DENTAL FLUORIDE VARNISH INCORPORATED WITH TULSI EXTRACT

Abstract

Aspect of the present invention relates to a dental fluoride varnish formulation comprising of resin, solvent, fluoride source, tulsi extract, solubilizing agent, adhesion promoting agent and distilled water for prevention and control from dental caries. The invention includes a method for preparation thereof using the constituents. The formulation for dental fluoride varnish is suitable for use in the treatment of hypersensitive teeth, and prevents from demineralization of teeth.

Specification

Description:FIELD OF THE INVENTION
The present invention relates to the field of dentistry. Particularly, the present invention relates to a fluoride varnish formulations and its application in dental care. More particularly, the present invention relates to an antimicrobial dental fluoride formulation with Tulsi extract and the method of preparation thereof for prevention and control from dental caries.

BACKGROUND OF THE INVENTION
Tooth decay, or dental caries, stands as a prevalent condition affecting people of all ages. Its onset results from a multifaceted interplay involving acid-producing bacteria, fermentable carbohydrates, and various host factors such as teeth and saliva. This condition manifests in both the visible parts (crowns) and roots of teeth, potentially beginning early in life with aggressive decay affecting infants' and toddlers' primary teeth.
In dentistry, fluoride treatments are commonly used during routine cleanings to protect teeth from decay. These fluoride compositions are frequently applied to children's teeth, and they are also used for adults. Fluoride application is widely acknowledged as an effective strategy for preventing tooth decay.
The patients undergoing radiotherapy for oral malignancies require regular fluoride application to prevent radiation caries. The high cost of the commercially available Fluoride Varnish and its limited availability restricts its use among economically compromised patients. To overcome the above drawback, the present invention provides a low-cost dental fluoride formulation with Tulsi extracts for prevention from dental caries and as a prophylactic measure in patients with increased risk of dental caries susceptibility.
A need therefore exists to for the enhancement in the performance of fluoride varnish by providing an additional antimicrobial effect by added Tulsi extract, to reduce total microbial load in patient's mouth.
The fluoride varnishes described in the present invention can be quickly applied to the tooth using a brush or small sponge. Once the solvents evaporate, they form a clear, colourless, and strongly adhering film. This film releases fluoride rapidly and significantly, resulting in a calcium fluoride-like layer on the enamel.

OBJECTIVES OF THE INVENTION
An objective of the present invention is to provide a formulation for dental fluoride varnish containing Tulsi extract.
Another objective of the present invention is to provide a formulation for dental fluoride varnish having antimicrobial properties for prevention from dental caries.
Yet another objective of the present invention is to provide a method for the preparation of dental fluoride varnish.
Another objective of the present invention is to provide a low-cost formulation for dental fluoride varnish.

SUMMARY OF THE INVENTION
This summary is provided to introduce a selection of concepts in a simplified format that are further described in the detailed description of the invention. These and other features, aspects, and advantages of the present invention will become better understood with reference to the following description and appended claims. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter. The invention will be described and explained with additional specificity and detail in detailed description.
Accordingly, the present invention provides an antimicrobial formulation for dental fluoride varnish containing Tulsi extract for prevention from dental caries, wherein the formulation comprising of:
a. resin in the range of 40-60% w/v;
b. solvent in the range of 10-40% v/v;
c. fluoride source 5% w/v;
d. tulsi extract 4% w/v;
e. solubilizing agent in the range of 0.1-0.5% v/v;
f. adhesion promoting agent in the range of 1-10% v/v; and
g. distilled water in the range of 2-15% v/v,
wherein each constituent is relative to the total volume of the varnish formulation.
Although the formulation specifies particular quantities, it's important to note that variations in the percentages of each ingredient are anticipated. The percentage adjustments or the inclusion of small amounts of additional ingredients fall within the scope of this disclosure.
In an embodiment of the present invention, there is provided an antimicrobial formulation for dental fluoride varnish, wherein the resin used is Gum Resin. The resin acts as a carrier which advantageously allows the composition to temporarily adhere to tooth tissue, which later naturally goes away as a result of the action of the tongue, saliva and/or other factors including removal by brushing the teeth.
In another embodiment of the present invention, there is provided an antimicrobial formulation for dental fluoride varnish, wherein the solvent used is ethanol with >99% purity, the solvent used in the formulation helps in drying the composition and later on the solvent gets evaporated.
In an embodiment of the present invention, there is provided an antimicrobial formulation for dental fluoride varnish, wherein the fluoride source is sodium fluoride, wherein sodium fluoride provides an antimicrobial activity specially against Streptococcus mutans, which is one of the most common causes of dental caries.
In a preferred embodiment of the present invention, there is provided an antimicrobial formulation for dental fluoride varnish which includes Tulsi extracts, wherein the Tulsi extract is prepared from fresh Tulsi leaves.
In another embodiment, of the present invention, there is provided an antimicrobial formulation for dental fluoride varnish, wherein the solubilizing agent is tween 80, wherein tween 80 is added to prevent phase separation of the different carrier components.
In another embodiment, of the present invention, there is provided an antimicrobial formulation for dental fluoride varnish, wherein an adhesion promoting agent is dimethyl phosphate that promotes adhesion of the varnish to the tooth surface.
The present invention further relates to a method for preparation of the dental fluoride varnish formulation, wherein the method comprising the steps of:
a. adding gum resin and ethanol in a glass beaker and magnetic stirring for 24 hours at 500 RPM to form a mixture;
b. subsequently, adding sodium fluoride to the mixture prepared above in step (a) and solubilizing it completely using magnetic stirring at 1000 RPM to form a mixture;
c. adding Tulsi extract to the mixture prepared above in step (b) and keeping it for magnetic stirring until the Tulsi extract gets completely dissolved; and
d. finally, adding Tween 80 and Dimethyl phosphate followed by distilled water to the magnetic stirred mixture prepared in step (c) and solubilizing the mixture completely under magnetic stirrer at 1000 RPM and keeping the mixture undisturbed for 10 hrs. to obtain the fluoride varnish formulation.
The formulation for dental fluoride varnish according to the invention are suitable in particular for use in the treatment of hypersensitive teeth, in the prophylaxis of caries in patients undergoing head and neck radiotherapy, in the treatment of incipient caries lesions, inhibition of demineralization of teeth, for protection of fissures and pits and for fluoridation of enamel.

