Evaluation of Mouthwash Containing Citrus hystrix DC., Moringa oleifera Lam. and Azadirachta indica A. Juss. Leaf Extracts on Dental Plaque and Gingivitis

Watunyoo Buakaew, Rungnapa Pankla Sranujit, Chanai Noysang, Supaporn Sangouam, Nungruthai Suphrom, Yordhathai Thongsri, Pachuen Potup, Kanchana Usuwanthim, Watunyoo Buakaew, Rungnapa Pankla Sranujit, Chanai Noysang, Supaporn Sangouam, Nungruthai Suphrom, Yordhathai Thongsri, Pachuen Potup, Kanchana Usuwanthim

Abstract

Oral hygiene and control of microbial plaque biofilm formation are effective methods for preventing gingivitis. Mouthwashes containing leaf extracts of the medicinal plants Citrus hystrix DC. (KL), Moringa oleifera Lam. (MO) and Azadirachta indica A. Juss. (NE) were assessed for oral healthcare and gingivitis adjunctive treatment. Three types of mouthwash were developed; KL, a combination of KL and MO (KL + MO), and a combination of KL, and NE (KL + NE). The mouthwashes were tested in vivo on 47 subjects with gingivitis who were allocated into five groups as (i) placebo, (ii) KL, (iii) KL + MO, (iv) KL + NE, and (v) 0.12% chlorhexidine gluconate (CHX). Participants were instructed to rinse with herbal mouthwash twice daily for two weeks. Gingival index (GI), plaque index (PI), and oral microbial colonies were measured at baseline and 15 days. Results showed that GI and PI of groups (ii)-(iv) significantly decreased over the placebo group, while accumulative reduction percentages of both Staphylococcus spp. and Candida spp. were found in groups (iii) and (iv). Findings indicated that the herbal mouthwashes reduced GI and PI, and showed potential as oral healthcare products.

Keywords: Azadirachta indica A. Juss; Citrus hystrix DC.; Moringa oleifera Lam.; gingivitis; mouthwash.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Trolox equivalent antioxidant capacity and total phenolic content of Citrus hystrix DC. (KL), Moringa oleifera Lam. (MO) and Azadirachta indica A. Juss. (NE) crude leaf ethanolic extracts. Results are expressed as Trolox equivalent (µM) and gallic acid equivalent (mg) for antioxidant and total phenolic contents, respectively. Data are presented as mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 compared to DMSO: dimethyl sulfoxide.
Figure 2
Figure 2
Chromatogram from GC-MS analysis. C. hystrix DC. (a), Moringa oleifera Lam. (b) and Azadirachta indica A. Juss. (c).
Figure 3
Figure 3
Flow chart of the study.
Figure 4
Figure 4
Intragroup comparison of gingival index (a) and plaque index (b) scores between baseline and day 15 after mouthwash usage by the Wilcoxon signed-rank test. Data are presented as mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

