Skip to content
2000
image of Novel Mouth Rinse Displaying Analgesic and Anti-inflammatory Properties: An in vitro Study

Abstract

Introduction

Periodontitis is a chronic inflammatory disease requiring effective anti-inflammatory treatments. Nano-silver and essential oils have shown potential due to their antimicrobial and anti-inflammatory properties. Combining these agents offers a promising therapeutic approach. This study investigated the cytotoxic and anti-inflammatory properties of a novel essential oil compound containing nanosilver using HaCaT and THP-1 human leukemia monocytic cell lines.

Materials and Methods

Neutral red uptake (NRU) assay was used to assess cytotoxicity and ELISA to evaluate the inflammatory cytokines. The test compound was compared to 0.12% chlorhexidine gluconate (CHX). Cytotoxicity was determined in HaCaT and THP-1 cell lines using NRU assay. TNF-α expression was measured using ELISA, and COX-2 inhibition assay was performed.

Results

Cytotoxicity of the test compound was nearly absent. TNF-α levels decreased in positive control (2.81 pg/ml) and test samples (1.30 pg/ml) compared to control (22.04 pg/ml). COX-2 inhibition assay revealed test compound (0-20%) and positive control (0-100%), with 25 μM celecoxib as a standard. IC for HaCaT cells was 0.6334% (positive control) and 0.6051% (test group). IC using THP-1 cells was not converged for the test and 424.6% for positive control. IC-50 for COX-2 inhibition was 1.469% in the test and 8.801% in the positive control.

Discussion

This study showed the possibility of novel essential oils and nano-silver-containing compounds as a medication material in preventing gingivitis. The cytotoxicity was negligible, while the level of TNF- α was much decreased, and COX-2 activity assays indicated its efficiency in anti-inflammatory properties. The results encourage the therapeutic potential of the compound for periodontitis, and further studies are required to demonstrate therapeutic efficiency and safety.

Conclusion

Results demonstrate the inhibitory effect of the test compound on COX-2 activity. The potential of a novel test compound containing essential oils and nano-silver as a promising anti-inflammatory agent warrants further investigation for its therapeutic applications in periodontitis.

© 2025 Bentham Science Publishers
Loading

Article metrics loading...

/content/journals/aiaamc/10.2174/0118715230384352250526103643
2025-06-11
2025-09-21

  • From This Site

    /content/journals/aiaamc/10.2174/0118715230384352250526103643
    dcterms_title,dcterms_subject,pub_keyword
    -contentType:Contributor -contentType:Concept -contentType:Institution
    10
    5
Loading full text...

Full text loading...

