CLIAMS:We Claim:

1. A tooth paste which is effective against precancerous lesions such as oral leukoplakia, oral lichen planus, oral submocous fibrosis, erythroplakia, periodontal diseases, dental caries and removal of dental plaque and it acts as antimicrobial and antifungal consisting of: (1) curcumin ; anti-cancerous , antioxidant, anti-inflammatory, antiviral, antibacterial, antifungal, and anticancer agents ; (2) curry leaves , Allium sativum and Tea tree oil; anti-microbial agent; (3) Elettaria cardamomum ; increase salivary secretion & antibacterial (4) Syzygium aromaticum ; suppressing toothache ; astringent (5) Foenieulum vulgare. ; promote salivary secretion & antibacterial (6) silica ; abrasive agent (7) Glycerol; Humectants agent (8) sodium carboxymethyl-cellulose; binder and thicker agents (9) sodium saccharin; sweetener agent; (10) Propyl Paraben ; preservative; ( 11) sodium lauroyl sarcosinate, foaming agent (12) calcium carbonate as abradant; (13) Flavoring agents such as coconut, grape fruit, orange, lime, lemon, mandarin, pineapple, strawberry, raspberry, mango, passion fruit, kiwi, apple, pear, peach, apricot, cherry, grapes, banana, cranberry, blueberry, black currant, red currant, gooseberry, lingon berries, thyme, basil, camille, valerian, fennel, parsley, spearmint, peppermint , chocolate etc

2. A tooth paste as claimed in claim 1, wherein percent amount ingredients by weight of Elettaria cardamomum is 0.01-0.1 %, Syzygium aromaticum is 0.01-0.1%, Foenieulum vulgare is 0.01-0.5%, Tea tree oil is 0.1-1%, Garlic extract is 0.1-1%, Eucalyptus oil is 0.01-2%, Aloe vera is 0.1-1%, Curcumin (yellow or white ) is 0.01-3% and the toothpaste matrix components comprises 15%-30% glycerol, 20%-40% calcium carbonate as abradant, 20%-40% silicon dioxide, 0.5%-1.5% sodium lauroyl sarcosinate, 0.2-1% sodium saccharin, 0.1-0.5% Propyl Paraben and other flavoring agents such as coconut, grape fruit, orange, lime, lemon, mandarin, pineapple, strawberry, raspberry, mango, passion fruit, kiwi, apple, pear, peach, apricot, cherry, grapes, banana, cranberry, blueberry, black currant, red currant, gooseberry, lingon berries, thyme, basil, camille, valerian, fennel, parsley, spearmint, peppermint , chocolate etc

3. Foaming anti-precancerous , antimicrobial, antiplaque tooth paste consisting of: a first anti-precancerous , cancerous with antioxidant, anti-inflammatory, antiviral, antibacterial, antifungal properties which also acts as a anti-inflammatory and an antiplaque agent, a second , third, fourth and fifth antimicrobial agents, a suppressing toothache and astringent agent, a sweetener , three salivary simulating agents, a suppressing toothache and astringent , a Humectants agent and a binder and thicker agents, one or more than one flavoring agents, one foaming agent , one bioavailability enhancer agent , composition being free of sugar and artificial sweetener, composition being free of any side effects.

4. A tooth paste as claimed in claim 3, wherein said first anti- precancerous, cancerous, antimicrobial, antioxidant, anti- inflammatory, antiviral, antibacterial, antifungal agent is curcumin.

5. A tooth paste as claimed in claim 3, wherein said one antimicrobial agent is curry leaves (Allium sativum) .

6. A tooth paste as claimed claim 3, wherein said other antimicrobial agent is Aloe vera.

7. A tooth paste as claimed in claim 3, wherein said bioavailability enhancer is Aloe vera.

8. A tooth paste as claimed in claim 3, wherein said other antimicrobial and anti-caries agent is tea tree oil.

9. A tooth paste as claimed in claim 3, wherein said Elettaria cardamomum is salivary simulating and antibacterial agent.

