Evaluation of Anti Bacterial Activity of Endodontic Sealers in Combination with Clove Oil

 

Hena Mariam Fathima1, R.V.Geetha2

1Graduate Student, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences, Saveetha University Chennai, Tamil Nadu, India.

2Associate Professor, Department of Microbiology, Saveetha Dental College, Saveetha Institute of Medical and Technical Sciences Saveetha University, Chennai, Tamil Nadu, India.

*Corresponding Author E-mail: rvgeetha2015@gmail.com

 

ABSTRACT:

Introduction: The main purpose of endodontic therapy is to clean and shape the root canal by means of instruments and chemical irrigation substances, in order to eliminate or reduce the amount of microorganisms, when the pulp is necrosed. However, infection may persist in canals with high anatomic complexity, with a great number of bacteria, especially facultatively anaerobic bacteria.In chronic pulpar necrosis, apical cement erosion may occur as well as the presence of bacteria in periradicular tissues, which are resistant against the organic defense of the host and the chemical solutions used in the root therapy. Clove essential oil, used as an antiseptic in oral infections, inhibits Gram-negative and Gram-positive bacteria. Clove oil has biological activities, such as antibacterial, antifungal, insecticidal and antioxidant properties. Materials and methods: The antimicrobial efficacy of commercially available root canal sealers namely zinc oxide eugenol and AH plus, were evaluated individually and also in combination with Clove oil against Enterococcus faecalis which was isolated from necrotic pulps and endodontic lesions using the disc diffusion technique. Results: Among the sealers tested AH plus showing the maximum effectiveness and the zone of inhibition was 18mm and for zinc oxide eugenol the zone of inhibition was 22mm in diameter. Conclusion: It can be concluded that the addition of clove oil to endodontic sealers increases the anti bacterial activity of the sealer. Clove oil is a natural, non toxic and affordable anti bacterial substance. Thus it can be widely used for dental practice involving endodontic sealers.

 

KEYWORDS: Antimicrobial, clove oil, endodontic sealers, Enterococcus faecalis, zinc oxide eugenol.

 

 


INTRODUCTION:

Endodontic sealers are used in conjunction with biologically acceptable semisolid or solid obturating materials to establish an adequate seal of the root canal system. There are some requirements for a material to be an endodontic sealer, namely: it should provide an excellent seal when set, radiopaque, dimensionally stable, non staining, insoluble in tissue fluids among others.

 

 

There are certain functions that are to be performed by endodontic sealers, some of which are: to act as a filler for canal irregularities and minor discrepancies between the root canal wall and the core filling material, to obturate the canal walls, act as a lubricant, radio opacity, microbial control assistance. The main purpose of endodontic therapy is to clean and shape the root canal by means of instruments and after chemical irrigation intra canal medicaments are used, to eliminate the microorganisms, when the pulp is necrosed1-3. After disinfection, the canal is sealed with a root filling, and one of main property of intra canal medicament is to have antimicrobial activity4. These include the elimination or reduction of microorganisms, rendering canal contents inert, prevention of post treatment pain, and to enhance anaesthesia.

 

Bacteria may survive after intra-canal medication for several reasons. Bacterial strains present in the root canal infection may be intrinsically resistant to the medicament, bacterial cells may be enclosed within anatomical variations inaccessible to the medicament, the medicament may be neutralized by tissue components and by bacterial cells or products, losing its antibacterial effects, medicaments may remain in the root canal system for insufficient time to reach and kill bacterial cells and bacteria may also alter their pattern of gene expression after changes in the environmental conditions. This alteration may allow them to survive in unfavorable environments5. Eugenol is the most important medicament seen in a dental office. Besides its pure form as sedative dressing and obtundent, it is used in varieties of zinc oxide eugenol cement formulations and liquid of endodontic sealers. It is also known as allyl guaiacol or eugenic acid, an odoriferous principle, the active ingredient of oil of cloves, comprising70-80% of its bulk6.

