Cytotoxicity of seven recent dentine bonding agents on mouse 3T3 fibroblast cells

Annette Olivier, Sias R. Grobler, Yusuf Osman

Oral and Dental Research Institute, Faculty of Dentistry, University of the Western Cape, Cape Town, South Africa.

DOI: 10.4236/ojst.2012.24043                

Abstract

Today it is generally accepted that most bonding agents are cytotoxic. In this study the relative cytotoxicity of seven recent dentine bonding agents on mouse 3T3 fibroblast cells were investigated. Materials and Methods. Near-confluent mouse 3T3 fibroblast cells were exposed to Dulbecco Modified Eagle’s Medium containing extractions from the seven different bonding agents. The cell survival rate was then determined using the standard MTT assay. Results. The cell survival rate ranking is: iBond (94%) < Gbond (78%) < Xeno V (71%) < Adper Easy Bond (63%) < Xeno V+ (61%) < Adper Scotchbond SE (33%) < XP Bond (32%). Part A of Adper Scotchbond SE had a survival rate of 35% and part B 38%. These two parts did not differ significantly. Adper Scotchbond SE and XP Bond do not differ significantly. While Xeno V+, Xeno V and Adper Easy Bond do not differ. (p < 5%; Tukey-Kramer Multiple-Comparison Test). Conclusion. All of the tested adhesive bonding agents were cytotoxic with survival rate of 3T3 cells between 94% to 31%. Of the 7 bonding agents tested iBond was found to be only slightly toxic and by far the least toxic. The two bonding agents (XP Bond and Adper Scotchbond SE) containing UDMA plus TEGDMA plus HEMA plus camphorquinone were found to be the most toxic.

