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Table of Contents   
ORIGINAL RESEARCH  
Year : 2022  |  Volume : 33  |  Issue : 2  |  Page : 169-173
Antibacterial efficacy of manuka honey, ocimum sanctum, curcuma longa and 0.2% chlorhexidine mouthwash on the level of streptococcus mutans and lactobacillus acidophilus - A randomized controlled trial


1 Department of Public Health Dentistry, Teerthanker Mahaveer Dental College and Research Centre, Moradabad, Uttar Pradesh, India
2 Dental College, JNIMS, Manipur, India
3 Department of Public Health Dentistry, Hazaribag, Jharkhand, India
4 Department of Biomedical Dental Sciences, College of Dentistry, Imam Abdulrahman Bin Faisal University, Moradabad, India
5 Department of Oral Pathology and Microbiology, Teerthanker Mahaveer Dental College and Research Centre, Moradabad, India

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Date of Submission21-Feb-2022
Date of Decision01-Jul-2022
Date of Acceptance01-Jul-2022
Date of Web Publication13-Oct-2022
 

   Abstract 


Objective: To assess the effects of Manuka honey, Ocimum sanctum, Curcuma longa, and 0.2% chlorhexidine mouthwash on Streptococcus mutans and Lactobacillus acidophilus levels. Material and Methods: A randomized controlled trial will be conducted on dental students of Teerthanker Mahaveer Dental College and Research Centre, Moradabad. The study participants will be divided into four groups. Each group will have a total of 20 individuals. By using a lottery system, Group A (Manuka honey mouthrinse), Group B (Ocimum sanctum mouthrinse), Group C (Curcuma longa mouthrinse), and Group D (0.2% chlorhexidine mouthrinse) will be chosen. To match the circadian cycle, saliva will be collected at baseline and again after 2 weeks between 10 and 10.30 a.m. The sterile container will subsequently be delivered to the microbiological laboratory and processed as soon as possible to measure Streptococcus mutans and Lactobacillus acidophilus count. For 2 weeks, participants were told to use 10 mL of mouthrinse twice daily. Results: The mean oral hygiene index-simplified (OHI-S) score of all the four groups showed reduction in their scores from baseline to after the study period. For both S. mutans and L. acidophilus, there was a substantial Percentage Reduction (PR) between the prerinse and postrinse samples in all four groups. Discussion: Because quantitative actions play a crucial part in the caries disease process, the changes in microbial activity before and after administration of experimental mouthwashes were examined. Conclusion: Essential oil aqueous extracts were as efficient antibacterial mouthwashes as chlorhexidine and iodine mouthwashes.

Keywords: Curcuma longa, Lactobacillus acidophilus, manuka honey, Ocimum sanctum, Streptococcus mutans

How to cite this article:
Jain A, Singh V, Lukram A, Chatterjee S, Khan AM, Dawar G. Antibacterial efficacy of manuka honey, ocimum sanctum, curcuma longa and 0.2% chlorhexidine mouthwash on the level of streptococcus mutans and lactobacillus acidophilus - A randomized controlled trial. Indian J Dent Res 2022;33:169-73

How to cite this URL:
Jain A, Singh V, Lukram A, Chatterjee S, Khan AM, Dawar G. Antibacterial efficacy of manuka honey, ocimum sanctum, curcuma longa and 0.2% chlorhexidine mouthwash on the level of streptococcus mutans and lactobacillus acidophilus - A randomized controlled trial. Indian J Dent Res [serial online] 2022 [cited 2022 Nov 29];33:169-73. Available from: https://www.ijdr.in/text.asp?2022/33/2/169/358438



   Introduction Top


Honey is defined as a sugary fluid ingredient manufactured by bees that have been used as a source of nutrients as well as remedy since antiquity. Because of its high osmotic pressure and antibacterial properties, honey can be kept for long periods without spoiling. It has been demonstrated to have wide-ranging antibiotic action, inhibiting the growth of a variety of bacteria, fungi, protozoa, and viruses. The slow dilution of untreated honey produces hydrogen peroxide. Honey has been found to contain a number of compounds that have antibacterial properties. It also stimulates blood lymphocytes, according to the current studies.[1],[2],[3],[4],[5]

