|Year : 2021 | Volume
| Issue : 5 | Page : 508-513
Gel formulation and evaluation of antimicrobial activity of turmeric (Curcuma longa L.) from Aceh, Indonesia against Aggregatibacter actinomycetemcomitans
Zaki Mubarak1, Wahyu E Sari2, Sunnati Sunnati1
1 Faculty of Dentistry, Universitas Syiah Kuala, Banda Aceh, Indonesia
2 Faculty of Veterinary Medicine, Universitas Syiah Kuala, Banda Aceh, Indonesia
|Date of Submission||01-Feb-2021|
|Date of Decision||22-May-2021|
|Date of Acceptance||02-Aug-2021|
|Date of Web Publication||11-Oct-2021|
Dr. Zaki Mubarak
Faculty of Dentistry, Universitas Syiah Kuala, Banda Aceh.
Source of Support: None, Conflict of Interest: None
Aim: Periodontitis is a chronic disease of dental tissue caused by a bacterial infection, for instance, Aggregatibacter actinomycetemcomitans is caused by dominant bacteria. Currently, there are many cases regarding bacterial resistance to some antibiotics, so an alternative search for new antibiotics from herbal plants is needed. Turmeric (Curcuma longa Linn.) is a plant that is known to contain bioactive compounds that can function as antimicrobials. So far, research on turmeric extract in the form of a gel formula used as a herbal medicine for periodontitis in Indonesia has not been reported. The aim of this research was to study the oral gel formula of the indigenous turmeric extract from Aceh, Indonesia and to evaluate its antimicrobial efficacy against A. actinomycetemcomitans. Materials and Methods: This research was designed to test the effectiveness of C. longa gel in vitro in inhibiting the growth of A. actinomycetemcomitan. Turmeric rhizome was extracted using the maceration method with 96% ethanol; then, the extract was formulated into a gel with concentrations of 0%, 25%, 50%, 75%, and 100%. The curcuma gel was then tested for antimicrobials against A. actinomycetemcomitans bacteria. Results: The in vitro antimicrobial test showed that a 50% concentration of turmeric rhizome extracts gel was effective in inhibiting the growth of A. actinomycetemcomitans with an inhibition zone of 10.8 mm, which is classified as a strong inhibition zone. Conclusion: The efficacy of the oral gel formula containing C. longa extracts as a natural antimicrobial agent has been confirmed, and the possibility of applying it as periodontitis herbal medicine has been suggested.
Keywords: Aggregatibacter actinomycetemcomitans, Antimicrobials, Herbal Medicine, Oral Gel, Turmeric Rhizome
|How to cite this article:|
Mubarak Z, Sari WE, Sunnati S. Gel formulation and evaluation of antimicrobial activity of turmeric (Curcuma longa L.) from Aceh, Indonesia against Aggregatibacter actinomycetemcomitans. J Int Oral Health 2021;13:508-13
|How to cite this URL:|
Mubarak Z, Sari WE, Sunnati S. Gel formulation and evaluation of antimicrobial activity of turmeric (Curcuma longa L.) from Aceh, Indonesia against Aggregatibacter actinomycetemcomitans. J Int Oral Health [serial online] 2021 [cited 2021 Dec 6];13:508-13. Available from: https://www.jioh.org/text.asp?2021/13/5/508/327864
| Introduction|| |
Periodontitis is a destructive inflammatory disease of the tooth-supporting tissue that is caused by specific microorganisms. Periodontitis causes permanent damage to the periodontium tissue, namely the gingiva, connective tissue, periodontal ligaments, and alveolar bone. The dominant bacteria in periodontitis are anaerobic Gram-negative bacteria, namely Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. Antibiotics are generally used in the treatment of periodontitis. Systemic administration of antibiotics has great potential to control the bacteria that cause periodontitis. However, long-term and inappropriate use of antibiotics can increase the incidence of antibiotic drug resistance. Currently, there are many cases regarding bacterial resistance to some antibiotics, including A. actinomycetemcomitans bacteria. Based on a previous study, A. actinomycetemcomitans bacteria was reported be resistant to amoxicillin, azithromycin, and metronidazole, penicillin, metronidazole, clindamycin, amoxicillin, ceftazidime, tetracycline, and amoxicillin/clavulanic acid. Due to the high number of cases of antibiotic resistance, including bacterial causes for periodontitis, currently, there are many searches for alternative sources of herbal medicine as an effort to deal with disease problems due to infection.
