ORIGINAL RESEARCH |
https://doi.org/10.5005/jp-journals-10015-2232 |
Anti-inflammatory Potential of a Mouthwash Formulated Using Clove and Ginger Mediated by Zinc Oxide Nanoparticles: An In Vitro Study
1,2,4Department of Prosthodontics, Saveetha Dental College and Hospital, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai, Tamil Nadu, India
3,5Department of Pharmacology, Saveetha Dental College and Hospital, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai, Tamil Nadu, India
6Department of Pedodontics, Saveetha Dental College and Hospital, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai, Tamil Nadu, India
Corresponding Author: Jerry Joe Chokkattu, Department of Prosthodontics, Saveetha Dental College and Hospital, Saveetha Institute of Medical and Technical Sciences (SIMATS), Chennai, Tamil Nadu, India, Phone: +91 9841026569, e-mail: dr.jerryjoe@gmail.com
Received on: 03 April 2023; Accepted on: 06 May 2023; Published on: 02 August 2023
ABSTRACT
Aim: The aim of the current study is to check for the anti-inflammatory activity of clove and ginger formulated along with zinc oxide (ZnO) nanoparticles (NPs) in a mouthwash.
Materials and methods: A mixture of 1 gm each of ginger and clove powder was dissolved in 100 mL distilled water, heated, and filtered. ZnO NPs were synthesized by mixing 20 mM zinc nitrate [Zn(NO3)2]with plant extract and centrifuging the resulting suspension. Mouthwash was prepared by dissolving 0.3 gm sucrose, 0.001 gm sodium benzoate, and 0.01 gm sodium lauryl sulfate in 9.5 mL distilled water and adding 50 µL peppermint oil and 500 µL NP suspension. The solution was shaken, and 10 mL of the resulting mouthwash was obtained. The standard used was diclofenac sodium with dimethyl sulfoxide (DMSO) as a control. The anti-inflammatory activity of the combination of clove and ginger formulation-mediated ZnO NP mouthwash was conducted through bovine serum albumin assay and egg albumin assay.
Results: In the bovine serum albumin assay, the various fixations of clove and ginger formulation from 10–50 μL obtained various results of inhibition of 45, 55, 65, 75, and 85%, respectively, in comparison to the standard diclofenac sodium. As a result, when the concentration of the ZnO NPs increased, the inflammatory activity also increased with significant (p < 0.05) values. In egg albumin assay, the various fixations of clove and ginger formulation from 10–50 μL obtained various results of inhibition 50, 65, 70, 70, and 80%, respectively, in comparison to the standard diclofenac sodium. Group stats and independent t-test reveal a significant correlation at 40 μL (p = 0.01) between the test group and the control group showing increased inflammatory activity.
Conclusion: The results of the study suggest that ZnO NPs strengthened with ginger and clove extracts may have anti-inflammatory characteristics and be utilized as an alternative formulation to commercially available mouthwash.
Clinical significance: Ongoing research and development in the field of dentistry has led to the emergence of alternative methods for improving the efficacy of dental varnishes in preventing dental caries. One promising approach involves incorporating herbal resources and NPs into the formulation. This development is clinically significant as it addresses the limitations of traditional agents and offers a potentially more effective way to prevent tooth decay.
How to cite this article: Selvaraj S, Chokkattu JJ, Shanmugam R, et al. Anti-inflammatory Potential of a Mouthwash Formulated using Clove and Ginger Mediated by Zinc Oxide Nanoparticles: An In Vitro Study. World J Dent 2023;14(5):394–401.
Source of support: The present study is funded by the following: Saveetha Institute of Medical and Technical Sciences, Saveetha Dental College and Hospitals, Saveetha University, M/s Trend Fashions India Pvt Ltd.