BRIEF DESCRIPTION OF THE DRAWING
The drawings described herein are for illustrative purposes only of selective embodiments and not all possible implementations and are not intended to limit the scope of the present disclosure. The invention itself, however, both as to organize and method of operation, may best be understood by reference to the detailed description which follows taken in conjunction with the accompanying drawings in which:
Fig. 1 shows Tulsi Powder mixed with ethanol.
Fig. 2 shows mixing of Tulsi extract with the varnish.
Fig. 3 shows mixing of all the compositions of varnish.
Fig. 4 shows Placement of paper with applied Fluoride Varnishes.
Fig. 5 shows Growth of bacteria after 48 hours.
Fig. 6 shows Measurement of zone of inhibition with the calibrated Vernier calliper
Fig. 7 shows Fluoride varnish with Tulsi extract (group 1)
Fig. 8 shows Fluoride varnish without Tulsi extract (group 2)
Fig. 9 shows VOCO-Profluorid (group 3)
Fig. 10 shows Representative Scanning Electron Microscopic (SEM) Micrographs of Group 1
Fig. 11 shows Representative Scanning Electron Microscopic (SEM) Micrographs of Group 2
Fig. 12 shows Representative Scanning Electron Microscopic (SEM) Micrographs of Group 3.

DETAILED DESCRIPTION OF THE INVENTION
In describing the invention, the following terminology will be used in accordance with the definitions set forth below. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and constituent similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described herein. As used herein, each of the following terms has the meaning associated with it in this section.
One or more constituents or components preceded by "comprises... a" does not, without more constraints, preclude the existence of constituents or components. Appearances of the phrase "in an embodiment", and similar language throughout this specification may, but not necessarily do, all refer to the same embodiment.
In the following specification and the claims, reference will be made to a number of terms, which shall be defined to have the following meanings. The singular forms "a", "an", and "the" include plural references unless the context clearly dictates otherwise.
The present invention relates to an antimicrobial dental fluoride varnish formulation containing Tulsi extract whose function when applied to the tooth is to prevent and control dental caries and tooth decay.
The present invention is cost effective, and consists of Tulsi extracts which is known for its antimicrobial properties specially against Streptococcus mutans, which is one of the most common causes of caries, additionally the present invention also aims to prevent from tooth decay.
In accordance with the present invention, an antimicrobial dental fluoride varnish formulation comprising of;
a. resin in the range of 40-60% w/v;
b. solvent in the range of 10-40% v/v;
c. fluoride source 5% w/v;
d. tulsi extract 4% w/v;
e. solubilizing agent in the range of 0.1-0.5% v/v;
f. adhesion promoting agent in the range of 1-10% v/v; and
g. distilled water in the range of 2-15% v/v,
wherein each constituent is relative to the total volume of the varnish formulation.
Although the formulation specifies particular quantities, it's important to note that variations in the percentages of each ingredient are anticipated. The percentage adjustments or the inclusion of small amounts of additional ingredients fall within the scope of this disclosure.
The present invention further includes a process to prepare Tulsi extracts, wherein the Tulsi extract is prepared from fresh Tulsi leaves and the process comprising the following steps:
a. collecting and washing fresh leaves of Tulsi with tap water to eliminate foreign particles from the collected fresh leaves;
b. drying the washed fresh leaves of Tulsi for at least 7 days at room temperature to remove moisture;
c. grinding the dehydrated leaves of Tulsi to fine powder form for further use;
d. mixing 2.5 g of the fine Tulsi powder as obtained in step (c) with 50 ml of ethanol and stored for 2 days to obtain an aqueous Tulsi;
e. stirring and filtering the aqueous Tulsi obtained in step (d) twice using Whatmann No. 1 filter paper to obtain a filtered aqueous Tulsi, and
f. drying the filtered aqueous Tulsi obtained in step (e) at room temperature to obtain dried solid residue of Tulsi extract, which can be stored in an airtight container till further use.