10. A tooth paste as claimed in claim 3, wherein said Syzygium aromaticum is suppressing toothache and astringent agent

11. A tooth paste as claimed in claim 3, wherein said Foenieulum vulgare is salivary simulating and antibacterial agent.

12. A tooth paste as claimed in claim 3, wherein said silica is abrasive agent .

13. The tooth paste as claimed in claim 3, wherein said sodium lauroyl sarcosinate is foaming agent.

14. A tooth paste as claimed in claim 3, wherein said Glycerol is humectants agent.

15. A tooth paste as claimed in claim 3, wherein said sodium carboxymethyl-cellulose is binder and thicker agent.

16. A tooth paste as claimed in claim 3, wherein said sweetener agent is sodium saccharin.

17. A tooth paste as claimed in claim 3, wherein said perseverated agent is propyl paraben.

18. A tooth paste as claimed in claim 3, wherein flavoring agents such as coconut, grape fruit, orange, lime, lemon, mandarin, pineapple, strawberry, raspberry, mango, passion fruit, kiwi, apple, pear, peach, apricot, cherry, grapes, banana, cranberry, blueberry, black currant, red currant, gooseberry, lingon berries, thyme, basil, camille, valerian, fennel, parsley, spearmint, peppermint , chocolate etc.

19. A tooth paste as claimed in claim 3, wherein calcium carbonate act as abradant.

20. A tooth paste as claimed in claim 3, wherein the tootrhpaste is having a pH of 6.5±0.3.

21. A tooth paste as claimed in claim 3, wherein curcumin act as antitumor, antioxidant, antiarthritic, anti-amyloid, anti-ischemic, and anti-inflammatory properties. In addition it may be effective in treating malaria, prevention of cervical cancer, and may interfere with the replication of the HIV virus multiple myeloma, pancreatic cancer, myelodysplastic syndromes, colon cancer, psoriasis, and Alzheimer's disease, oral cancer, hence, it can be used in prevention and treatment of all above diseases.

22. A toothpaste as claimed in claim 3, wherein anti oral cancer and antimicrobial toothpaste can be used in maintenance of oral hygiene in astronauts for long space missions as well as interplanetary mission .

23. A toothpaste as claimed in claim 3, wherein comprising salivary stimulating agents such as Elettaria cardamomum and Foenieulum vulgare , which can be used for maintaining oral hygiene in radiotherapy patients.

24. A tooth paste as claimed in claim 1 act as anti oral cancerous and precancerous lesions , antimicrobial, antioxidant, antibacterials, antifungal, anti-gingivitis, anti-periodontitis, antiplaque , anti-halitosis, anti-inflammatory , anti-xerostomia, anti-caries, teeth whiteness and anti-ulcers.

25. A tooth paste as claimed in claim 1 acts as tooth whitener.

26. A tooth paste as claimed in claim 1 acts as oral immunity enhancer.

27. A tooth paste as claimed in claim 1 act as wound healer.

28. A tooth paste as claimed in claim 1 acts as protection of total oral health for earth environments and interplanetary environments .
,TagSPECI:FIELD OF THE INVENTION –
The invention relates to dental and oral prophylactics and, more specifically anti-precancerous toothpastes and oral washes which exhibit medicinal, prophylactic, and hygienic effects. The composition consists of curcumin, curry leaves , Allium sativum, Aloe vera and Tea tree oil; Elettaria cardamomum, Syzygium aromaticum ; Foenieulum vulgare, silica, Glycerol, sodium carboxymethyl-cellulose; sodium saccharin, Propyl Paraben, sodium lauroyl sarcosinate, calcium carbonate, flavoring agents such as coconut, grape fruit, orange, lime, lemon, mandarin, pineapple, strawberry, raspberry, mango, passion fruit, kiwi, apple, pear, peach, apricot, cherry, grapes, banana, cranberry, blueberry, black currant, red currant, gooseberry, lingon berries, thyme, basil, camille, valerian, fennel, parsley, spearmint, peppermint , chocolate etc.

BACKGROUND OF THE INVENTION -

The invention resides in tooth paste containing mixture of curcumin, Aloe vera, beta carotene, allicin, coconut oil, Elettaria cardamomum, Foenieulum vulgare and Syzygium aromaticum, performing anti- cancerous and antimicrobial activity without any significant side effects. This toothpaste is used not only in prevention of oral pre-cancerous lesions as oral sub-mucous fibrosis, oral leukoplakia, oral ulcers , lichen planus, erythroplakia and cancerous lesions but also in periodontal diseases and dental caries. Furthermore, it increases the tooth whiteness.

The tooth paste has immense role in oral oncology and oral health to prevent oral precancerous, cancerous and periodontal diseases. It is a basically an inter-disciplinary work between oral oncology, periodontology and pharmacology.

