Journal of Dental Sciences
Volume 4, Issue 4 , Pages 178-186, December 2009

Evaluation of cytotoxicity of resin bonding materials toward human oral epithelial cells using three assay systems

  • Ming-Gene Tu

      Affiliations

    • School of Dentistry, China Medical University, Taichung, Taiwan
    • Department of Dentistry, China Medical University Hospital, Taichung, Taiwan
    • ,
  • Wen-Miin Liang

      Affiliations

    • Institute of Environmental Health, China Medical University College of Public Health, Taichung, Taiwan
    • Biostatistics Center, China Medical University, Taichung, Taiwan
    • ,
  • Tai-Chin Wu

      Affiliations

    • Institute of Environmental Health, China Medical University College of Public Health, Taichung, Taiwan
    • Biostatistics Center, China Medical University, Taichung, Taiwan
    • ,
  • San-Yue Chen

      Affiliations

    • School of Dentistry, China Medical University, Taichung, Taiwan
    • Corresponding Author InformationCorresponding author. School of Dentistry, China Medical University, 91, Hsueh-Shih Road, Taichung 40402, Taiwan

Received 11 July 2009; accepted 16 October 2009.

Article Outline

Background/purpose

Gingival tissue around teeth can be contacted by flowable photocured resin agents during operative procedures. The cytotoxicity of resin bonding agents toward human oral epithelial cells cannot be neglected. This study evaluates the cytotoxicity of resin bonding agents toward human oral epithelial cells using three assay systems.

Materials and methods

Nine commercial resin bonding agents (Clearfil Protect Bond, Clearfil SE Bond, Prime & Bond NT, Prisma Universal Bond 3, UniFil Bond, 3M Single Bond, CharmBond, Compobond, and ExciTE) and two resin monomers (methyl methacrylate and triethylene glycol dimethacrylate) were used. The detection methods for the cell survival rate included: (1) the CellTiter 96 AQueous One Solution Cell Proliferation Assay System, (2) PreMix WST-1 Cell Proliferation Assay System, and (3) CellTiter-Glo Luminescent Cell Viability Assay System. One-way analysis of variance and Scheffé test were used for the statistical analyses.

Results

Most of the components of the uncured bonding agents had cytotoxicity at 0.1 vol% (survival rates, 21.4% ± 1.4% to 113.1% ± 19.1%), except for UniFil Bond Primer (survival rates, 101.8% ± 3.5% to 113.1% ± 19.1%). More severe cytotoxicity was found at the 1.0 vol% concentration (survival rates, 0.2% ± 0.02% to 147.3% ± 10.8%), except for the methyl methacrylate monomer (survival rates, 96.5% ± 1.3% to 147.3% ± 10.8%). In the post-cured group, most bonding agents had little or no cytotoxicity, except for 3M Single Bond (survival rates, 52.4% ± 7.9% to 97.4% ± 17.8%) and Compobond (survival rates, 29.1% ± 7.1% to 79.5% ± 6.7%).

Conclusion

The same material detected by different assay systems may give different cytotoxic effects, and examination by three methods may provide more-objective results. Furthermore, since most of the bonding agent components detected by the three assay systems showed cytotoxicity before curing (and some components even showed cytotoxicity after curing), the application of bonding materials in the oral cavity should be very carefully performed.

Key words:  cell culture , CellTiter-Glo , cytotoxicity , MTS , resin bonding agent , WST-1

