Assessment of antimicrobial activity of sodium hypochlorite, calcium hypochlorite and grape seed extract against Enterococcus faecalis

  • Larissa Taís Soligo University of Passo Fundo
  • Doglas Cecchin University of Passo Fundo
  • Matheus Albino Souza University of Passo Fundo
  • Ezequiel Santin Gabrielli State University of Campinas
  • Huriel Scartazzini Palhano University of Passo Fundo
  • Ana Paula Farina University of Passo Fundo
Keywords: calcium hypochlorite, sodium hypochlorite, grape seed extract, Enterococcus faecalis.


OBJECTIVE: This study aimed to assess the antimicrobial action of calcium hypochlorite [Ca(OCl)2] at concentrations of 2.5% and 6%, and of grape seed extract (GSE) at concentrations of 10%, 30%, and 50%, against Enterococcus faecalis, comparing it to the action of 6% sodium hypochlorite (NaOCl).
METHODS: Saline solution was used as negative control. The inhibition halos of microbial growth were verified by the agar disk diffusion method. Twelve Petri plates were used for seeding with culture medium of approximately 5 mm in thickness. In each plate, 5 disks of pure and sterile antibiogram, soaked in the substances to be tested, were used and taken to the plate containing the seeded
bacterial strain. The plates remained in aerobic bacteriological incubator for 24 h at 37°C temperature. After 24 h, the inhibition halos were measured with a digital caliper. Analysis of Variance (ANOVA) was used for statistical analysis followed by Tukey's complementary test, at 5% significance.
RESULTS: The 6% Ca(ClO)2 presented inhibition halo statistically higher than the other solutions (p<0.05), followed by 2.5% Ca(ClO)2, which was statistically similar to 6% NaOCl (p>0.05). The GSE concentrations resulted in lower inhibition halos of active substances and the different concentrations were similar to each other. Lastly, saline solution presented the same inhibition halos in all groups (p<0.05).
CONCLUSION: It may be concluded that 6% Ca(OCl)2 presented higher antimicrobial activity than 6% NaOCl. On the other hand, all GSE concentrations were lower than NaOCl and Ca(OCl)2.


Stuart CH, Schwartz SA, Beeson TJ, Owatz CB. Enterococcus faecalis: its role in root canal treatment failure and current concepts in retreatment. J Endod. 2006;32:93-8.

Sundqvist G, Figdor D. Life as an endodontic pathogen. Ecological differences between the untreated and root-filled root canals. Endod Topics. 2003;6:3-28.

Byström A, Sundqvist G. Bacteriologic evaluation of the efficacy of mechanical root canal instrumentation in endodontic therapy. Scand J Dent Res. 1981;89:321-8.

Zehnder M. Root canal irrigants. J Endod. 2006;32:389-98.

Motta MV, Chaves-Mendonca MA, Stirton CG, Cardozo HF. Accidental injection with sodium hypochlorite: report of a case. Int Endod J. 2009;42:175-82.

Moreira DM, Almeida JF, Ferraz CC, Gomes BP, Line SR, Zaia AA. Structural analysis of bovine root dentin after use of different endodontics auxiliary chemical substances. J Endod. 2009;35:1023-7. http://dx.doi. org/10.1016/j.joen.2009.04.002

Ghisi AC, Kopper PM, Baldasso FE, Stürmer CP, Rossi-Fedele G, Steier L, de Figueiredo JA, Morgental RD, Vier-Pelisser FV. Effect of superoxidized water and sodium hypochlorite, associated or not with EDTA, on organic and inorganic components of bovine root dentin. J Endod. 2015;41: 925-30.

Santos JN, Carrilho MR, De Goes MF, Zaia AA, Gomes BP, Souza-Filho FJ, Ferraz CC. Effect of chemical irrigants on the bond strength of a selfetching adhesive to pulp chamber dentin. J Endod. 2006;32:1088-90.

Farina AP, Cecchin D, Barbizam JV, Carlini-Júnior B. Influence of endodontic irrigants on bond strength of a self-etching adhesive. Aust Endod J. 2011;37:26-30.

Dutta A, Saunders WP. Comparative evaluation of calcium hypochlorite and sodium hypochlorite on soft-tissue dissolution. J Endod. 2012;38:1395-8.

Leonardo NG, Carlotto IB, Luisi SB, Kopper PM, Grecca FS, Montagner F.