In a further aspect, the present invention provides a method of preparation of dental fluoride varnish containing Tulsi extract comprising the steps of:
a. adding 2.3 mg of gum resin and 2.15 ml of ethanol in a glass beaker and magnetic stirring for 24 hours at 500 RPM to form a mixture;
b. subsequently adding 0.25 mg of sodium fluoride to the mixture prepared above in step (a) and solubilizing it completely using magnetic stirring at 1000 RPM to form a mixture;
c. adding 0.20 mg of the prepared Tulsi extract to the mixture prepared above in step (b) and keeping it for magnetic stirring until the Tulsi extract gets completely dissolved;
d. adding 0.005 ml Tween 80 and 0.038 ml of Dimethyl phosphate, followed by 0.30 ml of distilled water to the magnetic stirred mixture prepared above in step (c) and solubilizing the mixture completely under magnetic stirrer at 1000 RPM for 2 hrs.; and
e. keeping the mixture obtained above in step (d) undisturbed for 10 hrs. to obtain the fluoride varnish, which later can be kept in an airtight container for future use.

In accordance with the present invention, the dental fluoride varnish formulation can take the form of formulations which is suitable for tooth and may comprise of solution of a liquid spray, or simple liquid form and the like.
Six TYC (Tryptone Yeast Cysteine) agar media plates were prepared and each plate was divided into 4 parts Figure 4. Bacterial suspension of Streptococcus mutans at the turbidity of 0.5 McFarland standard was spread over all the four parts with the help of a cotton swab (lawn culture method). Respective Fluoride varnishes Fluoride varnish with Tulsi extract (group 1), Fluoride varnish without Tulsi (group 2), VOCO-Profluorid (group 3) and control-plain paper disc (group 4) were applied over both the sides of the paper disc which were then carefully placed over the designated part on the culture plate. Plates were placed in an incubating chamber (Thermo Scientific) at CO2 5% and 37º C overnight and were checked for the growth of bacteria Figure 5. The width of the zone of inhibition around the paper disc was measured with the calibrated digital Vernier calliper in four different directions and the average of measurements was recorded Figure 6. All the three Fluoride varnishes, showed significant remineralization potential and antimicrobial properties against salivary Streptococcus mutans Table 1.

Table 1: Intergroup comparison of Antimicrobial property by Disc Diffusion method.
Zone of Inhibition
(Mean ± SD) F value P value
Group 1 13.93 ± 3.33 .080 .924
Group 2 13.01 ± 4.17
Group 3 13.46 ± 4.38

