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ORIGINAL ARTICLE
Year : 2018  |  Volume : 36  |  Issue : 2  |  Page : 142-150
 

Comparative evaluation of the antimicrobial susceptibility and cytotoxicity of husk extract of Cocos nucifera and chlorhexidine as irrigating solutions against Enterococcus Faecalis, Prevotella Intermedia and Porphyromonas Gingivalis – An in-vitro study


1 Department of Pedodontics and Preventive Dentistry, KLE University's KLE VK Institute of Dental Sciences, Belagavi, Karnataka, India
2 Dr. Prabhakar Kore Basic Science and Research Center, KLE University, Belagavi, Karnataka, India

Date of Web Publication2-Jul-2018

Correspondence Address:
Divyata Kohli
Department of Pedodontics and Preventive Dentistry, KLE University's KLE VK Institute of Dental Sciences, Nehru Nagar, Belagavi - 590 010, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/JISPPD.JISPPD_1176_17

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   Abstract 


Aim and Background: The aim of the present study is to evaluate and compare the antimicrobial susceptibility and cytotoxicity of Cocos nucifera and chlorhexidine (CHX) as irrigating solutions against Enterococcus faecalis, Prevotella intermedia, and Porphyromonas gingivalis. Materials and Methods: The ethanolic extract of husk of C. nucifera was prepared. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract were determined using the serial broth dilution method and its cytotoxicity was evaluated against human periodontal fibroblasts using 3-(4,5-dimethyl-thiazole-2-yl)-2,5-diphenyl tetrazolium bromide assay. Antibacterial susceptibility for two irrigating solutions, namely 2% CHX gluconate irrigant (Group I) and 1.5% C. nucifera husk irrigant (Group II), was tested against P. gingivalis, P. intermedia, and E. faecalis. Results: The MIC and MBC of C. nucifera husk extract for P. gingivalis were 468.75 μg/ml and 1562.5 μg/ml, for P. intermedia were 48.8 μg/ml and 1875 μg/ml, and for E. faecalis were 1562.5 μg/ml and 3750 μg/ml, respectively. The extract was nontoxic to the human periodontal fibroblast. Both the materials have shown similar antibacterial susceptibility and no difference was observed at baseline, 10, 30, and 60 min using two-way repeated measures of ANOVA. However, a statistically significant difference was observed between different time points for P. gingivalis and P. intermedia using Bonferroni multiple comparison test (f = 826.1390, P ≤ 0.05). Conclusion: 1.5% of ethanolic husk extract of C. nucifera has a significant antibacterial action against polymicrobial dental biofilm and its activity is comparable to that of 2% CHX which validates its use as a future irrigating solution for overcoming bacterial resistance with synthetic agents.


Keywords: Antibacterial, children, chlorhexidine, Cocos nucifera, herbals in dentistry, irrigating solution


How to cite this article:
Kohli D, Hugar SM, Bhat KG, Shah PP, Mundada MV, Badakar CM. Comparative evaluation of the antimicrobial susceptibility and cytotoxicity of husk extract of Cocos nucifera and chlorhexidine as irrigating solutions against Enterococcus Faecalis, Prevotella Intermedia and Porphyromonas Gingivalis – An in-vitro study. J Indian Soc Pedod Prev Dent 2018;36:142-50

How to cite this URL:
Kohli D, Hugar SM, Bhat KG, Shah PP, Mundada MV, Badakar CM. Comparative evaluation of the antimicrobial susceptibility and cytotoxicity of husk extract of Cocos nucifera and chlorhexidine as irrigating solutions against Enterococcus Faecalis, Prevotella Intermedia and Porphyromonas Gingivalis – An in-vitro study. J Indian Soc Pedod Prev Dent [serial online] 2018 [cited 2019 Jul 21];36:142-50. Available from: http://www.jisppd.com/text.asp?2018/36/2/142/235673





   Introduction Top


Despite modern advances in the prevention of dental caries, many teeth are lost prematurely in children. One of the primary objectives of pediatric dentistry is to preserve the health and integrity of primary teeth until they naturally exfoliate. When the infectious and inflammatory process is advanced, it is not possible to carry out conservative pulpal treatment rendering pulpectomy as the ideal treatment to eliminate microorganisms through chemo-mechanical disinfection of the root canal system.[1]

The peculiar morphology of the deciduous dentition with apical delta and significantly abundant lateral and accessory canals makes the infectious process difficult to ward off, especially in necrotic canals, abscesses, and sinus tract infections.[2] Hence, for complete elimination of infection, irrigation is an important step for the success of pulpectomy.[3]

Studies have demonstrated that despite the use of antimicrobial agents, microorganisms such as Enterococcus faecalis, Prevotella intermedia, and Porphyromonas gingivalis still persist in the root canal of the primary teeth.

