Cytotoxicity of two fluoride-releasing adhesive tapes

Sodium fluoride-polyvinyl alcohol (NaF-PVA) tape was developed to deliver fluoride to teeth by adding fluoride to polymer tape. Previous studies have demonstrated that tapes are effective and have antimicrobial properties. This study aimed to evaluate the cytotoxicity of two fluoride-releasing adhesive tapes. We investigated two polyvinyl alcohol (PVA) tapes: (i) a fluoride-PVA (F-PVA) tape, and (ii) a pullulan-incorporated F-PVA (PF-PVA) tape. The cytotoxicity test was conducted on human gingival fibroblasts (HGF) and human periodontal ligament (PDL) cells. Using an adhesive tape containing fluoride, we performed the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay on these cells. Genetic analysis of the cells was performed to conduct a stability test on humans. In the MTT assay, PF-PVA had 66% greater cytotoxicity than control by PDL and 69% by HGF. F-PVA showed less cytotoxicity than PF-PVA by 29% in PDL and 33% in HGF. Gene ontology (GO) analysis and gene set enrichment analysis (GSEA) were performed as gene expression analyses. GO analysis indicated that PF-PVA displayed more expression changes of genes related to cytotoxicity than F-PVA. In addition, GSEA found more inflammatory response associations in PF-PVA than in F-PVA. MTT and genetic testing yielded comparable results.

Fluoride; Polyvinyl alcohol tape; Pullulan; Cytotoxicity

[1] Dayo AF, Wolff MS, Syed AZ, Mupparapu M. Radiology of dental caries. Dental Clinics of North America. 2021; 65: 427–445.

[2] Featherstone JDB, Crystal YO, Alston P, Chaffee BW, Doméjean S, Rechmann P, et al. Evidence-based caries management for all ages-practical guidelines. Frontiers in Oral Health. 2021; 2: 657518.

[3] de Oliveira PRA, Barreto LSDC, Tostes MA. Effectiveness of CPP‐ACP and fluoride products in tooth remineralization. International Journal of Dental Hygiene. 2022; 20: 635–642.

[4] Zhang Q, Guan L, Guo J, Chuan A, Tong J, Ban J, et al. Application of fluoride disturbs plaque microecology and promotes remineralization of enamel initial caries. Journal of Oral Microbiology. 2022; 14: 2105022.

[5] Schiffner U. Use of fluorides for caries prevention. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz. 2021; 64: 830–837. (In German)

[6] Knutson JW. Sodium fluoride solutions: technic for application to the teeth. The Journal of the American Dental Association. 1948; 36: 37–39.

[7] Zendron MP, Rocha ADO, Simões MSDS, Santana CM, Bolan M, Cardoso M. Fluoride varnish in dentistry: a bibliometric analysis of the 100 most-cited papers. Caries Research. 2023; 57: 189–196.

[8] Noble J. Fluoride varnish for the teeth. Vital. 2008; 5: 39.

[9] Benedito LCB, Pedro Ricomini-Filho A, Tabchoury CPM, Cury JA. Anticaries potential of a fluoride foam. Brazilian Dental Journal. 2023; 34: 89–98.

[10] Mascarenhas AK. Is fluoride varnish safe? The Journal of the American Dental Association. 2021; 152: 364–368.

[11] Sousa GP, Lima CCB, Braga MM, Moura L, Lima MDM, Moura MS. Early childhood caries management using fluoride varnish and neutral fluoride gel: a randomized clinical trial. Brazilian Oral Research. 2022; 36: e099.

[12] Kim MJ, Lee SH, Lee NY, Lee IH. Evaluation of the effect of PVA tape supplemented with 2.26% fluoride on enamel demineralization using microhardness assessment and scanning electron microscopy: in vitro study. Archives of Oral Biology. 2013; 58: 160–166.

[13] Lee KY LS, Lee NY. Evaluation of fluoride-relasing capacity from polyvinyl alcohol polymer tape supplemented with NaF in oral cavity. Journal of the Korean Academy of Pediatric Dentistry. 2013; 40: 89–97.

[14] Jih MK, Lee SH, Lee NY. Physical and chemical properties of double-layers adhesive polymer film supplemented with 5% NaF. The Jounal of the Korean Dental Association. 2021; 59: 204–212.

[15] Jih M, Lee S, Lee N. Effect of polymer adhesive film supplemented 5% NaF on enamel remineralization. The Journal of the Korean Academy of Pedtatric Dentistry. 2014; 41: 218–224.

[16] Lee SH, Lee NY, Lee IH. Clinical evaluation of the efficacy of fluoride adhesive tape (F-PVA) in reducing dentin hypersensitivity. The American Journal of Dentistry. 2013; 26: 143–148.

[17] Oh YJ, Hong J. Application of the MTT-based colorimetric method for evaluating bacterial growth using different solvent systems. Lebensmittel-Wissenschaft & Technologie. 2022; 153: 112565.

