Mode of cell death of co-cultured dental mesenchymal stem cells following exposure to endodontic irrigating solutions
DOI:
https://doi.org/10.5195/d3000.2022.146Palabras clave:
Co-culture, Apoptosis, Necrosis, Irrigants, Cells, Cultured.Resumen
Aims: This in vitro study aimed to determine the mode of cell death of co-cultured dental mesenchymal stem cells following exposure to different irrigating solutions.
Methods and Material: Mesenchymal stem cells of the apical papilla (SCAPs) and periodontal ligament stem cells (PDLSCs) were isolated from the periapical region of a third molar of an 18-year-old patient and cultured. Adequate number of cells in each well, 6-well plate inserts were placed over the wells. The cells were then randomly exposed to 17% ethylenediaminetetraacetic acid (EDTA), 1.5% sodium hypochlorite (NaOCL), 2% chlorhexidine (CHX) and saline. After 1, 5, and 10 minutes cells were trypsinized and underwent flow cytometry. Apoptotic and necrotic cells were quantified. Data were analyzed using the One-way ANOVA and Tukey’s post-hoc test.
Results: With increasing exposure time, cell viability is decrease and the mode of cells death was necrosis in CHX and NaOCL groups and necrosis plus late apoptosis in 1 and 10 minutes in EDTA group.
Conclusion: Maximum cell death occurred following exposure to EDTA while minimum cell death occurred following exposure to CHX. Necrosis was the dominant mode of cell death in all groups.
Citas
References
Tait JF. Imaging of apoptosis. J Nucl Med. 2008;49:1573-6.
Hashemi M, Karami-Tehrani F, Ghavami S. Cytotoxicity effect of Cladribine on the MCF-7 human breast cancer cell line. Iran Biomed J. 2004;8(1):7-12.
Faria G, Celes MR, De Rossi A, Silva LAB, Silva JS, Rossi MA. Evaluation of chlorhexidine toxicity injected in the paw of mice and added to cultured l929 fibroblasts. J Endod. 2007;33:715-22.
Hashemi M, Karami‐Tehrani F, Ghavami S, Maddika S, Los M. Adenosine and deoxyadenosine induces apoptosis in oestrogen receptor‐positive and‐negative human breast cancer cells via the intrinsic pathway. Cell Prolif. 2005;38:269-85.
Zhang W, Torabinejad M, Li Y. Evaluation of cytotoxicity of MTAD using the MTT-tetrazolium method. J Endod. 2003;29:654-7.
Mollashahi NF, Saberi E, Karkehabadi H. Evaluation of cytotoxic effects of various endodontic irrigation solutions on the survival of stem cell of human apical papilla. Iran Endod J. 2016;11:293.
Barnhart BD, Chuang A, Dalle Lucca JJ, Roberts S, Liewehr F, Joyce AP. An in vitro evaluation of the cytotoxicity of various endodontic irrigants on human gingival fibroblasts. J Endod. 200531:613-5..
Kirkpatrick CJ, Fuchs S, Hermanns MI, Peters K, Unger RE. Cell culture models of higher complexity in tissue engineering and regenerative medicine. Biomaterials. 2007;28:5193-8.
Zhang W, Zhuang A, Gu P, Zhou H, Fan X. A review of the three-dimensional cell culture technique: Approaches, advantages and applications. Curr Stem Cell Res Ther. 2016;11:370-80.
Karkehabadi H, Yousefifakhr H, Zadsirjan S. Cytotoxicity of endodontic irrigants on human periodontal ligament cells. Iran Endod J. 2018;13:390.
AlKahtani A, Alkahtany SM, Mahmood A, Elsafadi MA, Aldahmash AM, Anil S. Cytotoxicity of QMix™ endodontic irrigating solution on human bone marrow mesenchymal stem cells. BMC oral health. 2014;14:27
Tu Y-Y, Yang C-Y, Chen R-S, Chen M-H. Effects of chlorhexidine on stem cells from exfoliated deciduous teeth. Taiwan Yi Xue Hui Za Zhi. 2015;114:17-22.
Bhatia SN, Yarmush ML, Toner M. Controlling cell interactions by micropatterning in co‐cultures: Hepatocytes and 3T3 fibroblasts. J Biomed Mater Res A. 1997;34:189-99.
Gerstenfeld L, Barnes G, Shea C, Einhorn T. Osteogenic differentiation is selectively promoted by morphogenetic signals from chondrocytes and synergized by a nutrient rich growth environment. Connect Tissue Res. 2003;44:85-91.
Faria G, Cardoso CR, Larson RE, Silva JS, Rossi MA. Chlorhexidine-induced apoptosis or necrosis in L929 fibroblasts: A role for endoplasmic reticulum stress. Toxicol Appl Pharmacol. 2009;234:256-65.
Essner MD, Javed A, Eleazer PD. Effect of sodium hypochlorite on human pulp cells: an in vitro study. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2011;112:662-6.