Sodium hypochlorite (NaOCl) and chlorhexidine (CHX) are one of the most popular irrigating solutions. NaOCl has antibacterial activity and the ability to dissolve necrotic tissue remnants. However, it is cytotoxic and has potential risk of injuring the permanent tooth germ. The constant increase in antibiotic resistance and side effects caused by the synthetic agents has led to search for other natural alternatives.[4]

Among the many herbal products that are being used in dentistry, Cocos nucifera is one such herb that has not been explored yet. Furthermore, very few studies have been carried out in Indian scenario utilizing this herb as an irrigant.

The beneficial medicinal effects of C. nucifera including the antibacterial activity result from the secondary products present in the plant, although it is usually not attributed to a single compound but a combination of the metabolites.[5]

Phytochemical screening of C. nucifera conducted by Alviano et al. has reported that this plant material is rich in alkaloids, flavonoids, catechin, and epicatechin together with condensed tannins, which confers on its potent antimicrobial properties.[6]

The use of these agents in primary teeth and their therapeutic effect in this treatment modality as irrigating solutions need to be further evaluated. Hence, in this study, we compared and evaluated the antimicrobial activity of C. nucifera and CHX as irrigating solutions against E. faecalis, P. intermedia, and P. gingivalis.


   Materials and Methods Top


The study was conducted in the Department of Pediatric and Preventive Dentistry at KLE University's KLE VK Institute of Dental Sciences, Belagavi and KLE University's Dr. Prabhakar Kore Basic Science Research Center, Belagavi, Karnataka, India.

Outpatients reporting to the Department of Pediatric and Preventive Dentistry for the purpose of extraction of primary and permanent teeth at the KLE University's VK Institute of Dental Sciences, Belagavi, and satisfying the inclusion criteria were selected for the study. Ethical clearance for the study was obtained from the Institutional review board of the college.

A total of twenty freshly extracted primary and permanent teeth in children were included for isolation of fibroblasts. Teeth with draining sinus or fistula, cyst, and granuloma were excluded from the study.

Preparation of extract

The 4 kg husk of C. nucifera used in the study was obtained from the local market and verified by the KLE University's Shri. BMK Ayurveda Mahavidyalaya, Belagavi, and Regional Medical Research Center branch of Indian Council of Medical Research, Belagavi. The husk was dried in hot air oven at 45°C until a constant weight of the sample is reached. This was ground into fine powder using a husk grinder available in the local market. Exactly 1.5 kg of the powdered sample was extracted with ethanol in the ratio of 3:2 (v/v) for 4 days.[7] The yield obtained was dark brown in color and weighed 110.5 g [Figure 1].
Figure 1: (a) Photograph showing 4 kg of husk of Cocos nucifera collected from the local market and (b) 110.5 g of brown ethanolic extract of husk of Cocos nucifera

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Determination of minimum inhibitory concentration and minimum bactericidal concentration

The ethanolic extract of C. nucifera was weighed and dissolved in 0.5 ml of dimethyl sulfoxide (MERCK, Specialty Pvt., Ltd.) present in microcentrifuge (Eppendorf) tube (Tarsons Products Pvt., Ltd., West Bengal). The minimum inhibitory concentration (MIC) was determined using broth dilution method [8] and, to confirm the inhibitory concentration, each of these serial dilutions was plated on blood agar culture plates under laminar air flow for both Group I (control group) and Group II (experimental group) in triplicates. The culture plates were incubated for 48 h for P. gingivalis and P. intermedia and 24 h for E. faecalis. Colony-forming units (CFUs) were recorded after the respective times of incubation.

