Analysis of an Herbal Ayurdantham Medicated Tooth liquid on Dentinal Tubule Occlusion using Scanning Electron Microscopy.

BACKGROUND: Dentin hypersensitivity is described as "short sharp pain arising from exposed dentin in response to various stimuli typically thermal, evaporative, tactile, osmotic or chemical and cannot be ascribed to any other form of dental defect or disease." Dentine hypersensitivity can be treated with various desensitizing agents ,where oral rinse has a promising effect in delivering the therapeutic effect all over the tooth surface including interproximal areas, as they can flow and reach areas where toothpastes are not much effective, hence the occlusion of dentinal tubules using Ayurdhantam medicated an herbal mouthwash was evaluated.
METHODOLOGY: Twenty such dentinal blocks were obtained; the surfaces of each dentin block were polished using a silicon carbide disc for 30 s to create a basic smear layer. The smear layer is finally detached by applying 17% EDTA solution onto the dentin blocks for 5 min to ensure the patency of the dentinal tubules, following which they were rinsed with distilled water. The specimens were placed into the Ayurdhantam solution (medicated) for 2 min every day for 7 days. The specimens were stored in artificial saliva during the study. The specimens were analyzed under scanning electron microscope (SEM) for the occlusion of dentinal tubules.
RESULTS: SEM images showed the partial occlusion of the tubules and a large amount of smear layer.
CONCLUSIONS: Based on this in vitro experiment carried out, it is shown that Ayurdhantam-medicated mouthwash can be used for treating dentine hypersensitivity effectively. Copyright:

INTRODUCTION

Dentin hypersensitivity (DH) is defined as “short sharp pain arising from exposed dentin in response to various stimuli typically thermal, evaporative, tactile, osmotic or chemical and cannot be ascribed to any other form of dental defect or disease.”[12]

While 4%–74% of the population is affected by DH, it is more prevalent in the age group of 20–40 years.[3]

The etiology of DH depends on several factors and results from the opening of dentinal tubules, due to loss of enamel by erosion, abrasion, abfraction or corrosion, or loss of cementum by brushing. Application of topical agents for DH will occlude the exposed dentinal tubules or have an instantaneous impact on the pulpal nerve fibers for desensitization.[4]

The most accepted method of delivering the desensitizing agents after toothpaste is mouthwashes. Mouth rinses can deliver the therapeutic effect all over the tooth surface including interproximal areas, as they can flow and reach areas where toothpastes are not much effective.

Natural products have been extensively used for medicinal purposes for thousands of years. These products have pharmacological properties such as antimicrobial, anti-inflammatory, astringent, and cytostatic effects. Besides maintaining oral hygiene, mouthwashes can also be used for many gum disorders and DH.[5]

However, a new product by name Ayurdhantam medicated as an herbal mouthwash which is commercially available claims to reduce DH. Hence, the study aimed to check the occlusion of the dentinal tubules.

METHODOLOGY

Specimen selection and preparation

Twenty well-formed, noncarious, sound, extracted human permanent molars were chosen for the study. Teeth having developmental defects, fractures, craze lines, wasting diseases, caries, or restorations are not included. Patient consent was obtained for the use of the extracted teeth.

The teeth were thoroughly disinfected and stored in 0.5% thymol at 4°C before their use. The teeth were sectioned with a diamond disc mounted on a slow-speed water-cooled handpiece. The first cut was made perpendicular to the long axis of the tooth above the cementoenamel junction, and the second cut was made parallel to the first to remove all the coronal enamel to expose the flat dentin surfaces to obtain 3-mm thick dentinal blocks. The surrounding enamel of the blocks was cut off creating specimens measuring 5 mm × 5 mm × 3 mm [Figure 1].

Figure 1

Schematic presentation of the preparation of a dentin block (Courtesy article 2)

Twenty such dentinal blocks were obtained free of enamel and pulpal exposure, which were subsequently embedded in acrylic resin. The surfaces of each dentin block were polished with a 600-grit silicon carbide disc for 30 s to create a definitive smear layer. The smear layer was finally removed by applying 17% EDTA solution onto the dentin blocks for 5 min to ensure the patency of the dentinal tubules, following which they were rinsed with distilled water. The specimens were placed into the Ayurdhantam solution (medicated) for 2 min every day for 7 days. During the study, the specimens were stored in artificial saliva.

Data analysis

The specimens were analyzed under a scanning electron microscope (SEM) for occlusion of the dentinal tubule. Samples were mounted on the aluminum holder stubs using a dual viscous carbon tape and coated with Au/Pd in an SPI-MODULE E5400 high-resolution sputter coater and examined in a ZEISS (EVO LS 15, Germany) scanning microscope at 15 kV.

RESULTS

The photomicrographs were obtained at different magnifications: ×500 and ×1000. Figures 2 and 3, respectively, show SEM images of the sample treated with Ayurdhantam-medicated mouthwash. The images showed the partial occlusion of the tubules and a large amount of smear layer.

