In vitro evaluation of the antibacterial potential of benzalkonium chloride in root canals infected by Enterococcus faecalis

doi:10.3877/cma.j.issn.1674-1366.2017.06.002

Abstract

Abstract:

Objective

To evaluate the antibacterial potential of benzalkonium chloride in root canals infected by Enterococcus faecalis (E.faecalis) .

Methods

Sixty-four human premolars extracted for orthodontics were prepared and inoculated with E.faecalis. Two teeth were selected respectively using random lottery method before and after inoculation, then split in to two halves along the mesiodistal axis. One half was processed with light microscopic (Brown & Brenn stain) to check the bacteria in dentinal tubules, and the other was observed with SEM to investigate the bacterial status in infected root canals. The rest of the teeth were randomly assigned to the following groups: 0.05% benzalkonium chloride, 0.1% benzalkonium chloride, 3% hydrogen peroxide, 2.5% sodium hypochlorite, 5.25% sodium hypochlorite, 0.9% sodium chloride for endodontic irrigation. Before and after irrigating, microbiological samples were collected from root canals for bacteriological culture, and colony forming units (CFU) were counted. The t-test was used to analyze the difference of bacterial counts in root canals before and after irrigation. The difference of bacterial descent among the six groups were analyzed by ANOVA. Bacterial status after irrigation was assessed with SEM.

Results

(1) E.faecalis was found to penetrate into the dentinal tubules about 250~ 1050 μm. In the infected root canal samples, E.faecalis was attached to the root canal wall to form a biofilm structure. (2) The number of E.faecalis in root canal in each group was effectively reduced excepted 0.9% sodium chloride group (P<0.05) . The bacterial descent of 0.05%, 0.1% benzalkonium chloride groups (7.90 ± 2.18, 8.30 ± 2.21) was significantly more than that of 3% hydrogen peroxide group (5.30 ± 2.31) and 2.5% sodium hypochlorite group (6.10 ± 1.20, P<0.05) . There were no statistical difference among 0.05% benzalkonium chloride group, 0.1% benzalkonium chloride group and 5.25% sodium hypochlorite group (8.70 ± 1.89, P>0.05) . Root canal surfaces were cleaner in benzalkonium chloride and 5.25% sodium hypochlorite groups than 3% hydrogen peroxide group and 2.5% sodium hypochloritethe group with SEM observation.

Conclusion

It was suggested that benzalkonium chloride solution had potential antibacterial activity against E.faecalis when it was used for root canal irrigation.

Key words: Enterococcus faecalis, Root canal irrigants, Benzalkonium compounds, Dental models

Jing Dai, Yanbin Chen, Shan Chen, Jing Ren, Kunman Li, Junying Yang. In vitro evaluation of the antibacterial potential of benzalkonium chloride in root canals infected by Enterococcus faecalis[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2017, 11(06): 326-332.