References

  1. Van Dyke T.E. Bartold P.M. Reynolds E.C. The nexus between periodontal inflammation and dysbiosis. Front. Immunol. 2020 11 511 10.3389/fimmu.2020.00511 32296429
    [Google Scholar]
  2. Poppolo Deus F. Ouanounou A. Chlorhexidine in dentistry: Pharmacology, uses, and adverse effects. Int. Dent. J. 2022 72 3 269 277 10.1016/j.identj.2022.01.005 35287956
    [Google Scholar]
  3. Gawish A.S. ElMofty M.S. Jambi S. Felemban D. Ragheb Y.S. Elsayed S.A. Phytotherapy in periodontics as an effective and sustainable supplemental treatment: A narrative review. J. Periodontal Implant Sci. 2024 54 4 209 223 Advance online publication [http://dx.doi.org/10.5051/jpis.2301420071 8290997
    [Google Scholar]
  4. Little J.W. Complementary and alternative medicine: Impact on dentistry. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 2004 98 2 137 145 10.1016/j.tripleo.2004.05.011 15316539
    [Google Scholar]
  5. Van Leeuwen M.P.C. Slot D.E. Van der Weijden G.A. The effect of an essential‐oils mouthrinse as compared to a vehicle solution on plaque and gingival inflammation: A systematic review and meta‐analysis. Int. J. Dent. Hyg. 2014 12 3 160 167 10.1111/idh.12069 24720368
    [Google Scholar]
  6. Silvestri D.L. McEnery-Stonelake M. Chlorhexidine: Uses and adverse reactions. Dermatitis 2013 24 3 112 118 10.1097/DER.0b013e3182905561 23665831
    [Google Scholar]
  7. Braga A.S. Girotti L.D. de Melo Simas L.L. Pires J.G. Pelá V.T. Buzalaf M.A.R. Magalhães A.C. Effect of commercial herbal toothpastes and mouth rinses on the prevention of enamel demineralization using a microcosm biofilm model. Biofouling 2019 35 7 796 804 10.1080/08927014.2019.1662897 1514534
    [Google Scholar]
  8. Chi Y. Wang Y. Ji M. Li Y. Zhu H. Yan Y. Fu D. Zou L. Ren B. Natural products from traditional medicine as promising agents targeting at different stages of oral biofilm development. Front. Microbiol. 2022 13 955459 10.3389/fmicb.2022.955459 6033896
    [Google Scholar]
  9. Rodrigues R.M. Stinckens M. Ates G. Vanhaecke T. Neutral red uptake assay to assess cytotoxicity in vitro. In:Cell. Viability Assays Methods in Molecular Biology. Friedrich O. Gilbert D.F. New York, NY Humana 2023 2644 10.1007/978‑1‑0716‑3052‑5_15
    [Google Scholar]
  10. Vajrabhaya L.O. Korsuwannawong S. Cytotoxicity evaluation of Clinacanthus nutans through dimethylthiazol diphenyltetrazolium bromide and neutral red uptake assays. Eur. J. Dent. 2016 10 1 134 138 10.4103/1305‑7456.175701 27011752
    [Google Scholar]
  11. Cortelli S.C. Cortelli J.R. Holzhausen M. Franco G.C.N. Rebelo R.Z. Sonagere A.S. Queiroz C.S. Costa F.O. Essential oils in one‐stage full‐mouth disinfection: Double‐blind, randomized clinical trial of long‐term clinical, microbial and salivary effects. J. Clin. Periodontol. 2009 36 4 333 342 10.1111/j.1600‑051X.2009.01376.x 19426180
    [Google Scholar]
  12. Cavalca Cortelli S. Cavallini F. Regueira Alves M.F. Alves Bezerra A. Queiroz C.S. Cortelli J.R. Clinical and microbiological effects of an essential-oil-containing mouth rinse applied in the “one-stage full-mouth disinfection” protocol—A randomized doubled-blinded preliminary study. Clin. Oral Investig. 2009 13 2 189 194 10.1007/s00784‑008‑0219‑3 18716800
    [Google Scholar]
  13. Cosyn J. Princen K. Miremadi R. Decat E. Vaneechoutte M. De Bruyn H. A double‐blind randomized placebo‐controlled study on the clinical and microbial effects of an essential oil mouth rinse used by patients in supportive periodontal care. Int. J. Dent. Hyg. 2013 11 1 53 61 10.1111/idh.12000 22998386
    [Google Scholar]
  14. Nascimento P.F.C. Alviano W.S. Nascimento A.L.C. Santos P.O. Arrigoni-Blank M.F. De Jesus R.A. Azevedo V.G. Alviano D.S. Bolognese A.M. Trindade R.C. Hyptis pectinata essential oil: chemical composition and anti‐ Streptococcus mutans activity. Oral Dis. 2008 14 6 485 489 10.1111/j.1601‑0825.2007.01405.x 18826378
    [Google Scholar]
  15. Thaweboon S. Thaweboon B. Effect of an essential oil-containing mouth rinse on VSC-producing bacteria on the tongue. Southeast Asian J. Trop. Med. Public Health 2011 42 2 456 462 [PMID: 21710870