PRIOR ART
Oral hygiene products as toothpaste, typically comprises of abrasive dispersed in a gel or paste base, with detergents, anti-tooth decay agent, flavoring agent and other agents as per requirements .

Observably, there is a extensive need for the development of complete oral health which is effective against anti-periodontitis, anti-gingivitis , oral ulcers, oral pre- cancerous and cancerous lesions & conditions, bad breath and antiplaque , dental caries etc . It should have following properties non toxic, anti- cancerous, antimicrobials, antioxidant , antifungal , antimicrobials , teeth whiteness etc activity in toothpaste.
Different formulation of tooth pastes and mouth wash formulations have been proposed in the patents and studies for the use in oral health. These formulations generally include a multitude of active ingredients such as fluoride agents, abrasives, and antibacterial agents as in these patents, U.S. Pat. No. 5,374,418, U.S. Pat. No. 5,597,553, U.S. Pat. No. 6,086,372, U.S. Pat. No. 6,123,925, and U.S. Pat. No. 6,331,291, U.S. Pat. No. 5,292,527, U.S. Pat. No. 5,284,648, U.S. Pat. No. 5,145,664, U.S. Pat. No. 4,839,158, U.S. Pat. No. 4,254,101 , U.S. Pat. No. 3,981,989, U.S. Pat. No. 4,122,163 , U.S. Pat. No. 4,414,199 and U.S. Pat. No. 7087219. This new formulation enhances the bioavailability , fast removal of the plaque and tooth whiteness action as compared to the formulation taught by patent application no.(1840/DEL/2010). Even, none of the conventional formulation has been directed to a composition that would act as both a prophylactic and as treatment for oral diseases including oral pre cancerous and cancerous lesions.

STATEMENT OF THE INVENTION –
It is now possible, using the methods of the invention, to prevent and treatment to patients suffering from oral leukoplakia, oral lichen planus, oral submocous fibrosis, erythroplakia, periodontal disease and dental caries, conditions of the mouth in a manner that is effective, efficient, and results in minimal undesirable side effects. Treatment using the method of the invention is far more acceptable to patients than the use of creams, gels, or ointments. Furthermore, treatment using a paste as swish results is far better than exposure of the entire oral cavity to the active agents.
An object of the present invention is to provide a safe and effective toothpaste for complete oral health care.
In a broad aspect, the present invention relates to an anti-precancerous, anti-microbial and anti-caries toothpaste comprising: (1) curcumin ; anti-cancerous , antioxidant, anti-inflammatory, antiviral, antibacterial, antifungal, and anti-pre-cancer agents ; (2) curry leaves , Allium sativum, Aloe vera and Tea tree oil; anti-microbial agent; (3) Elettaria cardamomum ; increases salivary secretion and is antibacterial (4) Syzygium aromaticum ; suppressing toothache ; astringent (5) Foenieulum vulgare. ; promotes salivary secretion & antibacterial (6) silica ; abrasive agent (7) Glycerol; Humectants agent (8) sodium carboxymethyl-cellulose; binder and thicker agents (9) sodium saccharin; sweetener agent; (10) Propyl Paraben ; perseverated ; ( 11) sodium lauroyl sarcosinate, foaming agent (12) calcium carbonate as abradant; (13) Flavoring agents such as coconut, grape fruit, orange, lime, lemon, mandarin, pineapple, strawberry, raspberry, mango, passion fruit, kiwi, apple, pear, peach, apricot, cherry, grapes, banana, cranberry, blueberry, black currant, red currant, gooseberry, lingon berries, thyme, basil, camille, valerian, fennel, parsley, spearmint, peppermint , chocolate etc.
Curcumin acts as anti-cancerous, anti-precancerous, antioxidant, anti-inflammatory, antiviral, antibacterial and antifungal agent. It is yellow or white curcumin as well as turmeric extract as disclosed in the earlier patent application(1840/DEL/2010).
The fresh curry leaves, Elettaria cardamomum and Garlic extract was process as disclosed in the earlier patent application(1840/DEL/2010).

