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ORIGINAL ARTICLE  
Year : 2019  |  Volume : 22  |  Issue : 5  |  Page : 479-482
An in vitro determination of antibacterial effect of silver nanoparticles gel as an intracanal medicament in combination with other medicaments against Enterococcus fecalis


1 Department of Conservative Dentistry and Endodontics, Dr. H.S.J.I.D.S and Hospital, Panjab University, Chandigarh, India
2 Department of Microbiology, Dr. H.S.J.I.D.S and Hospital, Panjab University, Chandigarh, India

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Date of Submission30-Mar-2020
Date of Decision28-Jun-2020
Date of Acceptance04-Jul-2020
Date of Web Publication04-Aug-2020
 

   Abstract 

Aim: The main aim of the study is to compare the antibacterial effect of Silver nanoparticle gel alone and combination of silver nanoparticle gel with various medicaments.
Materials and Methods: Intracanal dressings: Group 1 –Silver Nanocure gel, Group 2 - Silver Nanocure gel+ Cavisept gel(1:1),Group 3- Silver Nanocure gel+ Aveu-Cal gel(1:1) ,Group 4 – Silver Nanocure gel +Cavisept gel +Aveu-Cal gel(1:1:1) were taken on a culture plate inoculated with E.faecalis. Antibacterial activity was assessed using Agar diffusion test and results were noted as diameter of growth inhibition zone.
Statistical Analysis: Student t –test was used to analyse results.
Results: The diameter of combination of Silver nanocure gel+Cavisept +Aveu-Cal gel(1:1:1) was highest in comparison to other medicaments tested.
Conclusion: Intracanal dressing with a combination of all the three {Silver nanocure gel+Cavisept +Aveu-Cal gel(1:1:1) } is the best treatment for elimination of highly resistant Enterococcus faecalis in root canals.

Keywords: Agar diffusion test, inhibition zone, medicaments

How to cite this article:
Jhamb S, Singla R, Kaur A, Sharma J, Bhushan J. An in vitro determination of antibacterial effect of silver nanoparticles gel as an intracanal medicament in combination with other medicaments against Enterococcus fecalis. J Conserv Dent 2019;22:479-82

How to cite this URL:
Jhamb S, Singla R, Kaur A, Sharma J, Bhushan J. An in vitro determination of antibacterial effect of silver nanoparticles gel as an intracanal medicament in combination with other medicaments against Enterococcus fecalis. J Conserv Dent [serial online] 2019 [cited 2022 May 26];22:479-82. Available from: https://www.jcd.org.in/text.asp?2019/22/5/479/291316

   Introduction Top


Bacteria and their by-products are responsible for various pulpal and periapical diseases.[1],[2] Endodontic infections can be classified as biofilm-mediated infections. Enterococcus faecalis is the most dominant biofilm forming bacteria found in teeth with periradicular pathologies.[3] It has an intrinsic resistance to irrigant solutions, intracanal medicaments, several antibiotics, and highly alkaline pH. It has the ability to tolerate starvation, harsh environment, and invade deeply into the dentinal tubules.[3],[4] To achieve complete success in root canal treatment, suppression of the biofilm is necessary. Cleaning and shaping of the root canal along with irrigation protocol have been shown to effectively decrease the number of microorganisms in the root canal. However, these procedures are unable to completely eliminate bacteria from lateral canals, isthmuses, and apical deltas. The augmentation of the above antibacterial protocol is achieved by utilizing intracanal medicaments.[4],[5],[6]

Calcium hydroxide and chlorhexidine have been used since a long time for the removal of E. faecalis. Recently, nanoparticles have come into limelight. Nanoparticles are a class of newer medicaments which are hypothesized to have antibacterial effect. They cause disruption of the biofilm due to their nano size and structure. The nano size provides increased surface area which can absorb other medicaments and exert antimicrobial effect.[7],[8] Silver nanoparticles are commonly used as they show strong bactericidal potential against Gram-positive, Gram-negative, and multidrug-resistant bacteria.[9],[10] Silver has an ability to interact with bacterial cell wall leading to structural changes and then damaging the tissue protein. In this study, we compared and evaluated the antibacterial effect of silver nanoparticles alone and combination of silver nanoparticles with calcium hydroxide and chlorhexidine against E. faecalis. The study will help us evaluate and find the medicament which is highly efficacious in the removal of E. faecalis and hence will lessen the number of endodontic failures.


