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REAL–TIME CELL ANALYSIS OF CYTOTOXICITY OF DENTAL BONDING AGENTS ON HUMAN DENTAL PULP CELLS

Year 2020, Volume: 30 Issue: 2, 219 - 225, 15.04.2020
https://doi.org/10.17567/ataunidfd.658109

Abstract

Aim: To compare the cytotoxicity of dentin bonding agents on dental pulp cells through RTCA XCELLigence® impedance method, in order to investigate whether the self-etch adhesive would be more cytotoxic than two-step etch-and-rinse adhesive due to its higher acidity.
Materials and Methods: Human dental pulp cells were obtained from healthy third molars extracted during routine clinical treatment. Two self-etching(Tokuyama Bond Force“Tb”;Shofu Beauti Bond“Bb”), and two total-etching(Dentsply Prime&Bond NT“Pb”;3M Adper Single Bond“Ab”) dentin-bonding agents(DBA) were diluted serially with culture medium at a ratio of 1:250, 1:500, 1:1000(v:v) and filtered. Cytotoxicity was identified by plating the pulp cells into the disposable E-plate 96 of xCELLigence® system.
Results: Except for the 1:1000 dilution of Ab, each dilution of Ab was lower than the cell index of the control group at the 24th, and 48th hours. Furthermore, multiple comparison tests showed statistically significant differences between the control and each dilution of Ab (P = 0.000), except 1:1000 dilution of Ab at the 24th hour (P = 0.126). On the other hand, the cell indexes of 1:1000 dilution of Bb were higher than the cell index of the control at 24th and 48th hours, and there were statistically significant differences between them (P =0,000). At the 48th hour, 1:500 (P = 0.179) and 1:250 (P = 0.003) dilutions of Bb were lower than the control.
Conclusion: Among the DBAs we used in our study, total-etch Ab showed the most cytotoxic effect. Conversely, self-etch Bb containing phosphoric acid monomer, showed the least cytotoxic effect. So, it may be more appropriate for clinical practices.
Clinical relevance: Despite manufacturers’ claims, not all dentin bonding agents are biocompatible. Two-step etch-and-rinse adhesive may be less biocompatible than self-etch adhesive.
Keywords: Human dental pulp cell, cell culture, cytotoxicity, dental bonding agent, xCELLigence® system.