Trevino EG, Patwardhan AN, Henry MA, Perry G, Dybdal-Hargreaves N, Hargreaves KM, et al. Effect of irrigants on the survival of human stem cells of the apical papilla in a platelet-rich plasma scaffold in human root tips. J Endod. 2011;37:1109-15.
Longo JPF, Valois CA, Tapajós ÉCC, Santos MdFMA, de Azevedo RB. Cytotoxicity and genotoxicity of endodontic irrigants on human cells. Arch Oral Res. 2010;6.135-40
Mantellini M, Botero T, Yaman P, Dennison J, Hanks C, Nör J. Adhesive resin induces apoptosis and cell-cycle arrest of pulp cells. J Dent Res. 2003;82:592-6.
Trump BE, Berezesky IK, Chang SH, Phelps PC. The pathways of cell death: oncosis, apoptosis, and necrosis. Toxicol Pathol. 1997;25:82-8.
Cimpan MR, Matre R, Skaug N, Lie SA, Lygre H. The coinitiator DMABEE induces death by apoptosis and necrosis in human monoblastoid cells. Clin Oral Investig. 2005;9:168-72.
Hernandez E, Botero T, Mantellini M, McDonald N, Nör J. Effect of ProRoot® MTA mixed with chlorhexidine on apoptosis and cell cycle of fibroblasts and macrophages in vitro. Int Endod J. 2005;38:137-43.
Newton APN, Cadena SMS, Rocha MEM, Carnieri EGS, De Oliveira MBM. New data on biological effects of chlorhexidine: Fe2+ induced lipid peroxidation and mitochondrial permeability transition. Toxicol Lett. 2004;151:407-16.
Li YC, Kuan YH, Lee SS, Huang FM, Chang YC. Cytotoxicity and genotoxicity of chlorhexidine on macrophages in vitro. Environ Toxicol. 2014;29:452-8.
Giannelli M, Chellini F, Margheri M, Tonelli P, Tani A. Effect of chlorhexidine digluconate on different cell types: a molecular and ultrastructural investigation.
Toxicol In Vitro. 2008;22:308-17.
Meier P, Finch A, Evan G. Apoptosis in development. Nature. 2000;407:796.
Sakabe I, Paul S, Dansithong W, Shinozawa T. Induction of apoptosis in Neuro-2A cells by Zn2+ chelating. Cell Struct Funct. 1998;23:95-9.
Ostby N. Chelating in root canal therapy. Ethylene-diamine tetraacetic acid for cleansing and widening of root canals. Odontol Tidskr. 1957;65:3-11.
Amaral K, Rogero M, Fock R, Borelli P, Gavini G. Cytotoxicity analysis of EDTA and citric acid applied on murine resident macrophages culture. Int Endod J. 2007;40:338-43.
Descargas
Publicado
Número
Sección
Licencia
Authors who publish with this journal agree to the following terms:
- The Author retains copyright in the Work, where the term “Work” shall include all digital objects that may result in subsequent electronic publication or distribution.
- Upon acceptance of the Work, the author shall grant to the Publisher the right of first publication of the Work.
- The Author shall grant to the Publisher and its agents the nonexclusive perpetual right and license to publish, archive, and make accessible the Work in whole or in part in all forms of media now or hereafter known under a Creative Commons Attribution 4.0 International License or its equivalent, which, for the avoidance of doubt, allows others to copy, distribute, and transmit the Work under the following conditions:
- Attribution—other users must attribute the Work in the manner specified by the author as indicated on the journal Web site;
- The Author is able to enter into separate, additional contractual arrangements for the nonexclusive distribution of the journal's published version of the Work (e.g., post it to an institutional repository or publish it in a book), as long as there is provided in the document an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post online a prepublication manuscript (but not the Publisher’s final formatted PDF version of the Work) in institutional repositories or on their Websites prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work. Any such posting made before acceptance and publication of the Work shall be updated upon publication to include a reference to the Publisher-assigned DOI (Digital Object Identifier) and a link to the online abstract for the final published Work in the Journal.
- Upon Publisher’s request, the Author agrees to furnish promptly to Publisher, at the Author’s own expense, written evidence of the permissions, licenses, and consents for use of third-party material included within the Work, except as determined by Publisher to be covered by the principles of Fair Use.
- The Author represents and warrants that:
- the Work is the Author’s original work;
- the Author has not transferred, and will not transfer, exclusive rights in the Work to any third party;
- the Work is not pending review or under consideration by another publisher;
- the Work has not previously been published;
- the Work contains no misrepresentation or infringement of the Work or property of other authors or third parties; and
- the Work contains no libel, invasion of privacy, or other unlawful matter.
- The Author agrees to indemnify and hold Publisher harmless from Author’s breach of the representations and warranties contained in Paragraph 6 above, as well as any claim or proceeding relating to Publisher’s use and publication of any content contained in the Work, including third-party content.
Revised 7/16/2018. Revision Description: Removed outdated link.