Synergistic Antimicrobial and Stain-Removal Effects of Multiherbal–Sodium Hexametaphosphate Mouthwash - An in-vitro study

Harsini Harsini; Hendrawati Hendrawati; Mutiara Annisa

doi:10.7126/cumudj.1777957

EN

Synergistic Antimicrobial and Stain-Removal Effects of Multiherbal–Sodium Hexametaphosphate Mouthwash - An in-vitro study

Abstract

Objectives: This study aims to test a mouthwash made with cashew bark and bay leaf extract, along with sodium hexametaphosphate (SHMP) as stain removal and antimicrobial agent. Materials and methods: A 7% cashew bark extract and 10% bay leaf extract, both prepared through maceration. The mouthwash was split into three groups (n=6): FA (multiherbal extract), FB (multiherbal extract with SHMP), and FC (SHMP only). Teeth were stained in a tea solution made from a 2g black tea bag in 100mL of water for seven days, refreshing the solution daily. After staining, the teeth were soaked in each mouthwash for 90 minutes. Color changes were measured with a chromameter before and after soaking, color difference (ΔE) was calculated. The antimicrobial ability was tested against Staphylococcus aureus and Pseudomonas aeruginosa. The one-way ANOVA and post-hoc LSD test were used as statistical analysis (P=0.05). Results: FB exhibited the highest ΔE value for extrinsic stain removal (24.29±2.19). There are significant differences in ΔE values among the three groups, with FB demonstrating the most pronounced effect (P=0.01). FB produced the largest inhibition zones for both S. aureus (20.25±0.5) and P. aeruginosa (11.25±0.5). The one-way ANOVA and post-hoc LSD test confirmed that FB had the most significant inhibition zone diameter compared to FA and FC. Conclusions: The multi-herbal extract containing cashew bark and bay leaf extract combined with SHMP showed effectiveness in removing tea-extrinsic stained teeth and inhibiting the growth of S. aureus and P. aeruginosa.

Keywords

Ethical Statement

This experimental study was conducted with ethical approval from the Ethics and Advocacy Unit, Faculty of Dentistry, Universitas Gadjah Mada, Indonesia (Approval No. 50/UN1/KEP/FKG-RSGM/EC/2024).

References

1. Powers JM, Sakaguchi RL. Craig’s restorative dental materials. 12th ed. St. Louis: Mosby 2006:514–516.
2. Cho MJ. The tooth whitening effect of toothpaste containing high cleaning silica and sodium hexametaphosphate and the preventive effect of staining by coffee, tea and wine. Int J Clin Prev Dent 2020;19:192-199.
3. Eachempati P, Nagraj SK, Krishanappa SKK, Gupta P, Yaylali IE. Home-based chemically induced whitening (bleaching) of teeth in adults. Cochrane Database Syst Rev 2018;12:CD006202.
4. McGrath C, Clarkson J, Glenny AM, Walsh LJ, Hua F. Effectiveness of mouthwashes in managing oral diseases and conditions: do they have a role? Int Dent J 2023;73:69-73.
5. Epple M, Meyer F, Enax J. A critical review of modern concepts for teeth whitening. Dent J (Basel) 2019;7:79.
6. De Medeiros RFSB, da Silva RCM, Marinho LCN, Silva DNA, Tavares ILL, de Lima CACNRA, et al. Microbiological analysis of oral biofilms in patients admitted to intensive care unit: pilot study. Oral Dis 2023;29:2293-2296.
7. Joiner A, Luo W. Tooth colour and whiteness: a review. J Dent 2017;67:3-10.
8. Annísa M, Harsini H, Murti YB. Evaluation of herbal gel toothpaste formulated using bay leaf essential oil on physicochemical characteristics and extrinsic stain removal. J Herbmed Pharmacol 2023;12:425-431.