Tea tree oil (TTO) ie 100 percent pure oil (Imported from Australia by Thursday plantations Ltd.) was analyzed through gas chromatography. Oil was diluted to 0.2% using vehicle solution and 0.5% Tween 80. The toothpaste composition also comprises abrasive silica. Sorbitol acts as humectants. Sodium saccharin acts as sweeteners. Propyl Paraben acts as preservative. Aloe vera powder acts as bioavailability enhancer of curcumin.
This composition of the present invention may be formulated in a paste concentrate form.
Manufacture of the toothpaste as described in present invention is illustrated in the following example:
The following ingredients were assembled together in their respective amounts.

A toothpaste formulation according to the present invention was prepared as follows:

Elettaria cardamomum: 0.01-0.1 %, Syzygium aromaticum: 0.01-0.1%, Foenieulum vulgare: 0.01-0.5%: Tea tree oil: 0.1-1% ,Garlic extract: 0.1-1%
Eucalyptus oil: 0.01-2%, Aloe vera: 0.1-1%,Curcumin ([Curcuma longa] yellow or white or extract of Curcuma longa): 0.01-3% and the toothpaste matrix components comprises 15%-30% glycerol, 20%-40% calcium carbonate as abradant, 20%-40% silicon dioxide, 0.5%-1.5% sodium lauroyl sarcosinate, sodium saccharin: 0.2-1%,Propyl Paraben: 0.1-0.5%
Flavoring agents such as coconut, grape fruit, orange, lime, lemon, mandarin, pineapple, strawberry, raspberry, mango, passion fruit, kiwi, apple, pear, peach, apricot, cherry, grapes, banana, cranberry, blueberry, black currant, red currant, gooseberry, lingon berries, thyme, basil, camille, valerian, fennel, parsley, spearmint, peppermint , chocolate etc.
The composition of the powders and oils was mixed together to form a smooth paste. The paste was then packed into a tube.

The tooth paste (TP) had a pleasingly fresh, pleasant taste with a pH of about 6.5.
Example: 1

Anti-cancer testing of the Toothpaste
Cell Lines
To examine the effects of the present composition extract of TP on inhibition of cancer/tumor cells, various human cell lines were used including one normal cell line and eight cancer/tumor cell lines:

1. RPTEC (Cambrex): Normal renal proxima tubule epithelial cell, Human
2. A-498 (ATCC HTB-44): Kidney carcinoma, Human
3. HCT 116 (ATCC CCL-247): Colorectal carcinoma, Human
4. Hep 3B (ATCC HB-8064): Hepatocellular carcinoma, Human
5. LNCap clone FGC (ATCC CRL-1740): Prostate carcinoma, Human
6. MCF-7 (ATCC HTB-22): Mammary gland adenocarcinoma, Human
7. MKN45 (FDSC JCRB0245): Stomach carcinoma, Human
8. NCI-H226 (ATCC CRL-5826): Lung carcinoma, Human
9. NPC-TW01 (School of Medicine, NTU): Oral-Nasal- pharyngeal carcinoma, Human
10. KB: Oral cancer
All of the cell lines used were negative for mycoplasma test. Among these cell lines, RPTEC, the only normal cell line was maintained in REGM™ Bullet Kit at 37° C in a humidified atmosphere of 5% CO 2 /95% air, in the absence of antibiotics according to the manufacture's recommendation. On the other hand, the eight tumor cell lines were maintained in DulBeccco's modified essential medium (DMEM) supplied with 10% fetal bovine serum at 37° C, in a humidified atmosphere of 5% CO 2 /95% air in the absence of antibiotics.

Reagents

Fetal calf serum was purchased from Bio Whittaker (Walkersville, Md.), MTS was obtained from Promega (Madison, Wis.). All of the other chemicals were from Sigma Chemical (St. Louis, Mo.) and were standard analytic grade or higher. TP was dissolved in the complete culture medium at final concentration of 10mg/ml and was filter sterilized. Doxorubicin and Taxol were prepared in DMSO at concentration of 20 mM and were used as the positive controls.

