Cytotoxicity of V-Prep Versus Phosphoric Acid Etchant on Oral Gingival Fibroblasts

Victor Ghoubril, Sylvie Changotade, Didier Lutomski, Joseph Ghoubril, Carole Chakar, Maher Abboud, Louis Hardan, Naji Kharouf, Elie Khoury, Victor Ghoubril, Sylvie Changotade, Didier Lutomski, Joseph Ghoubril, Carole Chakar, Maher Abboud, Louis Hardan, Naji Kharouf, Elie Khoury

Abstract

The most used etchant in dental daily practice is the phosphoric acid (P.A.; 37%). However, acid etchants can induce necrosis on the oral mucosa and cause the ulceration of periodontal tissue when a rubber dam is not used. V-prep is a new practical alternative, and it has satisfactory results. It is used as a preparation before the application of a resin-modified glass ionomer composite (RMGIC) to bond the orthodontic brackets. The aim of this study was to investigate the effect of the V-prep on oral gingival fibroblasts cells by comparing the cell damage and cell viability after the use of V-prep and a conventional phosphoric acid etchant with different application times and concentrations. Therefore, Gingival fibroblasts passage 6 (GFP6) was grown and treated with an acid etchant and V-prep at three different concentrations (1:1, 1:2 and 1:10) for two different application durations (30 s and 1 min). The morphological changes, cell death and cell viability were assessed. Pyknosis, karyolysis, nucleus reversible and irreversible damages and membrane destruction were observed for both of the etchants at the higher concentrations and longer application durations. Mann-Whitney U-tests were used for the statistical analyses. The application of the V-prep for 30 s showed better values than the acid etchant did in the cell damage analysis and cell viability analysis (p = 0.03). V-prep at a 1:10 concentration applied for a 30 s duration can preserve the viability of gingival fibroblasts cells up to 100%. The toxicity of V-prep is equal or lower than the toxicity of the acid etchant that is commonly used in dentistry. Thus, the V-prep can be used with precautions intra-orally, and it should be applied on the enamel as a gel for 30 s only before it is rinsed and removed.

Keywords: V-prep; cell viability; dental acid etchants; oral mucosa; phosphoric acid.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Microscope observations of oral epithelial cells stained by Giemsa under 100-fold magnification. (A) Control group. (B) V-prep, 30 s, undiluted. (C) V-prep, 30 s, 1:2. (D) V-prep, 30 s, 1:10. (E) Acid etchant, 30 s, undiluted. (F) Acid etchant, 30 s, 1:2. (G) Acid etchant, 30 s, 1:10.
Figure 2
Figure 2
Microscope observations of oral epithelial cells stained by Giemsa under 250-fold magnification. (A) Control group. (B) V-prep, 30 s, undiluted. (C) V-prep, 30 s, 1:2. (D) V-prep, 30 s, 1:10. (E) Acid etchant, 30 s, undiluted. (F) Acid etchant, 30 s, 1:2. (G) Acid etchant, 30 s, 1:10.
Figure 3
Figure 3
Cell death percentage after 30 s of different etchant application.
Figure 4
Figure 4
Cell death percentage after 1 min of different etchant application.
Figure 5
Figure 5
Cell viability percentage after 30 s of different etchant application.
Figure 6
Figure 6
Cell viability percentage after 1 min of different etchant application.