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References 32

[1]	Pataky L, Iványi I, Grigár A，et al. Antimicrobial efficacy of various root canal preparation techniques：an in vitro comparative study[J]. J Endod，2002，28(8):603-605.
[2]	Carrotte P. 21st century endodontics：part 3[J]. Int Dent J，2005，55(4):247-253.
[3]	Haapasalo M, Endal U, Zandi H，et al. Eradication of endodontic infection by instrumentation and irrigation solutions[J]. Endod Topics，2005，10(1):77-102.
[4]	Zehnder M. Root canal irrigants[J]. J Endod，2006，32(5):389-398.
[5]	Ulusoy OI, Nayir Y, Celik K，et al. Apical microleakage of different root canal sealers after use of maleic acid and EDTA as final irrigants[J]. Braz Oral Res，2014，28(5):337-342.
[6]	Kobayashi Y, Hayashi M, Yoshino F，et al. Bactericidal effect of hydroxyl radicals generated from a low concentration hydrogen peroxide with ultrasound in endodontic treatment[J]. J Clin Biochem Nutr，2014，54(3):161-165.
[7]	Gjorgievska E, Apostolska S, Dimkov A，et al. Incorporation of antimicrobial agents can be used to enhance the antibacterial effect of endodontic sealers[J]. Dent Mater，2013，29(3):e29-e34.
[8]	Svensäter G, Bergenholtz G. Biofilms in endodontic infections[J]. Endod Topics，2004，9(1):27-36.
[9]	Stojanović N, Krunić J, Popović B，et al. Prevalence of Enterococcus faecalis and Porphyromonas gingivalis in infected root canals and their susceptibility to endodontic treatment procedures：a molecular study[J]. Srp Arh Celok Lek，2014，142(9-10):535-541.
[10]	Lu B, Zhang J, Huang X，et al. Expression of Interleukin-1β and Matrix Metalloproteinase-8 in Cytolytic and Noncytolytic Enterococcus faecalis-induced Persistent Apical Periodontitis：A Comparative Study in the Rat[J]. J Endod，2015，41(8):1288-1293.
[11]	Cheng X, Chen B, Qiu J，et al. Bactericidal effect of Er：YAG laser combined with sodium hypochlorite irrigation against Enterococcus faecalis deep inside dentinal tubules in experimentally infected root canals[J]. J Med Microbiol，2016，65(2):176-187.
[12]	Sedgley CM, Lennan SL, Appelbe OK. Survival of Enterococcus faecalis in root canals ex vivo[J]. Int Endod J，2005，38(10):735-742.
[13]	Distel JW, Hatton JF, Gillespie MJ. Biofilm formation in medicated root canals[J]. J Endod，2002，28(10):689-693.
[14]	郭惠杰，岳林.粪肠球菌在根管内定植模式的体外研究[J].北京大学学报（医学版），2009，41(6):699-701.
[15]	Berutti E, Marini R, Angeretti A. Penetration ability of different irrigants into dentinal tubules[J]. J Endod，1997，23(12):725-727.
[16]	Podar R, Kulkarni GP, Dadu SS，et al. In vivo antimicrobial efficacy of 6% Morinda citrifolia，Azadirachta indica，and 3% sodium hypochlorite as root canal irrigants[J]. Eur J Dent，2015，9(4):529-534.
[17]	Goztas Z, Onat H, Tosun G，et al. Antimicrobial effect of ozonated water，sodium hypochlorite and chlorhexidine gluconate in primary molar root canals[J]. Eur J Dent，2014，8(4):469-474.
[18]	Williamson AE, Cardon JW, Drake DR. Antimicrobial susceptibility of monoculture biofilms of a clinical isolate of Enterococcus faecalis[J]. J Endod，2009，35(1):95-97.
[19]	Gernhardt CR, Eppendorf K, Kozlowski A，et al. Toxicity of？concentrated sodium hypochlorite used as an endodontic irrigant[J]. Int Endod J，2004，37(4):272-280.
[20]	Ragnarsson KT, Rechenberg DK, Attin T，et al. Available chlorine consumption from NaOCl solutions passively placed in instrumented human root canals[J]. Int Endod J，2015，48(5):435-440.
[21]	Sirtes G, Waltimo T, Schaetzle M，et al. The effects of temperature on sodium hypochlorite short-term stability，pulp dissolution capacity，and antimicrobial efficacy[J]. J Endod，2005，31(9):669-671.
[22]	del Carpio-Perochena A, Bramante CM, de Andrade FB，et al. Antibacterial and dissolution ability of sodium hypochlorite in different pHs on multi-species biofilms[J]. Clin Oral Investig，2015，19(8):2067-2073.
[23]	Pawar R, Alqaied A, Safavi K，et al. Influence of an apical negative pressure irrigation system on bacterial elimination during endodontic therapy：a prospective randomized clinical study[J]. J Endod，2012，38(9):1177-1181.
[24]	Al-Ali M, Sathorn C, Parashos P. Root canal debridement efficacy of different final irrigation protocols[J]. Int Endod J，2012，45(10):898-906.
[25]	Haapasalo M, Shen Y, Qian W，et al. Irrigation in endodontics[J]. Dent Clin North Am，2010，54(2):291-312.
[26]	Wessels S, Ingmer H. Modes of action of three disinfectant active substances：a review[J]. Regul Toxicol Pharmacol，2013，67(3):456-467.
[27]	da Costa LF, Amaral CDSF, Barbirato DDS，et al. Chlorhexidine mouthwash as an adjunct to mechanical therapy in chronic periodontitis：A meta-analysis[J]. J Am Dent Assoc，2017，148(5):308-318.
[28]	Lamont T. Lower concentration of chlorhexidine and cetyl-pyridinium chloride mouthwash demonstrates some efficacy[J]. Evid Based Dent，2012，13(2):52-53.
[29]	邢飞鹏，孙立洲，王欣梅.苯扎氯铵溶液根管冲洗液抗菌效果研究[J].医药论坛杂志，2011，32(12):168-169.
[30]	Baron A, Lindsey K, Sidow SJ，et al. Effect of a Benzalkonium Chloride Surfactant-Sodium Hypochlorite Combination on Elimination of Enterococcus faecalis[J]. J Endod，2016，42(1):145-149.
[31]	Tarbox BB, Conroy BP, Malicky ES，et al. Benzalkonium chloride. A potential disinfecting irrigation solution for orthopaedic wounds[J]. Clin Orthop Relat Res，1998(346):255-261.
[32]	Liebert MA. 2 Final report on the safety assessment of benzalkonium chloride[J]. J Am Coll Toxicol，1989，8(4):589-625.

