Human Dental Pulp Stem Cells Exhibit Different Biological Behaviours in Response to Commercial Bleaching Products
Abstract
:1. Introduction
2. Materials and Methods
2.1. Specimen Selection and Preparation
2.2. Diffusion Evaluation
2.3. Biological Assays
2.3.1. Isolation and Culture of hDPSCs
2.3.2. Characterisation Assay
2.3.3. Conditioned Medium
2.3.4. Cell Viability Assay
2.3.5. Cell Migration
2.3.6. Cell Morphology in Presence of Extracts
2.3.7. Analysis of the Expression of Mesenchymal Stem Cell Surface Markers on hDPSCs Exposed to Bleaching Products Using Flow Cytometry
2.3.8. Detection of Apoptosis and Necrosis Using Flow Cytometry (Annexin-V/7-AAD Staining)
2.4. Statistical Analysis
3. Results
3.1. HP/CP Diffusion
3.2. Isolation and Characterisation of hDPSCs
3.3. MTT Assays
3.4. Cell Migration
3.5. Cell Morphology
3.6. Analysis of the Expression of Mesenchymal Stem Cell Surface Markers on hDPSCs Exposed to Bleaching Extracts Using Flow Cytometry
3.7. Apoptosis/Necrosis of hDPSCs in the Presence of Bleaching Extracts
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
- Cohen, S.C. Human pulpal response to bleaching procedures on vital teeth. J. Endod. 1979, 5, 134–138. [Google Scholar] [CrossRef]
- Pugh, G., Jr.; Zaidel, L.; Lin, N.; Stranick, M.; Bagley, D. High levels of hydrogen peroxide in overnight tooth-whitening formulas: Effects on enamel and pulp. J. Esthet. Restor. Dent. 2005, 17, 40–45, discussion 46–47. [Google Scholar] [PubMed]
- Kwon, S.R.; Li, Y.; Oyoyo, U.; Aprecio, R.M. Dynamic model of hydrogen peroxide diffusion kinetics into the pulp cavity. J. Contemp. Dent. Pract. 2012, 13, 440–445. [Google Scholar] [PubMed]
- Azer, S.S.; Machado, C.; Sanchez, E.; Rashid, R. Effect of home bleaching systems on enamel nanohardness and elastic modulus. J. Dent. 2009, 37, 185–190. [Google Scholar] [CrossRef] [PubMed]
- Sulieman, M. An overview of bleaching techniques: I. History, chemistry, safety and legal aspects. Dent. Update 2004, 31, 608–610, 612–604, 616. [Google Scholar] [CrossRef] [PubMed]
- De Oliveira Duque, C.C.; Soares, D.G.; Basso, F.G.; Hebling, J.; de Souza Costa, C.A. Influence of enamel/dentin thickness on the toxic and esthetic effects of experimental in-office bleaching protocols. Clin. Oral Investig. 2017, 21, 2509–2520. [Google Scholar] [CrossRef] [PubMed]
- Soares, D.G.; Basso, F.G.; Hebling, J.; de Souza Costa, C.A. Immediate and late analysis of dental pulp stem cells viability after indirect exposition to alternative in-office bleaching strategies. Clin. Oral Investig. 2015, 19, 1013–1020. [Google Scholar] [CrossRef] [PubMed]
- Ferreira, V.G.; Nabeshima, C.K.; Marques, M.M.; Paris, A.F.; Gioso, M.A.; dos Reis, R.S.; Machado, M.E. Tooth bleaching induces changes in the vascular permeability of rat incisor pulps. Am. J. Dent. 2013, 26, 298–300. [Google Scholar] [PubMed]
- Lima, A.F.; Marques, M.R.; Soares, D.G.; Hebling, J.; Marchi, G.M.; de Souza Costa, C.A. Antioxidant therapy enhances pulpal healing in bleached teeth. Restor. Dent. Endod. 2016, 41, 44–54. [Google Scholar] [CrossRef] [PubMed]
- Costa, C.A.; Riehl, H.; Kina, J.F.; Sacono, N.T.; Hebling, J. Human pulp responses to in-office tooth bleaching. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 2010, 109, e59–e64. [Google Scholar] [CrossRef] [PubMed]
- Cintra, L.T.A.; Ferreira, L.L.; Benetti, F.; Gastelum, A.A.; Gomes-Filho, J.E.; Ervolino, E.; Briso, A.L.F. The effect of dental bleaching on pulpal tissue response in a diabetic animal model. Int. Endod. J. 2017, 50, 790–798. [Google Scholar] [CrossRef] [PubMed]
