Restor Dent Endod.  2020 Nov;45(4):e47. 10.5395/rde.2020.45.e47.

Physicochemical properties, cytotoxicity and penetration into dentinal tubules of sodium hypochlorite with and without surfactants

• Coaguila-Llerena H
• Barbieri I
• Tanomaru-Filho M
• Leonardo RdT
• Ramos AP
• Faria G

• KMID: 2512038
• DOI: http://doi.org/10.5395/rde.2020.45.e47

Abstract

Objectives
The aim of this study was to assess the physicochemical properties, cytotoxicity and penetration into dentinal tubules of ChlorCid™ Surf (3% sodium hypochlorite [NaOCl] with surfactant) in comparison to ChlorCid™ (3% NaOCl without surfactant).
Materials and Methods
The physicochemical properties evaluated were pH, surface tension, free available chlorine (FAC) and contact angle. Cytotoxicity was evaluated in L929 fibroblasts exposed to the solutions by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and neutral red assays. Assessment of penetration into dentinal tubules was performed by staining single-rooted permanent human teeth with crystal violet (n = 9), which were irrigated with the solutions and analyzed in cervical, middle and apical segments. Data were analyzed by one-way analysis of variance (ANOVA) and Tukey's post-test, 2-way ANOVA and Bonferroni's post-test or t-test (α = 0.05).
Results
ChlorCid™ Surf and ChlorCid™ FAC values were close to those indicated by the manufacturer. ChlorCid™ Surf showed lower surface tension and contact angle on dentin, and higher pH than ChlorCid™ (p < 0.05). The penetration of ChlorCid™ Surf was higher in cervical and middle segments, compared with ChlorCid™ (p < 0.05). There was no difference in irrigant cytotoxicity (p > 0.05).
Conclusions
ChlorCid™ Surf showed lower surface tension, lower contact angle on root canal dentin, higher penetration into dentinal tubules and more alkaline pH, compared with ChlorCid™. However, both solutions showed similar cytotoxicity and FAC content.

Keyword

Dentin permeability; Materials testing; Physicochemical analysis; Sodium hypochlorite; Surface-active agents

Figure

• 

  Figure 1 Schematic representation of the contact angle evaluation of ChlorCid™ and ChlorCid™ Surf on root canal dentin. First, each tooth hemisection was mounted on PVC rings with acrylic resin. After resin curing, the rings were removed and the specimen was polished to avoid surface irregularities. Thereafter, 5 µL-drops of each solution were placed on the specimens to measure the contact angle using the OCA-20 system.PVC, poly vinyl chloride.

• 

  Figure 2 Mean and standard deviation of (A) surface tension in millinewton/meter (mN/m) and (B) contact angle (Ө) of ChlorCid™, ChlorCid™ Surf and deionized water (control). Different letters in columns indicate a significant difference between solutions. (C) Representative images of the contact angle on the dentin surface.

• 

  Figure 3 Viability of L929 fibroblasts after exposure to ChlorCid™ and ChlorCid™ Surf solutions at different doses by (A) MTT and (B) NR assays. No significant differences were founded between the solutions (p > 0.05).MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; NR, neutral red.

• 

  Figure 4 Penetration depth of ChlorCid™ and ChlorCid™ Surf solutions into dentinal tubules. (A) Mean and standard deviation in micrometers (µm) of penetration depth. Different lowercase letters in columns of each segment indicate a significant difference between the solutions. Different capital letters in columns indicate a significant difference between segments of each solution. (B) Representative images of penetration depth of ChlorCid™, ChlorCid™ Surf and water in cervical (B1-B3), middle (B4-B6) and apical (B7-B9) segments, respectively. The bleached crystal violet represents the penetration depth of irrigants into the dentin (arrow) (bar = 500 µm).De, dentin; RC, root canal.

Reference

1. Nair PN, Henry S, Cano V, Vera J. Microbial status of apical root canal system of human mandibular first molars with primary apical periodontitis after “one-visit” endodontic treatment. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2005; 99:231–252. PMID: 15660098.

2. Ricucci D, Siqueira JF Jr. Biofilms and apical periodontitis: study of prevalence and association with clinical and histopathologic findings. J Endod. 2010; 36:1277–1288. PMID: 20647081.