To evaluate the anti-tumoral activity of Toothpaste (extract) : Various human cancer cell lines was used to measure the potential cytotoxicity of extract of TP towards the NPC-TW01 and in vitro cytotoxicity assay was carried out. Tumor cells were seeded at a density of 2×10 3 to 8×10 3 cells/well in 96-well plate 16 hours prior to the chemicals treatment. After exposure to different concentrations of extract of TP (5.0 to 16.0 mg/ml), Doxorubixin (10 µM to 3.2 nM) or Taxol (2 µM to 0.02 nM) for 72 hours, cells were washed, replaced with medium containing 0.4mg/MTS [3-(4,5-dimethythiazol-2-yl)-5-(3-carboxymethosyphenyl)-2-(4sulfophenyl)-2H-tetrazolium] and further incubated for 2 hours. To quantify the metabolically viable cells, the conversion of MTS to formazan was measured by absorbance at 490 nm in a 96-well microtiter plate reader. The mock-treated control was used to evaluate the effect of the chemical on cell growth and to determine the concentration of chemical that inhibited 50% of cell growth (IC 50 ). The percentage of cytostasis was calculated by the formula:
Cytostasis(%)=[1-(B/A)]×100, where A is the absorbance of the untreated control and B is the absorbance of the treated cell.

To evaluate the effects of the tested drugs on cell growth, the concentration of chemical that caused 50% reduction of the treated cells (IC 50 ) was calculated from the dose response curve for each cell line. The IC 50 for both Doxorubicin and Taxol to various human tumor cellines were found between 800 to 11 nM and 11 to 0.003 nM respectively.. Our data showed that the cytotoxicity of TP toward cancer cell line depends on the nature of the tumor cells, showing the selectivity of extract of TP. Among all the tumor cell lines tested, MW was found to be most sensitive to KB, as the concentration of extract of TP as low as 0.59 mg/ml was able to cause 50% reduction of the treated cells, demonstrating the efficient anti-tumor activity of TP on KB and Oral-Nasal- pharyngeal carcinoma cells . In addition, cell growth stimulation was found when KB was cultured in higher concentration of TP. The results showed that TP has little or no toxicity to the normal cells and also further increase of conc.could inhibit growth of cancerous cell .

Example:2
The Toothpaste(TP) formulation set forth in Example I and placebos were evaluated in vivo against oral leukoplakia, oral lichen planus, submucous fibrosis and Erythroplakia etc. In the patients tested, response to present mouth compositions is excellent on precancerous lesions.

All diagnostic tests were evaluated for diagnosing oral pre-cancerous lesions. Each subject completed a medical and dental history questionnaire to determine the status of systemic diseases, chronic diseases, oral and dental diseases, smoking habit or alcohol and drug history.

TP and placebo were given four times in a day for 5 minutes. Pain control and lesion healing were the two main clinical variables for evaluating cure of oral leukoplakia, oral lichen planus and oral submucous fibrosis. To measure pain, we used a visual scale analog ranging from 0.5 (Very mild pain) to 5 (severe pain). For healing, we measured changes in lesion size, including ulcer size from baseline, while; in oral submucous fibrosis in addition to the above variables , change in mouth opening was considered. Final outcome was confirmed by clinical examination. Clinical and histopathological examinations were conducted along with collection of serum and salivary samples prior to treatment of TP. One week after clinical cure of diseases, clinical and histopathogical examinations were again performed along with collection of serum and salivary samples. Whole unstimulated saliva produced in a 5 min period ( 3ml) was collected, allowed to drain into a plastic container , and was centrifuged at 3000xg at 4 C for 5 min in order to remove bacterial and cellular debris. Saliva samples were stored at -80C until analysis. Blood was centrifuged at 1700xg for 10 min and plasma was separated. Plasma was stored at -80 C until analysis was performed. Serum and salivary IL-2, IL-1 beta, IL-6, IL-8, 8 OHdG ,MDA, vitamin C and E were measured These biomarkers were selected because these are salivary biomarkers of oral cancer

Results:

AVERAGE Lesion size ( Placebo)
- 20 oral leukoplakia: baseline: 23.4mm (5.4) : After study: 21.3mm (12.2)
- 20Lichen planus: baseline: 22.3mm (11.3) : after study : 21.3mm (13.2)
- 22 Submocous fibrosis; 22.8 (11.4): after study 21.4 (12.7)
AVERAGE pain score for patients
- 20 oral leukoplakia: baseline: 4.5 (1.5): after study: 4.0(1.5)
- 20 Lichen planus: baseline: 3.5(1.5): after study: 3.5(1.0)
- 20 oral submoucous fibrois: 4.5 (1.0): after study 4.0 (1.0)
Mouth opening only for submucous fibrosis
baseline : 28.4 (5.4) : after study 28.2 (5.9) mm


AVERAGE Lesion size (Our invention)