- Ferreira, L.L.; Gomes-Filho, J.E.; Benetti, F.; Carminatti, M.; Ervolino, E.; Briso, A.L.F.; Cintra, L.T.A. The effect of dental bleaching on pulpal tissue response in a diabetic animal model: A study of immunoregulatory cytokines. Int. Endod. J. 2018, 51, 347–356. [Google Scholar] [CrossRef] [PubMed]
- Rodríguez-Lozano, F.J.; Insausti, C.L.; Iniesta, F.; Blanquer, M.; Ramírez, M.D.; Meseguer, L.; Meseguer-Henarejos, A.B.; Marín, N.; Martínez, S.; Moraleda, J.M. Mesenchymal dental stem cells in regenerative dentistry. Med. Oral Patol. Oral Cir. Bucal 2012, 17, e1062–e1067. [Google Scholar] [CrossRef] [PubMed]
- Pisciotta, A.; Bertoni, L.; Riccio, M.; Mapelli, J.; Bigiani, A.; La Noce, M.; Orciani, M.; de Pol, A.; Carnevale, G. Use of a 3D Floating Sphere Culture System to Maintain the Neural Crest-Related Properties of Human Dental Pulp Stem Cells. Front. Physiol. 2018, 9, 547. [Google Scholar] [CrossRef] [PubMed]
- Cintra, L.T.; Benetti, F.; Ferreira, L.L.; Rahal, V.; Ervolino, E.; Jacinto Rde, C.; Gomes Filho, J.E.; Briso, A.L. Evaluation of an experimental rat model for comparative studies of bleaching agents. J. Appl. Oral Sci. 2016, 24, 171–180. [Google Scholar] [CrossRef] [PubMed]
- De Almeida, L.C.; Soares, D.G.; Gallinari, M.O.; de Souza Costa, C.A.; Dos Santos, P.H.; Briso, A.L. Color alteration, hydrogen peroxide diffusion, and cytotoxicity caused by in-office bleaching protocols. Clin. Oral Investig. 2015, 19, 673–680. [Google Scholar] [CrossRef] [PubMed]
- Dias Ribeiro, A.P.; Sacono, N.T.; Lessa, F.C.; Nogueira, I.; Coldebella, C.R.; Hebling, J.; de Souza Costa, C.A. Cytotoxic effect of a 35% hydrogen peroxide bleaching gel on odontoblast-like mdpc-23 cells. Oral Surg. Oral Med. Oral Pathol. Oral Radiol. Endod. 2009, 108, 458–464. [Google Scholar] [CrossRef] [PubMed]
- Hanks, C.T.; Fat, J.C.; Wataha, J.C.; Corcoran, J.F. Cytotoxicity and dentin permeability of carbamide peroxide and hydrogen peroxide vital bleaching materials, in vitro. J. Dent. Res. 1993, 72, 931–938. [Google Scholar] [CrossRef] [PubMed]
- Kinomoto, Y.; Carnes, D.L., Jr.; Ebisu, S. Cytotoxicity of intracanal bleaching agents on periodontal ligament cells in vitro. J. Endod. 2001, 27, 574–577. [Google Scholar] [CrossRef] [PubMed]
- Peters, O.A. Research that matters—Biocompatibility and cytotoxicity screening. Int. Endod. J. 2013, 46, 195–197. [Google Scholar] [CrossRef] [PubMed]
- Tomas-Catala, C.J.; Collado-Gonzalez, M.; Garcia-Bernal, D.; Onate-Sanchez, R.E.; Forner, L.; Llena, C.; Lozano, A.; Moraleda, J.M.; Rodriguez-Lozano, F.J. Biocompatibility of new pulp-capping materials neomta plus, mta repair hp, and biodentine on human dental pulp stem cells. J. Endod. 2018, 44, 126–132. [Google Scholar] [CrossRef] [PubMed]
- Marson, F.C.; Sensi, L.G.; Vieira, L.C.; Araujo, E. Clinical evaluation of in-office dental bleaching treatments with and without the use of light-activation sources. Oper. Dent. 2008, 33, 15–22. [Google Scholar] [CrossRef] [PubMed]
- Soares, D.G.; Ribeiro, A.P.; Lima, A.F.; Sacono, N.T.; Hebling, J.; de Souza Costa, C.A. Effect of fluoride-treated enamel on indirect cytotoxicity of a 16% carbamide peroxide bleaching gel to pulp cells. Braz. Dent. J. 2013, 24, 121–127. [Google Scholar] [CrossRef] [PubMed]
- Barja, G. The quantitative measurement of H2O2 generation in isolated mitochondria. J. Bioenerg. Biomembr. 2002, 34, 227–233. [Google Scholar] [CrossRef] [PubMed]
- Collado-Gonzalez, M.; Pecci-Lloret, M.R.; Tomas-Catala, C.J.; Garcia-Bernal, D.; Onate-Sanchez, R.E.; Llena, C.; Forner, L.; Rosa, V.; Rodriguez-Lozano, F.J. Thermo-setting glass ionomer cements promote variable biological responses of human dental pulp stem cells. Dent. Mater. 2018, 34, 932–943. [Google Scholar] [CrossRef] [PubMed]