 

Syzygium aromaticum commonly known as Clove belongs to the family Myrtacea. Oil ofcloves, also known as clove oil, is an essential oil from the clove plant, Syzygium aromaticum. It is a natural analgesic and antiseptic used primarily in dentistry for its main ingredient eugenol7. The germicidal properties of the oil make it very effective for relieving dental pain, toothache, sore gums, and mouth ulcers. As a result, clove oil is added to numerous dental products and medications, including mouthwash and toothpaste and filling material as a temporary alternative to a root canal. Eugenol isalso said to have several disadvantages. Sometimes the free eugenol in endodontic sealers can cause certain toxicity. Eugenol is also a wellknown allergen; it may also act as a tissue irritant and cause a burning sensation. The intracanal medicaments are generally removed after 5-7 days. Combining it with clove oil enhances its efficacy so that it can be removed earlier. The mean minimum inhibitory concentration of clove oil was found to be 0.62±0.458. Today, there is a renewed interest in the uses of traditional plant sources as medicine. This revival of interest is mainly due to the widespread belief that ‘green medicine’ is safe and more dependable than the costly synthetic drugs. This study was done to evaluate the antimicrobial activity of the few endodontic sealers in combination with clove oil. Hence, this study was done to evaluate the antibacterial activity of clove oil with intracanal medicaments and bring about its use in the field of dentistry. The aim of the study was to evaluate the anti bacterial activity of endodontic sealers in combination with clove oil.

 

MATERIALS AND METHOD:

The antimicrobial efficacy of commercially available root canal sealers namely zinc oxide eugenol and AH plus,were evaluated individually and also in combination with Clove oil against Enterococcus faecalis which was isolated from necrotic pulps and endodontic lesions using the disc diffusion technique.

 

Zinc oxide Eugenol:

Zinc oxide eugenol [ZOE] sets because of a combination of physical and chemical reaction, yielding hardening of the mixture due to formation of zinc eugenolate. The presence of free eugenol tend to weaken the set and has increased cytotoxicity. The advantages of ZOE are ease of manipulation-plasticity-radiopaque with some germicidal properties-minimal staining–accepted working time, low setting time in the absence of moisture and good sealing properties. But it causes irritation and not easily absorbedfrom periapical tissues. Zinc eugenolate has the disadvantage, however, of being decomposed by water through a continuous loss of the eugenol. This makes ZOE a weak, unstable material and precludes its use in bulk, such as for retrofillings placed apically through a surgical approach.

 

AH-Plus:

AH Plus consists of a paste-paste system, which is delivered in two tubes and in a double barrel syringe. In addition to the diepoxide, the epoxide paste contains radio opaque fillers and aerosil. The amine paste consists of three different types of amines, radio opaque fillers and aerosil. AH Plus is characterised by very good mechanical properties, high radio opacity, little polymerisation shrinkage, low solubility, and, not least, a high degree of stability on storage. The radio opaque fillers used in AH Plus ensure an exceptionally good radio opacity of the material, even when applied in very thin layers. Tightness and insolubility of the polymerised material are relevant for the function of a root canal sealer.

 

Preparation of disc:

Both sealers were mixed according to manufacturer instructions and 50µl of clove oil was mixed to it and loaded on to sterile filter paper discs. Discs were also prepared with Sealers without clove oil.

 

METHODOLOGY:

The antibacterial effect of the root canal sealers is tested against Enterococcus feacalis by disc diffusion technique. The broth culture of the bacterial strain compared to Mac Farland’s standard 0.5 was prepared. Lawn culture of the test organism, Enterococcus feacalis was made on the Muller Hinton agar [MHA-Hi media M1084] plates using sterile cotton swab and the plates were dried for 15 minutes. Filter paper discs loaded with root canal sealers alone and also in combination with clove oil were placed on the agar plates inoculated with the test organism. The inoculated plate with the sealers were kept at room temperature to allow the diffusion of the agents through the agar and were incubated at 37°C overnight and the zone of inhibition of growth was measured in millimeter diameter. All the tests were done in triplicate to minimize the test error.