References

  • 1. Cox CF, Suzuki S. Re-evaluating pulp protection: calcium hydroxide liners vs cohesive hybridization. J Am Dent Assoc 1994;125(7):823–831.
  • 2. Brannstrom M, Nyborg H. Pulp reaction to composite resin restorations. J Prosthet Dent 1972;27(2):181–189.
  • 3. Hilton TJ, Ferracane JL, Mancl L, Baltuck C, Barnes C, Baudry D, Shao J, Lubisich E, Gilbert A, Lowder L. Comparison of CaOH with MTA for direct pulp capping: a PBRN randomized clinical trial. J Dent Res 2013;92(7):16-22.
  • 4. Matsuo T, Nakanishi T, Shimizu H, Ebisu S. A clinical study of direct pulp capping applied to carious exposed pulps. J Endod 1996;22(10):551-556.
  • 5. Murray PE, Hafez A, Smith AJ, Cox CF. Hierarchy of pulp capping and repair activities responsible for dentin bridge formation. Am J Dent 2002;15(4):236-243.
  • 6. Schröder U. Effect of calcium hydroxide-containing pulp-capping agents on pulp cell migration, proliferation and differentiation. J Dent Res 1985:64;541-548.
  • 7. Tziafas D. Mechanisms controlling secondary initiation of dentinogenesis: a review. Int Endod J 1994;27(2):61-74.
  • 8. Bergenholtz G. Evidence for bacterial causation of adverse pulpal responses in resin-based dental restorations. Crit Rev Oral Biol Med 2000;11(4):467- 480.
  • 9. 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(3):223-229.
  • 10. Yevenes I, Baltra MO, Urzua I, Reyes J, Petrasic L. Chemical stability of two dentin single-bottle adhesives as a function solvent loss. Rev Odonto Cienc 2008;23:220-224.
  • 11. Eick JD, Gwinnett AJ, Pashley DH, Robinson SJ. Current concepts on adhesion to dentin. Crit Rev Oral Biol Med 1997;8(3):306-335.
  • 12. Banava S, Najibfard K, Garcia-Godoy F, Saghiri MA, Ghahremani MH, Ostad N. Impact of dilution and polymerization on cytotoxicity of dentin adhesives to human gingival fibroblasts: early exposure time. J Dent Res Dent Clin Dent Prospect 2015;9(3):151-158.
  • 13. Chang YC, Huang FM, ChengMH, Chou LSS, ChouMY. In vitro evaluation of the cytotoxicity and genotoxicity of root canal medicines on human pulp fibroblasts. J Endod 1998;24(9):604-606.
  • 14. Huang FM, Chang YC. Cytotoxicity of dentine-bonding agents on human pulp cells in vitro. Int Endod J 2002;35(11):905-909.
  • 15. Yalcin M, Barutcigil C, Umar I, Bozkurt BS, Hakki SS. Cytotoxicity of hemostatic agents on the human gingival fibroblasts. Eur Rev Med Pharmacol Sci 2013;17(7):984-988.
  • 16. Yoshida Y, Van meerbeeK B, Nakayama Y, Snauwaert J, Hellemans I, Lambrechts P, Vanherle H, Wasaka K. Evidence of chemical bonding at biomaterial-hard tissue interfaces. J Dent Res 2000;79(2):709-714.
