):3654-3659.
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purpose. To measure the effect of extracellular matrix substrate, pH, and O2 on Pseudomonas aeruginosa binding.
methods. Extracellular matrix substrates were prepared from human corneal epithelial cells cultured in 2% or 20% O2. P. aeruginosa strains ATCC 19660 or PAO1 (suspended in pH 7.0 or 7.5 buffer) were cultured on extracellular matrix substrates in 2% or 20% O2. The mean number of adherent bacteria per counted per field ± SEM (n = 15) was determined for combinations of bacteria, extracellular matrix substrate, pH, and O2. Binding in the presence of antibodies directed against laminin-5 was also measured.
results. Extracellular matrix substrates produced by cells cultured in 20% O2, combined with an environment of pH 7.0, provided the least favorable conditions for binding of strain 19660. In contrast, extracellular matrix substrates produced by cells cultured in 2% O2, combined with an environment of pH 7.0, provided the most favorable conditions for binding of strain 19660. Binding of PAO1, however, as a function of extracellular matrix substrate and pH, did not similarly compare with binding of strain 19660. Antibodies against laminin-5 chains served to increase the number of strain 19660 bacteria bound to extracellular matrix substrates compared with the control.
conclusions. The extracellular matrix secreted by hypoxic corneal epithelial cells is a substrate for binding of P. aeruginosa. Results in previous studies have shown that hypoxic extracellular matrix contains less laminin-5 protein than normoxic matrix. The antibody studies in this report suggest that the decrease in laminin-5 content in hypoxic matrix, relative to matrix secreted by normoxic corneal epithelium, may be responsible for increased bacterial adhesion.
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