 

RESULT:

Measuring the Size of the Zone of Inhibition

Table 1. Anti bacterial activity of sealers

S.No

Root canal sealants

Zone of inhibition

[In mm diameter]

Sealer

Sealer +clove oil

1

Zinc oxide Eugenol

19

22

2

AH plus

8

18

3

0.2% chlorhexidine

22

22

 

Growth inhibitory zones around each sealer were evidenced by the lack of bactericidal colonization adjacent to the sealer. The inhibitory zones were measured in millimeters. Wider zones of inhibition were indicative to greater antimicrobial activity of involving sealers. The results are given in Table 1. Among the sealers tested AH plus showing the maximum effectiveness and the zone of inhibition was 18mm and for zinc oxide eugenol the zone of inhibition was 22mm in diameter.

 

 

Figure 1. Zones of inhibition

 

DISCUSSION:

In this study, we used the agar diffusion test, which is the most widely used in vitro method for the evaluation of antimicrobial activity9. This method allows direct comparisons between materials and also indicates which sealers are more likely to have antimicrobial activity within the root canal system10. Besides that, its results are highly influenced by the diffusibility of the material across the medium11. Here, the addition of clove oil has made a substantial difference in the anti bacterial activity as can be seen. It is seen especially in the endodontic sealer AH plus.

 

Daniela et al12. results showed that the most resistant microorganism to the antimicrobial activity of all sealers tested was Enterococcus faecalis. This is one of the most resistant species usually found in root canal infections13. It is usually persistent and difficult to treat14. It is frequently associated with failure of the root canal treatment15. The AH-Plus sealer presented antimicrobial activity too because of the components of the epoxy resin and due to the discreet liberation of formaldehyde16. Sanjay et al17 conducted a study comparing the anti bacterial efficacy of intracanal medicaments in combination with clove oil against enterococcus faecalis where it was found that Group II was found to be more effective with a zone of inhibition of 23 mm, 22 mm, 24 mm, 26 mm, and 25 mm. Group IV was also found to be more effective against E. faecalis than Group III with a zone of inhibition of 32 mm, 37 mm, 34 mm, 32 mm, and 29 mm. Thanish et al18 found that among all the endodontic sealers, endomethasone shows the maximum zone of inhibitionof about 28mm. Zinc oxide eugenol shows zone of inhibition of about 19mm and MTA fillapex of about 20mm. Claudio et al19. found thatEndomethasone C, Argoseal, and Bioseal Normal exhibited the largest inhibition zones for all tested microorganisms. Sicura Seal and AH Plus had little antibacterial effect against E. faecalis. Sicura Seal also showed the least antibacterial effect against S. mutans, together with Acroseal. AH Plus and Acroseal had the least effect against S. aureus. Sahar et al20. According to this study, endo-fill had the highest inhibitory zone and adseal, the least. In summary the sequence on antimicrobial activity of studied sealers was EndoFill, AH-Plus, Adseal. From the studies conducted by other authors it can be seen that different endodontic sealers have different anti bacterial properties which can be enhanced with different essential oils. From our study we can conclude that endodontic sealers in conjunction with clove oil can be used to enhance the anti bacterial activity since clove oil is proved to have anti bacterial effects.

 

CONCLUSION:

It can be concluded that the addition of clove oil to endodontic sealers increases the anti bacterial activity of the sealer. Clove oil is a natural, non toxic and affordable anti bacterial substance. Thus it can be widely used for dental practice involving endodontic sealers.

 

REFERENCES:

1        Patel V, Santerre JP, Friedman S. Suppression of bacterial adherence by experimental root canal sealers. J Endod 2000;26 (1): 20-4.

2        Siqueira JF Jr, Batista MM, Fraga RC, de Uzeda M. Antibacterial effects of endodontic irrigants on black-pigmented gram-negative anaerobes and facultative bacteria. J Endod 1998;24 (6): 414-6.

3        Siqueira JF Jr, Lima KC, Magalhães FA, Lopes HP, de Uzeda M. Mechanical reduction of the bacterial population in the root canal by three instrumentation techniques. J Endod 1999;25 (5): 332-5.

4        Siqueira JF Jr, Favieri A, Gahyva SM, Moraes SR, Lima KC, Lopes HP. Antimicrobial activity and flow rate of newer and established root canal sealers. J Endod 2000;26 (5): 274-7.

