Characterization of minocycline transport by human neutrophils.
BACKGROUND
Tetracyclines are used in periodontal therapy as antimicrobial agents and as inhibitors of matrix metalloproteinases. Neutrophils appear to accumulate minocycline and other tetracyclines through a mechanism that has not been fully characterized.


METHODS
The transport of minocycline and other tetracyclines by isolated human neutrophils was characterized by measuring the increase in cell-associated fluorescence.


RESULTS
Quiescent neutrophils took up minocycline through a saturable, concentrative, sodium-dependent mechanism with a Michaelis constant (K(m)) of 153 micro g/ml (501 microM) and a maximal velocity of 240 ng/minute/10(6) cells. The efficiency of minocycline transport was not influenced significantly by a two-unit variation in extracellular pH and was not enhanced upon cell activation with phorbol myristate acetate. Neutrophil incubation in medium containing 10 micro g/ml minocycline, doxycycline, or tetracycline yielded steady-state intracellular/extracellular concentration ratios of approximately 64.0, 7.5, or 1.8, respectively. The dilution of extracellular minocycline or doxycycline triggered efflux from cells loaded with these antibiotics. Minocycline transport was competitively inhibited by the organic cations carnitine, diphenhydramine, and verapamil, but penicillin and other organic anions failed to produce inhibition.


CONCLUSIONS
Transport of tetracyclines by neutrophils could potentially enhance the effectiveness of these agents in periodontal therapy by enhancing or sustaining their therapeutic levels at inflammatory sites and by enhancing the killing of phagocytosed bacterial pathogens.
Walters
John D
JD
Section of Periodontology, College of Dentistry, The Ohio State University Health Sciences Center, Columbus, OH 43210, USA. walters2@osu.edu
D000328:Adult; D000900:Anti-Bacterial Agents / Q000493:pharmacokinetics*; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004318:Doxycycline / Q000493:pharmacokinetics; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007424:Intracellular Fluid / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D008911:Minocycline / Q000493:pharmacokinetics*; D009504:Neutrophils / Q000378:metabolism*; D012016:Reference Values; D013752:Tetracycline / Q000493:pharmacokinetics
D000900:Anti-Bacterial Agents; D013752:Tetracycline; D008911:Minocycline; D004318:Doxycycline