  • 17. Olmez A, Oztas N, Basak F, Sabuncuoglu B. A histopathologic study of direct pulp-capping with adhesive resins. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 1998;86(1):98-103.
  • 18. Costa CA, Oliveira MF, Giro EM, Hebling J. Biocompatibility of resin-based materials used as pulp-capping agents. Int Endod J 2003;36(12):831-839.
  • 19. Pereira JC, Segala AD, Costa CA. Human pulpal response to direct pulp capping with an adhesive system. Am J Dent 2000;13(3):139-147.
  • 20. Koliniotou-Koumpia E, Tziafas D. Pulpal responses following direct pulp capping of healthy dog teeth with dentine adhesive systems. J Dent 2005;33(8):639-647.
  • 21. Ke N, Wang X, Xu X, Abassi YA. The xCELLigence system for real-time and label-free monitoring of cell viability. Methods Mol Biol 2011;740;33-43.
  • 22. Limame R, Wouters A, Pauwels B, Fransen E, Peeters M, Lardon F, De Wever O, Pauwels P. Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays. Plos One 2012;7(10):e46536.
  • 23. Atienza JM, Yu N, Kirstein SL, Xi B, Wang X, Xu X, Abassi YA. Dynamic and label-free cell-based assays using the real-time cell electronic sensing system. Assay Drug Dev Technol 2006;4(5):597–607.
  • 24. Costa CAS, Vaerten MA, Edwards CA, Hanks CT. Cytotoxic effects of current dental adhesive systems on immortalized odontoblast cell line MDPC-23. Dent Mater 1999;15(6):434–441.
  • 25. Vajrabhaya LO, Pasasuk A, Harnirattisai C. Cytotoxicity evaluation of single component dentin bonding agents. Oper Dent 2003;28(4):440-444.
  • 26. Pashley DH, Matthews WG. The effects of outward forced convective flow on inward diffusion in human dentine in vitro. Arch Oral Biol 1993;38(7):577-582.
  • 27. Noda M, Wataha JC, Kaga M, Lockwooh PE, Volkmann KR, Sano H. Component of dentinal adhesives modulate heat shock protein 72 expression in heat stressed TPH-1 human monocytes at sublethal concentrations. J Dent Res 2002;81(4):265-269.
  • 28. Spagnuolo G, D’Anto V, Cosentino C, Schmalz G, Schweikl H, Rengo S. Effect of N-acetyl-L- cysteine on ROS production and cell death caused by HEMA in human primary gingival fibroblasts. Biomaterials 2006;27(9):1803-1809.
  • 29. Spagnuolo G, D’Anto V, Valletta R, Strisciuglio C, Schmalz G, Schweikl H, Rengo S. Effect of 2-hydroxyethyl methacrylate on human pulp cell survival pathways ERK and AKT. J Endod 2008;34(6):684-688.
  • 30. Geurtsen W, Leyhausen G. Chemical- biological interactions of the resin monomer triethyleneglycol-dimethacrylate (TEDGMA). J Dent Res 2001;80(12):2046-2050.
  • 31. Zorba YO, Bayındır YZ, Yıldız M. Geçmişten günümüze dentin bonding ajanlar. Atatürk Üniv Dis Hek Fak Derg Cilt: 14, Sayı:3, Sayfa:70-78, 2004
Year 2020, Volume: 30 Issue: 2, 219 - 225, 15.04.2020
https://doi.org/10.17567/ataunidfd.658109

