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Table of Contents   
ORIGINAL RESEARCH  
Year : 2020  |  Volume : 31  |  Issue : 6  |  Page : 830-834
Efficacy of T. Cordifolia (Guduchi) against plaque and gingivitis-A clinico-microbiological study


1 Department of Public Health Dentistry, Dr. D.Y. Patil Dental College and Hospital, Dr. D.Y. Patil Vidyapeeth, Pune, Maharashtra, India
2 Department of Rasayana and Bhaishyajya Kalpana, Dr. D.Y. Patil Ayurveda College and Hospital, Dr. D.Y. Patil Vidyapeeth, Pune, Maharashtra, India
3 Department of Dravyaguna, Dr. D.Y. Patil Ayurveda College and Hospital, Dr. D.Y. Patil Vidyapeeth, Pune, Maharashtra, India

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Date of Submission25-Jun-2018
Date of Decision30-Aug-2019
Date of Acceptance08-Apr-2020
Date of Web Publication22-Mar-2021
 

   Abstract 


Objectives: The present study was conducted to assess the clinical efficacy (against plaque and gingivitis) and antimicrobial efficacy (against gram negative, gram positive and S. mutans counts) of T. cordifolia (Guduchi). Methodology: A nonrandomized before and after trial for 21 days of intervention was performed involving 17 participants. Plaque index, gingival index were recorded and saliva samples were collected at baseline, 13th day and 22nd day. Repeated measures ANOVA test was performed to compare the before and after results of the clinical parameters, i.e. score of plaque index and gingival index. Freidman's test was applied for comparison between the three intervals (for microbiological evaluation of colony counts). Results: There was statistically significant reduction inplaque score, gingival score, and colony forming units of gram negative organisms and S. mutans from baseline to 22nd day (P = 0.001). There was no significant reduction with gram positive organisms. Conclusion: This clinico-microbiological study concluded that twice rinsing of T.cordifolia decoction as mouth rinse in adjunct to scaling and routine brushing of teeth for 21 days shows anti-plaque, anti-gingivitis, and antimicrobial action against gram negative organisms and S. mutans.

Keywords: Anti plaque agent, antimicrobial agent, dental plaque, gingivitis

How to cite this article:
Nair S, Kakodkar P, Shetiya SH, Dharkar N, Jayashree C, Rajpurohit L. Efficacy of T. Cordifolia (Guduchi) against plaque and gingivitis-A clinico-microbiological study. Indian J Dent Res 2020;31:830-4

How to cite this URL:
Nair S, Kakodkar P, Shetiya SH, Dharkar N, Jayashree C, Rajpurohit L. Efficacy of T. Cordifolia (Guduchi) against plaque and gingivitis-A clinico-microbiological study. Indian J Dent Res [serial online] 2020 [cited 2021 Apr 19];31:830-4. Available from: https://www.ijdr.in/text.asp?2020/31/6/830/311659



   Introduction Top


Gingivitis is a chronic inflammatory disease process which is confined to the gingiva of the teeth.[1] Dental plaque-”a thin tenacious film is considered as the predisposing factor in the development of gingivitis since it is composed of salivary glycoproteins and extracellular polysaccharide.[2]

Supragingival plaque is predominantly composed of gram positive organisms while subgingival plaque is mainly composed of gram negative organisms due to its anaerobic environment. One individual may harbor 150 or more different species. These microorganisms cause damage to the tissues of oral cavity by releasing various toxins, enzymes, and metabolic products which are regarded as causative factors for gingivitis.[2],[3]

Effective plaque control is necessary in order to arrest the progression of gingivitis into periodontitis. Numerous chemotherapeutic/antimicrobial agents have been evolved and are marketed in order to control plaque induced bacteria.[4] Chemotherapeutic agent like Chlorhexidine which is considered as the “gold standard mouthwash” have side effects like alteration of taste; staining of teeth, nausea and vomiting, and hence trials are being conducted to develop new alternative agents.[5]