Conflict of interest: None
Keywords: Anti-inflammatory agents, Ginger, Nanoparticle, Syzygium, Zinc oxide
INTRODUCTION
Nanotechnology is an important field in materials science that deals with creating and studying materials that are very small, around 100 nm in size. At this scale, materials can have unique properties that can be useful in a wide range of fields, from creating new fabrics and foods to advanced medical procedures.1 Nanoparticles (NP) are tiny particles that have a large surface area compared to their size, which can make them more effective in certain applications than larger materials. Nanotechnology offers exciting possibilities for developing new materials with enhanced properties and functionalities.2 NPs are referred to as controlled or modified particles.3 They have properties related to length that are remarkably distinct from those of bulk materials. Despite having shorter lengths than their counterparts, NPs have larger systems. Their practical initiatives in a variety of domains, including biosensors, nanomedicine, and bionanotechnology, are made possible by these precise resources.4 The length, composition, crystallinity, and shape of metallic NPs, such as ZnO (ZnO), titanium dioxide (TiO2), and silver, are frequently used to describe their inherent properties. Nanoscale properties can change when their dimensions are shrunk, including their chemical, mechanical, electrical, structural, morphological, and optical properties.5 These changed capabilities permit the NPs to interact uniquely with mobile biomolecules and therefore facilitate the bodily switch of NPs into the internal mobiles systems.6 Nanostructured substances have a bigger percentage of atoms on their floor, which result in excessive floor reactivity. Thus, nanomaterials have witnessed currently great significance in the field of dentistry involving formulations of various products like filling materials, anti-sensitive agents, surface coatings, toothpastes, mouthwashes, etc., targeted essentially towards balancing oral health. Among all these, mouthwash has been the most evolving dental product due to professional recommendations for the maintenance of oral biofilm and periodontal health.7 Bueczynska et al. claim that the oral microbiota associated with local infections may disperse and result in systemic, maybe lethal illnesses. Traditional methods are insufficient to identify the etiological agents of infections in dentistry because roughly 50% of mouth bacteria cannot be grown. In the search for better diagnosis, metagenomic analyses have proven to be giving greater insight into the heterogenicity of the bacteria.8 With the aid of metagenomic studies and the emergence of new nanotechnologies, given its contribution to the field of dentistry, demand has also increased for medicated, nonmedicated and naturally derived mouthwashes reinforced with nanomaterials. According to Carrouel et al., ZnO- and TiO2-NPs have been proven to have antimicrobial and anti-inflammatory properties, which are formulated in oral products.7
Zinc oxide (ZnO) in the nanometer range is remarkable for varied forms and has strong antibacterial activity against a broad range of bacterial species that have been studied by a large group of researchers. Currently, the antibacterial properties of ZnO in both micro and nanoscale formulations are being studied.9 When the size of a ZnO particle is reduced to the nanometer range, it exhibits strong antimicrobial effects. Nano-sized ZnO can interact with the bacterial middle or floor when it enters the cell.10 The interactions among those specific substances and microorganisms are often poisonous and have been exploited for antimicrobial programs, including within the meals industry.
Interestingly, numerous studies have revealed that ZnO NPs are not toxic to human cells; this has made it necessary to use them as antibacterial agents since they are toxic to germs and maintain biocompatibility with human cells.11 Although numerous proposals have been put forward and implemented to explain the various antibacterial effects of NPs, there is still ongoing debate regarding the most appropriate methods, particularly in relation to the physical–chemical properties of the particles.11,12
Exploration into the world of nanomaterials and the intricate mechanisms and phenomena that govern these nanostructured substances can be advanced through research on antibacterial nanomaterials, with a particular focus on ZnO NPs. Given the significant impact of bacterial infections on both human health and the socioeconomic landscape, the public has become increasingly attuned to the importance of finding effective ways to combat these diseases on a global scale. Thus, delving deeper into the realm of antibacterial nanomaterials not only holds immense potential for scientific advancement but also carries significant real-world implications for the betterment of society.13
Numerous experiments have been carried out to synthesize and characterize various properties of NPs, including but not limited to ZnO, titanium oxide, zirconium oxide, selenium, halloysite nanotubes, silymarin/hydroxyapatite, chitosan nanocomposites, plant-derived silver NPs, nanoemulsions, and oleoresins.14-26 These experiments have resulted in significant advancements in the development of nanostructured materials, which have multiple applications in various scientific fields.