Abstract

References

  • 1. Cox CF, Suzuki S. Re-evaluating pulp protection: calcium hydroxide liners vs cohesive hybridization. J Am Dent Assoc 1994;125(7):823–831.
  • 2. Brannstrom M, Nyborg H. Pulp reaction to composite resin restorations. J Prosthet Dent 1972;27(2):181–189.
  • 3. Hilton TJ, Ferracane JL, Mancl L, Baltuck C, Barnes C, Baudry D, Shao J, Lubisich E, Gilbert A, Lowder L. Comparison of CaOH with MTA for direct pulp capping: a PBRN randomized clinical trial. J Dent Res 2013;92(7):16-22.
  • 4. Matsuo T, Nakanishi T, Shimizu H, Ebisu S. A clinical study of direct pulp capping applied to carious exposed pulps. J Endod 1996;22(10):551-556.
  • 5. Murray PE, Hafez A, Smith AJ, Cox CF. Hierarchy of pulp capping and repair activities responsible for dentin bridge formation. Am J Dent 2002;15(4):236-243.
  • 6. Schröder U. Effect of calcium hydroxide-containing pulp-capping agents on pulp cell migration, proliferation and differentiation. J Dent Res 1985:64;541-548.
  • 7. Tziafas D. Mechanisms controlling secondary initiation of dentinogenesis: a review. Int Endod J 1994;27(2):61-74.
  • 8. Bergenholtz G. Evidence for bacterial causation of adverse pulpal responses in resin-based dental restorations. Crit Rev Oral Biol Med 2000;11(4):467- 480.
  • 9. 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(3):223-229.
  • 10. Yevenes I, Baltra MO, Urzua I, Reyes J, Petrasic L. Chemical stability of two dentin single-bottle adhesives as a function solvent loss. Rev Odonto Cienc 2008;23:220-224.
  • 11. Eick JD, Gwinnett AJ, Pashley DH, Robinson SJ. Current concepts on adhesion to dentin. Crit Rev Oral Biol Med 1997;8(3):306-335.
  • 12. Banava S, Najibfard K, Garcia-Godoy F, Saghiri MA, Ghahremani MH, Ostad N. Impact of dilution and polymerization on cytotoxicity of dentin adhesives to human gingival fibroblasts: early exposure time. J Dent Res Dent Clin Dent Prospect 2015;9(3):151-158.
  • 13. Chang YC, Huang FM, ChengMH, Chou LSS, ChouMY. In vitro evaluation of the cytotoxicity and genotoxicity of root canal medicines on human pulp fibroblasts. J Endod 1998;24(9):604-606.
  • 14. Huang FM, Chang YC. Cytotoxicity of dentine-bonding agents on human pulp cells in vitro. Int Endod J 2002;35(11):905-909.
  • 15. Yalcin M, Barutcigil C, Umar I, Bozkurt BS, Hakki SS. Cytotoxicity of hemostatic agents on the human gingival fibroblasts. Eur Rev Med Pharmacol Sci 2013;17(7):984-988.
  • 16. Yoshida Y, Van meerbeeK B, Nakayama Y, Snauwaert J, Hellemans I, Lambrechts P, Vanherle H, Wasaka K. Evidence of chemical bonding at biomaterial-hard tissue interfaces. J Dent Res 2000;79(2):709-714.
  • 17. Olmez A, Oztas N, Basak F, Sabuncuoglu B. A histopathologic study of direct pulp-capping with adhesive resins. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 1998;86(1):98-103.
  • 18. Costa CA, Oliveira MF, Giro EM, Hebling J. Biocompatibility of resin-based materials used as pulp-capping agents. Int Endod J 2003;36(12):831-839.
  • 19. Pereira JC, Segala AD, Costa CA. Human pulpal response to direct pulp capping with an adhesive system. Am J Dent 2000;13(3):139-147.
  • 20. Koliniotou-Koumpia E, Tziafas D. Pulpal responses following direct pulp capping of healthy dog teeth with dentine adhesive systems. J Dent 2005;33(8):639-647.
  • 21. Ke N, Wang X, Xu X, Abassi YA. The xCELLigence system for real-time and label-free monitoring of cell viability. Methods Mol Biol 2011;740;33-43.
  • 22. Limame R, Wouters A, Pauwels B, Fransen E, Peeters M, Lardon F, De Wever O, Pauwels P. Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays. Plos One 2012;7(10):e46536.
  • 23. Atienza JM, Yu N, Kirstein SL, Xi B, Wang X, Xu X, Abassi YA. Dynamic and label-free cell-based assays using the real-time cell electronic sensing system. Assay Drug Dev Technol 2006;4(5):597–607.
  • 24. Costa CAS, Vaerten MA, Edwards CA, Hanks CT. Cytotoxic effects of current dental adhesive systems on immortalized odontoblast cell line MDPC-23. Dent Mater 1999;15(6):434–441.
  • 25. Vajrabhaya LO, Pasasuk A, Harnirattisai C. Cytotoxicity evaluation of single component dentin bonding agents. Oper Dent 2003;28(4):440-444.
  • 26. Pashley DH, Matthews WG. The effects of outward forced convective flow on inward diffusion in human dentine in vitro. Arch Oral Biol 1993;38(7):577-582.
  • 27. Noda M, Wataha JC, Kaga M, Lockwooh PE, Volkmann KR, Sano H. Component of dentinal adhesives modulate heat shock protein 72 expression in heat stressed TPH-1 human monocytes at sublethal concentrations. J Dent Res 2002;81(4):265-269.
  • 28. Spagnuolo G, D’Anto V, Cosentino C, Schmalz G, Schweikl H, Rengo S. Effect of N-acetyl-L- cysteine on ROS production and cell death caused by HEMA in human primary gingival fibroblasts. Biomaterials 2006;27(9):1803-1809.
  • 29. Spagnuolo G, D’Anto V, Valletta R, Strisciuglio C, Schmalz G, Schweikl H, Rengo S. Effect of 2-hydroxyethyl methacrylate on human pulp cell survival pathways ERK and AKT. J Endod 2008;34(6):684-688.
  • 30. Geurtsen W, Leyhausen G. Chemical- biological interactions of the resin monomer triethyleneglycol-dimethacrylate (TEDGMA). J Dent Res 2001;80(12):2046-2050.
  • 31. Zorba YO, Bayındır YZ, Yıldız M. Geçmişten günümüze dentin bonding ajanlar. Atatürk Üniv Dis Hek Fak Derg Cilt: 14, Sayı:3, Sayfa:70-78, 2004
There are 31 citations in total.