3. Palazzi F, Blasi A, Mohammadi Z, Del Fabbro M, Estrela C. Penetration of sodium hypochlorite modified with surfactants into root canal dentin. Braz Dent J. 2016; 27:208–216. PMID: 27058386.

4. Zou L, Shen Y, Li W, Haapasalo M. Penetration of sodium hypochlorite into dentin. J Endod. 2010; 36:793–796. PMID: 20416421.

5. Faria G, Viola KS, Coaguila-Llerena H, Oliveira LRA, Leonardo RT, Aranda-García AJ, Guerreiro-Tanomaru JM. Penetration of sodium hypochlorite into root canal dentine: effect of surfactants, gel form and passive ultrasonic irrigation. Int Endod J. 2019; 52:385–392. PMID: 30220093.

6. Pécora JD, Guimarães LF, Savioli RN. Surface tension of several drugs used in endodontics. Braz Dent J. 1992; 2:123–127. PMID: 1290911.

7. Kontakiotis EG, Tzanetakis GN, Loizides AL. A comparative study of contact angles of four different root canal sealers. J Endod. 2007; 33:299–302. PMID: 17320719.

8. Abou-Rass M, Patonai FJ Jr. The effects of decreasing surface tension on the flow of irrigating solutions in narrow root canals. Oral Surg Oral Med Oral Pathol. 1982; 53:524–526. PMID: 6808432.

9. Giardino L, Ambu E, Becce C, Rimondini L, Morra M. Surface tension comparison of four common root canal irrigants and two new irrigants containing antibiotic. J Endod. 2006; 32:1091–1093. PMID: 17055914.

10. Bukiet F, Couderc G, Camps J, Tassery H, Cuisinier F, About I, Charrier A, Candoni N. Wetting properties and critical micellar concentration of benzalkonium chloride mixed in sodium hypochlorite. J Endod. 2012; 38:1525–1529. PMID: 23063229.

11. Palazzi F, Morra M, Mohammadi Z, Grandini S, Giardino L. Comparison of the surface tension of 5.25% sodium hypochlorite solution with three new sodium hypochlorite-based endodontic irrigants. Int Endod J. 2012; 45:129–135. PMID: 21906088.

12. Guastalli AR, Clarkson RM, Rossi-Fedele G. The effect of surfactants on the stability of sodium hypochlorite preparations. J Endod. 2015; 41:1344–1348. PMID: 25906921.

13. Stojicic S, Zivkovic S, Qian W, Zhang H, Haapasalo M. Tissue dissolution by sodium hypochlorite: effect of concentration, temperature, agitation, and surfactant. J Endod. 2010; 36:1558–1562. PMID: 20728727.

14. Iglesias JE, Pinheiro LS, Weibel DE, Montagner F, Grecca FS. Influence of surfactants addition on the properties of calcium hypochlorite solutions. J Appl Oral Sci. 2019; 27:e20180157. PMID: 30624467.

15. Giardino L, Cavani F, Generali L. Sodium hypochlorite solution penetration into human dentine: a histochemical evaluation. Int Endod J. 2017; 50:492–498. PMID: 27009869.

16. Clarkson RM, Kidd B, Evans GE, Moule AJ. The effect of surfactant on the dissolution of porcine pulpal tissue by sodium hypochlorite solutions. J Endod. 2012; 38:1257–1260. PMID: 22892746.

17. Wong DT, Cheung GS. Extension of bactericidal effect of sodium hypochlorite into dentinal tubules. J Endod. 2014; 40:825–829. PMID: 24862710.

18. del Carpio-Perochena A, Bramante CM, de Andrade FB, Maliza AG, Cavenago BC, Marciano MA, Amoroso-Silva P, Duarte MH. Antibacterial and dissolution ability of sodium hypochlorite in different pHs on multi-species biofilms. Clin Oral Investig. 2015; 19:2067–2073.

19. Yasuda Y, Tatematsu Y, Fujii S, Maeda H, Akamine A, Torabinejad M, Saito T. Effect of MTAD on the differentiation of osteoblast-like cells. J Endod. 2010; 36:260–263. PMID: 20113785.

