Clinical Oral Investigations

, Volume 18, Issue 1, pp 87–96 | Cite as

Genotoxicity in gingival cells of patients undergoing tooth restoration with two different dental composite materials

  • Antonija Tadin
  • Nada Galic
  • Marin Mladinic
  • Danijela Marovic
  • Ivan Kovacic
  • Davor Zeljezic
Original Article

Abstract

Objectives

Dental composite materials come into direct contact with oral tissue, especially gingival cells. This study was performed to evaluate possible DNA damage to gingival cells exposed to resin composite dental materials.

Materials and methods

Class V restorations were placed in 30 adult patients using two different composite resins. The epithelial cells of the gingival area along the composite restoration were sampled prior to and after 7, 30, and 180 days following the restoration of the tooth. DNA damage was analysed by comet and micronucleus assays in gingival exfoliated epithelial cells.

Results

The results showed significantly higher comet assay parameters (tail length and % DNA in the tail) within periods of 30 and 180 days. The micronucleus test for the same exposure time demonstrated a higher number of cells with micronuclei, karyolysis, and nuclear buds. Results did not reveal any difference between the two composite materials for the same duration of exposure.

Conclusion

Based on the results, we can conclude that the use of composite resins causes cellular damage. As dental composite resins remain in intimate contact with oral tissue over a long period of time, further research on their possible genotoxicity is advisable.

Clinical relevance

Long-term exposure of gingival cells to two different composite materials demonstrated certain DNA damage. However, considering the significant decline in micronuclei frequency after 180 days and efficiency in the repair of primary DNA damage, the observed effects could not be indicated as biologically relevant.

Keywords

Comet assay Dental composite DNA damage Genotoxicity test Micronucleus assay in gingival epithelial cells 