Considering the above-mentioned data, the aim of the current study focuses on the anti-inflammatory activity of mouthwash prepared using clove and ginger formulation mediated ZnO NPs with a null hypothesis that there is a significant difference in anti-inflammatory action due to ZnO NP mediated herbal formulation across different concentrations.
MATERIALS AND METHODS
Study Setting
The study was held in the Nanomedicine Laboratory, Department of Pharmacology, Saveetha Dental College, after obtaining approval from the Scientific Review Board (IHEC/SDC/UG-1921/21/130). The experiment was initiated in October 2022 and was conducted within a time period of three weeks from acquiring the raw materials, synthesis of NPs, formulation of mouth, and anti-inflammatory tests.
Preparation of Plant Extract
A total of 10 gm of raw clove (Syzygium aromaticum) and ginger (Zingiber officinale) were obtained from a local market. The ginger was peeled and subsequently sun-dried until it was sufficiently dehydrated for processing into a fine powder using a mixer grinder. Subsequently, 1 gm of ginger and 1 gm of clove powder were dissolved in 100 mL of distilled water. The resulting solution was heated on a hotplate for 10 minutes at a temperature of 60°C until it began to bubble. The resulting plant extract was collected in a flask using a Whatman filter placed on a funnel. The extract was subsequently transferred to an airtight storage container and chilled overnight.
Synthesis of ZnO NPs
Zinc oxide (ZnO) NPs were synthesized by dissolving 20 mM of Zinc nitrate [Zn(NO3)2] in 50 mL of deionized water. Then, 50 mL of this solution was mixed with 50 mL of plant extract. After a period of 2 days, the resulting NP suspension was subjected to centrifugation using a Lark refrigerator centrifuge at 8000 rpm for 10 minutes. The resultant pellets were separated from the supernatant and transferred to Eppendorf tubes for further analysis. This process was conducted in accordance with established scientific protocols for NP synthesis and characterization.
Preparation of Mouthwash
The mouthwash was prepared by taking 0.3 gm of sucrose, 0.001 gm of sodium benzoate, and 0.01 gm of sodium lauryl sulfate and dissolving them in 9.5 mL of distilled water. Around 50 µL of peppermint oil and 500 µL of the pellet were added and kept in the orbital shaker. A total of 10 mL of clove and ginger formulation-mediated ZnO NP mouthwash was obtained.
Visual Identification
During the experiment, changes in color were observed in a reaction mixture at various time intervals during incubation. The objective of the experiment was to reduce Zn(NO3)2 into ZnO NPs, which could be identified by a color change.
Anti-inflammatory Activity
The anti-inflammatory activity of the combination of clove and ginger formulation-mediated ZnO NP mouthwash was conducted in reference to the study conducted by Dharmadeva et al.27 The study was designed to evaluate the anti-inflammatory activity using two standard albumin denaturation methods. Egg albumin assay, a cheaper denaturation technique, was validated further by bovine serum albumin assay.27 A standard control group and test group each received three samples per concentration (10–50 µL) during the in vitro trial, which was conducted utilizing in vitro sampling. The standard used was diclofenac sodium with dimethyl sulfoxide (DMSO) as a control.
