Evaluation of the Cytotoxicity and Anti-biofilm Activity Of A Novel Dentifrice Containing 4-Hydroxycinnamic Acid: An In-Vitro Study Original Article
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Abstract
INTRODUCTION: Evaluating biocompatibility is critical for assessing the safety and therapeutic potential of compounds, often serving as a preliminary step in product development. This study aims to evaluate the biocompatibility and anti-biofilm activity of a novel dentifrice containing 4-hydroxycinnamic acid (4-HCA) against Streptococcus mutans (SM) and Lactobacillus acidophilus (LA).
MATERIALS AND METHODS: The present in vitro study involved three groups- Group 1- commercially available dentifrice as control, Group 2- 0.2% 4-HCA dentifrice, Group 3- 0.4% 4-HCA dentifrice. The antibiofilm efficacy of the test dentifrices was assessed using colony-forming unit (CFU) counts and the Crystal Violet assay. The cytotoxicity was evaluated using the MTT assay and microscopic examination of human gingival fibroblast (HGF) cells.
RESULTS:
ANTIBIOFILM ACTIVITY: The CFU counts for SM and LA were highest in the control group, followed by the 0.2% 4-HCA group, and lowest in the 0.4% 4-HCA group. Crystal Violet assay results showed significant inhibition of biofilm formation in both 4-HCA groups compared to the control.
BIOCOMPATIBILITY: The MTT assay indicated that Group 3 had the highest cell viability and proliferation rates 24 horse after a 2 minute exposure to the conditioned medium across the different dilutions (1:40, 1:100, and 1:1000), as compared to Groups 2 and 1, indicating better biocompatibility. Morphological analysis of HGF cells showed normal spindle-shaped morphology in cells treated with Groups 2 and 3.
CONCLUSION: The novel dentifrice containing 0.4% 4-HCA demonstrates significant anti-biofilm activity against SM and LA and is biocompatible with HGF cells. This study supports the development of 4-HCA-based dentifrices as safe and effective oral hygiene products.
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References
Rieshy V, Balakrishnan N, Arvind TP. Awareness and practice pertaining to the use of digital imaging for orthodontic purposes among undergraduate dental students. J Adv Pharm Technol Res. 2022 Dec 1;13(Suppl 2):S568-72.
Barros Pa, De Lima Cb, Custodio W et al. Use of dentifrices and mouthwashes during orthodontic treatment in the clinical management of orthodontists. RGO-Revista Gaúcha de Odontologia. 2022;70:e20220020.
Rajendran R, Antony S DP, Ashik P M et al. Remineralization potential of strontium-doped nano-hydroxyapatite dentifrice and casein phosphopeptide-amorphous calcium phosphate cream on white spot lesions in enamel following orthodontic debonding – A randomized controlled trial. SAGE Open Med. 2024 Jan;12:20503121231221634.
Rajendran R, Hussain MS, Sandhya R et al. Effect of remineralization agents on white spot lesions: a systematic review. J Pharm Bioallied Sci. 2022 Jul 1;14(Suppl 1):S7-12.
Reichardt E, Geraci J, Sachse S et al. Qualitative and quantitative changes in the oral bacterial flora occur shortly after implementation of fixed orthodontic appliances. Am J Orthod Dentofac Orthop. 2019 Dec 1;156(6):735-44.
Chhabria DR, Ramadoss R, Sundar S et al. Understanding the spatial and topographic characteristics of enamel white spot lesions for targeted remineralization. J Oral Biol Craniofac Res. 2024 Sep 1;14(5):594-9.
Ahn SJ, Lim BS, Lee SJ. Prevalence of cariogenic streptococci on incisor brackets detected by polymerase chain reaction. Am J Orthod Dentofacial Orthop 2007 Jun 1;131(6):736-41.
Jha AK, Chandra S, Shankar D et al. Evaluation of the prevalence of white spot lesions during fixed orthodontic treatment among patients reporting for correction of malocclusion: a prevalence study. Cureus. 2023 Jul 19;15(7).
Tufekci E, Dixon JS, Gunsolley JC et al. Prevalence of white spot lesions during orthodontic treatment with fixed appliances. Angle Orthod. 2011 Mar 1;81(2):206-10.
Thorat SU, Jain RK, Ramalingam K et al. Evaluation of Cytotoxicity of 4-Hydroxycinnamic Acid Using Tetrazolium Bromide Assay and Zebrafish Embryotoxicity: An In-Vitro Study. Cureus. 2024 Mar 10;16(3).
Alam MA, Subhan N, Hossain H et al. Hydroxycinnamic acid derivatives: a potential class of natural compounds for the management of lipid metabolism and obesity. Nutr Metab. 2016 Dec;13:1-3.
Laverty G, McCloskey AP, Gorman SP et al. Anti‐biofilm activity of ultrashort cinnamic acid peptide derivatives against medical device‐related pathogens. J Pept Sci. 2015 Oct;21(10):770-8.
Ramsundar K, Jain RK, Pitchaipillai SG. Inhibition of quorum sensing controlled virulence factors and biofilm formation of Streptococcus mutans isolated from orthodontic subjects by 4-hydroxycinnamic acid. Cureus. 2023 Oct 22;15(10).
Kozubek M, Serbian I, Hoenke S, Kraft O, Csuk R. Synthesis and cytotoxic evaluation of hydroxycinnamic acid rhodamine B conjugates. Results Chem. 2020 Jan 1;2:100057.
Khoroushi M, Kachuie M. Prevention and treatment of white spot lesions in orthodontic patients. Contemp Clin Dent. 2017 Jan 1;8(1):11-9.
Setvaji, Nisshitha & Kumar Subramanian, Aravind. (2024). Evaluation of Remineralisation Potential of an Indigenously Developed Dentifrice – An In Vitro Study. Texila Int J Pub Health. 12. 1-12.
Syed Va, Pavithra A, Rajasekar A et al. Preparation of Nutmeg Gel and Evaluation of its Efficacy as Local Drug Delivery in Stage II Grade A Periodontitis Patients: A Prospective Interventional Study. J Clin Diagn Res. 2025 Feb 1;19(2).
Dharini S, Pandiar D, Rajeshkumar S et al. Evaluation of Anti-inflammatory and Antioxidant Properties of Persea Americana and Syzygium Aromaticum-based Herbal Mouthwash Formulation: An In-vitro Study. J Clin Diagn Res. 2024 Oct 1;18(10).
Parekar D, Selvaganesh S, Nesappan T. Comparative Evaluation of Anti-bacterial, Anti-inflammatory Efficacy and Cytotoxicity of Triple Antibiotic Paste Modified Soft Liners with Conventional Soft Liners: An In-vitro Study. J Clin Diagn Res. 2024 Aug 1;18(8).
Korenaga A, Miyaoka T, Asami H et al. Synergetic inhibitory effect of isopropyl methyl phenol-based agents on biofilm formation by Streptococcus mutans. Plos One. 2024 Sep 23;19(9):e0310926.
Tabatabaei MH, Mahounak FS, Asgari N et al. Cytotoxicity of the ingredients of commonly used toothpastes and mouthwashes on human gingival fibroblasts. Front Dent. 2019 Dec 20;16(6):450.
Kasi SR, Roffel S, Özcan M et al. In vitro cytotoxicity (irritant potency) of toothpaste ingredients. Plos One. 2025 Jan 30;20(1):e0318565.