Indian Journal of Dental Research

: 2010  |  Volume : 21  |  Issue : 3  |  Page : 380--384

Chitosan-based polyherbal toothpaste: As novel oral hygiene product

Nitin C Mohire, Adhikrao V Yadav 
 Department of Pharmacy, Government College of Pharmacy, Vidyanagar, Karad, District- Satara, Maharashtra, India

Correspondence Address:
Nitin C Mohire
Department of Pharmacy, Government College of Pharmacy, Vidyanagar, Karad, District- Satara, Maharashtra


Objective: The objective of the present work was to develop chitosan-based polyherbal toothpaste and evaluate its plaque-reducing potential and efficacy in reduction of dental pathogens. Materials and Methods: Antimicrobial activity of herbal extracts against dental pathogens were performed by using disk diffusion method. The pharmaceutical evaluation of toothpaste was carried out as per the US Government Tooth Paste Specifications. A 4-week clinical study was conducted in patients with oro-dental problems to evaluate the plaque removing efficacy of chitosan-based polyherbal toothpaste with commercially available chlorhexidine gluconate (0.2% w/v) mouthwash as positive control. Total microbial count was carried out to determine the percentage decrease in the oral bacterial count over the period of treatment. Result: Herbal extracts were found to possess satisfactory antimicrobial activity against most of the dental pathogens. Chitosan-containing polyherbal toothpaste significantly reduces the plaque index by 70.47% and bacterial count by 85.29%, and thus fulfills the majority of esthetic and medicinal requirements of oral hygiene products. Conclusion: Chitosan-based polyherbal toothpaste proves itself as a promising novel oral hygiene product as compared with currently available oral hygiene products. A further study to confirm the exact mechanism and active constituents behind antiplaque and antimicrobial activity of chitosan-based polyherbal toothpaste and its efficacy in large number of patient population is on high demand.

How to cite this article:
Mohire NC, Yadav AV. Chitosan-based polyherbal toothpaste: As novel oral hygiene product.Indian J Dent Res 2010;21:380-384

How to cite this URL:
Mohire NC, Yadav AV. Chitosan-based polyherbal toothpaste: As novel oral hygiene product. Indian J Dent Res [serial online] 2010 [cited 2021 Aug 3 ];21:380-384
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Full Text

Dentifrices and mouthwashes that are generally produced by cosmetic manufacturers are really not cosmetics. They fall more properly into the field of hygienic products just as soap. Both products have more or less the same function, which are to cleanse, to counteract bad breath, and to leave a refreshing clean taste in the mouth. It may also be argued that properly cleaned teeth add to personal appearance and good health. [1] Good oral hygiene results in a mouth that looks and smells healthy. This means, teeth are clean and free of debris, gums are pink and do not hurt or bleed when you brush or floss, and bad breath is not a constant problem. [2]

Oral hygiene is of utmost importance as it contributes in esthetic expectations of an individual and of society. The strong positive association that exists between poor oral hygiene and gingival and periodontal diseases makes poor hygiene the primary etiologic agent. [3]

We have already reported antimicrobial activity of chitosan. [4] Chitosan, an abundant natural polymer, is obtained by alkaline N-deacetylation of chitin. Chitosan being a binding agent, bio-adhesive, bio-compatible, bio-degradable, and non-toxic polymer also possessing medicinal activities, such as antifungal, antibacterial, antiprotozoal, anticancer, antiplaque, antitartar, hemostatic, wound healing, and potentiates anti-inflammatory response, inhibits the growth of cariogenic bacteria, immunopotentiation, antihypertensive, serum cholesterol lowering, absorption enhancer, increases salivary secretion (anti-xerostomial), and helps in the formation of bone substitute materials. [5],[6],[7]

The adherence of oral bacteria on the tooth surface leads to plaque formation. It is believed that the adhesion between the bacteria and the tooth surface is due to electrostatic and hydrophobic interactions. These interactions are disrupted by chitosan derivatives because of competition by the positively charged amine group. [8],[9] The antibacterial activity of chitosan could be due to the electrostatic interactions between the amine groups of chitosan and the anionic sites on bacterial cell wall because of the presence of carboxylic acid residues and phospholipids. [10]

Use of most of the currently used gelling agents, such as tragacanth, Irish moss, and sodium alginate mucilage, in the toothpaste was limited only to their gelling capacity and also require antimicrobial preservatives due to their carbohydrate nature, whereas chitosan being a good gelling agent, does not require any preservatives as chitosan possess antimicrobial activities.