The MIC was taken as the lowest concentration that prevented the growth of the bacteria [Figure 2]. The minimum bactericidal concentration (MBC) was taken as the lowest concentration required to kill the bacteria [Figure 3].
Figure 2: Photograph showing minimum inhibitory concentration of 2% chlorhexidine gluconate (Group I) and Cocos nucifera husk extract (Group II) for Porphyromonas gingivalis, Prevotella Intermedia, and Enterococcus faecalis

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Figure 3: Photograph showing minimum bactericidal concentration (MBC) of 2% chlorhexidine gluconate (Group I) and Cocos nucifera husk extract (Group II) for Porphyromonas gingivalis, Prevotella intermedia, and Enterococcus faecalis

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Cytotoxicity assay

The extracted teeth were transferred to the phosphate-buffered saline transfer media and sent to KLE University's Dr. Prabhakar Kore Basic Science Research Center. The MTT 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (HiMedia laboratories Pvt., Limited, Mumbai; CAS no.: 298-93-1) assay was then carried out on human periodontal fibroblasts isolated from extracted teeth.[9] Serial dilutions of the extract were prepared in Dulbecco's modified Eagle medium (Genetix biotech Asia Pvt., Ltd., India). 100 μl of the extract of different concentrations was added to the wells and incubated for 24 h in the presence of 5% CO2, at 37°C into CO2 incubator (Yorco, York Scientific Industries, India). The absorbance of this colored solution can be quantified by measuring at a certain wavelength (usually between 500 and 600 nm) by a microplate absorbance reader (Bio-Rad Laboratories India Pvt., Ltd., Haryana, India). The degree of light absorption depends on the solvent [Figure 4]. The optical density was measured at a wavelength of 570 nm. The study was performed in triplicates. The result represents the mean of three readings.
Figure 4: Photograph showing microscopic picture of the surviving fibroblasts at concentrations of (a) 3.12%, (b) 6.25%, (c) 12.5%, (d) 25%, (e) 50%, (f) and 100% of the Cocos nucifera husk extract

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Preparation of Cocos nucifera husk extract irrigant

Weighed amount of C. nucifera husk extract (1.5 gm) was solubilized with 3.0 g of (2-Hydroxypropyl)-β-cyclodextrin (Sigma-Aldrich Co., USA) in the ratio of 1:2 with water to increase the solubility. Weighed quantity of complex (4.025 gm) was dissolved in 100 ml of purified water and other preservatives such as methyl paraben and propyl paraben were added to the above solution to get the final irrigant [Figure 5].
Figure 5: Photograph showing (a and b) Preparation of the Cocos nucifera husk irrigant and (c) 1.5% Cocos nucifera irrigant stored in a sterile container

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Antibacterial susceptibility testing

Antibacterial susceptibility for two irrigating solutions, namely 2% CHX gluconate irrigant (Vishal Dentocare Pvt., Ltd., Mumbai) (Group I) and 1.5% C. nucifera husk irrigant (Group II), was tested by direct contact test on 96-well microplates (SPL Life Sciences Co., Ltd., Korea). Direct contact test [10] is based on determining the turbidity of microbial growth in microplates. Facultative strains of E. faecalis (ATCC BAA-2128), P. gingivalis (BAA-308/W83), and P. intermedia (ATCC-25611) were grown on brain–heart infusion (BHI) agar media. Microorganisms were subcultured in appropriate culture media and under gaseous conditions to confirm their purity. Facultative strain was inoculated individually into tube containing 5 ml of sterile saline. Biofilms were established using the 96-well microtiter plates (SPL Life Sciences Co., Ltd., Korea). 0.01% serum albumin was coated for 1 h and discarded. The bacterial suspension of P. gingivalis, P. intermedia, and E. faecalis was adjusted to 0.5 McFarland scale = 1.5 × 108 CFU spectrophotometrically at 590 nm and 50 μL (approximately 107) and was placed in the 96-well microplate in triplicates and incubated in anaerobic jar (HiMedia laboratories Pvt., Ltd., Mumbai) for 72 h (Column A, Column B, and Column C). After incubation for 1 h in humidity at 37°C, there was a direct contact between the respective organism and the test material. 240 μL of BHI media was added to all the three columns. The kinetics of bacterial outgrowth in each well was measured at 595 nm using a microplate absorbance reader (Bio-Rad Laboratories India Pvt. Ltd., Haryana, India) every 10, 30, and 60 min in triplicates [Figure 6].{Figure 6}