Recently it was proposed by Nagi (2015)[6] that herbal alternatives have gained popularity owing to the minimal side effects caused by its application. The global interest among researchers in plants with antimicrobial and anti-inflammatory activities has increased in order to overcome the consequence of current issues associated with the wide-scale misuse of chemotherapeutic agents that induce microbial drug resistance.[7]

Guduchi (Tinospora Cordifolia/T.cordifolia) has been mentioned in the literature of Ayurveda and Indian Pharmacopeia, as a constituent of several compound preparations used in medical conditions like general weakness, dyspepsia, pyrexia and urinary diseases. It is commonly known as Giloy and is a glabrous and succulent plant. It is deciduous climbing shrub which has greenish yellow typical flowers.[8] The phytochemical evaluation of T. cordifolia stem and whole plant extract has revealed that the plant contains flavanoids, glycosides, alkaloids, steroids, terpenes, saponins, and tannin.[9]

In-vitro studies have proven the antimicrobial activity of T. cordifolia against S. mutans.[10],[11] An, in-vivo study[12] has confirmed that T. cordifolia leaves have been effective as an antimicrobial and antifungal agent in patients having HIV. Further, studies[13],[14],[15]have reported the anti-inflammatory action of T. cordifolia.

As against this background, that literature supports the beneficial effects of T.cordifolia, a need was felt to evaluate if it can be used as an herbal alternative against the dental pathogens and gingivitis. The aim of the study was to assess the clinical efficacy of T. cordifolia against plaque and gingivitis and the antimicrobial efficacy against the gram positive, gram negative, and Streptococcus mutans organism count in the mouth.


   Materials and Methods Top


This study was a non-randomized, before and after study without control group. Ethical clearance was obtained from the institutional ethics committee (Ref no: DYPDCH/1227/2014/37). Written informed consent was obtained from all the participants of this study. The study site was Nursing college. The participants were included as per the following inclusion criteria: Participants who were willing to participate in the study; participants having gingival inflammation (Loe and Sillness index,[16] score 1.12) and minimum of 28 natural teeth in the mouth. The exclusion criteria were: the presence of any systemic diseases, history of antibiotic therapy or mouthwash used in the previous 1 month, malocclusion, periodontitis, history of smoking and the presence of grossly carious teeth. Using the sample size calculator with reference of the previous study,[13] a sample of 14 was calculated. In addition, 3 more were added to compensate for any drop outs and hence, the total sample was 17. The study participants were second year nursing students who were recruited as per the above-mentioned criteria.

Preparation of decoction (mouthwash)

The T.cordifolia powder was commercially purchased from the Ayurveda pharmacy. Preparation of decoction was done at the Departmental Pharmacy of Rasashastra and Bhaishyajya Kalpana. 50 gms of coarse T.cordifolia powder along with freshly picked mint leaves (45 leaves) from the ayurveda nursery were boiled in 1800 ml of water till 1/16th proportion (200 ml) was obtained. The decoction was filtered and pH was recorded.[17]

Pilot study

Pilot study was conducted on 5 participants in order to establish the substantivity of the decoction. By means of sterile test tube, 1 ml saliva sample was collected from each participant five times, i.e. prerinse, postrinse (after 10 min), after 6 h, after 12 h and after 24 h. Agar disc diffusion method was performed for evaluation of growth of gram positive and gram negative bacilli and cocci on agar plates. The results showed that that there was 12 h substantivity which indicated that the decoction should be given twice a day to the participants.

Study procedure

Baseline clinico-microbiological data (plaque score, gingival score, gram positive, gram negative and S. mutans organism count) was collected from the recruited 17 participants and were followed by oral prophylaxis of the teeth. The investigator herself administered the mouthwash to the participants twice daily, every day for 21 days. In total 10 ml of the mouthwash was rinsed for 30 s. During the study period the participants followed their routine toothbrushing method and brushed their teeth twice daily. Clinico-microbiological data was recorded again on 13th day and 22nd day. Gingival score was recorded using the gingival index[16] and plaque index[18] was used to record the plaque.