References

    1. Newton K., Dixit V.M. Signaling in innate immunity and inflammation. Cold Spring Harb. Perspect. Biol. 2012;4:a006049. doi: 10.1101/cshperspect.a006049.
    1. Zhang J.-M., An J. Cytokines, inflammation, and pain. Int. Anesthesiol. Clin. 2007;45:27–37. doi: 10.1097/AIA.0b013e318034194e.
    1. Schett G., Neurath M.F. Resolution of chronic inflammatory disease: Universal and tissue-specific concepts. Nat. Commun. 2018;9:3261. doi: 10.1038/s41467-018-05800-6.
    1. Hasturk H., Kantarci A., Van Dyke T.E. Oral inflammatory diseases and systemic inflammation: Role of the macrophage. Front. Immunol. 2012;3:118. doi: 10.3389/fimmu.2012.00118.
    1. Löe H. The Gingival Index, the Plaque Index and the Retention Index Systems. J. Periodontol. 1967;38:610–616. doi: 10.1902/jop.1967.38.6_part2.610.
    1. Kinane D.F., Stathopoulou P.G., Papapanou P.N. Periodontal diseases. Nat. Rev. 2017;3:17–38. doi: 10.1038/nrdp.2017.38.
    1. Murakami S., Mealey B.L., Mariotti A., Chapple I.L.C. Dental plaque–induced gingival conditions. J. Clin. Periodontol. 2018;45:17–27. doi: 10.1111/jcpe.12937.
    1. Adams D., Addy M. Mouthrinses. Adv. Dent. Res. 1994;8:291–301. doi: 10.1177/08959374940080022401.
    1. Safiaghdam H., Oveissi V., Bahramsoltani R., Farzaei M.H., Rahimi R. Medicinal plants for gingivitis: A review of clinical trials. Iran J. Basic Med. Sci. 2018;21:978–991.
    1. Buakaew W., Pankla Sranujit R., Noysang C., Thongsri Y., Potup P., Nuengchamnong N., Suphrom N., Usuwanthim K. Phytochemical Constituents of Citrus hystrix DC. Leaves Attenuate Inflammation via NF-κB Signaling and NLRP3 Inflammasome Activity in Macrophages. Biomolecules. 2021;11:105. doi: 10.3390/biom11010105.
    1. Luetragoon T., Pankla Sranujit R., Noysang C., Thongsri Y., Potup P., Suphrom N., Nuengchamnong N., Usuwanthim K. Bioactive Compounds in Moringa oleifera Lam. Leaves Inhibit the Pro-Inflammatory Mediators in Lipopolysaccharide-Induced Human Monocyte-Derived Macrophages. Molecules. 2020;25:191. doi: 10.3390/molecules25010191.
    1. Kooltheat N., Kamuthachad L., Anthapanya M., Samakchan N., Sranujit R.P., Potup P., Ferrante A., Usuwanthim K. Kaffir lime leaves extract inhibits biofilm formation by Streptococcus mutans. Nutrition. 2016;32:486–490. doi: 10.1016/j.nut.2015.10.010.
    1. Lee J.-W., Ryu H.W., Park S.-Y., Park H.A., Kwon O.-K., Yuk H.J., Shrestha K.K., Park M., Kim J.H., Lee S., et al. Protective effects of neem (Azadirachta indica A. Juss.) leaf extract against cigarette smoke- and lipopolysaccharide-induced pulmonary inflammation. Int. J. Mol. Med. 2017;40:1932–1940. doi: 10.3892/ijmm.2017.3178.
    1. Jiang K., Chen L.-L., Wang S.-F., Wang Y., Li Y., Gao K. Anti-inflammatory Terpenoids from the Leaves and Twigs of Dysoxylum gotadhora. J. Nat. Prod. 2015;78:1037–1044. doi: 10.1021/np5010196.
    1. Zhao Q.-Q., Wang S.-F., Li Y., Song Q.-Y., Gao K. Terpenoids with anti-inflammatory activity from Abies chensiensis. Fitoterapia. 2016;111:87–94. doi: 10.1016/j.fitote.2016.04.002.
    1. Costa J.P., Islam T., Santos P.S., Ferreira P.B., Oliveira G.L.S., Alencar M.V.O.B., Paz M.F.C.J., Ferreira L.F., Feitosa C.M., Citó A.M.G.L., et al. Evaluation of Antioxidant Activity of Phytol Using Non-and Pre-Clinical Models. Curr. Pharm. Biotechnol. 2016;17:1278–1284. doi: 10.2174/1389201017666161019155715.
    1. Silva R.O., Sousa F.B.M., Damasceno S.R.B., Carvalho N.S., Silva V.G., Oliveira F.R.M.A., Sousa D.P., Aragão K.S., Barbosa A.L., Freitas R.M., et al. Phytol, a diterpene alcohol, inhibits the inflammatory response by reducing cytokine production and oxidative stress. Fundam. Clin. Pharmacol. 2014;28:455–464. doi: 10.1111/fcp.12049.
    1. Lee W., Woo E.-R., Lee D.G. Phytol has antibacterial property by inducing oxidative stress response in Pseudomonas aeruginosa. Free Radic. Res. 2016;50:1309–1318. doi: 10.1080/10715762.2016.1241395.