Keywords

Cytotoxicity; Bonding Agents; Mouse 3T3 Fibroblast

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Chen, R. S., Liu, C.C., Tseng, W. Y., Jeng , J. H. and Lin, C.P. (2003) Cytotoxicity of three dentin bonding agents on human dental pulp cells. Journal of Dentistry, 31, 223-229. doi:10.1016/S0300-5712(02)00088-X
[2]	Ratanasathien, S., Wataha, J.C., Hanks, C.T. and Dennison, J.B. (1995) Cytotoxic interactive effects of dentin bonding components on mouse fibroblasts. Journal of Dental Research, 74, 1602-1606. doi:10.1177/00220345950740091601
[3]	Tuncer, S., Demirci, M., Schweikl, H., Erguven, M., Bilir, A. and Tuncer, A.K. (2012) Inhibition of cell survival, viability and proliferation by dentin adhesives after direct and indirect exposure vitro.
[4]	Cardoso, M.V. (2011) Current aspects on bonding effecttiveness and stability in adhesive dentistry. Australian Dental Journal, 56, 31-44. doi:10.1111/j.1834-7819.2011.01294.x
[5]	Grobler, S.R., Olivier, A., Moodley, D, and van W Kotze, T.W. (2004) Cytotoxicity of two concentrations of a dentine bonding agent on mouse 3T3 and human pulp fibro-blast cell-lines. South African Dental Journal, 59, 368- 370.
[6]	Wataha, J.C. (2012) Predicting clinical biological responses to dental materials. Dental Materials, 28, 23-40. doi:10.1016/j.dental.2011.08.595
[7]	Szep, S., Kunkel, A., Ronge, K. and Heideman D. (2002) Cytotoxicity of modern dentin adhesives—in vitro testing on gingival fibroblasts. Journal of Biomedical Materials Research, 63, 53-60. doi:10.1002/jbm.10083
[8]	Hashieh, I.A., Franquin, J.C., Cosset, A. and Dejou, J. (1998) Relationship between dentin hydraulic conductance and the cytotoxicity of four dentin bonding resins in vitro. Journal of Dentisty, 26, 473-477.
[9]	Hashieh, I.A., Cosset, A., Franquin, J.C. and Camps, J. (1999). In vitro cytotoxicity of one-step dentin bonding systems. Journal of Endodontics, 25, 89-92. doi:10.1016/S0099-2399(99)80003-X
[10]	Vajrabhaya, L., Pasasuk, A. and Harnirattisai, C. (2003) Cytotoxicity evaluation of single component dentin bonding agents. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology and Endodontology, 28, 440-444. doi:10.1016/j.tripleo.2008.11.008
[11]	Rathbun, M.A., Craig, R.G., Hanks, C.T. and Filisko, F.E. (1991) Cytotoxicity of a Bis-GMA dental composite before and after leaching in organic solvents. Journal Bio-medical material Research, 25, 443-457. doi:10.1002/jbm.820250403
[12]	Geurtsen, W., Lehman, F., Spahl, W. and Leyhausen, G. (1998) Cytotoxicity of 35 dental resin composite monomers/additives in permanent 3T3 and three human primary fibroblast cultures. Journal of Biomedical Materials Research, 41, 474-480.
[13]	Schmalz, G. (1998) The biocompatibility of non-amalgam dental filling materials. European Journal of Oral Sciences, 2, 696-706. doi:10.1046/j.0909-8836.1998.eos10602ii05.x
[14]	Hanks, C.T., Strawn, S.E., Watahai, J.C. and Craig, R.G. (1991) Cytotoxic effects of resin components on cultured mammalian fibroblasts. Journal of Dental Research, 70, 1450-1455.
[15]	Trubiani, O., Caputi, S., Di Iorio, D., et al. (2010) The cytotoxic effect of resin-based sealers on dental pulp stem cells. International Endodontic Journal, 43, 664-653. doi:10.1111/j.1365-2591.2010.01729.x
[16]	Harorli, O.T., Bayindir, Y.A., ltunkaynak, Z. and Tatar, A. (2009) Cytotoxic effects of TEGDMA on THP-1 cells in vitro. Medicina Oral Patologia Oral Cirugia Bucal, 14, e489-493.
[17]	Van Landuyt, K.L., Snauwaert, J., De Munck, J., et al. (2007) Systematic review of the chemical composition of contemporary dental adhesives. Biomaterials, 28, 3757-3785. doi:10.1016/j.biomaterials.2007.04.044
[18]	Yeh, H., Chang, M., Lin, C., et al. (2009) Comparative cytotoxicity of five current dentin bonding agents: Role of cell cycle deregulation. Acta Materialia, 5, 3404-3410. doi:10.1016/j.actbio.2009.05.036
[19]	Issa, Y., Watts, D.C., Brunton, P.A., Waters, C.M. and Duxbury, A.J. (2004) Resin composite monomers alter MTT and LDH activity of human gingival fibroblasts in vitro. Dental Materials, 20, 12-20. doi:10.1016/S0109-5641(03)00053-8
[20]	Goldberg, M. (2008) In vitro and in vivo studies on the toxicity of dental resin components: A review. Clinical Investigations, 12, 1-8.
[21]	Peutzfeldt, A. (1997) Retention of propanal and diacetyl in experimental resins. Acta Odontologica Scandinavica, 55, 94-100. doi:10.3109/00016359709115399