The antimicrobial activity of Pterocarpus marsupium aqueous extract (PM), Stevia rebaudiana aqueous extract (SR), Glycyrrhiza glabra aqueous extract (GG) against selected dental pathogens was carried out at our research center to support the study.

Chlorhexidine gluconate (CHX) 0.2% solution is still the golden standard as mouthrinse for the prevention of plaque formation and development of gingivitis. CHX has good antimicrobial activity against the microbes responsible for oral infections. [11] Although antimicrobial effect of CHX is generally undisputed and well acknowledged, the mode of treatment and delivery system for maximal effectiveness is not yet fully clear. The side effects associated with CHX, such as extrinsic tooth staining, poor taste, taste disturbance, sensitivity changes in tongue, pain, and the content of alcohol led us to search for a novel oral hygiene product.

Considering the limitations of the presently available products and the promising properties of chitosan and selected herbs suitable to address oral care problems, present invention was directed to design and develop chitosan-based polyherbal toothpastes with enhanced performance in oral care as chitosan inhibits the growth of Streptococcus mutans [12] and Porphyromonas gingivalis; [13] microorganisms responsible for caries and gingivitis, respectively. Chitosan also potentiates the effectiveness of active ingredients of toothpaste for antimicrobial and anti-inflammatory activities.

 Materials and Methods


All the chemicals used are of analytical grade. Precipitated calcium carbonate, Dicalcium phosphate, Gum tragacanth, Sodium lauryl sulfate, Sorbitol, and Eugenol were purchased from Loba chemie, Mumbai. Silicon dioxide was received as a gift sample from Okasa Pharma, Satara. Chitosan was received as a gift sample from India See Food, Cochin, Kerala.


PM, GG, and SR, were prepared in various strengths: 50, 100, 500 μg/mL and their antimicrobial activity against selected dental pathogens, such as Pseudomonas aueroginosa, Proteus vulgaris, Escherichia coli, Bacillus subtilis, Lactobacillus acidophilus, Candida albicans were determined by using disk diffusion method as mentioned in [Figure 1]. Gentamycin 100 μg/mL was used as standard for evaluation of antimicrobial activity and fluconazole 10 μg/mL, was used as standard for evaluation of antifungal activity.{Figure 1}

Two toothpaste formulations (Test (Polyherbal), Placebo) were prepared and compared with CHX (0.2%) mouthwash for its clinical efficacy.

Test (Polyherbal) toothpaste: Chitosan gel and Sorbitol were mixed thoroughly. Abrasives, such as Precipitated calcium carbonate, Dicalcium phosphate, and other excipients were added and mixed in mortar and pestle. Finely ground herbal extracts and Eugenol were added and after thorough mixing, the toothpastes were filled in the tubes.

Placebo toothpaste was prepared by the same method but do not contain chitosan, herbal extracts, and eugenol. Gum tragacanth mucilage was used as gelling agent instead of chitosan gel. Purpose of adding different ingredients in the toothpaste is mentioned in [Table 1].{Table 1}

Evaluation of toothpastes

(A) Pharmaceutical evaluation

The formulated toothpastes were evaluated as per the US Government Tooth Paste Specifications. [20] The results obtained are presented in [Table 2].{Table 2}

(B) Clinical evaluation

The clinical evaluation was carried out as per the protocol approved by Institutional Ethics Committee. Eighteen healthy human volunteers, whose informed consent was first obtained, were selected for the study.

The groups were treated as follows:

Group-I: Placebo: Toothpaste without chitosan and herbal ingredients.

Group-II: Positive control: CHX (0.2% w/v) mouthwash.

Group-III: Test (Polyherbal): Toothpaste with chitosan, eugenol, and extracts of PM, SR, and GG.