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References 

  1. Hensten-Pettersen A , Jacobsen N . Toxic effects of dental materials . Int Dent J . 1991;41:265–273
  2. Ratanasathien S , Wataha JC , Hanks CT , Dennison JB . Cytotoxic interactive effects of dentin bonding components on mouse fibroblasts . J Dent Res . 1995;74:1602–1606
  3. Lefebvre CA , Schuster GS . Biocompatibility of visible lightcured resin systems in prosthodontics . J Prosthet Dent . 1994;71:178–185
  4. Wiltshire WA , Ferreira MR , Ligthelm AJ . Allergies to dental materials . Quintessence Int . 1996;27:513–520
  5. Chigira H , Manabe A , Hasegawa T , et al.   Efficacy of various commercial dentin bonding systems . Dent Mater . 1994;10:363–368
  6. Chen RS , Liu CC , Tseng WY , Jeng JH , Lin CP . Cytotoxicity of three dentin bonding agents on human dental pulp cells . J Dent . 2003;31:223–229
  7. Ozen J , Atay A , Toksoy Topcu F , Ural AU , Dalkiz M , Tunca YM . Analysis of the cytotoxicity of four dentin bonding agents on gingival fibroblasts . Turk J Med Sci . 2005;35:395–399
  8. Schedle A , Franz A , Rausch-Fan X , et al.   Cytotoxic effects of dental composites, adhesive substances, compomers and cements . Dent Mater . 1998;14:429–440
  9. Caughman WF , Caughman GB , Dominy WT , Schuster GS . Glass ionomer and composite resin cements: effects on oral cells . J Prosthet Dent . 1990;63:513–521
  10. Vajrabhaya LO , Pasauk A , Harnirattisal C . Cytotoxicity evaluation of single component dentin bonding agent . Oper Dent . 2003;28:440–444
  11. Grobler SR , Olivier A , Moodley D , van W Kotze TW . Cytotoxicity of two concentrations of a dentin bonding agent on mouse 3T3 and human pulp fibroblast cell-lines . SADJ . 2004;59:368–372
  12. Yoshida K , Greener EH . Effects of two amine reducing agents on the degree of conversion and physical properties of an unfilled light-cured resin . Dent Mater . 1993;9:246–251
  13. Asmussen E , Peutzfeldt A . Influence of UEDMA BisGMA and TEGDMA on selected mechanical properties of experimental resin composites . Dent Mater . 1998;14:51–56
  14. Anseth KS , Goodner MD , Reil MA , Kannurpatti AR , Newman SM , Bowman CV . The influence of comonomer composition on dimethacrylate resin properties for dental composites . J Dent Res . 1996;75:1607–1612
  15. Johnson GH , Powell LV , Gordon GE . Dentin bonding systems: a review of current products and techniques . J Am Dent Assoc . 1991;122:34–41
  16. Full CA , Hollander WR . The composite resin restoration: a literature review, part I: proper cavity preparation and placement techniques . ASDC J Dent Child . 1993;60:48–51
  17. Schweikl H , Schmalz G , Rackebrandt K . The mutagenic activity of unpolymerized resin monomers in Salmonella typhimurium and V 79 cells . Mutat Res . 1998;451:119–130
  18. Kaga M , Noda M , Ferrracane JL , Nakamura W , Oguchi H , Sano H . The in vitro cytotoxicity of elutes from dentin bonding resins and their effect on tyrosine phosphorylation of L929 cells . Dent Mater . 2001;17:333–339
  19. Geurtsen W , Lehmann F , Spahl W , Leyhausen G . Cytotoxicity of 35 dental resin composite monomer/additives in permanent 3T3 and three human primary fibroblast culture . J Biomed Mater Res . 1998;41:474–480
  20. Issa Y , Watts DC , Brunton PA , Waters CM , Duxbury AJ . Resin composite monomers alter MTT and LDH activity of human gingival fibroblasts in vitro . Dent Mater . 2004;20:12–20
  21. Steele C , Leigh J , Swoboda R , Fidel PL . Growth inhibition of Candida by human oral epithelial cells . J Infect Dis . 2000;182:1479–1485
  22. Yamagata N , Oshima H . Cytotoxic effects of restorative materials on early passage cultured cells derived from human gingival . Shika Zairyo Kikai . 1990;9:541–554 [In Japanese, English abstract]
  23. Schweikl H , Schmalz G . Toxicity parameters for cytotoxicity testing of dental materials in two different mammalian cell lines . Eur J Oral Sci . 1996;104:292–299
  24. Livny  , Kaplan I , Reifen R , Polak-Charcon S , Madar Z , Schwartz B . Oral cancer cells differ from normal oral epithelial cells in tissue like organization and in response to lycopene treatment: an organotypic cell culture study . Nutr Cancer . 2003;47:195–209
  25. Njoroge T , Genco RJ , Sojar HT , Hamada N , Genco CA . A role for fimbriae in Porphyromonas gingivalis invasion of oral epithelial cells . Infect Immun . 1997;65:1980–1984
  26. Dorn BR , Leung KP , Fox AP . Invasion of human oral epithelial cells by Prevotella intermedia . Infect Immun . 1998;66:6054–6057
  27. Chen XS , Fang TL , Cai RB , Guo GL . Development of MTS/pms colorimetric assay in the proliferation of leukemic cells . Zhongguo Shi Yan Xue Ye Xue Za Zhi . 2002;10:438–440 [In Chinese, English abstract]
  28. Berridge MV , Tan AS , McCoy KA , Wang R . The biochemical and cellular basis of cell proliferation assays that use tetrazolium salts . Biochemica . 1996;4:15–19
  29. Crouch SPM , Kozlowski R , Slater KJ , Fletcher J . The use of ATP bioluminescence as a measure of cell proliferation and cytotoxicity . J Immunol Methods . 1993;160:81–88
  30. Maehara Y , Anai H , Tamada R , Sugimachi K . The ATP assay is more sensitive than the succinate dehydrogenase inhibition test for predicting cell viability . Eur J Cancer Clin Oncol . 1987;23:273–276
  31. Wataha JC , Lockwood PE , Bouillaguet S , Noda M . In vitro biological response to core and flowable dental restorative materials . Dent Mater . 2003;19:25–31
  32. Riberio DA , Duarte MAH , Matsumoto MA , Marque MEA , Salvadori DMF . Biocompatibility in vitro tests of mineral trioxide aggregate and regular and white Portland cements . J Endod . 2005;31:605–607
  33. Boland EJ , MacDougall M , Carnes DL , Dickens SH . In vitro cytotoxicity of a remineralizing resin-based calcium phosphate cement . Dent Mater . 2006;22:338–345
  34. Thonemann B , Schmalz G , Hiller KA , Schweikl H . Responses of L929 mouse fibroblasts, primary and immortalized bovine dental papilla-derived cell lines to dental resin components . Dent Mater . 2002;18:318–323
  35. Yoshii E . Cytotoxic effects of acrylates and methacrylates: relationships of monomer structures and cytotoxicity . J Biomed Mater Res . 1997;37:517–524
  36. Costa CA , Vaerten MA , Edwards CA , Hanks CT . Cytotoxic effects of current dental adhesive systems on immortalized odontoblast cell line MDPC-23 . Dent Mater . 1999;15:434–441
  37. Szep S , Kunkel A , Ronge K , Heidemann D . Cytotoxicity of modern dentin adhesives—in vitro testing on gingival fibroblasts . J Biomed Mater Res . 2002;63:53–60
  38. Huang FM , Chang YC . Cytotoxicity of dentin-bonding agents on human pulp cells in vitro . Int Endod J . 2002;35:905–909

PII: S1991-7902(09)60024-6

doi:10.1016/S1991-7902(09)60024-6

Journal of Dental Sciences
Volume 4, Issue 4 , Pages 178-186, December 2009

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