Laboratory analysis

Unstimulated saliva sample (5ml) was collected from the participants. It was stored in the ice-box and transferred to the laboratory. Standard laboratory procedure was followed[19],[20] for cultivating the organisms. Pour plate method was used with serial dilution. Selective agar, i.e.,: Mitis agar for S mutans, PEA agar for gram positive organisms, and MacConkey's agar for gram negative organisms were used. Bio-chemical analysis and gram staining were done for identification and confirmation. The colony counter was used to calculate the colony forming units per sample.

Statistical analysis

The data were analyzed using statistical package for social sciences (SPSS) V.21. Repeated measures ANOVA test with Post hoc Bonferroni test was performed to compare the before and after results of the clinical parameters, i.e. score of plaque index and gingival index. Freidman's test was applied to compare the microbiological data. Differences were considered significant if the P values were <0.05.

For the ease of analyzing, a scale was prepared for the organism count per sample as given below:

Gram negative count: Negligible- code 4, 101-code 3, 102-code 2,103-code 1

S. mutans the coding was- <104-code 4,104- code 3,105-code 2,106-107-code 1

Gram positive count was 105-code 2,106-107-code 1.


   Results Top


In total 17 participants (15 females and 2 males) aged 18-22 years completed the study.

Clinical parameters

The baseline, intermediate (13th day) and end score (22nd day) for the plaque and gingival index are presented in [Table 1]. There was a statistically significant reduction in the plaque score from 3.48 to 2.44. Similarly, the gingival score also showed a significant reduction from 1.09 to 0.76. For both, the plaque and gingival index, intergroup comparison (between baseline, 13th day and 22nd day) showed a significant difference with Post hoc Bonferroni test.
Table 1: Comparison of plaque index and gingival index at three different intervals

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Microbiological parameters

[Table 2] and [Table 3] shows the distribution of the participants based on their gram negative and S. mutans colony counts. Significant difference was found from baseline scores to 22nd day scores. [Table 4] shows the distribution of the participants based on their gram positive colony count. No significant difference was found between baseline to 22nd day. However mild reduction in the number of colonies was noted from baseline to 22nd day.
Table 2: Comparison of counts of total gram negative colony between three intervals

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Table 3: Comparison of colony counts of S mutans between three different intervals

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Table 4: Comparison of the counts of gram positive colony forming units at three different intervals

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   Discussion Top


The foundation for prevention of gingivitis is maintenance of good oral hygiene. Dental plaque is considered as one of the etiological factor for both dental caries and gingival diseases.[21] The result of the present study: Twice rinsing of T. cordifolia decoction as mouthrinse in adjunct to scaling and routine tooth brushing for 21 days shows anti plaque, anti gingivitis and antimicrobial action against gram negative organisms and S. mutans.

Overall, the intervention was able to retard the plaque formation by 29.89%. There was a statistically significant difference at three different time intervals [Table 1].The probable plaque reducing action of T. cordifoliacan be by the virtue of its physical action (rinsing action) and chemical action (chemotherapeutic interaction of the secondary metabolites like alkaloids, flavanoids, tannins and other compounds of phenolicnature).[22]

Only one report, from the literature was retrieved where T. cordifolia was evaluated for its anti plaque property in chronic periodontitis patients,[13] where it showed negative results. However, this difference can be attributed to the mode of intervention. The latter used T. cordifolia in the form of Septillin capsules which yielded systemic action as versus the present study where T. cordifolia decoction as mouthwash was used which yielded local action and positive result.

Overall, the intervention was able to retard the gingivitis by 30.28% and there was a statistically significant difference at three different time intervals [Table 1]. The result of this study was in consensus with the previous studies.[13],[14],[15]

Antigingival action is due to the anti-inflammatory action of the T. cordifolia. Anti-inflammatory action of T. cordifolia has been proved in animal models. One study[23] has suggested that aqueous extract of T. cordifolia was effective in the suppression of oedema formation. The mechanism of action was by inhibiting the role of histamine and 5 HT during the initial phase of inflammation. The stem of T. cordifolia has various phytochemicals which are responsible for anti-inflammatory action. In humans, alkaloids, glycosides, diterpenoid lactones, steroids and aliphatic compounds play a key role in reducing inflammation. Steroid inhibits Interleukin 1, β.Interleukin 6 and COX-2 inhibitors.[9]