[18] Li S, Zhang H, Chen K, Jin M, Vu SH, Jung S, et al. Application of chitosan\/alginate nanoparticle in oral drug delivery systems: prospects and challenges. Drug Delivery. 2022; 29: 1142–1149.

[19] Singh RS, Kaur N, Singh D, Purewal SS, Kennedy JF. Pullulan in pharmaceutical and cosmeceutical formulations: a review. International Journal of Biological Macromolecules. 2023; 231: 123353.

[20] Suharyani I, Fouad Abdelwahab Mohammed A, Muchtaridi M, Wathoni N, Abdassah M. Evolution of drug delivery systems for recurrent aphthous stomatitis. Drug Design, Development and Therapy. 2021; 15: 4071–4089.

[21] Park DY, Lee NY, Lee SH. Preventive effect of fluoride-containing adhesive film made by nano technology on dental caries. Journal of the Korean Academy of Pediatric Dentistry. 2008; 35: 272662–272670.

[22] Netalkar PP, SR M, YM K, Natarajan S, Gadipelly T, Bhat P D, et al. Effect of nano‐hydroxyapatite incorporation on fluoride‐releasing ability, penetration, and adaptation of a pit and fissure sealant. International Journal of Paediatric Dentistry. 2022; 32: 344–351.

[23] Sun L, Chow LC. Preparation and properties of nano-sized calcium fluoride for dental applications. Dental Materials. 2008; 24: 111–116.

[24] Gamal AY, Bayomy MM. Effect of cigarette smoking on human PDL fibroblasts attachment to periodontally involved root surfaces in vitro. Journal of Clinical Periodontology. 2002; 29: 763–770.

[25] Holliday RS, Campbell J, Preshaw PM. Effect of nicotine on human gingival, periodontal ligament and oral epithelial cells. a systematic review of the literature. Journal of Dentistry. 2019; 86: 81–88.

[26] Lai Y-L, Chen Y-T, Lee S-Y, Shieh T-M, Hung S-L. Cytotoxic effects of dental resin liquids on primary gingival fibroblasts and periodontal ligament cells in vitro. Journal of Oral Rehabilitation. 2004; 31: 1165–1172.

[27] Maru VP, Madkaikar M, Sattar S, Chauhan R, Devi RKS. Response of intra canal medicaments on viability and survival of SHEDs. Journal of Clinical Pediatric Dentistry. 2022; 46: 65–71.

[28] Angius F, Floris A. Liposomes and MTT cell viability assay: an incompatible affair. Toxicology in Vitro. 2015; 29: 314–319.

[29] Gomez Perez M, Fourcade L, Mateescu MA, Paquin J. Neutral Red versus MTT assay of cell viability in the presence of copper compounds. Analytical Biochemistry. 2017; 535: 43–46.

[30] Maru V, Shetty O, Dixit U, Warke VG, Khande M, Mane S. SHED’s Response to various pulpotomy materials: cytotoxicity and gene expression analysis. Journal of Clinical Pediatric Dentistry. 2021; 45: 406–413.

[31] Nam OH, Oh TJ, Lee J, Hwang Y, Choi SC. Differential gene expression profiles of human periodontal ligament cells preserved in Hank’s balanced salt solution and milk. Dental Traumatology. 2020; 36: 58–68.

[32] Rathinam E, Govindarajan S, Rajasekharan S, Declercq H, Elewaut D, De Coster P, et al. Transcriptomic profiling of human dental pulp cells treated with tricalcium silicate-based cements by RNA sequencing. Clinical Oral Investigations. 2021; 25: 3181–3195.

[33] Katsuda T. Sub-GOFA: a tool for Sub-Gene Ontology function analysis in clonal mosaicism using semantic (logical) similarity. Bioinformation. 2022; 18: 53–60.

[34] Tickotsky N, Moskovitz M. Protein activation in periapical reaction to iodoform containing root canal sealer. Journal of Clinical Pediatric Dentistry. 2017; 41: 450–455.

[35] Zhang C, Sun Y, Xu M, Shu C, Yue Z, Hou J, et al. Potential links between COVID-19 and periodontitis: a bioinformatic analysis based on GEO datasets. BMC Oral Health. 2022; 22: 520.

[36] Monkley S, Overed-Sayer C, Parfrey H, Rassl D, Crowther D, Escudero-Ibarz L, et al. Sensitization of the UPR by loss of PPP1R15A promotes fibrosis and senescence in IPF. Scientific Reports. 2021; 11: 21584.

[37] Song Y, Liu S, Zhao Z, Zhang Y, Yang Y, Luo B. Relationship between PPP1R15A gene polymorphism (rs611251) and Epstein-Barr virus-associated tumors. Acta Virologica. 2017; 61: 445–452.

[38] Dovey CM, Diep J, Clarke BP, Hale AT, McNamara DE, Guo H, et al. MLKL requires the inositol phosphate code to execute necroptosis. Molecular Cell. 2018; 70: 936–948.e937.