Anti-inflammatory Activity Using Bovine Serum Albumin
A 0.5% of w/v aqueous solution of bovine serum albumin was prepared. A phosphate buffer saline was prepared by dissolving sodium chloride (8 gm), potassium chloride (0.2 gm), disodium hydrogen phosphate (1.44 gm), and potassium dihydrogen phosphate (0.24 gm) in 800 mL of distilled water. The pH of the mixture was adjusted to 6.3 using 1 N hydrochloric acid. A test solution of 0.5 mL was prepared using 0.45 mL of prepared bovine serum albumin (0.5% aqueous solution) with 0.05 mL of formulated mouthwash of various concentrations (10, 20, 30, 40, 50 µg/mL). And a standard solution of 0.5 mL was also prepared using 0.45 mL of bovine serum albumin and 0.05 mL of diclofenac sodium at varying concentrations (10, 20, 30, 40, and 50 µg/mL). Once the test and standard solutions were prepared, they were heated to 55°C in a water bath for 30 minutes after being incubated at room temperature for 20 minutes. After cooling, 0.25 mL of prepared phosphate buffer was added to the samples, followed by measuring the absorbance at 660 nm using a ultraviolet (UV) visible spectrophotometer. The standard solution represents 100% of protein denaturation; hence the results were compared with diclofenac sodium.28
The percentage of protein denaturation was determined utilizing the following equation:
Anti-inflammatory Activity Using Egg Albumin
The egg albumin assay was conducted using a reaction mixture of 5 mL containing 0.2 mL of egg albumin obtained from fresh hen’s egg, 2.8 mL of phosphate-buffered saline (PBS) with a pH of 6.3, and 2 mL of a test sample (formulated mouthwash) with varying concentrations of 10, 20, 30, 40, and 50 µg/mL. A standard solution was also prepared with a mixture of 0.2 mL of egg albumin from a fresh hen’s egg, 2.8 mL of PBS with a pH of 6.3, and 2 mL of diclofenac sodium drug at concentrations ranging from 10 to 50 µg/mL. To serve as a negative control, a control group was also prepared using a mixture of 0.2 mL of egg albumin, 2.8 mL of PBS with a pH of 6.3, and distilled water to make up a total volume of 5 mL. All sample tubes were incubated at 37 ± 2°C for 15 minutes, followed by heating to 70°C for 5 minutes in an incubator. After cooling, the absorbance of each sample was measured at 660 nm using a UV-visible spectrophotometer.
The percentage of protein denaturation was determined utilizing the following equation:
The data from the bovine serum albumin assay and egg albumin assay was collected in an Excel sheet, followed by group statistics, independent t-tests among the test group and control formulation using Statistical Package for the Social Sciences software (version 25, IBM, India).
RESULTS
Visual Observation
The mixture was observed for any color changes and was recorded during the incubation period. As the reaction progressed, the dark brown color of the mixture gradually transformed into an amber-colored liquid, indicating the formation of ZnO NPs. Furthermore, it was observed that the viscosity of the liquid increased as the incubation time increased. Finally, after 24 hours of incubation, the mixture reached a point where no further color changes were observed, indicating that the synthesis of ZnO NPs was complete.
Anti-inflammatory Activity
The anti-inflammatory activity of clove and ginger formulation-mediated ZnO NPs was assessed using bovine serum albumin assay and egg albumin assay. In the bovine serum albumin assay (Fig. 1 and Table 1), the various fixations of clove and ginger formulation from 10–50 μL obtained various results of inhibition 45, 55, 65, 75, and 85%, respectively, in comparison to the standard diclofenac sodium. As a result, when the concentration of the ZnO NPs increased, the inflammatory activity also increased with significant (p < 0.05) values in descriptive statistics and independent t-test when compared between test and control groups at all concentrations (Tables 2 and 3).