Medicinal plants are commonly employed by medical practitioners in their daily practice to treat a variety of ailments. Modern medicines may incorporate herbal extracts or synthetic substances derived from herbs, but the natural form has fewer side effects and is a less expensive alternative to pharmaceutical treatments. Ocimum sanctum is one such natural ingredient that has largely gone unrecognized despite its promise for treating a wide range of ailments. Oral disorders such as caries and periodontal disease are still widespread issues that have been linked to a variety of oral health issues. This is especially true in both emerging and wealthy countries' underprivileged populations.[6],[7],[8]

Due of its extensive traditional usage and negligible side effects, Curcuma longa L. (Zingiberaceae family) and its polyphenolic component curcumin have been subjected to a range of antimicrobial research. Curcumin and C.[9],[10] longa rhizome extract have been shown to exhibit antimicrobial activity against a variety of bacteria, viruses, fungi, and parasites. Curcumin's potential antibacterial action made it a good option for synergistically enhancing the inhibitory effect of existing antimicrobial drugs. The rhizome of Curcuma longa has long been used as an antibacterial and insect repellant.[11],[12],[13],[14],[15]

Mouthrinses are commonly employed in the administration of active substances to the teeth and gums as a supplement to dental hygiene. By limiting bacterial attachment and eliminating bacterial biofilm, such products may be able to affect plaque accumulation. Controlling the bacterial biofilm on teeth is critical for maintaining oral health, and it can be accomplished through adequate oral hygiene, the use of fluoride treatments, and frequent dental check-ups. Another strategy is to use antimicrobial-active chemical agents on tooth surfaces to reduce biofilm formation. These compounds are commonly used as adjuvants in the prevention and treatment of oral illnesses because they can limit bacterial colonisation, development, and metabolism, interrupting the formation of mature biofilm and causing biochemical and ecological changes.[16],[17]

Several chemicals, including bisbiguanides like chlorhexidine gluconate, have been researched for their effectiveness on supragingival plaque and gingivitis. Chlorhexidine is the best studied and effective antiseptic for plaque inhibition and gingivitis prevention when used twice daily as mouthrinse. Many of the currently available mouthrinses, such as chlorhexidine mouthwash, have disadvantages such as altered taste sensations, tooth discoloration, and so on. Furthermore, evidence from a number of studies suggests that its effect on caries prevention is inconclusive. The World Health Organization (WHO) has recommended researchers to look into the usage of natural goods such as herb and plant extracts to combat such negative effects.[18],[19]

As a result, the current study will assess the effects of Manuka honey, Ocimum sanctum, Curcuma longa, and 0.2% chlorhexidine mouthwash on  Streptococcus mutans Scientific Name Search obacillus acidophilus levels.


   Material and Methods Top


For a period of 2 weeks, students at Teerthanker Mahaveer Dental College and Research Centre, Moradabad will participate in a randomised controlled study. The study's ethical approval will come from the Institutional Ethical Committee (letter No. TMDCRC/SM/002). Clinical Trial Registry in India (CTRI) has been done (REF/2022/07/055890).

Students who agreed to participate in the study, provided written consent, and who met the inclusion criteria were considered for the study. Individuals with systemic disorders, a history of antibiotic medication, or who used mouthrinses before were excluded from the study.

The sample size was estimated using the difference between the two studies, with a standard deviation of 2.563 and a 5% acceptable mean error at the 95% confidence interval. A total of 20 people were expected to participate in the study. Because there would be four groups in the present study, 20 people were assigned to each group, for a total of 80 people who participated in the study.

Each group will have a total of 20 individuals. By using a lottery system, Group A (Manuka honey mouthrinse), Group B (Ocimum anctum mouthrinse), Group C (Curcuma longa mouthrinse), and Group D (0.2% chlorhexidine mouthrinse) will be chosen.

Curcuma longa and Ocimum sanctum extract preparation

Leaves of Ocimum sanctum and Curcuma longa will be collected from courtyards and dried in the sun. The dried leaves will then be finely pulverised. The maceration of 200 gm of finely powdered Ocimum sanctum and Curcuma longa in 100% ethanol followed. It will then be filtered using Whatman filter paper to get a clear filtrate. The resulting filtrate will be reduced to a solid residue of Ocimum sanctum extract and Curcuma longa at a low temperature of 60°C. Around 12 gm of residue (extract) will be obtained from 200 gm of tulsi dissolved in 1 L of ethanol, yielding a 4% w/w yield.

