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Table of Contents
ORIGINAL ARTICLE
Year : 2021  |  Volume : 9  |  Issue : 1  |  Page : 8-10

Comparative evaluation of antimicrobial efficacy of herbal v/s synthetic irrigating solutions: An in vitro study


1 Department of Pediatric and Preventive Dentistry, College of Dental Sciences and Research Center, Bopal, Ahmedabad, Gujarat, India
2 Dentistry Department, Government Medical College & Hospital, Surat, Gujarat, India
3 Krishnam Dental Clinic, Vadodara, Gujarat, India
4 Consultant Pedodontist, Delhi, India
5 Private Practitioner, Ahmedabad, Gujarat, India
6 Private Practitioner, Sai Krupa Hitech Dental Clinic, Surendranagar, Gujarat, India

Date of Submission09-Jan-2021
Date of Acceptance25-Jan-2021
Date of Web Publication25-Feb-2021

Correspondence Address:
Dr. Himani Gandhi
Department of Pediatric and Preventive Dentistry, College of Dental Sciences and Research Center, Bopal, Ahmedabad, Gujarat.
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/INJO.INJO_2_21

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  Abstract 

The major objective in root canal treatment is to disinfect the entire root canal system. Despite adequate treatment, failure occurs due to the presence of pathogenic residual bacteria in sites within the canal. In dentistry, because of the cytotoxic reactions of most of the commercial intracanal medicaments used and their inability to eliminate bacteria from dentinal tubules, there is a quest for novel bioactive materials/ herbal alternatives that are popular, increased shelf life, and low toxicity. This study is designed to explore the antimicrobial efficacy of pomegranate, garlic, and aloe vera extracts as a root canal irrigant against Enterococcus faecalis. Out of the three medicaments used in this study to test their antimicrobial activity against Enterococcus faecalis in comparison to 0.2% Chlorhexidine, it was found that none of the medicaments had a mean zone of inhibition more than 0.2% Chlorhexidine.

Keywords: Herbal, irrigation, medicament


How to cite this article:
Gandhi H, Dave DS, Shah N, Pathak A, Patel KR, Martin P. Comparative evaluation of antimicrobial efficacy of herbal v/s synthetic irrigating solutions: An in vitro study. Int J Oral Care Res 2021;9:8-10

How to cite this URL:
Gandhi H, Dave DS, Shah N, Pathak A, Patel KR, Martin P. Comparative evaluation of antimicrobial efficacy of herbal v/s synthetic irrigating solutions: An in vitro study. Int J Oral Care Res [serial online] 2021 [cited 2021 Jun 17];9:8-10. Available from: https://www.ijocr.org/text.asp?2021/9/1/8/310182




  Introduction Top


The major objective in root canal treatment is to disinfect the entire root canal system. Despite adequate treatment, failure occurs due to the presence of pathogenic residual bacteria in sites within the canal.[1] Several chemicals and therapeutic agents have been used to disinfect the root canal. The most effective among them are Sodium hypochlorite, Chlorhexidine, and Calcium hydroxide, which possess varying degrees of antibacterial activity.[2]Enterococcus faecalis is the most commonly isolated species from the canals of teeth presenting post-treatment diseases, accounting for up to 77% of therapeutic failures.[3]

Considering the ineffectiveness, potential side effects, and safety concerns of synthetic drugs, the herbal alternatives for endodontic usage might prove to be advantageous.[2] Various irrigating agents have been used during root canal procedures. However each one of them has their own demerits. As a result, there is a quest for novel bioactive materials that are safer and biodegradable.

Pomegranate (Punica granatum Linn) extract is found to inhibit Staphylococcal enterotoxin A production and is active against several highly pathogenic and sometimes antibiotic-resistant organisms.[4] Garlic (Allium sativum L.) has many properties, such as antimicrobial features; the common organisms inhibited by garlic include Streptococcus mutans, Staphylococcus aureus, and Escherichia coli.[1] It has been proven in several studies that aloe vera shows considerable antimicrobial activity against various species such as Streptococcus pyogen, Enterococcus faecalis, Candida albicans, and Staphylococcus aureus. It has been shown that E. faecalis may tolerate the antibacterial effects of calcium hydroxide treatment.[5]

This study is designed to explore the antimicrobial efficacy of pomegranate, garlic, and aloe vera extracts as a root canal irrigant against E. faecalis.


