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Accumulation of Ciprofloxacin and Minocycline by Cultured Human Gingival Fibroblasts

¹ Sections of Oral Biology and
² Periodontology, College of Dentistry, The Ohio State University Health Sciences Center, 305 West 12th Avenue, PO Box 182357, Columbus, OH 43218-2357;

View larger version (23K): [in this window] [in a new window]   Figure 1. Temperature-dependence of ciprofloxacin and minocycline accumulation by gingival fibroblasts. After incubation of cell monolayers in HBSS at 4° (•) and 37°C (•), 20 µg/mL ciprofloxacin or minocycline was added, and uptake was monitored over the indicated time intervals. The data represent the mean + SEM of 4 experiments. Insets: Lineweaver-Burk plots of ciprofloxacin (•) and minocycline () transport observed over 3 min at 37°C.

View larger version (22K): [in this window] [in a new window]   Figure 2. The pH-dependence and reversibility of ciprofloxacin and minocycline transport by gingival fibroblasts. Upper panel: Confluent cell monolayers were washed and overlaid with HBSS adjusted to the indicated pH. The kinetics of transport was assayed at 37°C, analyzed by the Lineweaver-Burk method and expressed as a percentage of the Vmax/Km ratio observed at pH 7.2. Data are presented as the mean + SEM of at least 3 experiments. Changes in pH produced a significant overall effect on ciprofloxacin transport (P < 0.02, ANOVA). Treatments that produced a significant difference from pH 7.2 are indicated by * (P < 0.05, Dunnett's test). Lower panel: Cell monolayers were loaded to steady-state by incubation for 20 min at 37°C in HBSS containing 50 µg/mL ciprofloxacin or minocycline. To trigger antibiotic efflux from the loaded cells, we diluted extracellular antimicrobial solutions 1:20 with 37°C HBSS. Efflux was monitored by the decrease in cell-associated fluorescence.

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