Calcium Hypochlorite Solutions: Evaluation of Surface Tension and Effect

of Different Storage Conditions and Time Periods over pH and Available Chlorine Content. J Endod. 2016;42:641-5.


de Almeida AP, Souza MA, Miyagaki DC, Dal Bello Y, Cecchin D, Farina AP.

Comparative evaluation of calcium hypochlorite and sodium hypochlorite associated with passive ultrasonic irrigation on antimicrobial activity of a root canal system infected with Enterococcus faecalis: an in vitro study. J Endod. 2014;40:1953-7.

Blattes GB, Mestieri LB, Böttcher DE, Fossati AC, Montagner F, Grecca FS. Cell migration, viability and tissue reaction of calcium hypochlorite based-solutions irrigants: An in vitro and in vivo study. Arch Oral Biol. 2017;73:34-39.

Wu CD. Grape products and oral health. J Nutr 2009;139:1818S-23S.

Sarni-Manchado P, Cheynier V, Moutounet M. Interactions of grape seed

tannins with salivary proteins. J Agric Food Chem. 1999;47:42-7. http://

Al-Ammar A, Drummond JL, Bedran-Russo AK. The use of collagen crosslinking agents to enhance dentin bond strength. J Biomed Mater Res

B Appl Biomater. 2009;91:419-4.

Bagchi D, Bagchi M, Stohs SJ, Das DK, Ray SD, Kuszynski CA, Joshi SS, Pruess HG. Free radicals and grape seed proanthocyanidin extract: importance in human health and disease prevention. Toxicology. 2000;148:187-97.

Cecchin D, Farina AP, Souza MA, Albarello LL, Schneider AP, Vidal CM, Bedran-Russo AK. Evaluation of antimicrobial effectiveness and dentine

mechanical properties after use of chemical and natural auxiliary irrigants. J Dent. 2015;43:695-702. doi: 10.1016/j.jdent.2015.03.013.

Su YS, Morrison DT, Ogle RA. Chemical kinetics of calcium hypochlorite

decomposition in aqueous solution. Chem Health Saf. 2009;16:21-5.

Dumani A, Guvenmez HK, Yilmaz S, Yoldas O, Kurklu ZG.Antibacterial Efficacy of Calcium Hypochlorite with Vibringe Sonic Irrigation System on

Enterococcus faecalis: An In Vitro Study. Biomed Res Int. 2016:8076131.

Baumgartner JC, Cuenin PR. Efficacy of several concentrations of sodium

hypochlorite for root canal irrigation. J Endod. 1992;18:605-12. http://

Vianna ME, Gomes BP, Berber VB, Zaia AA, Ferraz CC, de Souza-Filho FJ. In vitro evaluation of the antimicrobial activity of chlorhexidine and sodium hypochlorite. Oral Surg Oral Med Oral Pathol Oral Radiol Endod.


Whittaker HA, Mohler BM.The Sterilization of Milk Bottles With Calcium Hypochlorite. Am J Public Health (NY)1912;2:282-7.

Dornelles-Morgental R, Guerreiro-Tanomaru JM, de Faria-Júnior NB, Hungaro-Duarte MA, Kuga MC, Tanomaru-Filho M. Antibacterial efficacy of endodontic irrigating solutions and their combinations in root canals contaminated with Enterococcus faecalis. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2011;112:396-400.


Camps J, Pommel L, Aubut V, Verhille B, Satoshi F, Lascola B, About I. Shelf life, dissolving action, and antibacterial activity of a neutralized 2.5% sodium hypochlorite solution. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009;108:e66-73.

Mercade M, Duran-Sindreu F, Kuttler S, Roig M, Durany N. Antimicrobial

efficacy of 4.2% sodium hypochlorite adjusted to pH 12, 7.5, and 6.5 in

infected human root canals. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009;107:295-8.

Saito M, Hosoyama H, Ariga T, Kataoka S, Yamaji N. Antiulcer activity of

grape seed extract and procyanidins. J Agric Food Chem 1998;46:1460-4.

Baydar NG, Sagdic O, Ozkan G, Cetin S. Determination of antibacterial effects and total phenolic contents of grape (Vitis vinifera L.) seed extracts. Int J Food SCI Tech 2006;41:799-804.

Vidal CM, Aguiar TR, Phansalkar R, McAlpine JB, Napolitano JG, Chen SN, Araújo LS, Pauli GF, Bedran-Russo A. Galloyl moieties enhance the dentin biomodification potential of plant-derived catechins. Acta Biomater. 2014;10:3288-94. doi: 10.1016/j.actbio.2014.03.036

Original Article