   Results Top


The mean of MIC of CHX gluconate (Group I) for P. gingivalis is 0.20%/ml, for P. intermedia is 0.026%/ml, and for E. faecalis is 0.03%/ml. The mean of MIC of C. nucifera husk extract (Group II) for P. gingivalis is 468.75 mg/ml, for P. intermedia is 48.8 mg/ml, and for E. faecalis is 1562.5 mg/ml. The mean of MBC of CHX gluconate (Group I) for P. gingivalis is 0.5%/ml, for P. intermedia is 0.15%/ml, and for E. faecalis is 1%/ml [Table 1]. The mean of MBC of C. nucifera husk extract (Group II) for P. gingivalis is 1562.5 mg/ml, for P. intermedia is 1875 mg/ml, and for E. faecalis is 3750 mg/ml [Table 2]. The C. nucifera husk extract was nontoxic to the human periodontal fibroblast against which the cytotoxic activity has been tested [Table 3]. On comparison of antibacterial susceptibility of two groups at baseline, 10, 30, and 60 min against P. gingivalis, P. intermedia, and E. faecalis using two-way repeated measures of ANOVA, no statistical difference was observed at different time interval points [Table 4]. However, a statistically significant difference was observed between different time points (f = 826.1390, P ≤ 0.05) for P. gingivalis and P. intermedia when pair-wise comparison was done using Bonferroni multiple comparison test.
Table 1: Table showing Mean and Standard error of Minimum inhibitory concentration (MIC) of the two groups namely Chlorhexidine gluconate (Group I) and Cocos nucifera husk extract (Group II) against Porphyromonas gingivalis, Prevotella intermedia and Enterococcus faecalis

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Table 2: Table showing Mean and Standard error of Minimum Bactericidal concentration (MBC) the two groups namely Chlorhexidine gluconate (Group I) and Cocos nucifera husk extract (Group II) against Porphyromonas gingivalis, Prevotella intermedia and Enterococcus faecalis

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Table 3: Table showing mean of optical densities (OD) of surviving cells of two study groups namely Chlorhexidine gluconate (Group I) and Cocos nucifera husk extract (Group II) at a wavelength of 570 nm for different concentrations

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Table 4: Table showing the comparison of two groups namely 2% Chlorhexidine gluconate irrigant (Group I) and 1.5% Cocos nucifera husk irrigant (Group II) against Porphyromonas gingivalis, Prevotella intermedia, Enterococcus faecalis at Baseline, 10 minutes, 30 minutes and 60 minutes using two-way repeated measures of ANOVA

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   Discussion Top


The oral cavity is an affluent playground of microorganisms harboring more than 500 species of bacteria amassing almost 1010 bacteria. Figdor and Sundquist stated that “Leaving behind the nutritionally rich and different environment of the oral cavity, micro-organisms that establish in the root canal must breach enamel, invade dentin, overwhelm the immune response of the pulp and settle in the remaining necrotic tissue.”[11]

In the present study, ATCC strains of P. gingivalis, P. intermedia, and E. faecalis were used based on the observation made in a study by Dhariwal et al. that the most predominant organisms isolated from the infected, abscessed primary teeth were the black pigmented colonies of obligatory anaerobic bacilli such as P. intermedia, Porphyromonas species, Bacteroides species, and Fusobacterium species followed by the obligatory Gram-positive cocci Peptostreptococcus species.[12]

The primary objectives in pulpectomy for primary teeth are the maintenance of the teeth in the mouth in a healthy state, biomechanically disinfect and adequately fill the root canals with a resorbable material, promote physiologic root resorption, and fulfill its role as a space maintainer for succedaneous teeth.[13] The tortuous, bizarre, ribbon-shaped anatomy of the primary root canals and the multiple accessory canals can make biomechanical preparation difficult and lead to perforations.[13],[14],[15] The chemical treatments are used along with the limited mechanical debridement, to disinfect and remove necrotic material within the somewhat inaccessible canals, rather than “shaping” of the canals.[13] Herbal products have been used since ancient times in folk medicine involving both eastern and western medical traditions. The husk fiber of C. nucifera (Palmae) is rich in catechin and epicatechin together with condensed tannins, which confers to its aqueous extract a potent antioxidant characteristic. Antibacterial, antiviral, antileishmanial, anti-lymphoproliferative, and anti-neoplastic activities were also observed.[6] Phytochemical studies of the ethanolic extract of coconut fiber (mesocarp) revealed that there is the presence of phenols, tannins, leucoanthocyanidins, flavonoids, triterpenes, steroids and alkaloids, while a butanol extract recovered triterpenes, saponins, and condensed tannins.[16] Extract was obtained using ethanol as the main solvent as difficulties were encountered while attempting to dissolve it in distilled water.