Fig. 1: Anti-inflammatory activity tested in bovine serum albumin assay at various concentrations from 10–50 μL and the percentage of inhibition; the X-axis shows the concentration of the NP, and the Y-axis shows the percentage inhibition; values are done in triplicate n = 3, mean ± standard deviation (SD)
| 10 μL | 20 μL | 30 μL | 40 μL | 50 μL | |
|---|---|---|---|---|---|
| Clove and ginger (ZnO) | 42 | 54 | 68 | 73 | 81 |
| Standard | 47 | 60 | 72 | 78 | 84 |
| Group statistics | |||||
|---|---|---|---|---|---|
| Groups | N | Mean | SD | Standard error mean | |
| 10 µL | Test group | 3 | 42.0000 | 3.00000 | 1.73205 |
| Control group | 3 | 47.0000 | 0.00000 | 0.00000 | |
| 20 µL | Test group | 3 | 54.0000 | 1.00000 | 0.57735 |
| Control group | 3 | 60.0000 | 0.00000 | 0.00000 | |
| 30 µL | Test group | 3 | 68.0000 | 2.00000 | 1.15470 |
| Control group | 3 | 72.0000 | 0.00000 | 0.00000 | |
| 40 µL | Test group | 3 | 73.0000 | 2.00000 | 1.15470 |
| Control group | 3 | 78.0000 | 0.00000 | 0.00000 | |
| 50 µL | Test group | 3 | 81.0000 | 1.00000 | 0.57735 |
| Control group | 3 | 84.0000 | 0.00000 | 0.00000 | |
| Mean difference | 95% confidence interval of the difference | Degree of freedom (df) | Significance | |||||
|---|---|---|---|---|---|---|---|---|
| Concentration and groups | Mean | SD | Lower | Upper | ||||
| 10 µL | Test group | 42.00 | 3.00 | −5.00 | −9.81 | −0.19 | 4.00 | 0.04 |
| Control group | 47.00 | 0.00 | ||||||
| 20 µL | Test group | 54.00 | 1.00 | −6.00 | −7.60 | −4.40 | 4.00 | 0.00 |
| Control group | 60.00 | 0.00 | ||||||
| 30 µL | Test group | 68.00 | 2.00 | −4.00 | −7.21 | −0.79 | 4.00 | 0.03 |
| Control group | 72.00 | 0.00 | ||||||
| 40 µL | Test group | 73.00 | 2.00 | −5.00 | −8.21 | −1.79 | 4.00 | 0.01 |
| Control group | 78.00 | 0.00 | ||||||
| 50 µL | Test group | 81.00 | 1.00 | −3.00 | −4.60 | −1.40 | 4.00 | 0.01 |
| Control group | 84.00 | 0.00 | ||||||
An independent sample t-test was used for significance testing; a p-value of <0.05 is considered significant
In egg albumin assay (Fig. 2 and Table 4), the various fixations29 of clove and ginger formulation from 10–50 μL obtained various results of inhibition 50, 65, 70, 70, and 80%, respectively on comparison to the standard diclofenac sodium. Group stats and independent t-test reveal a significant correlation at 40 μL (p = 0.01) between the test group and control group showing an increased inflammatory activity (Tables 5 and 6). Overall, both assays have shown good anti-inflammatory properties of the mouthwash formulation, especially at higher concentrations.
Fig. 2: Anti-inflammatory activity tested in egg albumin assay at various concentrations from 10–50 μL and the percentage of inhibition; the X-axis shows the concentration of the NP, and the Y-axis shows the percentage inhibition. Values are done in triplicate n = 3, mean ± SD
| 10 μL | 20 μL | 30 μL | 40 μL | 50 μL | |
|---|---|---|---|---|---|
| Clove and ginger (ZnO) | 53 | 61 | 67 | 69 | 78 |
| Standard | 55 | 64 | 69 | 72 | 81 |
| Group statistics | |||||
|---|---|---|---|---|---|
| Groups | N | Mean | SD | Standard error mean | |
| 10 µL | Test group | 3 | 53.0000 | 3.00000 | 1.73205 |
| Control group | 3 | 55.0000 | 0.00000 | 0.00000 | |
| 20 µL | Test group | 3 | 61.0000 | 2.00000 | 1.15470 |
| Control group | 3 | 64.0000 | 0.00000 | 0.00000 | |
| 30 µL | Test group | 3 | 67.0000 | 2.00000 | 1.15470 |
| Control group | 3 | 69.0000 | 0.00000 | 0.00000 | |
| 40 µL | Test group | 3 | 69.0000 | 1.00000 | 0.57735 |
| Control group | 3 | 72.0000 | 0.00000 | 0.00000 | |
| 50 µL | Test group | 3 | 78.0000 | 2.00000 | 1.15470 |
| Control group | 3 | 81.0000 | 0.00000 | 0.00000 | |
| Mean difference | 95% confidence interval of the difference | df | Significance | |||||
|---|---|---|---|---|---|---|---|---|
| Concentration and groups | Mean | SD | Lower | Upper | ||||
| 10 µL | Test group | 53.00 | 3.00 | −2.00 | −6.81 | 2.81 | 4.00 | 0.31 |
| Control group | 55.00 | 0.00 | ||||||
| 20 µL | Test group | 61.00 | 2.00 | −3.00 | −6.21 | 0.21 | 4.00 | 0.06 |
| Control group | 64.00 | 0.00 | ||||||