    [Google Scholar]
  16. Jones C.G. Chlorhexidine: Is it still the gold standard? Periodontol. 2000 1997 15 1 55 62 10.1111/j.1600‑0757.1997.tb00105.x 9643233
    [Google Scholar]
  17. Baratakke S.U. Raju R. Kadanakuppe S. Savanur N.R. Gubbihal R. Kousalaya P.S. Efficacy of triphala extract and chlorhexidine mouth rinse against plaque accumulation and gingival inflammation among female undergraduates: A randomized controlled trial. Indian J. Dent. Res. 2017 28 1 49 54 10.4103/0970‑9290.203622 28393817
    [Google Scholar]
  18. Kairey L. Agnew T. Bowles E.J. Barkla B.J. Wardle J. Lauche R. Efficacy and safety of Melaleuca alternifolia (tea tree) oil for human health—A systematic review of randomized controlled trials. Front. Pharmacol. 2023 14 1116077 10.3389/fphar.2023.1116077 7033604
    [Google Scholar]
  19. Brookes Z. Teoh L. Cieplik F. Kumar P. Mouthwash effects on the oral microbiome: Are they good, bad, or balanced? Int. Dent. J. 2023 73 Suppl. 2 S74 S81 10.1016/j.identj.2023.08.010 7867065
    [Google Scholar]
  20. 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 10 978 991 10.22038/IJBMS.2018.31997.7690 30524670
    [Google Scholar]
  21. Sparabombe S. Monterubbianesi R. Tosco V. Orilisi G. Hosein A. Ferrante L. Putignano A. Orsini G. Efficacy of an all-natural polyherbal mouthwash in patients with periodontitis: A single-blind randomized controlled trial. Front. Physiol. 2019 10 632 10.3389/fphys.2019.00632 31191341
    [Google Scholar]
  22. Liu F.L. Rong Y. Zhou H. Yu T. Liu L. Cao Q. Qin Z. Qu L. Liao X. Jiang Q. Zhang N. Xu X. Cineole inhibits the biosynthesis of leukotrienes and prostaglandins to alleviate allergic rhinitis: Insights from metabolomics. J. Pharm. Biomed. Anal. 2023 234 115574 10.1016/j.jpba.2023.115574 37481900
    [Google Scholar]
  23. Batista A.L.A. Diógenes Alves Uchôa Lins R. de Souza Coelho R. do Nascimento Barbosa D. Moura Belém N. Alves Celestino F.J. Clinical efficacy analysis of the mouth rinsing with pomegranate and chamomile plant extracts in the gingival bleeding reduction. Complement. Ther. Clin. Pract. 2014 20 1 93 98 10.1016/j.ctcp.2013.08.002 24439653
    [Google Scholar]
  24. Pereira S.L. de Oliveira J.W. Angelo K.K. da Costa A.M. Costa F. Clinical effect of a mouth rinse containing Ocimum gratissimum on plaque and gingivitis control. J. Contemp. Dent. Pract. 2011 12 5 350 355 [PMID: 22269195
    [Google Scholar]
  25. Salehi B. López-Jornet P. Pons-Fuster López E. Calina D. Sharifi-Rad M. Ramírez-Alarcón K. Forman K. Fernández M. Martorell M. Setzer W. Martins N. Rodrigues C. Sharifi-Rad J. Plant-derived bioactives in oral mucosal lesions: A key emphasis to curcumin, lycopene, chamomile, aloe vera, green tea and coffee properties. Biomolecules 2019 9 3 106 106 10.3390/biom9030106 30884918
    [Google Scholar]
  26. Mehta S. Pesapathy S. Joseph M. Tiwari P. Chawla S. Comparative evaluation of a herbal mouthwash (Freshol) with chlorhexidine on plaque accumulation, gingival inflammation, and salivary Streptococcus mutans growth. J. Int. Soc. Prev. Community Dent. 2013 3 1 25 28 10.4103/2231‑0762.115717 24478976
    [Google Scholar]
  27. Kamath S. Hegde R.J. Comparison of the Streptococcus mutans and Lactobacillus colony count changes in saliva following chlorhexidine (0.12%) mouth rinse, combination mouth rinse, and green tea extract (0.5%) mouth rinse in children. J. Indian Soc. Pedod. Prev. Dent. 2017 35 2 150 155 10.4103/JISPPD.JISPPD_13_17 28492194
    [Google Scholar]
  28. Lakade L. Shah P. Shirol D. Comparison of antimicrobial efficacy of chlorhexidine and combination mouth rinse in reducing the Mutans streptococcus count in plaque. J. Indian Soc. Pedod. Prev. Dent. 2014 32 2 91 96 10.4103/0970‑4388.130780 24739906
    [Google Scholar]
  29. Lu M. Ge Y. Qiu J. Shao D. Zhang Y. Bai J. Zheng X. Chang Z. Wang Z. Dong W. Tang C. Redox/pH dual-controlled release of chlorhexidine and silver ions from biodegradable mesoporous silica nanoparticles against oral biofilms. Int. J. Nanomedicine 2018 13 7697 7709 10.2147/IJN.S181168 30538453