    1. Carvalho A.M.S., Heimfarth L., Pereira E.W.M., Oliveira F.S., Menezes I.R.A., Coutinho H.D.M., Picot L., Antoniolli A.R., Quintans L., Quintans-Júnior L.J. Phytol, a Chlorophyll Component, Produces Antihyperalgesic, Anti-inflammatory, and Antiarthritic Effects: Possible NFκB Pathway Involvement and Reduced Levels of the Proinflammatory Cytokines TNF-α and IL-6. J. Nat. Prod. 2020;83:1107–1117. doi: 10.1021/acs.jnatprod.9b01116.
    1. Abbas M.H., Al-Yaseen A., Alhamadi W. Prevalence of Staphylococcus Aureus among gingivitis in patient with orthodontic wires in Kufa City/Iraq. Pak. J. Biotechnol. 2017;14:91–96.
    1. Rams T.E., Feik D., Slots J. Staphylococci in human periodontal diseases. Oral. Microbiol. Immunol. 1990;5:29–32. doi: 10.1111/j.1399-302X.1990.tb00222.x.
    1. McCarney R., Warner J., Iliffe S., van Haselen R., Griffin M., Fisher P. The Hawthorne Effect: A randomised, controlled trial. BMC Med. Res. Methodol. 2007;7:30. doi: 10.1186/1471-2288-7-30.
    1. Marsh P.D. Dental plaque as a biofilm and a microbial community—Implications for health and disease. BMC Oral Health. 2006;6(Suppl. S1):S14. doi: 10.1186/1472-6831-6-S1-S14.
    1. Patil S., Rao R.S., Majumdar B., Anil S. Clinical Appearance of Oral Candida Infection and Therapeutic Strategies. Front. Microbiol. 2015;6:1391. doi: 10.3389/fmicb.2015.01391.
    1. Alzohairy M.A. Therapeutics Role of Azadirachta indica (Neem) and Their Active Constituents in Diseases Prevention and Treatment. Evid. Based Complement. Altern. Med. 2016;2016:7382506. doi: 10.1155/2016/7382506.
    1. Hossain M.A., Al-Toubi W.A.S., Weli A.M., Al-Riyami Q.A., Al-Sabahi J.N. Identification and characterization of chemical compounds in different crude extracts from leaves of Omani neem. J. Taibah Univ. Sci. 2013;7:181–188. doi: 10.1016/j.jtusci.2013.05.003.
    1. Chatterjee A., Saluja M., Singh N., Kandwal A. To evaluate the antigingivitis and antipalque effect of an Azadirachta indica (neem) mouthrinse on plaque induced gingivitis: A double-blind, randomized, controlled trial. J. Indian Soc. Periodontol. 2011;15:398–401. doi: 10.4103/0972-124X.92578.
    1. Elgamily H., Moussa A., Elboraey A., El-Sayed H., Al-Moghazy M., Abdalla A. Microbiological Assessment of Moringa Oleifera Extracts and Its Incorporation in Novel Dental Remedies against Some Oral Pathogens. Open Access Maced J. Med. Sci. 2016;4:585–590. doi: 10.3889/oamjms.2016.132.
    1. Kooltheat N., Sranujit R.P., Chumark P., Potup P., Laytragoon-Lewin N., Usuwanthim K. An ethyl acetate fraction of Moringa oleifera Lam. Inhibits human macrophage cytokine production induced by cigarette smoke. Nutrients. 2014;6:697–710. doi: 10.3390/nu6020697.
    1. Agouillal F., Moghrani H., Nasrallah N., El Enshasy H. A Review of Genetic Taxonomy, Biomolecules Chemistry and Bioactivities of Citrus hystrix DC. Biosci. Biotechnol. Res. Asia. 2017;14:285–305. doi: 10.13005/bbra/2446.
    1. Singh A., Negi M.S., Rajagopal J., Bhatia S., Tomar U.K., Srivastava P.S., Lakshmikumaran M. Assessment of genetic diversity in Azadirachta indica using AFLP markers. Theor. Appl. Genet. 1999;99:272–279. doi: 10.1007/s001220051232.
    1. Rufai S., Hanafi M.M., Rafii M.Y., Ahmad S., Arolu I.W., Ferdous J. Genetic Dissection of New Genotypes of Drumstick Tree (Moringa oleifera Lam.) Using Random Amplified Polymorphic DNA Marker. BioMed Res. Int. 2013;2013:604598. doi: 10.1155/2013/604598.
    1. Ainsworth E.A., Gillespie K.M. Estimation of total phenolic content and other oxidation substrates in plant tissues using Folin–Ciocalteu reagent. Nat. Protoc. 2007;2:875–877. doi: 10.1038/nprot.2007.102.
    1. Rahman M.M., Islam M.B., Biswas M., Khurshid Alam A.H.M. In vitro antioxidant and free radical scavenging activity of different parts of Tabebuia pallida growing in Bangladesh. BMC Res. Notes. 2015;8:621. doi: 10.1186/s13104-015-1618-6.
    1. Kadam P., Bhalerao S. Sample size calculation. Int. J. Ayurveda Res. 2010;1:55–57.

Source: PubMed

Sponsors et collaborateurs

Les conditions médicales

Interventions en matière de drogue

3
S'abonner