The MIC was tested using the broth dilution method and the MBC was detected by subculturing these on antibiotic-free media. The advantages of this technique include generation of a quantitative result (the MIC) and the examination of a substantial number of bacterial cells because of a large initial inoculum provided by the relatively large volume of broth in each of the tubes. The principal disadvantages include the tedious manual task of preparing the twofold solutions for each individual test, relatively large amount of reagents and space required for such tests, and the possibility of making errors in the preparation of antibiotic concentrations.[17]

The inhibition produced by the plant extracts against a particular organism depends on various extrinsic and intrinsic factors. The results of this study were comparable with that of a similar study done by Jose et al.[18] where they determined the antibacterial activity of the ethanolic C. nucifera husk against various cariogenic organisms and P. intermedia. However, our study showed a lower MIC and MBC for P. intermedia when compared to the study by Jose et al.[18] As per the literature search done by us, there were no other studies that could be used to compare our results.

Many types of biomaterials are utilized in dental practice procedures. Assessing cytotoxicity based on several cytotoxicity testing methods is a necessary step in evaluating the biocompatibility of all biomaterials. Therefore, in the present study, the IC50 value which is the half maximal inhibitory concentration of substance was recorded for CHX gluconate at 2% and C. nucifera husk extract at 100%, 50%, 25%, 12.5%, 6.25%, and 3.12% [Table 3]. The IC50 value for C. nucifera ethanolic husk extract is beyond the concentrations tested. Therefore, the C. nucifera ethanolic husk extract appears to be nontoxic to the human periodontal fibroblast against which the cytotoxic activity had been tested. According to the literature search done, no studies were found that evaluated the cytotoxicity of ethanolic extract of C. nucifera husk; however, there was a study done by Lima EB et al.[16] who has conducted toxicity studies in animal models and found that ethyl acetate and methanolic fractions of C. nucifera husk extracts are nontoxic.

The crude extract could have been used for the final preparation of the C. nucifera husk extract irrigant. However, an entirely miscible solution was not obtained when mixed with other preservatives. To overcome this problem, a complex of C. nucifera husk extract was prepared with beta-cyclodextrin in the ratio of 1:2.

Microorganisms undergo certain genetic and phenotypic variations in biofilm growth when compared to their planktonic counterparts. This results in increased resistance of the microorganisms to topical antimicrobials. Therefore, it is more prudent to check the efficacy of antimicrobial agents against the microorganisms in their biofilm mode of growth rather than in their planktonic form.[19] Thus, in the present study, a biofilm of P. gingivalis, P. intermedia, and E. faecalis was prepared to evaluate the antibacterial effectiveness of the two irrigants.

The results showed that the antibacterial susceptibility of the two groups, namely 2% CHX gluconate (Group I) and 1.5% C. nucifera husk irrigant (Group II) for P. gingivalis, P. intermedia, and E. faecalis, was comparable [Figure 7]. On comparison of the antibacterial effectiveness of the two groups against P. gingivalis and P. intermedia and E. faecalis at baseline, 10, 30, and 60 min [Table 4], no difference was observed between the two study groups with antibacterial susceptibility at different time interval points (P ≥ 0.05). However, a statistically significant difference was found between different time points when pair-wise comparison of antibacterial susceptibility was done against P. gingivalis and P. intermedia [Table 5], indicating that the antibacterial susceptibility increased over time (P ≤ 0.05).
Figure 7: Graphical representation of intergroup comparison of antibacterial susceptibility in two groups, namely 2% chlorhexidine gluconate irrigant (Group I) and 1.5% Cocos nucifera husk irrigant (Group II) against (a) Porphyromonas gingivalis, (b) Prevotella intermedia, and (c) Enterococcus faecalis at baseline, 10, 30, and 60 min