Manuka honey mouthrinse preparation

Nearly 10 gm of commercial Manuka honey will be added to 100 mL of boiling distilled water, and the mixture will be cooked for another 30 min. To achieve a concentration of 100% (w/v), the solution will be decreased to 10 mL. Before usage, the Manuka honey solution will be cooled to room temperature and stored in sterile individual containers.

Saliva sampling and microbiological testing

For the next 2 weeks, everyone will be required to follow the same oral hygiene routines. For oral hygiene index-simplified (OHI-S), clinical examinations will be conducted out. Before collecting saliva and after using the mouthrinse, a simplified oral hygiene index will be recorded. Individuals conducting investigations will be given explicit instructions to fast for 1 h before saliva collection. About 1 mL of unstimulated saliva will be collected by instructing the children to allow saliva to gather in the floor of their mouth for at least 1 min without swallowing it, then expectorate into a sterile container.

To match the circadian cycle, saliva will be collected at baseline and again after 2 weeks between 10 and 10.30 a.m. The sterile container will subsequently be delivered to the microbiological laboratory and processed as soon as possible. For 2 weeks, participants were told to use 10 mL of mouthrinse twice daily.

Saliva will be collected in a sterile container after 2 weeks and promptly transferred to a microbiological laboratory, where it will be processed.

A stirrer will be used to manually homogenise the saliva sample. Approximately 1 mL of normal saline will be used to dilute 100 μL of saliva (1:10 dilution). Nearly 5 of the 1:10 dilution sample will be streaked on mitis salivarius bacitracin (MSB) agar, a selective medium for Streptococcus mutans and Lactobacillus acidophilus, using an inoculation loop (2 mm inner diameter). MSB agar plates will be incubated at 37°C for 48 h. The microbiologist will do the microbial count and record the counts of Streptococcus mutans and Lactobacillus acidophilus. The counts of Streptococcus mutans and Lactobacillus acidophilus will be expressed as number of colony forming units/mL and will be done with a digital colony counter.

The study will be blinded in order to avoid bias. When the study was over, the pharmacist from the pharmaceutical college who handled the blinding and gave the mouthrinses in coded assignments decoded the mouthrinses to disclose the following contents: Two examiners will be chosen to ensure blind evaluation of the study participants for Group A (Manuka honey mouthrinse), Group B (Ocimum sanctum mouthrinse), Group C (Curcuma longa mouthrinse), and Group D (0.2% chlorhexidine mouthrinse). The statistician was kept in the dark about who was assigned to which of the four groups.

To reduce intra and inter examiner variability, both examiners will be educated prior to the start of the study under the supervision of a senior expert. Throughout the clinical examination, they will be reviewed for satisfactory agreement at regular intervals. The value of the Cronbach's alpha coefficient for inter examiner and intra examiner variability will be evaluated. The study will be done in accordance with the declaration of Helsinki's guidelines for human trials because it is a randomised control experiment.

Each group will receive 10 mL of the research solutions in sterile disposable cups, which will be distributed to the individuals in their respective groups at baseline and after 2 weeks.

On a weekly basis, each participant will be given their own mouthrinse to use at home. The sodium salt of benzoic acid is sodium benzoate. It has a long history of usage as a preservative in foods and other items, and its safety has been shown. To make the mouthrinse with Ocimum sanctum and Curcuma longa extract taste better, an artificial sweetening agent will be added. In Ocimum sanctum, Curcuma longa extract, and Manuka honey mouthrinses, sodium benzoate is utilised at a concentration of 0.03% to 0.1%.

The methyl ester of phydroxybenzoic acid is methylparaben. It is a nonvolatile, stable chemical that has been utilised in foods as an antibacterial preservative. Ocimum sanctum, Curcuma longa extract, and honey mouthrinses will all contain this methylparaben.

In the morning, when participants report to the Department, the examiner will supervise mouthrinsing.