Several studies in Indonesia report that there are types of medicinal plants that have antimicrobial properties, and one of them is the turmeric plant (Curcuma domestica Val.). The benefits of turmeric is that it includes essential oils, fatty oils, curcuminoid compounds, and other derivative compounds. This content makes turmeric a medicinal plant that has anti-inflammatory, antiviral, antibacterial, anti-protozoa, antineoplastic, antioxidant, and antinematoside effects. Based on a previous study, curcumin showed a similar antibacterial effect of A. actinomycemcomitans, which was reported to be sensitive at a lower concentration. Research conducted by Mohammed showed that curcumin expressed antibacterial activity against Gram-positive bacteria, namely Streptococcus mutans and Streptococcus pyogenes, respectively, with inhibition zone diameters of 9.7 mm and 10.2 mm. This research is based on the theory that states that curcumin extract exhibits antibacterial activity against various microbes and also against bacteria such as Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus, and Losteria monocytogenes., In addition, curcumin is also a strong inhibitor of growth against Gram-positive bacteria (Staphylococcus aureus and Streptococcus mutans), Gram-negative bacteria (E. coli and Pseudomonas aeruginosa), and pathogenic fungi (Candida albicans). A study conducted by Yuliati on the antibacterial effectiveness of turmeric extract on Bacillus sp. (Gram-positive) and Shigella dysentriae (Gram-negative) showed that the inhibition zone of Gram-positive bacteria is larger than that of Gram-negative bacteria.
So far, research on turmeric extract in the form of a gel formula used as a herbal medicine for periodontitis in Indonesia has not been reported. Many pharmaceutical dosage forms can be made in the form of gels from bioactive medicinal plants, because the gel preparations are considered easier to clean from the skin surface due to polar solvents. Therefore, this research is very important to study oral gel formula of the indigenous turmeric extract from Aceh, Indonesia and to evaluate its antimicrobial efficacy against A. actinomycetemcomitans bacteria that cause periodontitis.
| Materials and Methods|| |
Setting and design
This experimental in vitro design was used to assess the oral gel formula of the indigenous turmeric extract from Aceh, Indonesia and to evaluate its antimicrobial efficacy against A. actinomycetemcomitans. This research was conducted in Laboratory Pharmacology and Microbiology, Faculty of Veterinary Medicine, Universitas Syiah Kuala, from April until August 2020.
The samples used in this study were rhizomes of indigenous turmeric originally from Aceh, Indonesia. Sampling was taken using the purposive sampling method, which is a method of selection by which samples that match the inclusion criteria are selected. The main criteria for the inclusion of turmeric rhizomes are that they are 1–2 cm in diameter, and 5–15 cm in length, their outer skin color is light brown, their inside is orange, and they are slightly fibery. In this study, an attempt was made to accumulate no bias data as much as possible, by means of antimicrobial tests conducted with three repetitions. This research was conducted through several steps, starting from the extraction of the turmeric rhizome, gel formulation of the turmeric rhizome extract, and finally an in vitro assay of turmeric rhizome gel as an antimicrobial against A. actinomycetemcomitans. Further, from the research processes conducted, the effectiveness of curcumin gel formula in inhibiting bacteria A. actinomycetemcomitans was known; thus, it came to be used as periodontitis herbal medicine.
Extraction of turmeric rhizome (Curcuma Longa Linn)
Turmeric rhizome extraction was carried out by the maceration method using ethanol as a solvent. A total of 120 g of turmeric rhizome extract simplicia that was washed was immersed in a 96% ethanol solution of 750 mL at room temperature for five days and it was then filtered using analytical filter paper. After five days, the soaked sample was filtered using filter paper to produce filtrate 1 and pulp 1. The waste was then macerated with 75 mL of 96% ethanol solution, covered with aluminum foil, and left for two days, stirring occasionally. After two days, the sample was filtered using filter paper or flannel to produce filtrate 2 and pulp 2. Filtrate 1 and 2 were combined, and then they were evaporated using a rotary evaporator at a temperature of 40°C–50°C, so that a thick turmeric extract was obtained. The resulting viscous extract was left at room temperature until all the ethanol solvent evaporated. The extract was weighed and stored in a closed glass container before being used for testing. From the extraction process, an ethanol extract of turmeric rhizome with a concentration of 100% was obtained and this was used as a stock sample. The turmeric rhizome extract used for further testing was a concentration of 0%, 25%, 50%, 75%, and 100%. To obtain this concentration, the extract was diluted in ddH2O, a mixture of both, namely 25 g of the extract was diluted in 100 mL of ddH2O (25%), 50 g of the extract was diluted in 100 mL of ddH2O (50%), and 75 g of the extract was diluted in 100 mL of ddH2O (75%).