Embodiment examples
Example 1
Preparation of a Fluoride Varnish with added Tulsi extract
A varnish having the following composition is prepared by mixing the components.
40% of resin, 30% of ethyl alcohol, 5% sodium fluoride, 4% tulsi extract, 0.1-0.5% of solubilizing agent, 1-10% of adhesion promoting agent, and 2-15% distilled water. All the constituents are dissolved, accompanied by stirring, in the solvent introduced. A stable, practically colourless, homogeneous, low-viscosity varnish is obtained.
Example 2
An in vitro randomized concurrent parallel design is conducted among 36 extracted premolars. The specimens are divided into three groups, i.e., group 1: Fluoride varnish with Tulsi extract, group 2: Fluoride varnish without Tulsi extract, and Group 3: VOCO-Profluorid, and an artificial caries-like lesion is created on the surface of specimen and respective Fluoride varnishes are applied. The varnish according to the invention is applied to the enamel with a small brush and dried. The specimens are then kept under pH cycling till 4 weeks.
SEM-EDX (Table 2) and Vickers Microhardness (Table 3) test of the specimens are done to evaluate specimens at Baseline, after remineralization, after 2 weeks of pH cycling and after 4 weeks of pH cycling. Antimicrobial potential against Streptococcus mutans is checked through disc diffusion method. Fluoride content of the varnishes and drying time are also evaluated. Statistical analysis is done using One-way ANOVA and Repeated Measures ANOVA with respective post hoc analysis. The level of significance was set at p = 0.05.
The fluoride ion measurement in respective group is found as: group 1 fluoride ion measurement is 16900 ± 4929 ppm, and in group 2 fluoride ion measurement is 16700 ± 12380 ppm. In group 3, it is 17050 ± 5736 ppm. The maximum time taken for drying is seen in Voco-Profluorid followed by Fluoride varnish with Tulsi extract and then Fluoride varnish without Tulsi extract. The highest zone of inhibition for Streptococcus mutans is seen in indigenously prepared Fluoride varnish with Tulsi extract (13.93 ± 3.33 Abstract 2 mm) and least is seen in indigenously prepared Fluoride varnish without Tulsi extract, however the difference in zone of inhibition between the three Varnishes is not statistically significant. All the three Fluoride varnishes, showed significant remineralization potential. There is significant increase in fluoride content from the baseline value in all the three groups. The increase in microhardness from the caries-like lesion to 4 weeks of pH cycling was found in the range of 55-60%.

Table 1: Intergroup comparison of Fluoride (F), Phosphorous (P), Calcium (Ca) content and Calcium/Phosphorous (Ca/P) ratio at Baseline (Normal surface), after demineralization (caries-like lesions), after 2 weeks of remineralization and after 4 weeks of remineralization.
Intervals Groups F Mean ± SD P Mean ± SD Ca Mean ± SD Ca/P Mean ± SD
Baseline Group 1 3.51 ± 2.09 29.02 ± 1.90 70.20 ± 3.12 2.52 ± 0.26
Group 2 3.71 ± 2.10 30.15 ± 1.90 67.78 ± 3.99 2.26 ± 0.29
Group 3 3.82 ± 1.55 29.30 ± 2.09 68.98 ± 4.28 2.36 ± 0.18
Demineralization Group 1 2.12 ± 1.39 27.94 ± 1.21 64.79 ± 1.17 2.33 ± 0.19
Group 2 3.12 ± 1.64 29.61 ± 1.22 62.52 ± 1.49 2.12 ± 0.12
Group 3 1.91 ± 1.18 26.20 ± 1.81 63.50 ± 1.57 2.43 ± 0.18
Oneway ANOVA (p value) 0.209 0.003* 0.008* 0.003*
2 weeks of pH cycling Group 1 3.75 ± 0.58 29.72 ± 0.87 66.21 ± 1.59 2.21 ± 0.10
Group 2 4.75 ± 1.78 29.21 ± 1.94 66.99 ± 3.19 2.28 ± 0.28
Group 3 4.48 ± 1.32 29.85 ± 0.72 65.19 ± 1.71 2.19 ± 0.11
Oneway ANOVA (p value) 0.146 0.553 0.648 0.612
4 weeks of pH cycling Group 1 5.68 ± 1.62 29.51 ± 1.37 69.19 ± 3.66 2.34 ± 0.22
Group 2 7.13 ± 1.81 29.72 ± 0.65 65.42 ± 1.60 2.20 ± 0.1
Group 3 6.46 ± 1.26 30.06 ± 0.45 66.69 ± 2.70 2.22 ± 0.1
Oneway ANOVA (p value) 0.209 0.728 0.039* 0.119

Table 2: Intergroup and intergroup comparison of microhardness through Vickers microhardness test.
Baseline
After Demineralization
2 weeks of remineralization
4 weeks of remineralization
Repeated Measures ANOVA
p value
Group 1 345.61 ± 13.42 163.73 ± 16.87 217.16 ± 15.31 254.35 ± 28.09 0.000*
Group 2 349.90 ± 18.09 164.35 ±5.86 227.21 ± 11.54 264.07 ± 15.58 0.000*
Group 3 342.28 ± 18.90 163.18 ± 13.59 227.38 ± 23.84 260.95 ± 22.12 0.000*
Oneway ANOVA p value 0.820 0.992 0.658 0.799