[39] zhong Y, Zhang ZH, Wang JY, Xing Y, Ri MH, Jin HL, et al. Zinc finger protein 91 mediates necroptosis by initiating RIPK1-RIPK3-MLKL signal transduction in response to TNF receptor 1 ligation. Toxicology Letters. 2022; 356: 75–88.

[40] Sun G, Zhao T. Lung adenocarcinoma pathology stages related gene identification. Mathematical Biosciences and Engineering. 2020; 17: 737–746.

[41] Yap CC, Digilio L, McMahon L, Winckler B. The endosomal neuronal proteins Nsg1/NEEP21 and Nsg2/P19 are itinerant, not resident proteins of dendritic endosomes. Scientific Reports. 2017; 7: 10481.

[42] Bi H, Wang G, Li Z, Zhou L, Zhang M. MEG3 regulates CSE-induced apoptosis by regulating miR-421/DFFB signal axis. International Journal of Chronic Obstructive Pulmonary Disease. 2023; 18: 859–870.

[43] Ma T, Li H, Liu H, Peng Y, Lin T, Deng Z, et al. Neat1 promotes acute kidney injury to chronic kidney disease by facilitating tubular epithelial cells apoptosis via sequestering miR-129-5p. Molecular Therapy. 2022; 30: 3313–3332.

[44] Zhu Y, Han Q, Wang L, Wang B, Chen J, Cai B, et al. Jinhua Qinggan granules attenuates acute lung injury by promotion of neutrophil apoptosis and inhibition of TLR4/MyD88/NF-κB pathway. Journal of Ethnopharmacology. 2023; 301: 115763.

[45] Dai Y, Sun X, Wang C, Li F, Zhang S, Zhang H, et al. Gene co-expression network analysis reveals key pathways and hub genes in Chinese cabbage (Brassica rapa L.) during vernalization. BMC Genomics. 2021; 22: 236.

[46] López-Otín C, Blasco MA, Partridge L, Serrano M, Kroemer G. The hallmarks of aging. Cell. 2013; 153: 1194–1217.

[47] Chovatiya G, Ghuwalewala S, Walter LD, Cosgrove BD, Tumbar T. High‐resolution single‐cell transcriptomics reveals heterogeneity of self‐renewing hair follicle stem cells. Experimental Dermatology. 2021; 30: 457–471.

[48] Fan J, Shi S, Qiu Y, Liu M, Shu Q. Analysis of signature genes and association with immune cells infiltration in pediatric septic shock. Frontiers in Immunology. 2022; 13: 1056750.

[49] Pistis G, Vázquez-Bourgon J, Fournier M, Jenni R, Cleusix M, Papiol S, et al. Gene set enrichment analysis of pathophysiological pathways highlights oxidative stress in psychosis. Molecular Psychiatry. 2022; 27: 5135–5143.

[50] Capece D, Verzella D, Flati I, Arboretto P, Cornice J, Franzoso G. NF-κB: blending metabolism, immunity, and inflammation. Trends in Immunology. 2022; 43: 757–775.

[51] DiDonato JA, Mercurio F, Karin M. NF‐κB and the link between inflammation and cancer. Immunological Reviews. 2012; 246: 379–400.

[52] Lauper K, Iudici M, Mongin D, Bergstra SA, Choquette D, Codreanu C, et al. Effectiveness of TNF-inhibitors, abatacept, IL6-inhibitors and JAK-inhibitors in 31 846 patients with rheumatoid arthritis in 19 registers from the ‘JAK-pot’ collaboration. Annals of the Rheumatic Diseases. 2022; 81: 1358–1366.

[53] Go GW, Mani A. Low-density lipoprotein receptor (LDLR) family orchestrates cholesterol homeostasis. Yale Journal of Biology and Medicine. 2012; 85: 19–28.

[54] Kusuyama J, Komorizono A, Bandow K, Ohnishi T, Matsuguchi T. CXCL3 positively regulates adipogenic differentiation. Journal of Lipid Research. 2016; 57: 1806–1820.

[55] Bimstein E. Periodontal health and disease in children and adolescents. Pediatric Clinics of North America. 1991; 38: 1183–1207.

[56] Gebistorf M, Mijuskovic M, Pandis N, Fudalej PS, Katsaros C. Gingival recession in orthodontic patients 10 to 15 years posttreatment: a retrospective cohort study. American Journal of Orthodontics and Dentofacial Orthopedics. 2018; 153: 645–655.

[57] Singh RS, Kaur N, Rana V, Kennedy JF. Pullulan: a novel molecule for biomedical applications. Carbohydrate Polymers. 2017; 171: 102–121.

[58] Le NN, Le-Vinh B, Friedl JD, Jalil A, Kali G, Bernkop-Schnürch A. Polyaminated pullulan, a new biodegradable and cationic pullulan derivative for mucosal drug delivery. Carbohydrate Polymers. 2022; 282: 119143.