Details

Primary Language English
Subjects Dentistry
Journal Section Araştırma Makalesi
Authors

Seyda Ersahan

Elif Aybala Oktay This is me

Fidan Alakus Sabuncuoglu This is me

Hüseyin Tort This is me

Publication Date April 15, 2020
Published in Issue Year 2020 Volume: 30 Issue: 2

Cite

APA Ersahan, S., Oktay, E. A., Alakus Sabuncuoglu, F., Tort, H. (2020). REAL–TIME CELL ANALYSIS OF CYTOTOXICITY OF DENTAL BONDING AGENTS ON HUMAN DENTAL PULP CELLS. Atatürk Üniversitesi Diş Hekimliği Fakültesi Dergisi, 30(2), 219-225. https://doi.org/10.17567/ataunidfd.658109
AMA Ersahan S, Oktay EA, Alakus Sabuncuoglu F, Tort H. REAL–TIME CELL ANALYSIS OF CYTOTOXICITY OF DENTAL BONDING AGENTS ON HUMAN DENTAL PULP CELLS. Ata Diş Hek Fak Derg. April 2020;30(2):219-225. doi:10.17567/ataunidfd.658109
Chicago Ersahan, Seyda, Elif Aybala Oktay, Fidan Alakus Sabuncuoglu, and Hüseyin Tort. “REAL–TIME CELL ANALYSIS OF CYTOTOXICITY OF DENTAL BONDING AGENTS ON HUMAN DENTAL PULP CELLS”. Atatürk Üniversitesi Diş Hekimliği Fakültesi Dergisi 30, no. 2 (April 2020): 219-25. https://doi.org/10.17567/ataunidfd.658109.
EndNote Ersahan S, Oktay EA, Alakus Sabuncuoglu F, Tort H (April 1, 2020) REAL–TIME CELL ANALYSIS OF CYTOTOXICITY OF DENTAL BONDING AGENTS ON HUMAN DENTAL PULP CELLS. Atatürk Üniversitesi Diş Hekimliği Fakültesi Dergisi 30 2 219–225.
IEEE S. Ersahan, E. A. Oktay, F. Alakus Sabuncuoglu, and H. Tort, “REAL–TIME CELL ANALYSIS OF CYTOTOXICITY OF DENTAL BONDING AGENTS ON HUMAN DENTAL PULP CELLS”, Ata Diş Hek Fak Derg, vol. 30, no. 2, pp. 219–225, 2020, doi: 10.17567/ataunidfd.658109.
ISNAD Ersahan, Seyda et al. “REAL–TIME CELL ANALYSIS OF CYTOTOXICITY OF DENTAL BONDING AGENTS ON HUMAN DENTAL PULP CELLS”. Atatürk Üniversitesi Diş Hekimliği Fakültesi Dergisi 30/2 (April 2020), 219-225. https://doi.org/10.17567/ataunidfd.658109.
JAMA Ersahan S, Oktay EA, Alakus Sabuncuoglu F, Tort H. REAL–TIME CELL ANALYSIS OF CYTOTOXICITY OF DENTAL BONDING AGENTS ON HUMAN DENTAL PULP CELLS. Ata Diş Hek Fak Derg. 2020;30:219–225.
MLA Ersahan, Seyda et al. “REAL–TIME CELL ANALYSIS OF CYTOTOXICITY OF DENTAL BONDING AGENTS ON HUMAN DENTAL PULP CELLS”. Atatürk Üniversitesi Diş Hekimliği Fakültesi Dergisi, vol. 30, no. 2, 2020, pp. 219-25, doi:10.17567/ataunidfd.658109.
Vancouver Ersahan S, Oktay EA, Alakus Sabuncuoglu F, Tort H. REAL–TIME CELL ANALYSIS OF CYTOTOXICITY OF DENTAL BONDING AGENTS ON HUMAN DENTAL PULP CELLS. Ata Diş Hek Fak Derg. 2020;30(2):219-25.

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