Analysis of antimicrobial efficacy of sodium hypochlorite and ozonated water against biofilm in oval canals

Merçon, Ivy Rodrigues; Campos, Francisco Ubiratan Ferreira de; Fontana, Carlos Eduardo; Pelegrine, Rina Andréa; Martin, Alexandre Sigrist De; Bueno, Carlos Eduardo da Silveira

doi:10.1590/0103-6440202305318

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Article • Braz. Dent. J. 34 (3) • May-Jun 2023 • https://doi.org/10.1590/0103-6440202305318 linkcopy

Analysis of antimicrobial efficacy of sodium hypochlorite and ozonated water against biofilm in oval canals

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This in vitro study compared the antimicrobial efficacy of 2.5% sodium hypochlorite (NaOCl) and 8 µg/mL ozonated water agitated by passive ultrasonic irrigation (PUI) or PUI combined with EndoActivator (EA) against mature multispecies biofilm. One hundred and five oval-shaped mandibular premolars were instrumented, sterilized, and inoculated with Enterococcus faecalis, Candida albicans, and Staphylococcus aureus, divided into: control group - saline; O₃ group - ozonated water; O₃ PUI group - ozonated water with PUI agitation; O₃ PUI+EA group - ozonated water with PUI+EA agitation; NaOCl group - NaOCl; NaOCl PUI group - NaOCl with PUI agitation; and NaOCl PUI+EA group - NaOCl with PUI+EA agitation. Microbiological samples were collected before (S1) and after (S2) the disinfection procedures and the data were statistically analyzed using the Kruskal-Wallis test. In the culture method, there was significant disinfection in the O₃ PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). The combination of NaOCl with PUI+EA reduced microbial counts to zero (p˂0.05). In the qPCR method, there was a significant reduction in the total count of viable microorganisms in the O₃ PUI, O₃ PUI+EA, NaOCl, NaOCl PUI, and NaOCl PUI+EA groups (p˂0.05). It can be concluded that 2.5% NaOCl with and without agitation, as well as 8 µg/mL ozonated water with its action enhanced by the agitation techniques, were effective in root canal disinfection, and their antimicrobial efficacy is related to the microorganisms present in the biofilm.

Key Words:
biofilm; apical periodontitis; disinfection; sodium hypochlorite; ozone

	Saline (control)	O₃	O₃ PUI	O₃ PUI + EA	NaOCl	NaOCl PUI	NaOCl PUI + EA	(p)
Before	0.39 (0.61)^Aa	0.39 (0.68)^Aa	0.64 (0.27)^Aa	0.59 (0.35)^Aa	0.36 (0.31)^Aa	0.42 (0.52)^Aa	0.44 (0.35)^Aa	>0.05
After	0.64 (0.54)^Aa	0.20 (0.40)^A1	0.23 (0.15)^A1	0.20 (0.16)^B1	0.00 (0.30)^Bb	0.00(0.15)^Bb	0.00 (0.00)^Bb2	<0.05
(p)	>0.05	>0.05	>0.05	<0.05	<0.05	<0.05	<0.05

	Saline (control)	O₃	O₃ PUI	O₃ PUI + EA	NaOCl	NaOCl PUI	NaOCl PUI + EA	(p)
%	3.08 (20.69)^A	21.01 (35.9)^A	51.39 (41.4)^A	54.59 (33)^A	100 (10.5)^B	100 (9.2)^B	100 (0)^B	<0.05

	Saline (control)	O₃	O₃ PUI	O₃ PUI + EA	NaOCl	NaOCl PUI	NaOCl PUI + EA	(p)
E. faecalis
Before	3.24 (1.96)Aa	5.37 (1.44)Ba	8.62 (6.92)Ba	8.36 (4.59)Ba	9.76 (12)Ba	9.05 (5.51)Ba	9.34 (8.38)Ba	<0.05
After	4.01 (0.82)Aa	2.02 (5.36)Aa	3.66 (2.18)Ab	1.61 (4.73)Ab	2.70 (1.39)Ab	1.97 (8.87)Ab	3.01 (2.68)Ab	>0.05
(p)	>0.05	>0.05	<0.05	<0.05	<0.05	<0.05	<0.05
S. aureus
Before	4.31 (3.27)Aa	3.81 (3.29)Aa	4.09 (4.29)Aa	2.97 (3.52)Aa	4.23 (6.07)Aa	2.19 (2.16)Aa	3.96 (2.24)Aa	>0.05
After	4.66 (2.46)Aa	1.24 (3.01)Ba	0.00 (0.00)Bb	0.00 (0.00)Bb	0.00 (0.00)Bb	0.00 (0.00)Bb	0.00 (0.00)Bb	<0.05
(p)	>0.05	>0.05	<0.05	<0.05	<0.05	<0.05	<0.05
C. albicans
Before	3.24 (2.6)Aa	7.56 (4.2)ABa	10.04 (8.3)Ba	9.83 (3.95)Ba	9.33 (5.25)Ba	9.31 (2.95)Ba	10.76 (9.50)Ba	<0.05
After	4.44 (2.07)Aa	3.68 (3.31)Aa	3.83 (5.55)Ab	2.65 (4.79)Ab	3.64 (5.73)Ab	3.37 (3.43)Ab	2.09 (3.64)Ab	>0.05
(p)	>0.05	>0.05	<0.05	<0.05	<0.05	<0.05	<0.05

%	Saline (Control)	O₃	O₃ PUI	O₃ PUI + EA	NaOCl	NaOCl PUI	NaOCl PUI + EA	(p)
E. faecalis	0.00 (2.96)^A	63.61 (28.78)^B	64.36 (29.95)^B	76.31 (33.65)^B	65.39 (22.65)^B	77.12 (24.83)^B	77.69 (19.62)^B	<0.05
S. aureus	0.00 (4.23)^A	95.89 (63.77)^B	100.00 (0.00)^B	100.00 (0.00)^B	100.00 (0.00)^B	100.00 (0.00)^B	100.00 (0.00)^B	<0.05
C. albicans	0.00 (3.77)^A	47.80 (37.33)^B	65.80 (36.34)^B	62.54 (47.75)^B	61.92 (45.18)^B	58.11 (29.45)^B	70.66 (19.08)^B	<0.05

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[TFN1] CFU: colony-forming unit; O₃: ozone; PUI: passive ultrasonic irrigation; EA: EndoActivator; NaOCl: sodium hypochlorite.

[TFN2] Data presented as median (interquartile deviation) and compared by the Kruskal-Wallis test, followed by Dunn’s test.

[TFN3] Different uppercase letters in the same column and different lowercase letters and numbers in the same row indicate statistically significant differences.