  Materials and Methods Top


The bacterial stock culture of E. faecalis strain (ATCC 29212) was obtained from JP Laboratories, Davangere, India. The extracts of pomegranate, garlic, and aloe vera were obtained from Bapuji Pharmacy College, Davangere, Karnataka, India.

The standard strain of E. faecalis (ATCC 29212) was grown on Brain Heart Infusion (BHI) broth overnight; turbidity was adjusted to 0.5 Mc Farland scale to obtain a cell density of 1.5 × 108 bacterial/mL; and it was inoculated in Mueller Hinton Agar plates. Four round wells measuring about 4 mm deep and 8 mm in diameter were punched by using a sterile stainless steel template; they were numbered as 1, 2, 3, and 4 consecutively for the different concentrations used for each of the test groups to be evaluated. As a control, 0.2% chlorhexidine gluconate solution was taken for the evaluation.

After making serial dilutions of each of the extracts and three round wells in each of the agar plates, 50 µL of specific concentration of each extract was dispensed into each well by using a sterile micropipette. All agar plates were then incubated at 37°C for 24h, according to Clinical Laboratory Standard Institute guidelines (CLSI). After 24h of incubation at 37ºC, zones of inhibition (that is areas where no growth of bacteria is present) around each well were examined. They appeared as a clear, circular halo surrounding the wells. Diameters of the bacterial growth inhibition zones or halos were measured by using an Hi Antibiotic Zone scale in millimeters, and this represented the inhibition value.


  Results Top


The mean zone of inhibition was measured for each sample, and the results obtained are listed in [Table 1]. The results were subjected to statistical analysis by applying analysis of variance and post hoc Tukey HSD tests for multiple comparisons.
Table 1: Mean Values of Zone of Bacterial Inhibition (mm) of Five medicaments at Four Different Concentrations and Control Chlorhexidine after 24 h

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On applying one-way analysis of variance, a statistically significant difference was seen between the zone of inhibition of five samples within groups and between groups, that is P < 0.001. Post hoc Tukey HSD tests were applied to make multiple comparisons between different groups, such as Group A (0.2% chlorhexidine), Group B (pomegranate), Group C (garlic), and Group D (aloe vera) with different concentrations of 5%, 25%, 50%, and 100%, respectively.

The mean zone of inhibition for positive control, that is Group A (0.2% chlorhexidine) was 18.36 mm. Comparisons between Group A (0.2% chlorhexidine), Group B (pomegranate), Group C (garlic), and Group D (aloe vera) were statistically significant with a P value of <0.001.

Out of the three medicaments used in this study to test their antimicrobial activity against E. faecalis in comparison to Group A (0.2% chlorhexidine), it was found that none of the medicaments had a mean zone of inhibition more than Group A (0.2% chlorhexidine).


  Discussion Top


Complete debridement and effective disinfection of the root canal space is an important prerequisite for achieving long-term success of nonsurgical endodontics.[6]

Various chemicals are used to disinfect the root canal. Sodium hypochlorite (NaOCl), chlorhexidine, and normal saline are the most commonly used root canal irrigants.[2] Owing to the potential side effects, safety concerns, and ineffectiveness of conventional allopathic formulations, the consumption of preparations from medicinal plants has increased over the past few decades.

Herbal or natural products have been used in dental and medical practice for thousands of years and have become even more popular today due to their high antimicrobial activity, biocompatibility, anti-inflammatory, and antioxidant properties.[7]

Enterococcous faecalis, a Gram-positive facultative anaerobe, was chosen as the primary test organism because it is the dominant species recovered in failed endodontic cases.[6]

Chlorhexidine gluconate has become recognized as an effective oral antimicrobial agent. Chlorhexidine also possesses many properties, such as broad-spectrum antimicrobial activity, property of substantivity, and a relative absence of toxicity, which suggest that it is useful as an endodontic disinfectant.[8],[9]