References

  1. 1.
    Van Noort R (2002) Introduction to dental materials, 2nd edn. Mosby Wolfe, LondonGoogle Scholar
  2. 2.
    Øilo G (1992) Biodegradation of dental composites/glass-ionomer cements. Adv Dent Res 6:50–54PubMedGoogle Scholar
  3. 3.
    Larsen IB, Munksgaard EC (1991) Effect of human saliva on surface degradation of composite resins. Scand J Dent Res 99:254–261PubMedGoogle Scholar
  4. 4.
    Michelsen VB, Moe G, Skålevik R, Jensen E, Lygre H (2007) Quantification of organic eluates from polymerized resin-based dental restorative materials by use of GC/MS. J Chromatogr B Anal Technol Biomed Life Sci 850:83–91CrossRefGoogle Scholar
  5. 5.
    Michelsen VB, Moe G, Strøm MB, Jensen E, Lygre H (2008) Quantitative analysis of TEGDMA and HEMA eluted into saliva from two dental composites by use of GC/MS and tailor-made internal standards. Dent Mater 24:724–731PubMedCrossRefGoogle Scholar
  6. 6.
    Michelsen VB, Kopperud HB, Lygre GB, Björkman L, Jensen E, Kleven IS, Svahn J, Lygre H (2012) Detection and quantification of monomers in unstimulated whole saliva after treatment with resin-based composite fillings in vivo. Eur J Oral Sci 120:89–95PubMedCrossRefGoogle Scholar
  7. 7.
    Di Pietro A, Visalli G, La Maestra S, Micale R, Baluce B, Matarese G, Cingano L, Scoglio ME (2008) Biomonitoring of DNA damage in peripheral blood lymphocytes of subjects with dental restorative fillings. Mutat Res 650:115–122PubMedCrossRefGoogle Scholar
  8. 8.
    Polydorou O, König A, Hellwig E, Kümmerer K (2009) Long-term release of monomers from modern dental-composite materials. Eur J Oral Sci 117:68–75PubMedCrossRefGoogle Scholar
  9. 9.
    Ferracane JL (1994) Elution of leachable components from composites. J Oral Rehabil 21:441–452PubMedCrossRefGoogle Scholar
  10. 10.
    Goldberg M (2008) In vitro and in vivo studies on the toxicity of dental resin components: a review. Clin Oral Investig 12:1–8PubMedCrossRefGoogle Scholar
  11. 11.
    Polydorou O, Trittler R, Hellwig E, Kummerer K (2007) Elution of monomers from two conventional dental composite materials. Dent Mater 23:1535–1541PubMedCrossRefGoogle Scholar
  12. 12.
    Manojlovic D, Radisic M, Vasiljevic T, Zivkovic S, Lausevic M, Miletic V (2011) Monomer elution from nanohybrid and ormocer-based composites cured with different light sources. Dent Mater 27:371–378PubMedCrossRefGoogle Scholar
  13. 13.
    Spahl W, Budzikiewicz H, Geurtsen W (1998) Determination of leachable components from four commercial dental composites by gas liquid chromatography/mass spectrometry. J Dent 26:137–145PubMedCrossRefGoogle Scholar
  14. 14.
    Moharamzadeh K, Van Noort R, Brook IM (2007) HPLC analysis of components released from dental composites with different resin compositions using different extraction media. J Mater Sci Mater Med 18:133–137PubMedCrossRefGoogle Scholar
  15. 15.
    Chauvel-Lebret DJ, Auroy P, Tricot-Doleux S, Bonnaure-Mallet M (2001) Evaluation of the capacity of the SCGE assay to assess the genotoxicity of biomaterials. Biomaterials 22:1795–1801PubMedCrossRefGoogle Scholar
  16. 16.
    Hanks CT, Wataha JC, Sun Z (1996) In vitro models of biocompatibility: a review. Dent Mater 12:186–193PubMedCrossRefGoogle Scholar
  17. 17.
    Pizzoferrato A, Ciapetti G, Stea S, Cenni E, Arciola CR, Granchi D, Savarino L (1994) Cell culture methods for testing biocompatibility. Clin Mater 15:173–190PubMedCrossRefGoogle Scholar
  18. 18.
    Müller BP, Eisenträger A, Jahnen-Dechent W, Dott W, Hollender J (2003) Effect of sample preparation on the in vitro genotoxicity of a light curable glass ionomer cement. Biomaterials 24:611–617PubMedCrossRefGoogle Scholar
  19. 19.
    Grover P, Danadevi K, Mahboob M, Rozati R, Banu BS, Rahman MF (2003) Evaluation of genetic damage in workers employed in pesticide production utilizing the Comet assay. Mutagenesis 18:201–205PubMedCrossRefGoogle Scholar
  20. 20.
    Westphalen GH, Menezes LM, Prá D, Garcia GG, Schmitt VM, Henriques JA, Medina-Silva R (2008) In vivo determination of genotoxicity induced by metals from orthodontic appliances using micronucleus and comet assays. Genet Mol Res 7:1259–1266PubMedCrossRefGoogle Scholar
  21. 21.
    Van Goethem F, Lison D, Kirsch-Volders M (1997) Comparative evaluation of the in vitro micronucleus test and the alkaline single cell gel electrophoresis assay for the detection of DNA damaging agents: genotoxic effects of cobalt powder, tungsten carbide and cobalt-tungsten carbide. Mutat Res 392:31–43PubMedCrossRefGoogle Scholar
  22. 22.
    Singh NP, McCoy MT, Tice RR, Schneider EL (1998) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191CrossRefGoogle Scholar
  23. 23.
    Tice RR, Strauss GH (1995) The single cell gel electrophoresis/comet assay: a potential tool for detecting radiation-induced DNA damage in humans. Stem Cells 13(1):207–214PubMedGoogle Scholar
  24. 24.
    Collins AR, Dobson VL, Dusinská M, Kennedy G, Stĕtina R (1997) The comet assay: what can it really tell us? Mutat Res 375:183–193PubMedCrossRefGoogle Scholar
  25. 25.
    Fenech M, Holland N, Chang WP, Zeiger E, Bonassi S (1999) The HUman MicroNucleus Project—an international collaborative study on the use of the micronucleus technique for measuring DNA damage in humans. Mutat Res 428:271–283PubMedCrossRefGoogle Scholar