- 50 oral leukoplakia: baseline: 22.4mm (4.5): end study: 3.2mm (1.3)
- 50 Lichen planus: baseline: 24.3mm (4.8) : end study: 3.4mm (1.6)
- 50 Submocous fibrosis; 24.6 (4.2) : end study 3.7 (1.3)
AVERAGE pain score for patients
- 50 oral leukoplakia: baseline: 3.5 (1.5): end study: 0.5(0.5)
- 50 Lichen planus: baseline: 4.0(1.5) :end study: 0.5(0.5)
- 50 oral submoucous fibrois: 4.5 (1.5) :end study 0.5 (0.5)
Mouth opening only for submucous fibrosis
baseline : 20.2 (4.6) : end study 45.2 (7.3) mm


Pain scores and size of lesion in oral leukoplakia, submucous fibrosis and lichen planus improved significantly ( P=0.0001) in TP as compared to placebo. In case of submucous fibrosis, mouth opening recovered significantly after TP usage as compared to placebo. Oral leukoplakia, submucous fibrosis and lichen planus were clinically cured after 234 (34), 267(47), and 242(42) days, respectively. Serum and salivary IL-2, IL-1 beta, IL-6, IL-8, 8 OHdG and MDA levels were significantly decreased ( P=0.0001, 95% CI) , while vitamin C and E levels were significantly increased after treatment with TP as compared to placebo.

Example 3.
The Toothpaste formulation set forth in Example 1 was evaluated in vivo against gingivitis & periodontitis and tooth whiteness.
ComparativeData