| 30 µL | Test group | 67.00 | 2.00 | −2.00 | −5.21 | 1.21 | 4.00 | 0.16 |
| Control group | 69.00 | 0.00 | ||||||
| 40 µL | Test group | 69.00 | 1.00 | −3.00 | −4.60 | −1.40 | 4.00 | 0.01 |
| Control group | 72.00 | 0.00 | ||||||
| 50 µL | Test group | 78.00 | 2.00 | −3.00 | −6.21 | 0.21 | 4.00 | 0.06 |
| Control group | 81.00 | 0.00 | ||||||
Independent sample t-test was used for significance testing; a p-value of <0.05 is considered significant
DISCUSSION
Zinc oxide (ZnO) is said to be an inorganic material having a wide range of applications that is functional, strategic, promising, and adaptable. Since Zn and O are categorized in groups 2 and 6 of the periodic table, respectively, it is known as an II–VI semiconductor.30,31 These qualities allow ZnO to have remarkable applications in a variety of industries. ZnO’s wide band gap significantly affects its characteristics, including optical absorption and electrical conductivity.32 When ZnO is doped with other metals, the excitonic emission can persist longer at normal temperatures, and the conductivity rises. Although ZnO has a weak covalent link, it has a very strong ionic bond in ZnO. In comparison to organic and inorganic materials, it has superior heat resistance, higher selectivity, and longer durability.33 The exploration of ZnO’s usage as a novel antibacterial agent has been prompted by the manufacture of nanoscale ZnO. The studies conducted by Hsu and Ozcano et al. have proved the unique antibacterial and antifungal capabilities of ZnO NPs that are complemented by their strong catalytic and strong photochemical activity, which is in correlation with the current study with a significant anti-inflammatory activity at 10–50 μL concentrations in bovine serum assay and at 40 μL concentrations in egg albumin assay.34,35
Plants have been used for their therapeutic properties for ages. However, only a small number of scientific studies have been conducted, and many plants still need to be investigated. The evaluation of medicinal plants is, therefore, particularly interesting due to their long history and widespread continued use.36 An evergreen plant with a height of 10–20 cm, cloves grow in hot, tropical areas. It originally came from tropical Asian countries. The family Myrtaceae includes the clove Syzygium aromaticum L. (also known as Eugenia caryophyllata, syn.).37 It tastes very peppery and has a potent phenolic odor. Dried flower buds are frequently used as spices. The oil is colorless to the light yellowish fluid that is heavier than water and freely soluble in alcohol. It is produced by steam distillation of clove buds.38 Clove has long been a food source for humans. It is renowned for its therapeutic benefits in addition to its flavorful capabilities. Clove has long been prized as a medicinal herb in Ayurveda, Chinese medicine, and Western herbalism.39 Clove has long been used as a carminative, antispasmodic, and to enhance peristalsis. Clove essential oil is primarily used to treat the symptoms of toothache and throat and mouth inflammation in dental crises.39,40 Clove oil has reportedly been employed in the creation of some toothpaste and mouthwashes in this regard. Clove is antimutagenic, antioxidant, antithrombotic, antiparasitic, and anti-inflammatory, according to current studies. Clove oil is also said to have antifungal, antiviral, and antibacterial effects.41 According to Lewis, it has been identified that some clove components, primarily eugenol (70–90%), tanene (13%), and fixed oil (10%), can act as physiological markers providing several health benefits.42 This is in correlation with the result of the current study proving the anti-inflammatory properties. Also supporting the present study results, eugenol displays anti-inflammatory and anti-arthritic properties, according to studies in the arthritic rat model. When interleukin injections made rabbits feverish, eugenol dramatically reduced fever. These results lend credence to several of the widely used medical applications of clove oil.43
