In vitro antibacterial action of Tetraclean, MTAD and five experimental irrigation solutions.
OBJECTIVE
To investigate the antibacterial effect of Tetraclean, MTAD and five experimental irrigants using both direct exposure test with planktonic cultures and mixed-species in vitro biofilm model.


METHODS
Tetraclean, MTAD and five experimental solutions that were modifications of existing formulae including MTAD + 0.01% cetrimide (CTR), MTAD + 0.1% CTR, MTAC-1 (Tween 80 replaced by 0.01% CTR in MTAD), MTAC-2 (Tween 80 replaced by 0.1% CTR) and MTAD-D (MTAD without the Tween 80 and no CTR added) were used as disinfectants in the experiments. In the direct exposure test, a suspension of Enterococcus faecalis was mixed with each of the solutions. After 0.5, 1, 3 and 10 min, an inactivator was added and the number of surviving bacteria was calculated. A mixed-species biofilm from subgingival plaque bacteria was grown in brain heart infusion broth in anaerobic conditions on synthetic hydroxyapatite discs. Two-week-old biofilms were exposed to the solutions for 0.5, 1 and 3 min. The samples were observed by confocal laser scanning microscopy after bacterial viability staining. The scans were quantitatively analysed, and the volume of killed cells of all cells was calculated for each medicament.


RESULTS
Tetraclean and MTAC-2 (0.1% CTR) killed planktonic E. faecalis in <30 s. Complete killing of bacteria required 1 min by MTAC-1, 3 min by MTAD + 0.1% CTR and 10 min by MTAD, MTAD-D and MTAD + 0.01% CTR. In the biofilm test, there were significant differences in microbial killing between the different solutions and times of exposure (P < 0.005). MTAC-2 showed the best performance, killing 71% of the biofilm bacteria in 3 min, followed by MTAC-1 and Tetraclean. MTAD and the three MTAD modifications demonstrated the lowest antibacterial activity.


CONCLUSIONS
Tetraclean was more effective than MTAD against E. faecalis in planktonic culture and in mixed-species in vitro biofilm. CTR improved the antimicrobial properties of the solutions, whereas Tween 80 seemed to have a neutral or negative impact on their antimicrobial effectiveness.
Department of Restorative Dentistry, Faculty of Dentistry of Araraquara, State University of São Paulo, Araraquara, São Paulo, Brazil.
F G
FG
Pappen
Y
Y
Shen
W
W
Qian
M R
MR
Leonardo
L
L
Giardino
M
M
Haapasalo
D000891:Anti-Infective Agents, Local / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects*; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000494:pharmacology; D019343:Citric Acid / Q000494:pharmacology; D015169:Colony Count, Microbial; D019551:Dental Disinfectants; D003786:Dental Pulp Cavity / Q000382:microbiology*; D004203:Disinfection / Q000379:methods*; D004318:Doxycycline / Q000494:pharmacology; D004338:Drug Combinations; D013293:Enterococcus faecalis / Q000187:drug effects / Q000502:physiology; D006801:Humans; D010485:Periapical Periodontitis / Q000517:prevention & control*; D011136:Polysorbates / Q000494:pharmacology; D012388:Root Canal Irrigants / Q000737:chemistry / Q000494:pharmacology*; D016369:Smear Layer
D000891:Anti-Infective Agents, Local; D002593:Cetrimonium Compounds; D019551:Dental Disinfectants; D004338:Drug Combinations; C475043:MTAD (intracanal irrigant); D011136:Polysorbates; D012388:Root Canal Irrigants; D019343:Citric Acid; D004318:Doxycycline; D000077286:Cetrimonium