Volume 4, Issue 4, Pages 178-186 (December 2009)

3 of 10

ABSTRACT

ABSTRACT + REFS

FULL-TEXT PDF (170 KB)

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

Received 11 July 2009; accepted 16 October 2009.

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

No full text is available. To read the body of this article, please view the PDF online.

Article Outline

• Abstract

• References

• Copyright

References

1. 1 Hensten-Pettersen A , Jacobsen N . Toxic effects of dental materials . Int Dent J . 1991;41:265–273 . MEDLINE

2. 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 . MEDLINE | CrossRef

3. 3 Lefebvre CA , Schuster GS . Biocompatibility of visible lightcured resin systems in prosthodontics . J Prosthet Dent . 1994;71:178–185 . Abstract | Full-Text PDF (842 KB) | CrossRef

4. 4 Wiltshire WA , Ferreira MR , Ligthelm AJ . Allergies to dental materials . Quintessence Int . 1996;27:513–520 . MEDLINE

5. 5 Chigira H , Manabe A , Hasegawa T , et al. Efficacy of various commercial dentin bonding systems . Dent Mater . 1994;10:363–368 . MEDLINE | CrossRef

6. 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 . Abstract | Full Text | Full-Text PDF (440 KB) | CrossRef

7. 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. 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 . Abstract | Full Text | Full-Text PDF (236 KB) | CrossRef

9. 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 . Abstract | Full-Text PDF (2260 KB) | CrossRef

10. 10 Vajrabhaya LO , Pasauk A , Harnirattisal C . Cytotoxicity evaluation of single component dentin bonding agent . Oper Dent . 2003;28:440–444 . MEDLINE

11. 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 . MEDLINE

12. 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 . MEDLINE | CrossRef

13. 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 . Abstract | Full Text | Full-Text PDF (160 KB) | CrossRef

14. 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 . MEDLINE | CrossRef

15. 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 . MEDLINE

16. 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. 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. 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 . Abstract | Full Text | Full-Text PDF (237 KB) | CrossRef

19. 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 . MEDLINE | CrossRef

20. 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 . Abstract | Full Text | Full-Text PDF (200 KB) | CrossRef

21. 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 . MEDLINE | CrossRef

22. 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] . MEDLINE

23. 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 . MEDLINE | CrossRef

24. 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 . MEDLINE | CrossRef

25. 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 . MEDLINE

26. 26 Dorn BR , Leung KP , Fox AP . Invasion of human oral epithelial cells by Prevotella intermedia . Infect Immun . 1998;66:6054–6057 . MEDLINE

27. 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] . MEDLINE

28. 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. 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 . MEDLINE | CrossRef

30. 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 . MEDLINE | CrossRef

31. 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 . Abstract | Full Text | Full-Text PDF (234 KB) | CrossRef

32. 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 . Abstract | Full Text | Full-Text PDF (185 KB) | CrossRef

33. 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 . Abstract | Full Text | Full-Text PDF (208 KB) | CrossRef

34. 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 . Abstract | Full Text | Full-Text PDF (105 KB) | CrossRef

35. 35 Yoshii E . Cytotoxic effects of acrylates and methacrylates: relationships of monomer structures and cytotoxicity . J Biomed Mater Res . 1997;37:517–524 . MEDLINE | CrossRef

36. 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 . Abstract | Full Text | Full-Text PDF (1329 KB) | CrossRef

37. 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 . MEDLINE | CrossRef

38. 38 Huang FM , Chang YC . Cytotoxicity of dentin-bonding agents on human pulp cells in vitro . Int Endod J . 2002;35:905–909 . MEDLINE | CrossRef

a School of Dentistry, China Medical University, Taichung, Taiwan

b Department of Dentistry, China Medical University Hospital, Taichung, Taiwan

c Institute of Environmental Health, China Medical University College of Public Health, Taichung, Taiwan

d Biostatistics Center, China Medical University, Taichung, Taiwan

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

PII: S1991-7902(09)60024-6

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

3 of 10