Ginger, or Zingiber officinale Roscoe, has been used medicinally for a very long time. Traditional Chinese and Indian medicine has traditionally utilized ginger to treat a range of ailments, including arthritis, asthma, stomach aches, diarrhea, and respiratory issues.44 Currently, herbalists mostly advise using ginger and its derivatives to treat dyspepsia and ward against motion sickness. According to Adebayo et al., research from a few recent trials has rekindled interest in ginger as a chronic inflammatory disease treatment.45 This interest can be linked to the early 1970s discovery that nonsteroidal anti-inflammatory medications (NSAIDs) work by impairing prostaglandin production (PGs).46 Nair mentioned that a good scientific justification for ginger’s anti-inflammatory benefits was later discovered to exist in the form of components that block PG production.47 Later research found that ginger also includes elements with pharmacological traits related to the unique family of dual-acting NSAIDs.48 The cyclooxygenase and lipoxygenase pathways are used by compounds in this family to suppress arachidonic acid metabolism.49,50 These drugs are being looked into as a unique class of anti-inflammatory drugs because they have considerably fewer side effects than traditional NSAIDs. The pharmacological effects of ginger on the inflammatory process go far beyond PG production inhibition.48,50 These investigations found that ginger has an impact on the cytokines that are created and released at inflammatory sites. These molecules have emerged as some of the most promising targets for the therapy of inflammatory illnesses with chronic onset.51 Recent clinical experiments that corroborate the conventional belief that ginger has analgesic and anti-inflammatory characteristics add to the preclinical data on mechanisms by which ginger generates its effects which correlates with the anti-inflammatory property mediated with ZnO NPs of the current study.52 Promising antimicrobial results have been seen with zinc NPs synthesized at 25, 50, and 100 uL concentrations along with clove and cinnamon due to a few special properties of NPs like the capacity to damage bacterial cell walls, tampering with the oxidative stress resistance genes etc. In the present study zone of inhibition was highest at 100 uL against the bacteria.53 With the above studies and references, the findings of the current study prove the hypothesis that the antibacterial efficacy of the clove, ginger and ZnO NPs mediated formulation can be used as an alternative to antimicrobial mouthwash.
Limitations
To evaluate the anti-inflammatory activity, this investigation was conducted in vitro, which may not be sufficient to demonstrate the effectiveness of the novel ZnO NP formulation and its clinical usefulness. The findings presented in the current study should be interpreted in view of certain limitations, including the need for an expanded control group, application of different sampling methods, examination of different activities such as antioxidant, microbial, cytotoxic, etc., and enhancement of statistical analyses.
Areas of Future Research
To ensure a favorable outcome, supplementary in vitro examinations such as membrane stabilization assays can be conducted. In due course, in vivo investigations may be feasible, utilizing diverse concentrations of the samples to perform in vivo methodologies such as the carrageenan-induced rat paw edema test and the egg albumin-induced rat paw edema test.27
CONCLUSION
The present study has exemplified an economical and environmentally safe way of ZnO NPs using a clove-ginger formulation. The clove and ginger formulation mediated ZnO NP mouthwash showed good anti-inflammatory at higher concentrations. Additional invasive studies may provide further support for the future use of the mouthwash. The upcoming challenge is to conduct in vivo investigations to identify the histopathological mechanism and various activities of the mouthwash.
ACKNOWLEDGMENT
The authors thank their sincere gratitude to Saveetha Dental College for their constant support in carrying out this research.
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