Toothpaste(TP) of the invention was compared for effectiveness with known commercial formulations. Each of the patients was first treated with the known formulations. In the patients tested, response to the prior art compositions was poor as compared to present formulation.
Patient #60 Diagnosis: Gingivitis
Patient # 60Diagnosis: Periodontits
Response Drug Pt.# Pt.#, Gingival index, periodontal index, probing depth , attachment lose and bleeding on probing were evaluated in before and after present TP and other tooth pastes. 0: no improvement, very slight improvement, : slight improvement, : moderate improvement, : good improvement, : excellent improvement.
The subjects, of age varying from 23 to 60 years (median age 45 ± 21), with generally good health (pregnant and breastfeeding excluded), who met the following inclusion criteria, were included into the study: a minimum of 20 sound, natural teeth; a mean plaque index (PI) of at least 1.5; a mean gingival index (GI) of at least 1.0. Subjects with orthodontic appliances or removable prosthetics, tumors of the soft or hard oral tissues, and advanced periodontal disease, receiving antibiotic therapy 2 weeks before the beginning of the study were excluded. Third molars and the teeth with cervical restorations or prosthetic crowns were not included in the tooth count. The selection of participants was made by convenience based especially on the availability to the study, while the study was conducted. All subjects read and signed informed consent forms before the start of the study. The protocol for the study was approved by the local ethical review committee.
The examination baseline consisted of a complete soft and hard tissues examination that was performed to register the condition of oral mucosa, so that any changes in the course of the study could be identified, making assessment to whether these changes could be related to the TP. The gingivitis of the mesiobuccal, midbuccal, distobuccal, mesiolingual, midlingual, and distolingual of all eligible teeth was scored using the Talbott modification Gingival Index of the Löe-Silness , in which the gum was scored on a four-point scale from 0 (absence of inflammation) to 3 (severe inflammation). The supragingival plaque of the mesiobuccal, midbuccal, distobuccal, mesiolingual, midlingual, and distolingual of all eligible teeth was scored using the Turesky modification of the Quigley-Hein Plaque Index . Previously, disclosing with erythrosine 3% solution, plaque was scored on a six-point scale from 0 (no plaque) to 5 (plaque covers two-thirds or more of the tooth surface). Each tooth was divided into six areas, three buccal (mesiobuccal, midbuccal, distobuccal) and three lingual (mesiolingual, midlingual, distolingual), and the plaque was quantified using the Turesky modification of the Quigley–Hein Plaque Index. The Severity Plaque Index and Severity Gingival Index were also evaluated. These indices measured the rate of the surface that had high count of plaque (count similar to 3, 4, 5) of the modification Quigley-Hein Plaque Index and high gingival index (count similar to 2, 3 modification Gingival Index of the Löe-Silness . These examinations were all repeated after 45 and 90 days of the use of the TP. After baseline examination, each subject received a complete oral prophylaxis, which included the removal of all supragingival plaque and calculus deposits. Soon after, the subjects received TP. They were instructed to brush their teeth as usual with TP, twice a day, for one minute, right after their meals in the morning and at night. Participants were required not to use toothpaste throughout the study. When new supplies were issued, subjects returned their used materials, so that the compliance to the product could be monitored.
All examinations were conducted by a single examiner trained to optimize the consistency of the study. Prior to the study, adviser trained the dental examiner, as a “gold standard,” directing him to introduce the periodontal probe, gently, into the gingival sulcus, keeping the instrument parallel to the long axis of the tooth, sliding it from the distal to the mesial so delicately in the buccal and lingual surface of each evaluated tooth. For calibration, there were examined nine subjects who were not included in the stud. For the plaque index there was performed a theoretical calibration. Soon after, photos were used to obtain a standardization intra-examiner. The photos were exhibited by the adviser to the examiner that noted the values of plaque index corresponding to each picture. After 15 days, the same pictures were exposed to the examiner that registered again the values of the plaque index. Next, the plaque index obtained in the first and second time was compared to verify the level of intra-examiner. Then, we obtained a kappa value of 0.73, considered a substantial estimate of reliability. The statistical package BioEstat version 4.0 was used for data analysis in this study. The average adjusted in the baseline for both scores of the modified Plaque Index of Quigley-Hein and modified Gingival Index of Löe-Silness as for the corresponding severity scores, for being a nonparametric distribution, was compared through covariance analysis, by Friedman test for data obtained at 45 and 90 days of the study. All statistical tests of hypotheses had two strands and a significance level of ?? <. 00 5 was considered.
The mean baseline scores of Gingival Index GI, 45 and 90 days, were recorded. The TP demonstrated a reduction in gingivitis over than 40%, being statistically significant, comparing to the scores of 45 and 90 days in the baseline scores (?? < . 00 5). Also, comparing to the scores of 90 days to 45 days, there was a statistically significant reduction in gingivitis. The TP showed a reduction in rate of the surface with scores 3, 4, and 5 of plaque index. This reduction was statistically significant, (over 70%), comparing the mean baseline scores to 45 and 90 days (?? < .0 0 5).
The mean baseline scores of plaque index (PI), 45 and 90 days, were obtained. Analysis of variance using the Friedman test was performed with the baseline scores as covariates, showing that the TP had an effect on the plaque in the examinations at 45 and 90 days, being statistically significant (?? < . 00 5), with a reduction of 76% and 74%, respectively. In Severity Plaque Index, there was observed a statistically significant difference comparing to the mean baseline scores of 45 and 90 days. There was a reduction of 71% in the second exam, remaining the same after the third exam (?? < . 0 05). It is also effective against gingivitis. The teeth whiteness was significantly increased after tooth brushing of this tooth paste

Example 4: Effect of Toothpaste(TP) on oral malodor , salivary bacteria and oral immunity