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Table 5: Table showing the pair wise comparisons of antibacterial susceptibility of two groups namely 2% Chlorhexidine gluconate irrigant (Group I) and 1.5% Cocos nucifera husk irrigant (Group II) against Porphyromonas gingivalis, Prevotella intermedia, Enterococcus faecalis at Baseline, 10 min, 30 min and 60 min by Bonferroni test

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According to the literature search done by us, there were no studies that compared the antibacterial effectiveness of CHX with C. nucifera husk extract irrigant. However, the results of the antibacterial effectiveness of CHX were similar as quoted by Mohammadi.[20] The antibacterial effectiveness of C. nucifera husk irrigant could be attributed to a lot of secondary metabolites present in the plant. One of the major constituents of the plant is the polyphenols which impart a hydrophobic activity to the extract. This, in turn, causes partitioning of the lipid cell membrane of the bacterial cell wall causing leakage of molecules and ions from the cells and its death ultimately.[21] The tannins present in the extract provide an astringent effect and an antibacterial effect by reacting with the proteins through nonspecific forces and lead to cell death. The flavonoids and the flavones act by binding with the extracellular and soluble proteins and forming complexes with the cell wall leading to cell lysis. It is a known fact that catechin and epicatechin provide antibacterial activity by promoting cell cycle arrest and causing apoptosis of cancerous cells. Since husk of C. nucifera contains catechin and epicatechin, it could be contemplated that these components could be responsible for its antibacterial activity.[18] The increased antibacterial effectiveness of C. nucifera husk extract could also be because of the complex of the extract with beta-cyclodextrin which increased its solubility, thus increasing the availability of the active constituents responsible for the antibacterial property.

This in vitro study has shown results that are very promising and can be effectively used for the management of infected primary root canals. As per the literature search, C. nucifera husk extract consists of astringent, anti-oxidant, and analgesic properties due to which it can be used for the preparation of an ideal pulpotomy agent.


   Conclusion Top


Ethanolic husk extract of C. nucifera has a significant inhibitory action against the endodontic pathogens, which indicates the presence of active compounds that can be incorporated into modern oral care systems for overcoming bacterial resistance with synthetic agents. As the mystery of the tree of heaven or the tree of life continues to unravel, the results of the present study can be further justified by a larger sample size and its use in a clinical scenario.

Acknowledgment

I would like to thank Mr. U.D. Bolmal for his help to carry out this study.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
   References Top

1.
Valdez-Gonzalez C, Mendez-Gonzalez V, Torre-Delgadillo G, Flores-Reyes H, Gaitan-Fonseca C, Pozos-Guillen AJ, et al. Effectiveness of oxidative potential water as an irrigant in pulpectomized primary teeth. J Clin Pediatr Dent 2012;37:31-5.  Back to cited text no. 1
    
2.
Jolly M, Singh N, Rathore M, Tandon S, Banerjee M. Propolis and commonly used intracanal irrigants: Comparative evaluation of antimicrobial potential. J Clin Pediatr Dent 2013;37:243-9.  Back to cited text no. 2
[PUBMED]    
3.
Ruviére DB, Leonardo MR, da Silva LA, Ito IY, Nelson-Filho P. Assessment of the microbiota in root canals of human primary teeth by checkerboard DNA-DNA hybridization. J Dent Child (Chic) 2007;74:118-23.  Back to cited text no. 3
    
4.
Prabhakar J, Senthilkumar M, Priya MS, Mahalakshmi K, Sehgal PK, Sukumaran VG, et al. Evaluation of antimicrobial efficacy of herbal alternatives (Triphala and green tea polyphenols), MTAD, and 5% sodium hypochlorite against Enterococcus faecalis biofilm formed on tooth substrate: An in vitro study. J Endod 2010;36:83-6.  Back to cited text no. 4
    
5.
Parekh J, Jadeja D, Chanda S. Efficacy of aqueous and methanol extracts of some medicinal plants for potential antibacterial activity. Turk J Biol 2005;29:203-10.  Back to cited text no. 5
    