  26. 26.
    Møller P (2005) Genotoxicity of environmental agents assessed by the alkaline comet assay. Basic Clin Pharmacol Toxicol 96(1):1–42PubMedCrossRefGoogle Scholar
  27. 27.
    Mladenov E, Iliakis G (2011) Induction and repair of DNA double strand breaks: the increasing spectrum of non-homologous end joining pathways. Mutat Res 711:61–72PubMedCrossRefGoogle Scholar
  28. 28.
    Tolbert PE, Shy CM, Allen JW (1992) Micronuclei and other nuclear anomalies in buccal smears: methods development. Mutat Res 271:69–77PubMedCrossRefGoogle Scholar
  29. 29.
    Vrzoc M, Petras ML (1997) Comparison of alkaline single cell gel (Comet) and peripheral blood micronucleus assays in detecting DNA damage caused by direct and indirect acting mutagens. Mutat Res 381:31–40PubMedCrossRefGoogle Scholar
  30. 30.
    Szeto YT, Benzie IF, Collins AR, Choi SW, Cheng CY, Yow CM, Tse MM (2005) A buccal cell model comet assay: development and evaluation for human biomonitoring and nutritional studies. Mutat Res 578:371–381PubMedCrossRefGoogle Scholar
  31. 31.
    Eren K, Ozmeriç N, Sardaş S (2002) Monitoring of buccal epithelial cells by alkaline comet assay (single cell gel electrophoresis technique) in cytogenetic evaluation of chlorhexidine. Clin Oral Investig 6:150–154PubMedCrossRefGoogle Scholar
  32. 32.
    Baričević M, Ratkaj I, Mladinić M, Želježić D, Kraljević SP, Lončar B, Stipetić MM (2012) In vivo assessment of DNA damage induced in oral mucosa cells by fixed and removable metal prosthodontic appliances. Clin Oral Investig 16:325–331PubMedCrossRefGoogle Scholar
  33. 33.
    Hafez HS, Selim EM, Kamel Eid FH, Tawfik WA, Al-Ashkar EA, Mostafa YA (2011) Cytotoxicity, genotoxicity, and metal release in patients with fixed orthodontic appliances: a longitudinal in-vivo study. Am J Orthod Dentofac Orthop 140:298–308CrossRefGoogle Scholar
  34. 34.
    Faccioni F, Franceschetti P, Cerpelloni M, Fracasso ME (2003) In vivo study on metal release from fixed orthodontic appliances and DNA damage in oral mucosa cells. Am J Orthod Dentofac Orthop 124:687–693CrossRefGoogle Scholar
  35. 35.
    Ahmed RH, Aref MI, Hassan RM, Mohammed NR (2010) Cytotoxic effect of composite resin and amalgam filling materials on human labial and buccal epithelium. Nat Sci 8:48–53Google Scholar
  36. 36.
    Belien JA, Copper MP, Braakhuis BJ, Snow GB, Baak JP (1995) Standardization of counting micronuclei: definition of a protocol to measure genotoxic damage in human exfoliated cells. Carcinogenesis 16:2395–2400PubMedCrossRefGoogle Scholar
  37. 37.
    Bloching M, Reich W, Schubert J, Grummt T, Sandner A (2008) Micronucleus rate of buccal mucosal epithelial cells in relation to oral hygiene and dental factors. Oral Oncol 44:220–226PubMedCrossRefGoogle Scholar
  38. 38.
    Rezende EF, Mendes-Costa MC, Fonseca JC, Ribeiro AO (2011) Nuclear anomalies in the buccal cells of children under dental treatment. RSBO 8:182–188Google Scholar
  39. 39.
    Erdemir EO, Sengün A, Ulker M (2007) Cytotoxicity of mouthrinses on epithelial cells by micronucleus test. Eur J Dent 1:80–85PubMedCentralPubMedGoogle Scholar
  40. 40.
    Carlin V, Matsumoto MA, Saraiva PP, Artioli A, Oshima CT, Ribeiro DA (2012) Cytogenetic damage induced by mouthrinses formulations in vivo and in vitro. Clin Oral Investig 16:813–820PubMedCrossRefGoogle Scholar
  41. 41.
    Laskaris G, Scully C (2003) Periodontal manifestations of local and systemic diseases: colour atlas and text. Springer, BerlinCrossRefGoogle Scholar
  42. 42.
    Martins RA, Gomes GA, Aguiar O Jr, Ribeiro DA (2009) Biomonitoring of oral epithelial cells in petrol station attendants: comparison between buccal mucosa and lateral border of the tongue. Environ Int 35:1062–1065PubMedCrossRefGoogle Scholar
  43. 43.
    Reichl FX, Esters M, Simon S, Seiss M, Kehe K, Kleinsasser N, Folwaczny M, Glas J, Hickel R (2006) Cell death effects of resin-based dental material compounds and mercurials in human gingival fibroblasts. Arch Toxicol 80(6):370–377PubMedCrossRefGoogle Scholar
  44. 44.
    Kleinsasser NH, Schmid K, Sassen AW, Harréus UA, Staudenmaier R, Folwaczny M, Glas J, Reichl FX (2006) Cytotoxic and genotoxic effects of resin monomers in human salivary gland tissue and lymphocytes as assessed by the single cell microgel electrophoresis (Comet) assay. Biomaterials 27(9):1762–1770PubMedCrossRefGoogle Scholar
  45. 45.
    Lima CF, Oliveira LU, Cabral LA, Brandão AA, Salgado MA, Almeida JD (2010) Cytogenetic damage of oral mucosa by consumption of alcohol, tobacco and illicit drugs. J Oral Pathol Med 39(6):441–446PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Antonija Tadin
    • 1
  • Nada Galic
    • 2
  • Marin Mladinic
    • 3
  • Danijela Marovic
    • 2
  • Ivan Kovacic
    • 1
  • Davor Zeljezic
    • 3
  1. 1.Study of Dental Medicine, School of MedicineUniversity of SplitSplitCroatia
  2. 2.Department of Endodontics and Restorative Dentistry, School of Dental MedicineUniversity of ZagrebZagrebCroatia
  3. 3.Division for MutagenesisInstitute for Medical Research and Occupational HealthZagrebCroatia

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