    [Google Scholar]
  30. Pletts M.W. Burrell R.E. Clinically relevant evaluation of the antimicrobial and anti-inflammatory properties of nanocrystalline and nanomolecular silver. Wound Repair Regen. 2025 33 1 e13249 10.1111/wrr.13249 9707715
    [Google Scholar]
  31. Martínez-Aguilar V.M. Carrillo-Ávila B.A. Sauri-Esquivel E.A. Guzmán-Marín E. Jiménez-Coello M. Escobar-García D.M. Pozos-Guillén A. Quantification of TNF- α in patients with periodontitis and type 2 diabetes. BioMed Res. Int. 2019 2019 1 6 10.1155/2019/7984891 1355282
    [Google Scholar]
  32. Zhang Z. Association between COX2 -765G/C polymorphism and periodontitis in Chinese population: A meta-analysis. BMC Oral Health 2018 18 1 32 10.1186/s12903‑018‑0483‑9 29514641
    [Google Scholar]
  33. Elgendy E. Ali S.M. Zineldeen D. Effect of local application of tea tree (Melaleuca alternifolia) oil gel on long pentraxin level used as an adjunctive treatment of chronic periodontitis: A randomized controlled clinical study. J. Indian Soc. Periodontol. 2013 17 4 444 448 10.4103/0972‑124X.118314 24174722
    [Google Scholar]
  34. Bhavikatti S.K. Zainuddin S.L.A. Ramli R.B. Nadaf S.J. Dandge P.B. Khalate M. Karobari M.I. Insights into the antioxidant, anti-inflammatory and anti-microbial potential of Nigella sativa essential oil against oral pathogens. Sci. Rep. 2024 14 1 11878 10.1038/s41598‑024‑62915‑1 8789533
    [Google Scholar]
  35. Van Leeuwen M.P.C. Slot D.E. Van der Weijden G.A. Essential oils compared to chlorhexidine with respect to plaque and parameters of gingival inflammation: A systematic review. J. Periodontol. 2011 82 2 174 194 10.1902/jop.2010.100266 21043801
    [Google Scholar]
  36. Wang S. Fang L. Zhou H. Wang M. Zheng H. Wang Y. Weir M.D. Masri R. Oates T.W. Cheng L. Xu H.H.K. Liu F. Silica nanoparticles containing nano-silver and chlorhexidine respond to pH to suppress biofilm acids and modulate biofilms toward a non-cariogenic composition. Dent. Mater. 2024 40 2 179 189 10.1016/j.dental.2023.11.006 7951751
    [Google Scholar]
  37. Van der Weijden F.A. Van der Sluijs E. Ciancio S.G. Slot D.E. Can chemical mouthwash agents achieve plaque/gingivitis control? Dent. Clin. North Am. 2015 59 4 799 829 10.1016/j.cden.2015.06.002 26427569
    [Google Scholar]
  38. Brookes Z. McGrath C. McCullough M. Antimicrobial mouthwashes: An overview of mechanisms-what do we still need to know? Int. Dent. J. 2023 73 Suppl. 2 S64 S68 10.1016/j.identj.2023.08.009 7867063
    [Google Scholar]
  39. Alshehri F.A. The use of mouthwash containing essential oils (LISTERINE®) to improve oral health: A systematic review. Saudi Dent. J. 2018 30 1 2 6 10.1016/j.sdentj.2017.12.004 30166864
    [Google Scholar]
  40. Charles C.A. Lisante T.A. Revankar R. Cortelli J.R. Cortelli S.C. Aquino D. Goyal C.R. Amini P. Early benefits with daily rinsing on gingival health improvements with an essential oil mouthrinse--post-hoc analysis of 5 clinical trials JDH 2014 88 40 50 (Suppl. 1) 25071149
    [Google Scholar]
  41. Fang L. Zhang Y. Cheng L. Zheng H. Wang Y. Qin L. Cai Y. Cheng L. Zhou W. Liu F. Wang S. Silica nanoparticles containing nano-silver and chlorhexidine to suppress Porphyromonas gingivalis biofilm and modulate multispecies biofilms toward healthy tendency. J. Oral Microbiol. 2024 16 1 2361403 10.1080/20002297.2024.2361403 38847000
    [Google Scholar]
/content/journals/aiaamc/10.2174/0118715230384352250526103643
Loading
Novel Mouth Rinse Displaying Analgesic and Anti-inflammatory Properties: An in vitro Study
Bentham Science Publishers ; https://doi.org/10.2174/0118715230384352250526103643
/content/journals/aiaamc/10.2174/0118715230384352250526103643
/content/journals/aiaamc/10.2174/0118715230384352250526103643
Loading

Data & Media loading...


  • Article Type:     Research Article
Keywords: inflammation ; Anti-inflammatory ; periodontal ; neutral red uptake assay ; cytotoxicity ; ELISA ; demonstrate

Most Cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error
Please enter a valid_number test