Comparative Study on the Efficacy of Commercially Available Chemical and Herbal Mouthwash against Oral Microorganisms

Aim: The present study was done to determine the activity of Naturally prepared licorice mouthwash in comparison to chlorhexidine mouthwash. Introduction: Maintenance of oral hygiene is very important in preventing the growth of a sticky film of bacteria and food particles that accumulates on teeth and hence Mouthwashes are prescribed in dentistry for treatment of several oral conditions and for prevention against various microorganisms. Licorice is derived from "liquiritiae” which is shrub or herb and the yellow colour of it is due to the flavonoids content of the plant, which includes liquiritin, isoliquiritin and other compounds. Herbal Mouthwashes (mouth rinses) are solutions or liquids intended to reduce the microbial load in the oral cavity. Materials and Methods: This study was conducted in Saveetha Dental Hospital, Chennai, Tamilnadu, India. The liquorice was extracted and formulated as mouth wash. Fig. 1 denotes the salivary samples were obtained from 20 volunteers (10 test group, 10 control group) and then, the participants were asked to rinse their mouth with freshly prepared licorice mouthwash(test group), chlorhexidine mouthwash (control group) and then paired saliva samples were obtained immediately after and before the mouthwash and finally the Colony forming units were recorded for pre and post usage of both the herbal and commercially available mouthwash. Original Research Article Reshawn and Muralidharan; JPRI, 33(58A): 162-169, 2021; Article no.JPRI.74419 163 Results: Study showed that there was a mean reduction in the colony-forming units with herbal mouthwash and Chlorhexidine mouth washes. Herbal mouthwash showed reduction in colony forming growth after usage of herbal mouthwash.


INTRODUCTION
Mouthwashes are prescribed in dentistry for treatment of several oral conditions and for prevention against various microorganisms [1]. Maintenance of oral hygiene is very important in preventing the growth of sticky film of bacteria and food particles that accumulates on teeth. Oral hygiene measures include mechanical aids like toothbrushes, floss, interdental cleansers and chemotherapeutic agents like mouthwashes, dentifrices and chewing gums. Mouthwashes (mouth rinses) are solutions or liquids intended to reduce the microbial load in the oral cavity [2,3]. Glycyrrhiza glabra: The genus includes nearly 20 species native to North , South America, Australia, Europe, Asia etc. The English name licorice is derived from "liquiritiae" which is shrub or herb and the yellow colour of it is due to the flavonoids content of the plant, which includes liquiritin, isoliquiritin and other compounds.Glycyrrhiza glabra is a commonly used herb in Ayurvedic medicine. Various Studies indicate that Glycyrrhiza glabra Linn possesses vast properties like antimalarial, antispasmodic ,antibacterial, antioxidant, antiinflammatory and anti hyperglycemic properties. Various others properties includes antiulcer, antiviral,antihepatotoxic, antifungal and herpes simplex have also been studied [4,5].
Chlorhexidine is regarded as the "gold standard" anti-plaque agent [6] However, it is not a "Magic Bullet" due to certain side effects like tooth staining, taste disturbance, etc [1,6,7]Most of the mouthwashes available in the market contain alcohol and other chemicals such as chlorhexidine gluconate and triclosan.These chemicals cause various side effects ranging from taste disturbance to allergic contact stomatitis. To overcome such side effects, nontoxic herbal mouthwashes using various herbs and plant extracts are introduced. Natural herbs like Glycyrrhiza glabra, triphala, tulsi patra, jyeshtamadh, neem, clove oil, pudina, ajwain, and lots of more used either alone or together are scientifically proven to be safe and effective against various oral health problems like bleeding gums, halitosis, mouth ulcers, and preventing cavity .Our team has extensive knowledge and research experience that has translate into high quality publications [8][9][10][11][12].
The major strength of those natural herbs is the absence of any side effects and they do not contain alcohol and sugar found in their counter products. The microorganisms prey on these ingredients, releasing by-products and causing halitosis. Thus, herbal mouth rinses promote better oral hygiene and health [7]. The present study compared the effectiveness of an herbal mouthwash with different brands of commercially available mouthwashes in reducing the oral bacterial count.

