Tetracycline at subcytotoxic levels inhibits matrix metalloproteinase-2 and -9 but does not remove the smear layer.
BACKGROUND
The antibacterial and anticollagenolytic properties of tetracycline (TCN) are valuable in periodontal therapy, and TCN treatment can remove the smear layer following root instrumentation. However, recent reports pointing to cytotoxic effects of several acids prompted this study to define TCN concentrations that are anticollagenolytic and remove the smear layer, but have low cytotoxicity.


METHODS
Human gingival (hGF) and periodontal ligament (hPDL) cells were treated short- (3 minutes) or long-term (24 hours) with TCN to determine concentrations yielding 50% (TD(50)) and 90% (TD(10)) cell survival. Activity assays measured TCN concentrations with half-maximal inhibition (IC(50)) of matrix metalloproteinase- 2 and -9 (MMP-2 and -9). Finally, we analyzed the effects of TCN with high (75 mg/ml) or low (1 mg/ml) cytotoxicity on the smear layer by scanning electron microscopy (SEM).


RESULTS
The TD(50) for TCN after short-term treatment was 4 mg/ml for both hGF and hPDL. Ninety percent of the cells survived 0.2 mg/ml. With long-term treatment, the TD(50) for hGF and hPDL was 70 and 30 microg/ml, respectively, and the TD(10) was 20 and 5 microg/ml. HGF and hPDL recovered from the 3-minute treatment with 1 mg/ml, but not from concentrations exceeding 3 and 9 mg/ml, respectively. The IC(50) was 25 microg/ml for both MMP-2 and MMP-9. Whereas 75 mg/ml TCN removed the smear layer, 1 mg/ml TCN had no effects.


CONCLUSIONS
Tetracycline has significant cytotoxicity on periodontal cells. Since non-cytotoxic concentrations of TCN inhibited MMP-2 and -9 but had no effects on the smear layer, TCN is not recommended for root surface treatment.
Wang
Yao
Y
Departments of Periodontics and Biochemistry, University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900, USA.
Morlandt
Anthony B
AB
Xu
Xiaoping
X
Carnes
David L
DL
Chen
Zhihua
Z
Steffensen
Bjorn
B
D000900:Anti-Bacterial Agents / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects; D005881:Gingiva / Q000166:cytology / Q000187:drug effects; D006801:Humans; D020128:Inhibitory Concentration 50; D061965:Matrix Metalloproteinase Inhibitors; D010513:Periodontal Ligament / Q000166:cytology / Q000187:drug effects*; D011480:Protease Inhibitors / Q000633:toxicity*; D016369:Smear Layer; D013752:Tetracycline / Q000633:toxicity*; D014092:Tooth Root / Q000187:drug effects*
D000900:Anti-Bacterial Agents; D061965:Matrix Metalloproteinase Inhibitors; D011480:Protease Inhibitors; D013752:Tetracycline