Figure 2

Scanning electron microscope image of dentinal tubule occlusion at ×500

Figure 3

Scanning electron microscope image of dentinal tubule occlusion at ×1000

DISCUSSION

Various treatment modalities were proposed for DH, in which the medication is self-managed by the patient at home or practiced by a dental professional in the dental office.[6] Numerous modalities were administered for the therapy of DHs such as over-the-counter desensitizing toothpaste, mouthwashes, lasers, iontophoresis, dentinal sealers, and soft tissue grafting.[4]

Treatment approaches to relieve the DH is to cut off the neural response to pain stimuli or to occlude open dentinal tubules to prevent the hydrodynamic mechanism.[7]

Due to an increased awareness of holistic medical practices, the usage of “herbal” medicine has raised the importance and promoted the growth of reciprocal and different therapies in the oral healthcare profession.[8] The natural oral rinse is an aqueous antibacterial solution used against the microbes in the oral cavity to counter infection and reduce halitosis which acts as an antiseptic and plays a vital role in curing diseases.[9] A new product Ayurdhantam-medicated mouthwash showed promising results due to antioxidant, anti-inflammatory, and analgesic properties [Table 1].

Table 1

Composition: each 10 ml liquid contains

Sanskrit name	Botanical name	Parts used	Anti-inflammatory	Antioxidant	Analgesic
Haritaki	Terminalia bellirica	Fr pulp	✓	✓	✓
Bibhitaki	Terminalia Chebula	Fr pulp	✓	✓
Amalaki	Emblica officinalis	Fr pulp	✓	✓	✓
Nimba	Azadirachta indica	Patra	✓	✓
Yestimadhu	Glycyrrhiza Glabra	Root	✓	✓
Pippali	Piper longum	Seed	✓		✓
Saindhava	Chloride of sodium	-	✓
Lavanga			✓	✓	✓
Ela			✓		✓

The term antioxidant is used for molecules that reduce the damaging capacity of free radicals using their scavenging property. They are stable enough for donating an electron to a rampaging free radical and neutralizing it. Lavaga (clove) acts as a scavenger and the presence of enzyme activator which adds up to antioxidant action.[10] It also has an anesthetic effect because essential oil helps in numbing the nerves and decreases the pain by soothing effect. The key oil of clove is also an antiseptic, which benefits in eliminating the oral bacteria.[11]

Terminalia bellirica has excessive phenolic, flavonoid content where gallic acid, and ferulic acids being the major phenolic compounds has demonstrated high antioxidant and radical scavenging activity.[12] Emblica officinalis has diverse phytochemical ingredients of the plant such as emblicanins A and B, gallic acid, and ellagic acids, which are powerful free radical scavengers. Recently, it has been described that the superoxide scavenging properties of E. officinalis approximates L-ascorbic acid, which is a well-endorsed antioxidant.[13] Azadirachta indica and Terminalia chebula also contribute to antioxidant property.

T. chebula, Glycyrrhiza glabra, Terminalia bellirica, and E. officinalis showed an anti-inflammatory action. It is also considered to be a powerhouse of bioactive agents and flavonoids. It is imperative to note that flavonoids present as a component are the main active and reactive agents capable of stimulating reparative dentin formation. This could be one of the factors that enable to reduction of dentin permeability efficiently. Based on the hydrodynamic theory, agents used for the treatment of DH should be capable of occluding the dentinal tubules and prevent nerve sensitivity. Thus, the above-mentioned components are shown to be capable of obliterating the dentinal tubules.[3]

Calcium phosphate technologies are other choice for treating hypersensitivity.[14] Saliva helps naturally in reducing DH; by transporting calcium and phosphate into dentin tubules to induce dentinal tubule plugging; and by producing a defensive layer of salivary glycoprotein. Alkaline pH favors both processes.[7] Ayurdhantam-medicated mouthwash has Saindhava lavana (Rock salt) which contains sodium chloride, calcium, phosphorus, potassium, chromium, zinc, and strontium[15] and G. glabra has flavonoids in which glycyrrhizin is a potassium and calcium salt of glycyrrhizic acid.[16] Potassium causes desensitizing effects directly on intradental nerves. It is said that the high extracellular concentration of potassium prevents the nerve cell's repolarization and the transmission of the pain impulse.[17] Amorphous calcium phosphate makes calcium and phosphate ions accessible in saliva to advance remineralization.

The dentin consists of the mineralization phase, which is mainly hydroxyapatite, and an organic matrix made up of collagen (type I, type III, and type V collagen). Odontoblasts are placed at the external layer of the pulp and form the dentin which protects the pulp.[1819] Odontoblastic process spreads into the dentinal tubule of dentin. Therefore, the pulp and the dentin form an intricate relationship and manipulate each other. Dentin is a permeation structure. The stimuli from the oral cavity may irritate the pulp and induce inflammation; studies have demonstrated the infiltration of the anti-inflammatory and antibacterial drugs through coronal dentin which improves the pulpal condition.[2021] Thus, it is feasible to utilize anti-inflammatory agents onto the open dentin to treat the pulp diseases.[22] The anti-inflammatory and analgesic effect of the medicated mouth rinse helps in reducing the sensitivity.

The presence of the smear layer which is seen on SEM images might add up to occlusion of the tubules, which decreases the DH. Further studies with comparison and the simulation of the oral environment are needed.

CONCLUSION

Our study critically examined the use of Ayurdhantam medicated as a natural medicated mouthwash for treating DH. While the composition of solution used in the study is unique, the efficacy of treatment is evaluated using SEM analysis of high resolution. One of the limitations of this study is that the in vivo oral cavity conditions could not be simulated. Based on this in vitro experiment carried out, it is shown that Ayurdhantam-medicated mouthwash can be used for treating DH effectively. Future in vivo studies, comparing Ayurdhantam to other available desensitizing products, can highlight its feasibility in clinical practice.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.