6.
Alviano WS, Alviano DS, Diniz CG, Antoniolli AR, Alviano CS, Farias LM, et al. In vitro antioxidant potential of medicinal plant extracts and their activities against oral bacteria based on Brazilian folk medicine. Arch Oral Biol 2008;53:545-52.  Back to cited text no. 6
[PUBMED]    
7.
Akinpelu DA, Alayande KA, Aiyegoro OA, Akinpelu OF, Okoh AI. Probable mechanisms of biocidal action of Cocos nucifera husk extract and fractions on bacteria isolates. BMC Complement Altern Med 2015;15:116.  Back to cited text no. 7
    
8.
Balouiri M, Sadiki M, Ibnsouda SK. Methods for in vitro evaluating antimicrobial activity: A review. J Pharm Anal 2016;6:71-9.  Back to cited text no. 8
[PUBMED]    
9.
Odabaş ME, Ertürk M, Çınar Ç, Tüzüner T, Tulunoğlu Ö. Cytotoxicity of a new hemostatic agent on human pulp fibroblasts in vitro. Med Oral Patol Oral Cir Bucal 2011;16:e584-7.  Back to cited text no. 9
    
10.
Zhang H, Shen Y, Ruse ND, Haapasalo M. Antibacterial activity of endodontic sealers by modified direct contact test against Enterococcus faecalis. J Endod 2009;35:1051-5.  Back to cited text no. 10
[PUBMED]    
11.
Figdor D, Sundqvist G. A big role for the very small – Understanding the endodontic microbial flora. Aust Dent J 2007;52:S38-51.  Back to cited text no. 11
[PUBMED]    
12.
Dhariwal NS, Hugar SM, Harakuni S, Sogi S, Assudani HG, Mistry LN, et al. A comparative evaluation of antibacterial effectiveness of sodium hypochlorite, curcuma longa, and Camellia sinensis as irrigating solutions on isolated anaerobic bacteria from infected primary teeth. J Indian Soc Pedod Prev Dent 2016;34:165-71.  Back to cited text no. 12
[PUBMED]  [Full text]  
13.
Finn SB. Clinical Pedodontics. 4th ed. Philadelphia: W.B. Saunders; 1998.  Back to cited text no. 13
    
14.
Ingle JI, Bakland LK. Endodontics. 5th ed. Hamilton, London: B.C. Decker Inc.; 2002.  Back to cited text no. 14
    
15.
Curzon ME, Roberts JF, Kennedy DB. Kennedy's Pediatric Operative Dentistry. 4th ed. Oxford: John Wright and Sons; 1997.  Back to cited text no. 15
    
16.
Lima EB, Sousa CN, Meneses LN, Ximenes NC, Santos Júnior MA, Vasconcelos GS, et al. Cocos nucifera (L.) (Arecaceae): A phytochemical and pharmacological review. Braz J Med Biol Res 2015;48:953-64.  Back to cited text no. 16
    
17.
Jorgensen JH, Ferraro MJ. Antimicrobial susceptibility testing: A review of general principles and contemporary practices. Clin Infect Dis 2009;49:1749-55.  Back to cited text no. 17
[PUBMED]    
18.
Jose M, Cyriac MB, Pai V, Varghese I, Shantaram M. Antimicrobial properties of Cocos nucifera (coconut) husk: An extrapolation to oral health. J Nat Sci Biol Med 2014;5:359-64.  Back to cited text no. 18
[PUBMED]    
19.
Mistry KS, Sanghvi Z, Parmar G, Shah S. Comparative evaluation of antimicrobial activity of herbal extracts with 5.25% sodium hypochlorite against multispecies dentinal biofilm. Saudi Endod J 2016;6:71-6.  Back to cited text no. 19
  [Full text]  
20.
Mohammadi Z. Chlorhexidine gluconate, its properties and applications in endodontics. Iran Endod J 2008;2:113-25.  Back to cited text no. 20
[PUBMED]    
21.
Prashant GM, Chandu GN, Murulikrishna KS, Shafiulla MD. The effect of mango and neem extract on four organisms causing dental caries: Streptococcus mutans, Streptococcus salivavius, Streptococcus mitis, and Streptococcus sanguis: An in vitro study. Indian J Dent Res 2007;18:148-51.  Back to cited text no. 21
[PUBMED]  [Full text]  


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 5], [Figure 7]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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  2005 - Journal of Indian Society of Pedodontics and Preventive Dentistry | Published by Wolters Kluwer - Medknow 
Online since 1st May '05