References

    1. Pickett K.L., Sadowsky P.L., Jacobson A., Lacefield W. Orthodontic In Vivo Bond Strength: Comparison with In Vitro Results. Angle Orthod. 2001;71:141–148. doi: 10.1043/0003-3219(2001);2.
    1. Pinho M., Manso M.C., Almeida R.F., Martin C., Carvalho Ó., Henriques B., Silva F., Pinhão Ferreira A., Souza J.C.M. Bond Strength of Metallic or Ceramic Orthodontic Brackets to Enamel, Acrylic, or Porcelain Surfaces. Materials. 2020;13:5197. doi: 10.3390/ma13225197.
    1. Grazioli G., Hardan L., Bourgi R., Nakanishi L., Amm E., Zarow M., Jakubowicz N., Proc P., Cuevas-Suárez C.E., Lukomska-Szymanska M. Residual Adhesive Removal Methods for Rebonding of Debonded Orthodontic Metal Brackets: Systematic Review and Meta-Analysis. Materials. 2021;14:6120. doi: 10.3390/ma14206120.
    1. Buonocore M.G. A Simple Method of Increasing the Adhesion of Acrylic Filling Materials to Enamel Surfaces. J. Dent. Res. 1955;34:849–853. doi: 10.1177/00220345550340060801.
    1. Sharma S., Tandon P., Nagar A., Singh G.P., Singh A., Chugh V.K. A Comparison of Shear Bond Strength of Orthodontic Brackets Bonded with Four Different Orthodontic Adhesives. J. Orthod. Sci. 2014;3:29–33. doi: 10.4103/2278-0203.132892.
    1. Hardan L., Bourgi R., Kharouf N., Mancino D., Zarow M., Jakubowicz N., Haikel Y., Cuevas-Suárez C.E. Bond Strength of Universal Adhesives to Dentin: A Systematic Review and Meta-Analysis. Polymers. 2021;13:814. doi: 10.3390/polym13050814.
    1. Flores A.R., Sáez E.G., Barceló F. Metallic Bracket to Enamel Bonding with a Photopolymerizable Resin-Reinforced Glass Ionomer. Am. J. Orthod. Dentofac. Orthop. 1999;116:514–517. doi: 10.1016/S0889-5406(99)70181-2.
    1. Mickenautsch S., Yengopal V., Banerjee A. Retention of Orthodontic Brackets Bonded with Resin-Modified GIC versus Composite Resin Adhesives--a Quantitative Systematic Review of Clinical Trials. Clin. Oral Investig. 2012;16:1–14. doi: 10.1007/s00784-011-0626-8.
    1. Summers A., Kao E., Gilmore J., Gunel E., Ngan P. Comparison of Bond Strength between a Conventional Resin Adhesive and a Resin-Modified Glass Ionomer Adhesive: An in Vitro and in Vivo Study. Am. J. Orthod. Dentofac. Orthop. 2004;126:200–206; quiz 254–255. doi: 10.1016/j.ajodo.2003.06.013.
    1. Ghoubril V., Ghoubril J., Khoury E. A Comparison between RMGIC and Composite with Acid-Etch Preparation or Hypochlorite on the Adhesion of a Premolar Metal Bracket by Testing SBS and ARI: In Vitro Study. Int. Orthod. 2020;18:127–136. doi: 10.1016/j.ortho.2019.07.003.
    1. Sofan E., Sofan A., Palaia G., Tenore G., Romeo U., Migliau G. Classification Review of Dental Adhesive Systems: From the IV Generation to the Universal Type. Ann. Stomatol. 2017;8:1–17. doi: 10.11138/ads/2017.8.1.001.
    1. Blomlöf J., Lindskog S. Periodontal Tissue-Vitality after Different Etching Modalities. J. Clin. Periodontol. 1995;22:464–468.
    1. Blomlöf J., Jansson L., Blomlöf L., Lindskog S. Long-Time Etching at Low PH Jeopardizes Periodontal Healing. J. Clin. Periodontol. 1995;22:459–463. doi: 10.1111/j.1600-051X.1995.tb00177.x.