9. Basheer RR, Abouelmagd DM, Alnefaie A, Baamer R. Effect of at-home versus over-the-counter bleaching agents on enamel color, roughness, and color stability. Cureus 2023;15:e39036.
10. Dionysopoulos D, Papageorgiou S, Papadopoulos C, Davidopoulou S, Konstantinidis A, Tolidis K. Effect of whitening toothpastes with different active agents on the abrasive wear of dentin following tooth brushing simulation. J Funct Biomater 2023;14:268.
11. Ghajari MF, Shamsaei MB, Galouyak MS. Abrasiveness and whitening effect of charcoal-containing whitening toothpastes in permanent teeth. Dent Res J (Isfahan) 2021;18:51.
12. Jamwal N, Rao A, Mc Shanka G, Shenoy RK, Bh MP, Jodalii P, et al. Effect of whitening toothpastes on the surface roughness and microhardness of human teeth: an in vitro study. Clin Oral Investig 2023;27:7889-7897.
13. Souza-Rodrigues RD, Ferreira S, D’Almeida-Couto RS, Lachowski KM, Sobral MÂP, Marques MM. Choice of toothpaste for the elderly: an in vitro study. Braz Oral Res 2015;29:94.
14. Alqahtani MQ. Tooth-bleaching procedures and their controversial effects: a literature review. Saudi Dent J 2014;26:33-46.
15. Harsini, Sutardjo I, Sudibyo, Sunarintyas S, Soedarsono. Effect of stem bark extract Anacardium occidentale Linn. as antiseptic towards expression of VEGF. Dentika Dent J 2014;18:134-139.
16. Pommerville JC. Fundamentals of microbiology. 10th ed. Burlington: Jones & Bartlett Learning 2014:218.
17. Salehi B, Gültekin-Özgüven M, Kirkin C, Özçelik B, Morais-Braga MFB, Carneiro JNP, et al. Antioxidant, antimicrobial, and anticancer effects of Anacardium plants: an ethnopharmacological perspective. Front Endocrinol (Lausanne) 2020;11:295.
18. Annisa M, Nuryanti A, Dewi AH. Effectivity of multifunction herbal toothpaste containing bay leaf (Eugenia polyantha Wight) extract as extrinsic stain removal on teeth and denture. Odonto Dent J 2022;9:40-50.
19. Hajizadeh M, Movahedi ZM, Sheibani N, Moosavi MAA. An outlook on suicide enzyme inhibition and drug design. J Iran Chem Soc 2021;18:15-25.
20. Marquillas CB, Procaccini R, Malmagro MV, Sanchez-Martin MJ. Breaking the rules: tooth whitening by means of a reducing agent. Clin Oral Investig 2019;23:2303-2310.
21. Luka B, Duerrschnabel A, Neumaier S, Schlueter N, Vach K. Interaction between hexametaphosphate, other active ingredients of toothpastes, and erosion-abrasion in enamel in vitro. Caries Res 2023;57:265-275.
22. Pentapati KC, Sabnis N, Shrikhande B, Kashyap R, Kukkamalla MA. Stain removal efficacy of herbal mouth rinses containing sodium hexametaphosphate. J Res Med Educ Ethics 2014;4:165-170.
23. Radzki D, Wilhelm-Węglarz M, Pruska K, Kusiak A, Ordyniec-Kwaśnica I. A fresh look at mouthwashes: what is inside and what is it for? Int J Environ Res Public Health 2022;19:3926.
24. Daniel WW, Cross CL. Biostatistics: A Foundation for Analysis in the Health Sciences. 10th ed. Hoboken, NJ: John Wiley & Sons 2013:189.
25. Wang C, Lucas R, Smith AJ, Cooper PR. An in vitro screening assay for dental stain cleaning. BMC Oral Health 2017;17:128.
26. Annisa M, Kanina PAR, Hamid NLB, Nuryanti A. Effectiveness of green tea, bay leaf, and lime peel extracts as toothpaste active agents for extrinsic stain removal. Padjadjaran J Dent 2022;34:47-56.
27. Bubonja-Šonje M, Knežević S, Abram M. Challenges to antimicrobial susceptibility testing of plant-derived polyphenolic compounds. Arh Hig Rada Toksikol 2020;71:300-311.
28. Tafazoli A, Tafazoli Moghadam E. Camellia sinensis mouthwashes in oral care: a systematic review. J Dent (Shiraz) 2020;21:249-262.
29. Harsini, Irnawati D, Widyasrini DA, Sunarintyas S, Siswomihardjo W. Glycerine addition in cashew stem bark extract mouthwash influences the inhibition of bacterial growth. Res J Pharm Technol 2024;17:156-162.
30. Banjarhor SDS, Artanti N. Antioxidant properties of flavonoids. Med J Indones 2014;23:239-244.
31. Hartanti L, Yonas SMK, Mustamu JJ, Wijaya S, Setiawan HK, Soegianto L. Influence of extraction methods of bay leaves (Syzygium polyanthum) on antioxidant and HMG-CoA reductase inhibitory activity. Heliyon 2019;5:e01485.
32. Li Y, Miao Y, Yang L, Zhao Y, Wu K, Lu Z, et al. Recent advances in the development and antimicrobial applications of metal-phenolic networks. Adv Sci (Weinh) 2022;9:2202684.
33. Yilmaz E, Erdilek AD. Effectiveness of antimicrobial mouthwashes in caries biofilm. AJ Health Sci 2023;5:167-175.
34. Dorr T, Moynihan PJ, Mayer C. Editorial: bacterial cell wall structure and dynamics. Front Microbiol 2019;10:2051.
35. Fux AC, Casonato Melo C, Michelini S, Swartzwelter BJ, Neusch A, Italiani P, et al. Heterogeneity of lipopolysaccharide as source of variability in bioassays and LPS-binding proteins as remedy. Int J Mol Sci 2023;24:8395.
36. Yan Y, Xia X, Fatima A, Zhang L, Yuan G, Lian F, et al. Antibacterial activity and mechanisms of plant flavonoids against gram-negative bacteria based on the antibacterial statistical model. Pharmaceuticals (Basel) 2024;17:292.