In the first test phase, the group 1 subjects (N = 30) were instructed to brush with 10 ml of the experimental TP ( 50%) as semisolid for 30 seconds twice per day (after waking and before sleeping) for 28 days, and Group 2 (N = 30 ) to tooth paste with the placebo. In the second phase, after a one week washout period, each group then used the opposite toothpaste for 28 days. At baseline, and after 28 days, oral malodor was evaluated by the organoleptic measurement (OM) and gas chromatographic(GC ) me t h o d s . C o n c e n t r a t i o n s of H2S, CH3SH and (CH3 )2S were evaluated with GC. Clinical outcome variables included plaque and gingival indices, tongue coating index (TCI) and tongue discoloration index ( TDI ). Subjects continued their u s u a l o r a l hygiene practices during the study, except for the morning of each clinical assessment. Samples of the resting whole saliva were collected into a sterile plastic tube over a 5 minutes period, prior and end of study for oral status assessments, and were immediately analyzed. There was no statistically significant difference between the control group and the experimental group at base line. Statistically significant improvements in reducing oral malodor occurred in experiment group which used TP for 28 days , compared to baseline scores (p < 0.01).
The placebo tooth paste used for 28 days, on the other hand, showed no statistically significant difference in oral malodor compared with baseline scores. After using the toothpaste for 28 days, especially, the mean concentrations (ng/10 mL) of H2S, CH3SH and (CH3)2S in the experimental group were significantly reduced compared to those in the control group. Oral status evaluations with the experimental TP used for 28 days, a statistically significant inhibition in plaque accumulation was evident compared to toothpaste used before (p < 0.05). With the placebo toothpaste used for 28 days on the other hand, no statistically significant inhibition was observed compared to toothpaste used before. Further, the mean score of the experimental group was not significantly lower than the mean plaque score of the control group after 28 days. There was statistically significant difference compared with the GI value before tooth brushing . With the experimental TP used for 28 days, statistically significant inhibition in tongue coating was evident compared to before tooth brushing (p < 0.01). For the placebo tooth brushing used for 28 days, no statistically significant inhibition in tongue coating occurred compared with levels before tooth brushing. The TCI scores of subjects within the experimental group were significantly lower than those of the control group after 28 days.
There were no statistically significant differences between numbers of total bacterial counts with experimental and the placebo tooth paste used over 2 8 days. The detection threshold for this method was 10 copies of bacterial DNA per reaction. AT baseline , Fusobacterium Nucleatum (FN) was detected in all subjects. With Experimental TP for 28 d a y s , mean counts of F.n. was reduced, showing statistically significant bacterial inhibition in saliva compared to the baseline (p < 0.01). The placebo tooth paste group after 28 days, on the other hand, showed no statistically significant inhibition compared to the baseline. There were statistically significant differences between the mean number of F.n. counts in the experimental and those in the control group after 28 days (p < 0.05).With the experimental paste (TP) used for 28 days, the numbers of Prophyromonas gingivalis (PG) count was reduced, however, there was no significant difference compared with the before brushing baseline. In the control group, no statistically significant inhibition was observed compared to the baseline. PG was detected in all subjects before and after tooth brushing in both experimental and control group. There was statistically significant difference between the numbers of PG counts in the experimental and control after 28 days. At baseline, Tannerella forsythensis ( T.F) were detected in all subjects, however, after using the experimental TP for 28 days. The experimental TP reduced the mean number of T.F counts after 21 days, also, there was statistically significant difference compared with baseline counts. The control TP showed no reduction in the mean number of T.F counts after 21 days, also, there was no statistically significant difference compared to baseline counts. Oral immunity was significantly increased after brushing this tooth paste by measuring salivary immunity biomarkers and decreased levels of salivary micriobials.

OBJECT OF THE INVENTION –
According to the present formulation, it contains curcumin which acts as anti-cancerous, antioxidant, anti-inflammatory, antiviral, antibacterial, antifungal, and anti-precancer teeth whiteness agent. According to present formulation, garlic and aleo vera acts as anti bacterial which inhibits growth of various gram-positive and gram-negative bacteria in the oral cavity. According to another embodiment of the invention, comprises tea tree oil attributed mainly to terpinen-4-ol acts as anti microbial.
According to another embodiment of the invention comprises Elettaria cardamomum, act as salivary secretion simulating and antibacterial agent.

According to another embodiment of the invention comprises Syzygium aromaticum, acts in suppressing toothache and act as astringent agent.
According to another embodiment of the invention comprises Foenieulum vulgare, act as salivary secretion simulating and antibacterial agent.

According to another embodiment of the invention silica, act as abrasive agent
According to another embodiment of the invention Glycerol, act as Humectants agent .

According to another embodiment of the invention sodium carboxymethyl-cellulose, act as binder and thicker agent.

Yet another embodiment of the invention comprises sodium saccharin, act as sweetener agent.

Yet another embodiment of the invention comprises Flavoring agents such as coconut, grape fruit, orange, lime, lemon, mandarin, pineapple, strawberry, raspberry, mango, passion fruit, kiwi, apple, pear, peach, apricot, cherry, grapes, banana, cranberry, blueberry, black currant, red currant, gooseberry, lingon berries, thyme, basil, camille, valerian, fennel, parsley, spearmint, peppermint , chocolate etc

Another embodiment of the invention comprises calcium carbonate as abradant .

Other embodiment of the invention comprises Aloe vera powder which acts as bioavailability enhancer of curcumin. It is also antimicrobials agent.
Other embodiment of this invention comprises sodium lauroyl sarcosinate, acts as foaming agent.

Yet another embodiment of the invention comprises Propyl Paraben, acts as preservative.