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Modulation of Gingival Fibroblast Minocycline Accumulation by Biological Mediators

Section of Periodontology, College of Dentistry, The Ohio State University Health Sciences Center, 305 West 12th Avenue, PO Box 182357, Columbus, OH 43218-2357, USA;

View larger version (22K): [in a new window]   Figure 1. Stimulation of fibroblast minocycline accumulation by FGF-2 and PDGF. Confluent fibroblast cultures were starved for 20 hrs and treated with the indicated growth factor concentrations for 1 or 6 hrs. Cell DNA content did not increase significantly under these experimental conditions. After brief incubation at 37°C, 40 µg/mL minocycline was added, and uptake was monitored for 3 min. The data represent the mean ± SEM of 5 individual experiments. Both agents produced a significant treatment effect after 1 and 6 hrs (P < 0.003, repeated-measures ANOVA). Conditions that produced a significant increase in minocycline accumulation compared with controls (P > 0.05, Dunnett’s test) are indicated by *.

View larger version (21K): [in a new window]   Figure 2. Effects of TGF-ß1 and TNF- on fibroblast minocycline accumulation. Experiments were conducted as described in the legend for Fig. 1. The data represent the mean ± SEM of 5 individual experiments. There were no significant effects by TGF at 1 hr (P = 0.24, repeated-measures ANOVA). However, incubation with TGF for 6 hrs produced a significant treatment effect (P < 0.001, repeated-measures ANOVA), and TNF- produced a significant treatment effect at 1 and 6 hrs (P < 0.001 and P 0.008, respectively). Conditions that produced a significant increase in minocycline accumulation compared with controls (P > 0.05, Dunnett’s test) are indicated by *.

View larger version (18K): [in a new window]   Figure 3. Enhanced accumulation of tetracyclines by PMA-activated fibroblasts. Confluent fibroblast cultures were starved for 20 hrs and treated with 3 to 100 nM PMA for 15 min prior to assaying transport of minocycline, doxycycline, and tetracycline. The data represent the mean ± SEM of 6 individual experiments. PMA produced a significant treatment effect on the accumulation of all 3 agents (P 0.002, repeated-measures ANOVA). PMA concentrations indicated by * induced a significant increase in antibiotic accumulation (P > 0.05, Dunnett’s test).