MATERIALS AND METHODS
This study was conducted in Saveetha Dental Hospital, Chennai, Tamilnadu, India and 28g of freshly prepared Glycyrrhiza glabra (athimathuram) and was mixed with 450ml of distilled water and were boiled for 10 minutes.Then the solution was strained and the strained liquid was stored in a sterilised vessel [13].

Test Group
Saliva samples were obtained from 10 volunteers and then, the participants were asked to rinse their mouth with freshly prepared licorice mouthwash and then two saliva samples were obtained immediately after and before the herbal mouthwash.

Control Group
Saliva samples were obtained from volunteers and then the participants were asked to rinse their mouth with chlorhexidine, the saliva samples were collected immediately after and before the mouthwash. After collecting saliva samples, the samples were sent to the microbiology laboratory immediately. Fig 1  denotes the collected samples before and after using mouthwash and saline was added in the test tubes and culture plates were used to see the reduction of oral pathogens before and after Licorice mouthwash and then colony forming units were recorded for pre and post usage of the mouthwash with the help of colony counter app.

RESULTS AND DISCUSSION
In our present study Fig. 3 represents Nutrient agar which is used in procedures commonly performed for the cultivation of microbes and supporting growth of a wide range of nonfastidious organisms and its popular because it can grow a variety of types of bacteria and fungi, and contains many nutrients needed for the bacterial growth and in our present study, the efficacy of the herbal mouthwash and Chlorhexidine mouthwash in reducing oral pathogens were assessed. In our present study Table 1 and Fig. 5 explains the total Colony forming unit before and after using herbal mouthwash where the mean value before using herbal mouthwash was 350.7 and the mean after using herbal mouthwash was 176.9.
In Fig. 6 the mean reduction percentage is estimated to be 49.56%.From our study Table 2 and Fig 5 explains the total Colony forming unit before and after using Chlorhexidine mouthwash which was considered as a negative control in our study where the mean value before using Chlorhexidine mouthwash was 295.86 and the mean value after using Chlorhexidine mouthwash was 11.714,in Fig. 5 mean reduction percentage is estimated to be 96.04%. While in a study Decker et al. improved antiplaque strategies by using Chlorhexidine mouthwash in combination with plaque. Chlorhexidine (0.1%) was used as the positive control, saline was the negative control, and two CHT derivatives were used against streptococcus sanguis. Their results proved that Chlorhexidine and CHT combination was more stronger than Chlorhexidine alone because of the bioadhesive properties of CHT and the antibacterial activity of Chlorhexidine [14].  5. Error graph depicts the CFU before and after usage of Herbal mouthwash.X axis represents the number of participants involved in this study, Y axis represents the CFU.Blue colour denotes the CFU before using mouthwash, green colour depicts the CFU after usage of mouthwash. From this it is inferred that the majority of the participants showed more CFU before mouthwash when compared to CFU after herbal mouthwash  [17]Limitation of the study is small sample size and in vitro.Our team has extensive knowledge and research experience that has translate into high quality publications [18][19][20][21][22][23][24][25][26][27][28][29], [30][31][32][33][34]. In future similar study in large scale productions for targeted drug delivery to treat and prevent a wide array of oral microbial infections.

CONCLUSION
The results of our study and also figure 6 shows a mean reduction in the colony-forming units with herbal mouthwash and Chlorhexidine mouthwashes and showed that both mouthwashes gave reduction in colony forming growth after its usage. Though chlorhexidine shows higher percentage reduction (98 %) the herbal mouthwash has reduced the count upto 50 percent.

FUNDING SOURCE
The present study was supported by the following agencies: .Saveetha Dental College .SIMATS, Saveetha University .V-care physiotherapy clinic Tuticorin CONSENT As per international standard or university standard, Participants' written consent has been collected and preserved by the author(s).

ETHICAL APPROVAL
As per international standard or university standard written ethical approval has been collected and preserved by the author(s).