[22]	Van Landuyt, K.L., Nawrot, T., Geebelen, B., et al. (2011) How much do resin based materials release? A meta-analytical approach. Dental Materials, 27, 723-474.
[23]	Koulaouzidou, E.A., Helvatjoglu-Antoniades, M., Palaghias, G., Karaniuka-Kouma, A. and Antoniades, D. (2009) Cytotoxicity of dental materials in vitro. European Journal of Dentistry, 3, 3-9.
[24]	Grobler, S.R., Basson, N.J. and Rossouw, R.J. (1996) Shear bond strength, microleakage and antimicrobial properties of Aelitebond. American Journal of Dentisty, 9, 120-124.
[25]	Grobler, S.R., Olivier, A., Moodley, D. and Van Wyk Kotze, T.J. (2008) Cytotoxicity of recent bonding agents on mouse fibroblast cells. Quintessence International, 39, 511-516.
[26]	Mosmann, T. (1983) Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 65, 55-63. doi:10.1016/0022-1759(83)90303-4
[27]	Koulaouzidou, E.A., Konstantinos, T.P., Yiannaki, M.D. and Papazisis, G. (2009) Effects of dentin bonding agents on the cell cycle of fibroblasts. Journal of Endodontics, 35, 275-279. doi:10.1016/j.joen.2008.11.013
[28]	American National Standards Institute and American Dental Association (2000) Revision, document no. 41 for recommended standard practices for biological evaluation of dental materials.
[29]	Wataha, J.C., Hanks, C.T. and Sun, Z. (1994) Effect of cell-line on in vitro metal ion cytotoxicity. Dental Materials, 10, 156-161.
[30]	Moharamzadeh, K., Van Noort, R., Brook, I.M. and Scutt, A.M. (2007) Cytotoxicity of resin monomers on human gingival fibroblasts and HaCaT keratinocytes. Dental Materials, 23, 40-44. doi:10.1016/j.dental.2005.11.039
[31]	Koliniotou-Koubia, E., Dionysopoulos, P., Koulaouzidou, E.A., Kortsaris, A.H. and Papadogiannis, Y. (2001) In vitro cytotoxicity of six dentin bonding agents. Journal of Oral Rehabilitation, 28, 971-975. doi:10.1046/j.1365-2842.2001.00769.x
[32]	Yasuda, Y., Inuyama, H., Maeda, H., Akamine, A., N?r, J.E. and Saito, T. (2008) Cytotoxicity of one step bonding agents toward dental pulp and odontoblast-like cells. Journal of Oral Rehabilitation, 35, 940-946. doi:10.1111/j.1365-2842.2008.01885.x
[33]	Sigusch, B.W., Pflaum, T., V?lpel, A., Schinkel, M. and Jandt, K.D. (2009) The influence of various light curing units on the cytotoxicity of dental adhesives. Dental Materials, 25, 1446-1452. doi:10.1016/j.dental.2009.06.016
[34]	Atsumi, T., Murata, J., Kamiyanagi, I., Fujisawa, S. and Ueha, T. (1998) Cytotoxicity of photosensitizers camphorquinone and 9-fluorenone with visible light irradiation on a human submandibularduct cell line in vitro. Archives of Oral Biology, 43, 73-81. doi:10.1016/S0003-9969(97)00073-3
[35]	Atsumi, T., Ishihara, M., Kadoma, Y., Tonosaki, K, and Fujisawa S. (2004) Comparative radical production and cytotoxicity induced by camphorquinone and 9-fluo-renone against human pulp fibroblasts. Journal of Oral Rehabilitation, 12, 1155-1164. doi:10.1111/j.1365-2842.2004.01357.x
[36]	Pagoria, D., Lee, A. and Geurtsen, W. (2005) The effect of camphorquinone(CQ) and cQ-related photosensitizers on the generation of reactive oxygen species and the production of oxidative DNA damage. Biomaterials, 26, 4091-4099. doi:10.1016/j.biomaterials.2004.10.022
[37]	Okada, N., Muraoka, E., Fujisawa, S. and Machino, M. (2008) Effects of visible light-irradiated camphorquinone and 9-fluorenone on murine oral mucosa. Dent Mater J., 27, 809-813. doi:10.4012/dmj.27.809
[38]	Porto, I.C.C.M., Oliveira, D.C., Raele, R.A., Ribas, K.H.S., Montes, M.A.J.R. and De Castro, C.M.M.B. (2011) Cytotoxicity of current adhesive systems: In vitro testing on cell cultures of primary murine macrophages. Dental Materials, 27, 221-228.
[39]	EI-Kholany, N.R., Abielhassan, M.H., Elembaby, A.E. and Maria, O.M. (2011) Apoptotic effect of different selfetch dental adhesives on odontoblasts in cell culture. Archives of Oral Biology. doi:10.1016/j.archoralbio.2011.11.019
[40]	Vajrabhaya, L.O., Korsuwannawong, S., Bosl, C., and Schmalz, G. (2009) The cytotoxicity of self-etching primer bonding agents in vitro. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology and Endodontology, 107, e86-90. doi:10.1016/j.tripleo.2008.11.008
[41]	Ito, S., Tay, F.R., Hashimoto, M., et al. (2005) Effects of multiple coatings of two all-in-one adhesives on dentin bonding. The Journal of Adhesive Dentistry, 7, 133-141.