In the present study, significant reduction of gram negative colonies has been observed [Table 2]. This may be attributed to the antimicrobial properties exhibited by the alkaloids and phenolic derivatives.[9]

The present study reported no significant difference in the colony forming units of gram positive organisms [Table 4]. This result was similar to that of an in vitro study[23] which suggested that aqueous extract of T.cordifolia did not effectively inhibit the growth of gram positive bacteria while, the ethanolic extract had greater antibacterial activity. This can be a limitation for mouthwash preparation, as alcoholic mouthwash are not recommended and have a risk for oral cancer. But a contrasting result has been presented[11] which demonstrated a positive antibacterial activity of T.cordifolia extract against selected gram positive organisms like S. aureus, S. sanguinis, S. salivarius, and L. acidophilus.

S. mutans is considered as one of the causative agents of dental caries. In the present study, significant reduction was found in the S. mutans count [Table 3], which was similar to the results of the in vitro studies.[10],[11],[12]

In total 5 mg of aqueous extract of T. cordifolia decoction which was administered as mouthwash in the present study was effective against S. mutans. However, Mistry, et al. (2014)[10] suggested that methanolic extract of T. cordifolia was most effective against S. mutans at a concentration of 3 mg. Peter et al. (2014)[12] suggests that it can be used as a potential additive to oral hygiene practice.

A major drawback of T. cordifolia is its bitter taste. To reduce its bitterness, 47 mint leaves were added which made the decoction palatable. Mint has germicidal property and a study[24] suggests it also has antimicrobial property against S. mutans. The overall antimicrobial effect on S. mutans in the present study may be a combined effect of T. cordifolia and mint. Further studies should be planned to prepare T. cordifolia formulation with flavouring and sweetening agent which can overcome the bitterness problem.

This study incorporates a preventive trial study design as explained by Chilton and Fleiss (1986),[25] where the trial begins by considering all the samples from “ Zero” as the baseline score. Preventive trials enquire the outcome of formulation on retarding patient plaque accumulation or the development of gingivitis; which has been followed in the present study. Further, the study subjects were placed on their routine toothbrushing regimen in order to conduct the study in the real life situation. Oral prophylaxis was done to begin the trial from ground zero. Because of scaling procedure, the local factors (plaque and calculus) were removed. Hence it can be postulated that the outcome of the study is a combined result of scaling and mouth rinsing. No side effects were noticed in the oral cavity of the participants after using the mouthwash. One of the drawback of this product is that it cannot be stored and used multiple times. Hence, the decoction used for rinsing was freshly prepared everyday.

The current study has some limitations. The 21 day intervention is relatively small. The study can be further improved with randomized controlled trial, larger sample, longer duration and follow-up to confirm the present results.


   Conclusion Top


This clinico-microbiological study concluded that twice rinsing of T. cordifolia decoction as mouthrinse as an adjunct to scaling and routine brushing of teeth for 21 days shows anti-plaque, anti-gingivitis, and antimicrobial action against gram negative organisms and S. mutans. T. cordifolia is effective as an antimicrobial agent and may aid as an herbal alternative to chemotherapeutic mouthrinses.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
   References Top

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Quirynen M, Quirynen M, Teughels W, Haake SK, Newman MG. Microbiology of periodontal diseases.In: Newman MG, et al., editors Carranza's Clinical Periodontology. 10th ed. Elsevier Health Sciences 2016.p. 243-72.  Back to cited text no. 1
    
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Gottschall JC, Harder W, Prins RA. Principles of enrichment, isolation, cultivation and preservation of bacteria. In: Balows A, et al. editors. The Prokaryocetes. 2nd ed. New York: Springer-Verlag; 1992. p. 149-96.  Back to cited text no. 19
    
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Correspondence Address:
Dr. Pradnya Kakodkar
Department of Public Health Dentistry, Dr. D.Y. Patil Dental College and Hospital, Pune - 411 018, Maharashtra
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijdr.IJDR_521_18

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