(a) Clinical evaluation to study antiplaque activity

The study involved 18 subjects who underwent inclusion/exclusion criteria were divided into 3 groups containing 6 members in each group. Subjects were distributed in such a way that the average baseline plaque index of each group remains fairly the same. The study consisted of assessing the baseline plaque index status according to the criteria given by Silness and Loe (1964). [21] The subjects were supplied with polyherbal toothpaste, chlorhexidine mouthwash, and placebo toothpaste for application for a period of 4 weeks. After 2 and 4 weeks of use of the assigned product, the examining dentist scored the subjects for plaque index. The same dentist was interpreted at each examination, to avoid inter-examiner variation. The scores of the plaque index are presented in [Table 3] and [Figure 2].{Table 3}{Figure 2}

(b) Oral bacterial count from subjects

The subjects of each group were asked to simulate chewing action with sterile cotton rolls and asked to swallow the saliva thus collected over the next 1 min. This procedure was carried out in order to clear the mouth of any residual saliva. The subjects were then made to chew the cotton roll for next 4 min and expectorate into sterile bottles. Aseptically collected sample was further diluted and incubated on nutrient agar media. The percentage decrease in the total bacterial count in the study groups is presented in [Table 4] and [Figure 3].{Table 4}{Figure 3}


Bacterial plaque accumulated on teeth surfaces and composed of native oral flora, is the primary etiologic agent for periodontal disease and dental caries, which may result in tooth loss if left untreated. [22],[23] S. mutans and lactobacilli are the microorganisms that are primarily associated with dental caries. [24]

S. mutans, P. gingivalis, P. aueroginosa, P. vulgaris, E. coli, B. subtilis, L. acidophilus, and C. albicans are the common dental pathogens found to be associated with most of the orodental problems. As chitosan possesses and potentiates inhibitory action against S. mutans and P. gingivalis, it is necessary to evaluate the antimicrobial activity of herbal extracts against the remaining dental pathogens. Thus antimicrobial activities of PM, SR, and GG at various strengths were determined against selected dental pathogens and found to be satisfactory for maintaining good oral hygiene.

Pharmaceutical evaluation of novel chitosan-based polyherbal toothpaste was found to be well complied with all the US Government Tooth Paste Specifications.

The clinical evaluation carried out in healthy human volunteers as per the protocol showed that human volunteers in the test (polyherbal) group had a significant decrease in both plaque index and decrease in bacterial count as compared with positive control and placebo group.

The reduction in plaque index is caused by the combined effect of the physical properties, such as mucoadhesion and chemical interactions of chitosan with bacterial cell wall and antimicrobial effect of herbal extracts and eugenol. The percentage decrease in total bacterial count in the study groups were again due to the use of chitosan and herbal extracts.

The slight decrease in the plaque index as well as bacterial count in the placebo group might be due to performing routine oral hygiene practices by the volunteers from this group. This indicates the importance of routine oral hygiene practices.

 Result and Conclusion

Several toothpastes are currently being used as potential means of oral hygiene products. However, none of these fulfill the comprehensive requirements of oral care. The results of the study suggest that extracts of PM, SR, and GG possess satisfactory antimicrobial activity, which ultimately results in reducing total microbial count. Thus incorporation of chitosan as the gelling agent as well as therapeutic agent with herbal extracts in toothpaste reduces the plaque index by 70.47% and bacterial count by 85.29%, and thus fulfills the majority of esthetic and medicinal requirements of oral hygiene products.

From the results obtained in this study, it is concluded that chitosan-based polyherbal toothpaste proves itself as a promising oral hygiene product.


Ultimate aim of every research is to benefit human beings. Thus further studies are needed to identify and purify active ingredients from herbal extracts for its future use in toothpastes, mouthrinse, and other oral hygiene products. In addition, long-term clinical studies in large number of patient population will be required to evaluate the usefulness of these materials more exactly. The actual mechanism involved behind antimicrobial activity of herbal extracts need to be researched so as to decide its dose and duration of therapy.


Authors are thankful to Dr. S.B. Bhise, Principal, Govt. College of Pharmacy, Karad, for providing all necessary facilities and Okasa Pharma, Satara, for providing gift samples. We kindly appreciate leading Dentist Dr. Jaiwant Sawant, Karad, and Vaishali Gaikwad for providing valuable support in the clinical evaluation. We are thankful to Rameshwardasji Birla Smarak Kosh for their financial support.


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