Details

Primary Language

English

Subjects

Dental Materials

Journal Section

Research Article

Authors

Harsini Harsini
0000-0002-1785-6812
Indonesia

Hendrawati Hendrawati
0000-0002-4360-0976
Indonesia

Mutiara Annisa ^*
0000-0003-2379-9532
Indonesia

Publication Date

July 1, 2026

Submission Date

September 6, 2025

Acceptance Date

March 24, 2026

Published in Issue

Year 2026 Volume: 29 Number: 2

DOI

https://doi.org/10.7126/cumudj.1777957

IZ

https://izlik.org/JA22XG63WW

Cite

RIS / Bibtex

APA

Harsini, H., Hendrawati, H., & Annisa, M. (2026). Synergistic Antimicrobial and Stain-Removal Effects of Multiherbal–Sodium Hexametaphosphate Mouthwash - An in-vitro study. Cumhuriyet Dental Journal, 29(2), 216-222. https://doi.org/10.7126/cumudj.1777957

AMA

1.Harsini H, Hendrawati H, Annisa M. Synergistic Antimicrobial and Stain-Removal Effects of Multiherbal–Sodium Hexametaphosphate Mouthwash - An in-vitro study. Cumhuriyet Dent J. 2026;29(2):216-222. doi:10.7126/cumudj.1777957

Chicago

Harsini, Harsini, Hendrawati Hendrawati, and Mutiara Annisa. 2026. “Synergistic Antimicrobial and Stain-Removal Effects of Multiherbal–Sodium Hexametaphosphate Mouthwash - An In-Vitro Study”. Cumhuriyet Dental Journal 29 (2): 216-22. https://doi.org/10.7126/cumudj.1777957.

EndNote

Harsini H, Hendrawati H, Annisa M (July 1, 2026) Synergistic Antimicrobial and Stain-Removal Effects of Multiherbal–Sodium Hexametaphosphate Mouthwash - An in-vitro study. Cumhuriyet Dental Journal 29 2 216–222.

IEEE

[1]H. Harsini, H. Hendrawati, and M. Annisa, “Synergistic Antimicrobial and Stain-Removal Effects of Multiherbal–Sodium Hexametaphosphate Mouthwash - An in-vitro study”, Cumhuriyet Dent J, vol. 29, no. 2, pp. 216–222, July 2026, doi: 10.7126/cumudj.1777957.

ISNAD

Harsini, Harsini - Hendrawati, Hendrawati - Annisa, Mutiara. “Synergistic Antimicrobial and Stain-Removal Effects of Multiherbal–Sodium Hexametaphosphate Mouthwash - An In-Vitro Study”. Cumhuriyet Dental Journal 29/2 (July 1, 2026): 216-222. https://doi.org/10.7126/cumudj.1777957.

JAMA

1.Harsini H, Hendrawati H, Annisa M. Synergistic Antimicrobial and Stain-Removal Effects of Multiherbal–Sodium Hexametaphosphate Mouthwash - An in-vitro study. Cumhuriyet Dent J. 2026;29:216–222.

MLA

Harsini, Harsini, et al. “Synergistic Antimicrobial and Stain-Removal Effects of Multiherbal–Sodium Hexametaphosphate Mouthwash - An In-Vitro Study”. Cumhuriyet Dental Journal, vol. 29, no. 2, July 2026, pp. 216-22, doi:10.7126/cumudj.1777957.

Vancouver

1.Harsini Harsini, Hendrawati Hendrawati, Mutiara Annisa. Synergistic Antimicrobial and Stain-Removal Effects of Multiherbal–Sodium Hexametaphosphate Mouthwash - An in-vitro study. Cumhuriyet Dent J. 2026 Jul. 1;29(2):216-22. doi:10.7126/cumudj.1777957