•  
  •  
 

Abstract

Objective: The objective of this study was to determine the DNA damage caused by ceramic bracket (Clear II, SIA Orthodontic Manufacturer Srl, Italy) on human buccal mucosal cells using comet assay and the mutagenicity of orthodontic bonding system (Transbond XT; 3M Unitek) by Ames test. Methods: In this study, twenty orthodontic patients were recruited from Specialist Orthodontic Clinic, Hospital Universiti Sains Malaysia. The Buccal mucosal cell sample was obtained from each patient at three time points-before (TO), after one month (T1) and after two months (T2) of ceramic bracket placement for performing comet assay. The spot test version of Ames test was performed using four Salmonella Typhimurium (S. Typhimurium) tester strains (TA 98, TA100, TA1535 and TA1537) for mutagenicity testing. Total comet score (TCS) and damage frequency (DF) were used to determine the DNA damage using non-parametric Friedman test followed by multiple pairwise comparison. The Ames test was analysed by a non-statistical method based on revertant growth ring formation. Results: There was no significant change of both TCS and DF between T0 and T1, but both parameters increased significantly from T0 to T1 and from T1 to T2. Non-statistical analysis was carried out to evaluate the results of Ames test based on the formation of revertant colony growth ring. None of the S. Typhimurium tester strains showed any revertant growth ring formation around the light cure adhesive primer. Conclusion: Ceramic bracket does not cause any DNA damage on human buccal mucosal cells and light cure adhesive primer is non mutagenic under the present test conditions.

References

  1. Mockers O, Deroze D, Camps J. Cytotoxicity of orthodontic bands, brackets and archwires in vitro. Dent Mater. 2002; 18(4):311-7.

  2. Ahrari F, Tavakkol Afshari J, Poosti M, Brook A. Cytotoxicity of orthodontic bonding adhesive resins on human oral fibroblasts. Eur J Orthod. 2010; 32(6):688-92.

  3. Contaldo M, Lucchese A, Lajolo C, Rupe C, Di Stasio D, Romano A, Petruzzi M, Serpico R. The oral microbiota changes in orthodontic patients and effects on oral health: An overview. J Clin Med. 2021; 10(4):780.

  4. Faccioni F, Franceschetti P, Cerpelloni M, Fracasso ME. In vivo study on metal release from fixed orthodontic appliances and DNA damage in oral mucosa cells. Am J Orthod Dentofacial Orthop. 2003; 124(6):687-93; discussion 693-4.

  5. Toy E, Yuksel S, Ozturk F, Karatas OH, Yalcin M. Evaluation of the genotoxicity and cytotoxicity in the buccal epithelial cells of patients undergoing orthodontic treatment with three light-cured bonding composites by using micronucleus testing. Korean J Orthod. 2014; 44(3):128-35.

  6. Tadin A, Galic N, Mladinic M, Marovic D, Kovacic I, Zeljezic D. Genotoxicity in gingival cells of patients undergoing tooth restoration with two different dental composite materials. Clin Oral Investig. 2014; 18(1):87-96.

  7. Kassie F, Parzefall W, Knasmüller S. Single cell gel electrophoresis assay: A new technique for human biomonitoring studies. Mutat Res. 2000; 463(1):13-31.

  8. Rao KS, Xu Y, Shaw E, Parton JW. Mutagenicity testing applied for regulation of developing products. Curr Sep. 2004; 20(4):141-4.

  9. Mortelmans K, Zeiger E. The Ames Salmonella/ microsome mutagenicity assay. Mutat Res. 2000; 455(1-2):29-60.

  10. Rezai-Basiri M, Samini M, Ghazi-Khansari M, Rezayat M, Sahebgharani M, Partoazar A. Monitoring ames assay on urine of clinical pathology laboratories technicians. J Pharmacol Toxicol. 2008; 3(3):230-5.

  11. Besaratinia A, Van Straaten HW, Godschalk RW, Van Zandwijk N, Balm AJ, Kleinjans JC, Van Schooten FJ. Immunoperoxidase detection of polycyclic aromatic hydrocarbon-DNA adducts in mouth floor and buccal mucosa cells of smokers and nonsmokers. Environ Mol Mutagen. 2000; 36(2):127-33.

  12. Szeto YT, Benzie IF, Collins AR, Choi SW, Cheng CY, Yow CM, Tse MM. A buccal cell model comet assay: development and evaluation for human biomonitoring and nutritional studies. Mutat Res. 2005; 578(1-2):371-81.

  13. Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, Miyamae Y, Rojas E, Ryu JC, Sasaki YF. Single cell gel/comet assay: Guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen. 2000; 35(3):206-21.

  14. Hafez HS, Selim EM, Kamel Eid FH, Tawfik WA, Al-Ashkar EA, Mostafa YA. Cytotoxicity, genotoxicity, and metal release in patients with fixed orthodontic appliances: A longitudinal invivo study. Am J Orthod Dentofacial Orthop. 2011; 140(3):298-308.

  15. Collins AR, Ma AG, Duthie SJ. The kinetics of repair of oxidative DNA damage (strand breaks and oxidised pyrimidines) in human cells. Mutat Res. 1995; 336(1):69-77.