Gel formulation of turmeric rhizome extracts
The process of making turmeric rhizome extract gel begins by weighing all the necessary ingredients according to the calculation. The next step involves the development of a gelling agent, and then adding additional ingredients (such as preservatives, penetration enhancers, etc.) to the gelling agent that is already in the finished form. The gelling agent used in this study was carboxymethylcellulose (CMC), and aquades served as a solvent. Further, the active substance of the turmeric rhizome ethanol extract was added to the gel preparation. Gel preparations were made with concentrations of 0%, 25%, 50%, 75%, and 100%.
In vitro test of turmeric rhizome gel as antimicrobial against A. actinomycetemcomitans
Bioactivity tests for microbial indicators as antibacterials use the Kirby-Bauer disk diffusion test on disk paper (6 mm diameter). The disk diffusion test method, as much as 100 μL of turmeric extract, with concentrations of 0%, 25%, 50%, 75%, and 100% is placed on the disk paper and placed on the Mueller Hinton Agar (MHA) media that has been grown by bacteria pathogen. The A. actinomycetemcomitans (ATCC 33384) bacteria used were diluted according to McFarland standard 0.5 (1.5 × 108 CFU/mL). Petri dishes were incubated and dried at 37°C for 24–48h, and three repetitions were carried out. The clear zone was then measured to determine the zone of inhibition of the ethanol extract of turmeric rhizome against the tested bacteria. The criteria for the strength of bacterial inhibition used were very strong (inhibition zone >20 mm), strong (inhibition zone 10–20 mm), moderate (inhibition zone 5–10 mm), and weak (<5 mm).
| Results|| |
The characteristics of turmeric rhizome extract gel are that it is semisolid, translucent, and colloid disperse; has a dark yellowish brown color; and has a distinctive aroma of turmeric. The difference in color is influenced by the concentration of the turmeric rhizome extract. This study was successfully carried out by producing gel preparations with various concentrations, namely 0%, 25%, 50%, 75%, and 100% [Figure 1]. The 0% formula was made without an extract, and the other four formulas (F2–F5) were made by adding extracts with different concentrations. In addition, the antimicrobial activity of turmeric from Aceh, Indonesia against A. actinomycetemcomitans has been tested in the range 0%–100%.
|Figure 1: Gel formula of Curcuma longa rhizome extracts (25%, 50%, 75%, 100%)|
Click here to view
The antimicrobial test was carried out with three replications, showing the inhibition zone diameter of 9.4 mm for a concentration of 25%, 10.8 mm for a concentration of 50%, 11.6 mm for a concentration of 75%, and 13.9 mm for a concentration of 100% [Table 1].
|Table 1: The diameter of the antimicrobial inhibition zone of the Curcuma longa extract gel formula against A. actinomycetemcomitans|
Click here to view
The in vitro antimicrobial test using the agar disk diffusion method showed that the turmeric rhizome extract gel was able to form a clear zone around the growth medium of the tested bacteria, and different diameters of inhibition were shown at different concentrations [Figure 2]. Thus, this indicates that the gel formula of the turmeric rhizome extract is able to inhibit the growth of the A. actinomycetemcomitans bacteria that causes periodontitis.
|Figure 2: Clear zone results of in vitro antimicrobial test for turmeric rhizome extract gel formulation using ethanol solvent (left to right: negative control, 25%, 50%, 75%, 100%; middle: positive control)|
Click here to view
| Discussion|| |
Gel formulation and evaluation of the antimicrobial activity of turmeric (C. longa L.) from Aceh, Indonesia against A. actinomycetemcomitans was determined in the present study. The A. actinomycetemcomitans bacteria used was the potent periodontal pathogen. In this study, the gel formulation of turmeric from Aceh, Indonesia was successfully formulated into five formulas with a concentration range of 0%–100%. Based on several previous studies, it was reported that many pharmaceutical dosage forms can be made from bioactive medicinal plants, because gel preparations are considered easier to clean from the skin surface due to polar solvents. The gel is a semisolid preparation that is clear, translucent and matches the active substance and is a colloidal spread that has strength caused by tissues binding to each other in a scattered phase. In addition, the gel has a structure that is resistant to environmental changes and has a flow of viscosity.
Some of the advantages of gel preparations include their non-stickiness, high water content so that a large amount of water can hydrate the horny layer, and changes in the permeability of the horny tissue to become more permeable to active ingredients that can increase the permeases of the active ingredients. In addition, a gel preparation is able to provide a comfortable feeling when applied to the target area; the preparation is easy to dry, thus forming a film layer that is easily washed with water. In this research, the gel formulation of turmeric extracts used CMC as a gelling agent. This was different from research by Cahyani and Putri, which used carbopol 940 as a gelling agent for a formulation of Curcuma heyneana Val.