- Dominici, M.; Le Blanc, K.; Mueller, I.; Slaper-Cortenbach, I.; Marini, F.; Krause, D.; Deans, R.; Keating, A.; Prockop, D.; Horwitz, E. Minimal criteria for defining multipotent mesenchymal stromal cells. The international society for cellular therapy position statement. Cytotherapy 2006, 8, 315–317. [Google Scholar] [CrossRef] [PubMed]
- Cavalcanti, B.N.; Rode, S.M.; Marques, M.M. Cytotoxicity of substances leached or dissolved from pulp capping materials. Int. Endod. J. 2005, 38, 505–509. [Google Scholar] [CrossRef] [PubMed]
- International Organization for Standardization. 10993-12. Biological evaluation of Medical Devices—Part 12: Sample Preparation and Reference Materials; International Organization for Standardization: Geneva, Switzerland, 2007. [Google Scholar]
- De Almeida, L.C.; Soares, D.G.; Azevedo, F.A.; Gallinari Mde, O.; Costa, C.A.; dos Santos, P.H.; Briso, A.L. At-home bleaching: Color alteration, hydrogen peroxide diffusion and cytotoxicity. Braz. Dent. J. 2015, 26, 378–383. [Google Scholar] [CrossRef] [PubMed]
- Jha, N.; Ryu, J.J.; Choi, E.H.; Kaushik, N.K. Generation and role of reactive oxygen and nitrogen species induced by plasma, lasers, chemical agents, and other systems in dentistry. Oxid. Med. Cell. Longev. 2017, 2017, 7542540. [Google Scholar] [CrossRef] [PubMed]
- Benetti, F.; Gomes-Filho, J.E.; Ferreira, L.L.; Ervolino, E.; Briso, A.L.F.; Sivieri-Araujo, G.; Dezan-Junior, E.; Cintra, L.T.A. Hydrogen peroxide induces cell proliferation and apoptosis in pulp of rats after dental bleaching in vivo: Effects of the dental bleaching in pulp. Arch. Oral Biol. 2017, 81, 103–109. [Google Scholar] [CrossRef] [PubMed]
- Vaz, M.M.; Lopes, L.G.; Cardoso, P.C.; Souza, J.B.; Batista, A.C.; Costa, N.L.; Torres, E.M.; Estrela, C. Inflammatory response of human dental pulp to at-home and in-office tooth bleaching. J. Appl. Oral Sci. 2016, 24, 509–517. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Rodríguez-Lozano, F.J.; Bueno, C.; Insausti, C.L.; Meseguer, L.; Ramírez, M.C.; Blanquer, M.; Marín, N.; Martínez, S.; Moraleda, J.M. Mesenchymal stem cells derived from dental tissues. Int. Endod. J. 2011, 44, 800–806. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wu, T.T.; Li, L.F.; Du, R.; Jiang, L.; Zhu, Y.Q. Hydrogen peroxide induces apoptosis in human dental pulp cells via caspase-9 dependent pathway. J. Endod. 2013, 39, 1151–1155. [Google Scholar] [CrossRef] [PubMed]
Bleaching Products | Applications | Diffusion (mM) |
---|---|---|
Pola Office | 1 × 30 | 38.4 ± 8.2 |
Pola Office | 3 × 15 | 39.3 ± 1.2 |
Perfect Bleach | 1 × 30 | 28.2 ± 6.5 |
Perfect Bleach | 3 × 15 | 48.6 ± 1.7 |
Polanight | 1 × 90 | 27.7 ± 0.4 |
© 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Llena, C.; Collado-González, M.; Tomás-Catalá, C.J.; García-Bernal, D.; Oñate-Sánchez, R.E.; Rodríguez-Lozano, F.J.; Forner, L. Human Dental Pulp Stem Cells Exhibit Different Biological Behaviours in Response to Commercial Bleaching Products. Materials 2018, 11, 1098. https://doi.org/10.3390/ma11071098
Llena C, Collado-González M, Tomás-Catalá CJ, García-Bernal D, Oñate-Sánchez RE, Rodríguez-Lozano FJ, Forner L. Human Dental Pulp Stem Cells Exhibit Different Biological Behaviours in Response to Commercial Bleaching Products. Materials. 2018; 11(7):1098. https://doi.org/10.3390/ma11071098
Chicago/Turabian StyleLlena, Carmen, Mar Collado-González, Christopher Joseph Tomás-Catalá, David García-Bernal, Ricardo Elías Oñate-Sánchez, Francisco Javier Rodríguez-Lozano, and Leopoldo Forner. 2018. "Human Dental Pulp Stem Cells Exhibit Different Biological Behaviours in Response to Commercial Bleaching Products" Materials 11, no. 7: 1098. https://doi.org/10.3390/ma11071098