   Materials and Methods Top


Culturing enterococcus faecalis

Pure culture of E. faecalis (ATCC 29212) was used as the test microorganism. Bacterial colonies were isolated after 24 h of incubation and then suspended in 5 ml of brain heart infusion broth. They were incubated at 37°C for 4 h. After culturing E. faecalis in the culture media, 0.5 McFarland standard of the bacterial suspension was prepared and cultured on Mueller–Hinton agar culture medium by a sterile swab in all directions. Then, in each culture plate, 5 wells measuring 6 mm in diameter and 2 mm in depth were created with a sterile pipette for placement of the material samples.

Division into groups

Four groups were made according to the medicament used:

  • Group 1 – Silver nanocure gel
  • Group 2 – Silver nanocure gel + Cavisept gel (1:1)
  • Group 3 – Silver nanocure gel + Aveu-Cal gel (1:1)
  • Group 4 – Silver Nanocure gel + Cavisept gel + Aveu-Cal gel (1:1:1).


Microbiologic testing

The wells were then filled with respective medicament from each group. The process was repeated five times and all microbial tests were performed under aseptic conditions. The plates were then incubated at 37°C for 1 week, and the diameter of the growth inhibition zone was evaluated at 24 h, 48 h, and 1 week after culture. The mean of the three values was calculated and reported as the diameter of zone of inhibition. The results were analyzed using student's t-test.


   Results Top


Antibacterial activity of all the three medicaments in different combinations is shown in [Table 1]. The results obtained revealed statistically significant difference in the antibacterial effect of combination of silver nanocure gel with various medicaments in comparison to silver nanocure gel alone. Silver nanocure gel in combination with calcium hydroxide (Group 2) was more efficacious as compared to silver nanocure gel alone (Group 1) (P < 0.00001). Antibacterial activity of silver nanocure gel in combination with chlorhexidine (Group 3) was also significantly higher than silver nanocure gel alone (P < 0.00001). When silver nanocure gel was combined with both calcium hydroxide and chlorhexidine (Group 4) the antibacterial activity was again higher in comparison to silver nanocure gel used alone (P < 0.00001). The zone of inhibition for combination of all the three medicaments was maximum [Figure 1].
Table 1: Diameter of zone of inhibition of different medicaments

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Figure 1: Average diameter

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   Discussion Top


Root canal disinfection is the mainstay of successful endodontic treatment.[11] Major challenges involved in achieving the above-mentioned goal are the microorganism-induced biofilms and the limitations associated with the currently used intracanal medicaments and irrigants in eliminating the same. Hence, in the present study, the effectiveness of four medicaments was studied and their efficacy against E. faecalis was estimated. The nanoparticles and their various combinations have been used as intracanal medicaments and their effect on E. faecalis is measured by observing the diameter of zone of inhibition in the agar diffusion test.

E. faecalis was used as a test organism in the study as it has a major role in the etiology of persistent endodontic infections. It is majorly responsible for root canal failures since it has the ability to thrive in the root canals in adverse situations and form biofilm and has high resistance against commonly used intracanal medicaments.[12],[13],[14]

When silver nanocure gel (Group 1) was used alone, it showed slight antibacterial activity against E. faecalis. This can be explained by the fact that silver ions released due to oxidation of nanoparticles bind to the cell membrane and penetrate inside the bacteria and react with specific proteins. This alters the bacterial metabolism and inhibits vital enzymatic systems, such as respiratory process and cellular division, resulting in cell death. The results in the present study are in agreement with the study done by Kim et al.[15]

In Group 2, where silver nanocure gel was used in combination with Cavisept gel showed a more significant antibacterial activity in comparison to silver nanocure gel alone, but it was significantly lesser than the other combinations tested (P < 0.0001). Chlorhexidine has a unique property of substantivity.[16],[17],[18],[19] However, recent studies have shown that biofilm formation by E. feacalis is a major hindrance in the working of chlorhexidine.[20] To increase the efficacy of cavisept gel, silver nanoparticles have been added to it. Its scientific basis is that silver gets ionized and destroys the cell membrane allowing the penetration of chlorhexidine leading to cell death.[21] This helps in increased removal of E. faecalis from the root canals and its efficient cleaning.