组别	样本量（例）	菌落计数		冲洗前后菌落差值	t值	P值
组别	样本量（例）	根管冲洗前	根管冲洗后	冲洗前后菌落差值	t值	P值
0.05%苯扎氯铵溶液组	10	8.50 ± 1.72	0.60 ± 1.07	7.90 ± 2.18	12.338	0.000
0.1%苯扎氯铵溶液组	10	8.80 ± 2.04	0.50 ± 0.85	8.30 ± 2.21	11.857	0.000
3%过氧化氢溶液组	10	8.20 ± 1.99	2.90 ± 0.99	5.30 ± 2.31	7.573	0.000
2.5%次氯酸钠溶液组	10	8.40 ± 1.96	2.30 ± 1.42	6.10 ± 1.20	7.789	0.000
5.25%次氯酸钠溶液组	10	8.70 ± 1.89	0	8.70 ± 1.89	14.568	0.000
0.9%氯化钠溶液组	10	8.50 ± 1.84	8.00 ± 2.11	0.50 ± 1.35	0.565	0.579

组别	样本量（例）	菌落计数		冲洗前后菌落差值	t值	P值
组别	样本量（例）	根管冲洗前	根管冲洗后	冲洗前后菌落差值	t值	P值
0.05%苯扎氯铵溶液组	10	8.50 ± 1.72	0.60 ± 1.07	7.90 ± 2.18	12.338	0.000
0.1%苯扎氯铵溶液组	10	8.80 ± 2.04	0.50 ± 0.85	8.30 ± 2.21	11.857	0.000
3%过氧化氢溶液组	10	8.20 ± 1.99	2.90 ± 0.99	5.30 ± 2.31	7.573	0.000
2.5%次氯酸钠溶液组	10	8.40 ± 1.96	2.30 ± 1.42	6.10 ± 1.20	7.789	0.000
5.25%次氯酸钠溶液组	10	8.70 ± 1.89	0	8.70 ± 1.89	14.568	0.000
0.9%氯化钠溶液组	10	8.50 ± 1.84	8.00 ± 2.11	0.50 ± 1.35	0.565	0.579

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