Based on the results, turmeric rhizome formula extracts gels with concentrations of 25%, 50%, 75%, and 100%, which can all inhibit A. actinomycetemcomitans with various categories of inhibition [Table 1]. Based on the indicated inhibition zone, it can be stated that the turmeric rhizome extract gel is effective in inhibiting the growth of the A. actinomycetemcomitans bacteria with the strong inhibition category. Tetracycline antibiotic as a positive control has shown a zone of inhibition of 21–22 mm diameter against A. actinomycetemcomitan, and 10% of ddH2O disks as a negative control has shown no zone of inhibition [Figure 1]. According to Davis and Stout, the criteria for the strength of the antibacterial power are as follows: The inhibition zone diameter of 5 mm or less is categorized as weak, the inhibition zone of 5–10 mm is categorized as moderate, the inhibition zone of 10–20 mm is categorized as strong, and the inhibition zone of 20 mm or more is categorized as very strong. Based on the results of this research, it can be stated that 50% concentration of oral gel formula turmeric extracts has been effective in inhibiting A. actinomycetemcomitan growth bacteria, with an inhibition zone of 10.8 mm, which is classified as a strong inhibition zone. Nevertheless, the maximum zone of inhibition was obtained at 100% concentration with an inhibition zone of 13.9 mm and it was also classified into a strong inhibition zone.
Based on a previous study, the formulation of curcumin gel has been reported for various applications, such as for anti-inflammatory ones. The formulation of medicinal plant extract gel preparations as antibacterial agents has been widely used, such as the preparation of arumanis mango leaf extract gel as an antibacterial agent for Staphylococcus aureus and Propionibacterium acnes, white turmeric extract gel preparation as an antiseptic, binahong gel preparation as an antibacterial agent, and turmeric rhizome extract gel preparation against Staphylococcus aureus bacteria.
A. actinomycetemcomitans was used in this study as a potential pathogen causing periodontitis. Periodontitis is inflammation that affects the supporting tissues of the teeth; is caused by microorganisms and can cause progressive damage to the periodontal ligament, alveolar bone; and is accompanied by pocket formation. The main cause of periodontal disease is the presence of microorganisms that colonize the dental plaque. The increase in the number of Gram-negative organisms in the subgingival plaque such as Porphiromonas gingivalis, Actinobacillus actinomycetemcomitans, Tannerela forsythia, and Treponema denticola initiates periodontal infection.,
Turmeric rhizome is a medicinal plant that holds a high position in dentistry. Rudhra et al. reported the antibacterial efficacy of the curcumin oral gel against Porphyromonas gingivalis and Prevotella intermedia with a diameter of clear zone of 10.3 mm and 11.4 mm, respectively. An in vitro evaluation of curcumin in situ gel as a stabilized formulation for periodontal application was also reported by Nasra et al. and Hosadurga et al., which states that about 2% curcumin gel from India was effective in the treatment of experimental periodontitis based on the results of the anti-inflammatory test. Further in vivo and further definitive randomization, clinical trials further microbiological research compared to commercial periodontitis drugs is an opportunity for further research. Oral gel as a local drug delivery agent in patients with periodontitis to find out its mechanism of action against periodontal pathogens and the host response in effect is complete.
Bomdyal et al. reported that curcumin inhibited the growth of A. actinomycetemcomitans; it acts a host modulatory agent in periodontitis with damaging of the bacterial membrane or perturbing the bacterial membrane integrity. The antibacterial activity mechanism of curcumin can be through the inhibition of bacterial cell proliferation by blocking the dynamics of FtsZ assembly in the Z ring., Curcumin has also been shown to have antibacterial activity through the effect of virulence, quorum sensing, and biofilm initiation.
We thank Faculty of Dentistry, Universitas Syiah Kuala, for the facility provided. We also thank Laboratory of Pharmacology and Laboratory of Microbiology, Faculty of Veterinary Medicine, who have helped in this research.
Financial support and sponsorship
The Ministry of Education and Culture of The Republic of Indonesia awarded to Prof. Dr. drg. Zaki Mubarak, M.Si with the contract no: 268/UN11/SPK/PNB/2020.
Conflicts of interest
There are no conflicts of interest.
ZM, WES, and S conceptualized and designed this research. The research was carried out by ZM, WES, and S. WES analyzed the data and result. ZM and WES drafted, revised, and finalized the article. All authors read and approved the final article.
Ethical policy and institutional review board statement
Not applicable as study did not involve any participants or patients.
Patient declaration of consent
Data availability statement
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[Figure 1], [Figure 2]