ADVANTAGES OF THE INVENTION
• The dental fluoride varnish with Tulsi extract provides antimicrobial activities specially against Streptococcus mutans.
• The dental fluoride varnish with Tulsi extract demineralizes early enamel lesions.
• The dental fluoride varnish with Tulsi extract led to microhardness to tooth.
• The dental fluoride varnish with Tulsi extract prevents and control tooth decay.
SPECIFIC EMBODIMENTS OF THE PRESENT DISCLOSURE:
The present disclosure relates to a dental fluoride varnish formulation, wherein said fluoride varnish formulation comprising of:
resin in the range of 40-60% w/v;
solvent in the range of 10-40% v/v;
fluoride source 5% w/v;
tulsi extract 4% w/v;
solubilizing agent in the range of 0.1-0.5% v/v;
adhesion promoting agent in the range of 1-10% v/v; and
distilled water in the range of 2-15% v/v,
wherein each constituent is relative to the total volume of the varnish formulation.

Such dental fluoride varnish formulation is disclosed, wherein the resin used is Gum Resin.
Such dental fluoride varnish formulation is disclosed, wherein the solvent is ethanol with >99% purity.

Such dental fluoride varnish formulation is disclosed, wherein the fluoride source is Sodium Fluoride.
Such dental fluoride varnish formulation is disclosed, wherein the Tulsi extract is prepared from fresh Tulsi leaves.
Such dental fluoride varnish formulation is disclosed, the solubilizing agent is tween 80.
Such dental fluoride varnish formulation is disclosed, wherein the adhesion promoting agent is dimethyl phosphate.
The present disclosure also relates to a method for preparation of the dental fluoride varnish formulation, wherein the method comprising the steps of:
e. adding gum resin and ethanol in a glass beaker and magnetic stirring for 24 hours at 500 RPM to form a mixture;
f. subsequently, adding sodium fluoride to the mixture prepared above in step (a) and solubilizing it completely using magnetic stirring at 1000 RPM to form a mixture;
g. adding Tulsi extract to the mixture prepared above in step (b) and keeping it for magnetic stirring until the Tulsi extract gets completely dissolved; and
h. finally, adding Tween 80 and Dimethyl phosphate followed by distilled water to the magnetic stirred mixture prepared in step (c) and solubilizing the mixture completely under magnetic stirrer at 1000 RPM and keeping the mixture undisturbed for 10 hrs. to obtain the fluoride varnish formulation.

Such method is disclosed, wherein the fluoride varnish formulation is in the form of a liquid spray or simple liquid form and applicable to teeth with the help of brush.
, Claims:WE CLAIM:
1. A dental fluoride varnish formulation, wherein said fluoride varnish formulation comprising of:
resin in the range of 40-60% w/v;
solvent in the range of 10-40% v/v;
fluoride source 5% w/v;
tulsi extract 4% w/v;
solubilizing agent in the range of 0.1-0.5% v/v;
adhesion promoting agent in the range of 1-10% v/v; and
distilled water in the range of 2-15% v/v,
wherein each constituent is relative to the total volume of the varnish formulation.

2. The dental fluoride varnish formulation as claimed in claim 1, wherein the resin used is Gum Resin.

3. The dental fluoride varnish formulation as claimed in claim 1, wherein the solvent is ethanol with >99% purity.

4. The dental fluoride varnish formulation as claimed in claim 1, wherein the fluoride source is Sodium Fluoride.

5. The dental fluoride varnish formulation as claimed in claim 1, wherein the Tulsi extract is prepared from fresh Tulsi leaves.

6. The dental fluoride varnish formulation as claimed in claim 1, the solubilizing agent is tween 80.
7. The dental fluoride varnish formulation as claimed in claim 1, wherein the adhesion promoting agent is dimethyl phosphate.

8. A method for preparation of the dental fluoride varnish formulation, wherein the method comprising the steps of:
i. adding gum resin and ethanol in a glass beaker and magnetic stirring for 24 hours at 500 RPM to form a mixture;
j. subsequently, adding sodium fluoride to the mixture prepared above in step (a) and solubilizing it completely using magnetic stirring at 1000 RPM to form a mixture;
k. adding Tulsi extract to the mixture prepared above in step (b) and keeping it for magnetic stirring until the Tulsi extract gets completely dissolved; and
l. finally, adding Tween 80 and Dimethyl phosphate followed by distilled water to the magnetic stirred mixture prepared in step (c) and solubilizing the mixture completely under magnetic stirrer at 1000 RPM and keeping the mixture undisturbed for 10 hrs. to obtain the fluoride varnish formulation.

9. The method for preparing the dental fluoride varnish formulation as claimed in claim 8, wherein the fluoride varnish formulation is in the form of a liquid spray or simple liquid form and applicable to teeth with the help of brush.

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