The Statistical Package for the Social Sciences (SPSS) version 22 will be used to enter and evaluate data (SPSS Inc., Chicago, IL, USA). The Chi-square test was performed to compare the counts of Streptococcus mutans and Lactobacilus acidophilus at baseline and after 2 weeks. The Kruskal–Wallis test and the Mann–Whitney U-test will be used to compare OHIS between research groups.


   Results Top


The mean OHI-S score of all the four groups showed reduction in their scores from baseline to after the study period but the maximum decline was in Chlorhexidine group and Manuka honey group followed by Ocimum sanctum group and Curcuma longa group [Table 1].
Table 1: Mean OHI-S score within group and between group at baseline and after 2 weeks

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The mean percentage reduction (PR) of S. mutans and L. acidophilus are shown in [Table 2] and [Table 3]. For both S. mutans and L. acidophilus, there was a substantial PR between the prerinse and postrinse samples in all four groups. When compared to Curcuma longa and Ocimum sanctum, Manuka Honey and chlorhexidine demonstrated highly substantial antibacterial action against S. mutans and L. acidophilus. There was no significant difference in antibacterial activity between Manuka honey (75.6%) and chlorhexidine (71.6%) against Streptococcus mutans. When compared to the other two materials, Curcuma longa and Ocimum sanctum had the lowest Percentage Reduction (PR) (65%) and (68%), respectively, against the species. There was no significant difference in antibacterial activity between Manuka honey (79.7%) and chlorhexidine (78%) against Lactobacillus acidophilus.
Table 2: Microbial analysis percentage of Streptococcus mutans

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Table 3: Microbial analysis percentage of Lactobacillus acidophilus

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When compared to the other two materials, Curcuma longa and Ocimum sanctum had the lowest PR (62%) and (64%), respectively, against the species.

After the completion of 2-week period of the study within comparison and in between comparison among all the four groups were found to be statistically significant in both Strepcoccus mutans and Lactobacillus acidophilus species [Table 4].
Table 4: In between and within comparison of four subject groups

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   Discussion Top


Because quantitative actions play a crucial part in the caries disease process, the changes in microbial activity before and after administration of experimental mouthwashes were examined. Chlorhexidine gluconate is positively charged and has a strong affinity for negative ions found in microbial cell membranes. It interferes with the enzymatic action of dehydrogenase and adenosine triphosphatase in bacterial cell walls, causing cell membrane disruption 8. The chlorhexidine demonstrated a significant reduction in microbial activity in this investigation, resulting in effective anticariogenicity. Chlorhexidine, on the other hand, has been linked to tooth discolouration 9 and an unpleasant taste 10. For people with a higher caries risk, the efficacy of synthetic antimicrobial agents as mouthwashes is still unknown. To compare its efficacy with chlorhexidine, two medicinal herbs and Manuka honey with proven and stronger inhibitory action against common cariogenic infections were chosen. Because of its ability to suppress S. mutans and L. Acidophilus, Manuka Honey, Ocimum sanctum, and Curcuma longa mouthrinse demonstrated to be an efficient anticaries mouthwash in this investigation. Manuka Honey had the best antibacterial efficiency, followed by chlorhexidine, Ocimum sanctum, and Curcuma longa. S. mutans and L. acidophilus are inhibited by the aqueous extracts Manuka honey, Curcuma longa, and Ocimum sanctum. As a result of their impact on Streptococcus mutans and Lactobacillus acidophilus, Manuka honey and herbal extract can be recommended for the treatment of dentinal caries.


   Conclusion Top


Essential oil aqueous extracts were as efficient antibacterial mouthwashes as chlorhexidine and iodine mouthwashes. Future research should focus on enhancing the practicality of herbal mouthrinses so that they can be utilised as a viable alternative to 0.2% chlorhexidine alone, with intense antibacterial activity and cost-effective caries prevention procedures.

Declaration of patient consent

The authors certify that they have obtained all appropriate patient consent forms. In the form the patient(s) has/have given his/her/their consent for his/her/their images and other clinical information to be reported in the journal. The patients understand that their names and initials will not be published and due efforts will be made to conceal their identity, but anonymity cannot be guaranteed.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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Correspondence Address:
Dr. Ankita Jain
Department of Public Health Dentistry, Teerthanker Mahaveer Dental College and Research Centre, Moradabad, Uttar Pradesh
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijdr.ijdr_159_22

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