  16. Kobayashi H, Sugiyama C, Morikawa Y, Hayashi M, Sofuni T. A comparison between manual microscopic analysis and computerized image analysis in the single cell gel electrophoresis assay. MMS Commun. 1995; 3(2):103-15.

  17. Maron DM, Ames BN. Revised methods for the Salmonella mutagenicity test. Mutat Res. 1983; 113(3-4):173-215.

  18. Fredericks HE. Mutagenic potential of orthodontic bonding materials. Am J Orthod. 1981; 80(3):316- 24.

  19. Fernández-Miñano E, Ortiz C, Vicente A, Calvo Guirado JL, Ortiz AJ. Metallic ion content and damage to the DNA in oral mucosa cells of children with fixed orthodontic appliances. Biometals. 2011; 24(5):935.

  20. Gingold EB. The Ames Test. In: Slater RJ, editor. Experiments in Molecular Biology. Humana Totowa, NJ: Springer; 1986. p. 265-378.

  21. Garhammer P, Schmalz G, Hiller KA, Reitinger T. Metal content of biopsies adjacent to dental cast alloys. Clin Oral Investig. 2003; 7(2):92-7.

  22. Vitral JC, Fraga MR, de Souza MA, Ferreira AP, Vitral RW. In-vitro study of cellular viability and nitric oxide production by J774 macrophages stimulated by interferon gamma with ceramic, polycarbonate, and polyoxymethylene brackets. Am J Orthod Dentofacial Orthop. 2010; 137(5):665- 70.

  23. Es-Souni M, Es-Souni M, Fischer-Brandies H. Assessing the biocompatibility of NiTi shape memory alloys used for medical applications. Anal Bioanal Chem. 2005; 381(3):557-67.

  24. Westphalen GH, Menezes LM, Prá D, Garcia GG, Schmitt VM, Henriques JA, Medina-Silva R. In vivo determination of genotoxicity induced by metals from orthodontic appliances using micronucleus and comet assays. Genet Mol Res. 2008; 7(4):1259-66.

  25. Quadras DD, Krishna Nayak US, Kumari NS, Priyadarshini, HR, Gowda S, Fernandes B, Pujari P. In vivo study on release of nickel, chromium, and zinc and dna damage in buccal mucosa cells from patients treated with fixed orthodontic appliances. J Indian Orthod Soc. 2018; 52(2):115-9.

  26. Zoroddu MA, Schinocca L, Kowalik-Jankowska T, Kozlowski H, Salnikow K, Costa M. Molecular mechanisms in nickel carcinogenesis: Modeling Ni(II) binding site in histone H4. Environ Health Perspect. 2002; 110 Suppl 5(Suppl 5):719-23.

  27. Grimsdottir MR, Gjerdet NR, Hensten-Pettersen A. Composition and in vitro cor rosion of orthodontic appliances. Am J Orthod Dentofacial Orthop. 1992; 101(6):525-32.

  28. Bakhtari A, Bradley TG, Lobb WK, Berzins DW. Galvanic corrosion between various combinations of orthodontic brackets and archwires. Am J Orthod Dentofacial Orthop. 2011; 140(1):25-31.

  29. Rongo R, Valletta R, Bucci R, Rivieccio V, Galeotti A, Michelotti A, D’Antò V. In vitro biocompatibility of nickel-titanium esthetic orthodontic archwires. Angle Orthod. 2016; 86(5):789-95.

  30. Tomakidi P, Koke U, Kern R, Erdinger L, Krüger H, Kohl A, Komposch G. Assessment of acute cyto- and genotoxicity of corrosion eluates obtained from orthodontic materials using monolayer cultures of immortalized human gingival keratinocytes. J Orofac Orthop. 2000; 61(1):2-19.

  31. Sjögren G, Sletten G, Dahl JE. Cytotoxicity of dental alloys, metals, and ceramics assessed by millipore filter, agar overlay, and MTT tests. J Prosthet Dent. 2000; 84(2):229-36.

  32. Milleding P, Haraldsson C, Karlsson S. Ion leaching from dental ceramics during static in vitro corrosion testing. J Biomed Mater Res. 2002; 61(4):541-50.

  33. Cross NG, Taylor RF, Nunez LJ. “Single-step” orthodontic bonding systems: Possible mutagenic potential. Am J Orthod. 1983; 84(4):344-50.

  34. Vande Vannet BM, Hanssens JL. Cytotoxicity of two bonding adhesives assessed by threedimensional cell culture. Angle Orthod. 2007; 77(4):716-22.

  35. Malkoc S, Corekci B, Botsali HE, Yalçin M, Sengun A. Cytotoxic effects of resin-modified orthodontic band adhesives. Are they safe? Angle Orthod. 2010; 80(5):890-5.

  36. Heravi F, Abbaszadegan MR, Merati M, Hasanzadeh N, Dadkhah E, Ahrari F. DNA damage in oral mucosa cells of patients with fixed orthodontic appliances. J Dent (Tehran). 2013; 10(6):494-500.

  37. Huang TH, Tsai CY, Chen SL, Kao CT. An evaluation of the cytotoxic effects of orthodontic bonding adhesives upon a primary human oral gingival fibroblast culture and a permanent, human oral cancer-cell line. J Biomed Mater Res. 2002; 63(6):814-21.

Download

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.