    1. Akman A.C., Demiralp B., Güncü G.N., Kiremitçi A., Sengün D. Necrosis of Gingiva and Alveolar Bone Caused by Acid Etching and Its Treatment with Subepithelial Connective Tissue Graft. J. Can. Dent. Assoc. 2005;71:477–479.
    1. Jurado C.A., Fischer N.G., Sayed M.E., Villalobos-Tinoco J., Tsujimoto A. Rubber Dam Isolation for Bonding Ceramic Veneers: A Five-Year Post-Insertion Clinical Report. Cureus. 2021;13:e20748. doi: 10.7759/cureus.20748.
    1. Hu H., Li C., Li F., Chen J., Sun J., Zou S., Sandham A., Xu Q., Riley P., Ye Q. Enamel Etching for Bonding Fixed Orthodontic Braces. Cochrane Database Syst. Rev. 2013;2013:CD005516. doi: 10.1002/14651858.CD005516.pub2.
    1. Kim D., Kwak J., Jo R., Jung D., Youn D., Oh N., Jang J. Effects of Dental Acid Etchants in Oral Epithelial Cells. Oral Biol. Res. 2019;43:299–305. doi: 10.21851/obr.43.04.201912.299.
    1. Terhune W.F., Sydiskis R.J., Davidson W.M. In Vitro Cytotoxicity of Orthodontic Bonding Materials. Am. J. Orthod. 1983;83:501–506. doi: 10.1016/S0002-9416(83)90249-X.
    1. Park J.-H., Shin H.-J., Park S.-H., Kim J.-W., Cho K.-M. Iatrogenic chemical burn on facial skin by 37% phosphoric acid etchant. J. Korean Acad. Conserv. Dent. 2009;34:38–41. doi: 10.5395/JKACD.2009.34.1.038.
    1. Nakajima A., Kurihara H., Yagita H., Okumura K., Nakano H. Mitochondrial Extrusion through the Cytoplasmic Vacuoles during Cell Death. J. Biol. Chem. 2008;283:24128–24135. doi: 10.1074/jbc.M802996200.
    1. Cummings B.S., Wills L.P., Schnellmann R.G. Measurement of Cell Death in Mammalian Cells. Curr. Protoc. Pharm. 2004;56:12.8.1–12.8.24. doi: 10.1002/0471141755.ph1208s25.
    1. Riss T.L., Moravec R.A., Niles A.L., Duellman S., Benink H.A., Worzella T.J., Minor L. Cell Viability Assays. In: Markossian S., Grossman A., Brimacombe K., Arkin M., Auld D., Austin C., Baell J., Chung T.D.Y., Coussens N.P., Dahlin J.L., et al., editors. Assay Guidance Manual. Eli Lilly & Company and the National Center for Advancing Translational Sciences; Bethesda, MD, USA: 2004.
    1. Fröb L., Rüttermann S., Romanos G.E., Herrmann E., Gerhardt-Szép S. Cytotoxicity of Self-Etch Versus Etch-and-Rinse Dentin Adhesives: A Screening Study. Materials. 2020;13:452. doi: 10.3390/ma13020452.
    1. Pupo Y.M., Bernardo C.F.d.F., de Souza F.F.d.F.A., Michél M.D., Ribeiro C.N.d.M., Germano S., Maluf D.F. Cytotoxicity of Etch-and-Rinse, Self-Etch, and Universal Dental Adhesive Systems in Fibroblast Cell Line 3T3. Scanning. 2017;2017:9650420. doi: 10.1155/2017/9650420.
    1. Ozer F., Blatz M.B. Self-Etch and Etch-and-Rinse Adhesive Systems in Clinical Dentistry. Compend. Contin. Educ. Dent. 2013;34:12–14, 16, 18; quiz 20, 30.
    1. Cecchin D., Farina A.P., Vidal C.M., Bedran-Russo A.K. A Novel Enamel and Dentin Etching Protocol Using α-Hydroxy Glycolic Acid: Surface Property, Etching Pattern, and Bond Strength Studies. Oper. Dent. 2018;43:101–110. doi: 10.2341/16-136-L.

Source: PubMed

Sponsors et collaborateurs

Les conditions médicales

Interventions en matière de drogue

3
S'abonner