The major advantages of using herbal alternatives are easy availability, cost-effectiveness, increased shelf life, low toxicity, and lack of microbial resistance reported so far.[10] Also, they are safer than synthetic alternatives, offering profound therapeutic benefits and more affordable treatment. Less activity of pomegranate might be due to less amount of pomegranate fruit used (100 gm); an aqueous extract of pomegranate was used. The next antimicrobial activity in a decreasing order was shown by aloe vera extract, with a mean zone of inhibition such as 5.43 mm at 100% concentration, which was in agreement with that of Subramanium et al.[10] This study revealed that aloe vera extract is not as effective as 0.2% chlorhexidine, whose mean zone of inhibition was 18.36 mm.

There are conflicting results with regard to the aloe vera extract because of the use of plants from different geographical locations with variations in their chemical composition and also because of different isolation techniques that are used to extract compounds from the aloe leaf pulp.[5] In our study also, aloe vera was effective only at 100% concentration, which is in accordance to an earlier report.[8]

In this study, the mean zone of inhibition of garlic extract is 1.40 mm at 100% concentration. One of the most biologically active compounds, allicin (diallyl thiosulfinate or diallyl disulfide) does not exist in garlic until it is crushed or cut; Allicin exerted antibacterial activity against Salmonella typhimurium, primarily by interfering with RNA synthesis.[9] According to Eswar et al.,[1] this could be the reason for the antibacterial efficacy of garlic extract.

However, there is scarce information on the quality, safety, and greater efficiency of these products for use in dentistry. As most of the studies are carried out ex vivo, more of these medicaments should be subjected to animal and human studies to determine their effectiveness, side effects, toxicity, and drug interactions.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Eswar K, Venkateshbabu N, Rajeswari K, Kandaswamy D Dentinal tubule disinfection with 2% chlorhexidine, garlic extract, and calcium hydroxide against enterococcus faecalis by using real-time polymerase chain reaction: In vitro study. J Conserv Dent 2013;16:194-8.  Back to cited text no. 1
    
2.
Pujar M, Makandar S Herbal usage in endodontics—a review. Int J Contemp Dent 2011;2:34-7.  Back to cited text no. 2
    
3.
Siqueira JF Jr, Rôças IN Exploiting molecular methods to explore endodontic infections: Part 2—redefining the endodontic microbiota. J Endod 2005;31:488-98.  Back to cited text no. 3
    
4.
Lansky EP Beware of pomegranates bearing 40% ellagic acid. J Med Food 2006;9:119-22.  Back to cited text no. 4
    
5.
Sureshchandra B, Kumar AJ Antibacterial efficacy of aloe vera extract on resistant antimicrobial strains in endodontics. J Endod 2011;23:59.  Back to cited text no. 5
    
6.
Bhardwaj A, Ballal S, Velmurugan N Comparative evaluation of the antimicrobial activity of natural extracts of morinda citrifolia, papain and aloe vera (all in gel formulation), 2% chlorhexidine gel and calcium hydroxide, against enterococcus faecalis: An in vitro study. J Conserv Dent 2012;15:293-7.  Back to cited text no. 6
    
7.
Prasannabalaji N, Muralitharan G, Sivanandan RN, Kumaran S, Pugazhvendan SR Antibacterial activities of some Indian traditional plant extracts. Asian Pac J Trop Dis 2012;2(Suppl 1):291-5.  Back to cited text no. 7
    
8.
Jeansonne MJ, White RR A comparison of 2.0% chlorhexidine gluconate and 5.25% sodium hypochlorite as antimicrobial endodontic irrigants. J Endod 1994;20:276-8.  Back to cited text no. 8
    
9.
Basrani B, Santos JM, Tjäderhane L, Grad H, Gorduysus O, Huang J, et al. Substantive antimicrobial activity in chlorhexidine treated human root dentin. Oral Surg Oral Med Oral Patho Oral Radiol Endod 2002;94:240-5.  Back to cited text no. 9
    
10.
Gulve NM, Gulve ND Comparison of antimicrobial efficacy of ginger extract and 2% sodium hypochlorite against Enterococcous faecalis using agar diffusion method. J Ind Dent Assoc 2010;4:347-9.  Back to cited text no. 10
    



 
 
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