20. Ultradent Products. ChlorCid® Surf [Internet]. South Jordan, UT: Ultradent Products;2018. cited 2020 Sep 3. Available from: https://assets.ctfassets.net/wfptrcrbtkd0/4TD20QySslMLYQ7krTGsHX/fcc08568844faa1a5ac6b149a927537f/ChlorCid-Surf-Sales-Sheet-1005815AR03.pdf.

21. European Directorate for the Quality of Medicines & HealthCare. European pharmacopoeia. Strasbourg Cedex: European Directorate for the Quality of Medicines & HealthCare;2014. p. 24–25.

22. Baird R, Eaton A, Rice E. Standard methods for the examination of water and wastewater. Washington, D.C.: American Public Health Association; American Water Works Association; Water Environment Federation;2017. p. 11–12.

23. Andrade MA, Favarin B, Derradi R, Bolean M, Simão AM, Millán JL, Ciancaglini P, Ramos AP. Pendant-drop method coupled to ultraviolet-visible spectroscopy: a useful tool to investigate interfacial phenomena. Colloids Surf A Physicochem Eng Asp. 2016; 504:305–311. PMID: 28190931.

24. Tartari T, Wichnieski C, Bachmann L, Jafelicci M Jr, Silva RM, Letra A, van der Hoeven R, Duarte MA, Bramante CM. Effect of the combination of several irrigants on dentine surface properties, adsorption of chlorhexidine and adhesion of microorganisms to dentine. Int Endod J. 2018; 51:1420–1433. PMID: 29862516.

25. Viola KS, Rodrigues EM, Tanomaru-Filho M, Carlos IZ, Ramos SG, Guerreiro-Tanomaru JM, Faria G. Cytotoxicity of peracetic acid: evaluation of effects on metabolism, structure and cell death. Int Endod J. 2018; 51(Suppl 4):e264–e277. PMID: 28134990.

26. Yilmaz Z, Basbag B, Buzoglu HD, Gümüsderelioglu M. Effect of low-surface-tension EDTA solutions on the wettability of root canal dentin. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2011; 111:109–114. PMID: 21176825.

27. Guneser MB, Arslan D, Dincer AN, Er G. Effect of sodium hypochlorite irrigation with or without surfactants on the bond strength of an epoxy-based sealer to dentin. Clin Oral Investig. 2017; 21:1259–1265.

28. Dragan O, Tomuta I, Casoni D, Sarbu C, Campian R, Frentiu T. Influence of mixed additives on the physicochemical properties of a 5.25% sodium hypochlorite solution: an unsupervised multivariate statistical approach. J Endod. 2018; 44:280–285.e3. PMID: 29153984.

29. Ballal NV, Ferrer-Luque CM, Sona M, Prabhu KN, Arias-Moliz T, Baca P. Evaluation of final irrigation regimens with maleic acid for smear layer removal and wettability of root canal sealer. Acta Odontol Scand. 2018; 76:199–203. PMID: 29126370.

30. Milosevic A. The influence of surface finish and in-vitro pellicle on contact-angle measurement and surface morphology of three commercially available composite restoratives. J Oral Rehabil. 1992; 19:85–97. PMID: 1533872.

31. Rossi-Fedele G, Guastalli AR, Doğramacı EJ, Steier L, De Figueiredo JA. Influence of pH changes on chlorine-containing endodontic irrigating solutions. Int Endod J. 2011; 44:792–799. PMID: 21658076.

32. Vogel A. A textbook of quantitative inorganic analysis. London: Longmans;1962.

33. Coaguila-Llerena H, Rodrigues EM, Tanomaru-Filho M, Guerreiro-Tanomaru JM, Faria G. Effects of calcium hypochlorite and octenidine hydrochloride on L929 and human periodontal ligament cells. Braz Dent J. 2019; 30:213–219. PMID: 31166398.

34. Zhang W, Torabinejad M, Li Y. Evaluation of cytotoxicity of MTAD using the MTT-tetrazolium method. J Endod. 2003; 29:654–657. PMID: 14606789.

35. Peters OA. Research that matters - biocompatibility and cytotoxicity screening. Int Endod J. 2013; 46:195–197. PMID: 23398041.

36. Fotakis G, Timbrell JA. In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett. 2006; 160:171–177. PMID: 16111842.

37. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65:55–63. PMID: 6606682.

38. Repetto G, del Peso A, Zurita JL. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nat Protoc. 2008; 3:1125–1131. PMID: 18600217.