In Group 3, silver nanocure gel in combination with Aveu cal gel was used and this also proved significantly more effective against E. faecalis. This finding is consistent with other studies.[22],[23],[24],[25] This finding was in agreement with the findings of other studies which suggested combination of silver nanoparticles and calcium hydroxide to be better in comparison to silver nanoparticle gel and calcium hydroxide gel used alone.[23] It has been observed that silver nanoparticles from silver nanocure gel create pits in the cell wall of microorganism which leads to disruption of the biofilm.[22],[23] The other mechanism of action is increased amount of calcium hydroxide delivery due to changes in bacterial permeability. The release of hydroxyl ions in the root canal environment kills bacteria by damaging the cytoplasmic membrane, denaturation of proteins, and damaging DNA.[24],[25] Hence, the above combination helps in effective elimination of E. faecalis. The Group 4 where silver nanocure gel was used in combination with Cavisept gel and Aveu cal gel, showed most significant reduction in the E. faecalis activity. All the three medicaments combined have been shown to have a synergistic effect and hence have been able to cause maximum eradication of E. faecalis from the microbial flora of root canals. This was observed in our study. As per our knowledge, this is the first study done so far in the English literature that has tested all three medicaments in combination.

Limitations

The limitation of the study is its in vitro nature. The above findings need to be tested in vivo. The strength of the study is the efficacy of all the three intracanal medicaments studied with latest scientific approach that is in combination with nanotechnology, which is the future of dentistry. We are hopeful of a bright future of these small particles.


   Conclusion Top


In this in vitro study, it may be concluded that silver nanocure gel, cavisept gel, and Aveu-cal gel combination is the best among for elimination of E. faecalis from the root canal.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
   References Top

1.
Kakehashi S, Stanley HR, Fitzgerald RJ. The effects of surgical exposures of dental pulps in germ-free and conventional laboratory rats. Oral Surg Oral Med Oral Pathol 1965;20:340-9.  Back to cited text no. 1
    
2.
Möller AJ, Fabricius L, Dahlén G, Ohman AE, Heyden G. Influence on periapical tissues of indigenous oral bacteria and necrotic pulp tissue in monkeys. Scand J Dent Res 1981;89:475-84.  Back to cited text no. 2
    
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Madhubala MM, Srinivasan N, Ahamed S. Comparative evaluation of propolis and triantibiotic mixture as an intracanal medicament against Enterococcus faecalis. J Endod 2011;37:1287-9.  Back to cited text no. 3
    
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Sabrah AH, Yassen GH, Gregory RL. Effectiveness of antibiotic medicaments against biofilm formation of Enterococcus faecalis and Porphyromonas gingivalis. J Endod 2013:39:1385-89.  Back to cited text no. 4
    
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Wu D, Fan W, Kishen A, Gutmann JL, Fan W. Evaluation of antibacterial efficacy of Silver nanoparticles against Enteroccocus faecalis biofilms. J Endod 2014:40:285-90.  Back to cited text no. 5
    
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Delgado RH, Gasparoto TH, Sipert CR, Pinheiro CR, Moraes IG, Garcia RB. Antimicrobial effects of calcium hydroxide and chlorhexidine on Enterococcus faecalis. J Endod 2010:36:1389-93.  Back to cited text no. 6
    
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Cheng Z, Al Zaki A, Hui JZ, Muzykantov VR, Tsourkas A. Multifunctional nanoparticles: Cost versus benefit of adding targeting and imaging capabilities. Science 2012;338:903-10.  Back to cited text no. 7
    
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Rai MK, Deshmukh SD, Ingle AP, Gade AK. Silver nanoparticles: The powerful neoweapon against multi-drug resistant bacteria. J Appl Microbiol. 2012:112:841-52.  Back to cited text no. 8
    