5        Siqueira JF Jr, Lopes HP. Mechanisms of antimicrobial activity of calcium hydroxide: A critical review. Int Endod J 1999;32 (5): 361-9.

6        Jyoti BB. Phytotherapeutics in conservative dentistry and endodontics: A review. J Conserv Dent 2005;8 (2): 31-9.

7        Thosar N, Basak S, Bahadure RN, Rajurkar M. Antimicrobial efficacy of five essential oils against oral pathogens: An in vitro study. Eur J Dent 2013;7 Suppl 1:S71-7.

8        Dhinahar S, Lakshmi T. Role of botanicals as antimicrobial agents in management of dental infections: A review. Int J Pharm Bio Sci 2011;2 (4): 690-704.

9        Leonardo MR, da Silva LAB, Tanomaru Filho M, Bonifácio KC, Ito IY. In vitro evaluation of antimicrobial activity of sealers and pastes used in endodontics. J Endod. 2000;26:391-4.

10      Siqueira JF Jr, Favieri A, Gahyva SMM, Moraes SR, Lima KC, Lopes HP. Antimicrobial activity and flow rate of newer and established root canal sealers. J Endod. 2000;26:274-7.

11      Fraga RC, Siqueira JF Jr, de Uzeda M. In vitro evaluation of antibacterial effects of photo-cured glass ionomer liners and dentin bonding agents during setting. J Prosthet Dent. 1996;76:483-6.

12      Daniela Cristina Miyagak, Elaine Manso Oliveira Franco de Carvalho, Carlos Roberto Colombo Robazza, Jorge Kleber Chavasco, Gustavo Labegalline LevoratoIn vitro evaluation of the antimicrobial activity of endo- dontic sealers. Braz Oral Res 2006;20 (4): 303-6.

13      Siqueira JF Jr, Batista MMD, Fraga RC, de Uzeda M. Antibacterial effects of endodontic irrigants on black-pig-mented gram-negative anaerobes and facultative bacteria. J Endod. 1998;24:414-6.

14      Engström B. The significance of enterococci in root canal treatment. Odontol Revy. 1964;15:87-106.

15      Sundqvist G, Figdor D, Persson S, Sjogren U. Micro- biologic analysis of teeth with failed endodontic treatment and the outcome of conservative re-treatment. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1998;85:86-93.

16      Siqueira JF Jr, Favieri A, Gahyva SMM, Moraes SR, Lima KC, Lopes HP. Antimicrobial activity and flow rate of newer and established root canal sealers. J Endod. 2000;26:274-7.

17      Sanjay Madhavan, Muralidharan, Comparing the antibacterial efficacy of intracanal medicaments in combination with clove oil against enterococcus faecalis, Asian J Pharm Clin Res, Vol 8, Issue 5, 2015, 136-138.

18      Thanish Ahamed S, Geetha R V, Comparative Effect of Commercially Available Endodontic Sealers Against Enterococcus faecalis, Int. J. Pharm. Sci. Rev. Res., 44 (2), May-June 2017; Article No. 36, Pages: 186-187

19      Claudio Poggio, Marco Lombardini , Marco Colombo, Alberto Dagna, Enrica Saino, Carla Renata Arciola, Livia Visai, Antibacterial effects of six endodontic sealers, Int J Artif Organs (2011; :9) 908-913.

20      Sahar Shakouie, Mahsa Eskandarinezhad, Shahriar Shahi, Hadi mokhtari, Mohammad FroughReihani, Mohammadhossein Soroush and Armin Gosili, Antimicrobial efficacy of AH-Plus, adseal and endofill against Enterococcus faecalis-An in vitro study, African Journal of Microbiology Research Vol. 6 (5), pp. 991-994, 9 February, 2012.

 

 

 

 

 

Received on 28.03.2018         Modified on 14.04.2018

Accepted on 24.06.2018       © RJPT All right reserved

Research J. Pharm. and Tech 2018; 11(8): 3355-3358.

DOI: 10.5958/0974-360X.2018.00616.9