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Morones JR, Elechiguerra JL, Camacho A, Holt K, Kouri JB, Ramírez JT, et al. The bactericidal effect of silver nanoparticles. Nanotechnology 2005;16:2346-53.  Back to cited text no. 9
    
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Ayala-Nunez NV, Villegas HH, Turrent LC, Padilla CR. Silver nanoparticles toxicity and bactericidal effect against methicillin resistant Staphylococcus aureus: Nanoscale does matter. J Nanobiotechnology 2009;5:2-9.  Back to cited text no. 10
    
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Siqueira JF Jr., Araújo MC, Garcia PF, Fraga RC, Dantas CJ. Histological evaluation of the effectiveness of five instrumentation techniques for cleaning the apical third of root canals. J Endod 1997;23:499-502.  Back to cited text no. 11
    
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Kishen A. Advanced therapeutic options for endodontic biofilms. Endod Topics 2010:22:99-123.   Back to cited text no. 12
    
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Siqueira JF Jr., Rocas IN, Ricucci D. Biofilm in endodontic infection. Endod Topics 2010:22:33-49.  Back to cited text no. 13
    
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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.  Back to cited text no. 14
    
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Kim JS, Kuk E, Yu KN, Kim JH, Park SJ, Lee HJ, et al. Antimicrobial effects of silver nanoparticles. Nanomedicine 2007;3:95-101.  Back to cited text no. 15
    
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Mohammadi Z, Abbott PV. The properties and applications of chlorhexidine in endodontics. Int Endod J 2009:42:288-302.  Back to cited text no. 16
    
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Basrani B, Tjäderhane L, Santos JM, Pascon E, Grad H, Lawrence HP, et al. Efficacy of chlorhexidine- and calcium hydroxide-containing medicaments against Enterococcus faecalis in vitro. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2003;96:618-24.  Back to cited text no. 17
    
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Gomes BP, Souza SF, Ferraz CC, Teixeira FB, Zaia AA, Valdrighi L, et al. Effectiveness of 2% chlorhexidine gel and calcium hydroxide against Enterococcus faecalis in bovine root dentin in vitro. Int Endod J 2003:36:267-75.  Back to cited text no. 18
    
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Shen Y, Qian W, Chung C, Olsen I, Haapasalo M. Evaluation of the effect of two chlorhexidine preparations on biofilm bacteria in vitro: A three-dimensional quantitative analysis. J Endod 2009;35:981-5.  Back to cited text no. 19
    
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Shen Y, Stojicic S, Haapasalo M. Antibacterial efficacy of chlorhexidine against bacteria in biofilms at different stages of development. J Endod 2011:37:657-61.  Back to cited text no. 20
    
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Castellano JJ, Shafii SM, Ko F, Donate G, Wright TE, Mannari RJ, et al. Comparative evaluation of silver-containing antimicrobial dressings and drugs. Int Wound J 2007;4:114-22.  Back to cited text no. 21
    
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de Souza-Filho FJ, Soares Ade J, Vianna ME, Zaia AA, Ferraz CC, Gomes BP. Antimicrobial effect and pH of chlorhexidine gel and calcium hydroxide alone and associated with other materials. Braz Dent J 2008;19:28-33.  Back to cited text no. 22
    
23.
Javidi M, Afkhami F, Zarei M, Ghazvini K, Rajabi O. Efficacy of a combined nanoparticle/calcium hydroxide root canal medication on elimination of Enterococcus faecalis. Aust Endod J 2014:40:61-5.  Back to cited text no. 23
    
24.
Morones JR, Elechiguerra JL, Camacho A, Holt K, Kouri JB, Ramirez JT, et al. The bactericidal effect of silver nanoparticles. Nanotechnology 2005:6:2346-53.  Back to cited text no. 24
    
25.
Mohammadi Z, Shalavi S, Yazdizadeh M. Antimicrobial activity of Calcium hydroxide in endodontics: A Review. Chonnam Med J 2012:48:133-40.  Back to cited text no. 25
    

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Correspondence Address:
Dr. Swaty Jhamb
Room No. 201, Department of Conservative and Endodontics, Dr. H.S.J.I.D.S, Panjab University, Chandigarh
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/JCD.JCD_113_20

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