Mode of cell death of co-cultured dental mesenchymal stem cells following exposure to endodontic irrigating solutions

Autores/as

  • Eshagh Ali Saberi Department of Endodontics, Faculty of Dentistry, Oral and Dental Diseases Research Center, Zahedan University of Medical Sciences, Zahedan, Iran
  • Narges Farhad Mollashahi Department of Endodontics, Faculty of Dentistry, Oral and Dental Diseases Research Center, Zahedan University of Medical Sciences, Zahedan, Iran
  • Mahboobe Ahmadi post graduate student, Zahedan University of Medical Sciences, Zahedan, Iran
  • Arezoo Pirhaji assistant Professor, Department of Endodontics, Faculty of Dentistry, Zahedan University of Medical Sciences, Zahedan, Iran

DOI:

https://doi.org/10.5195/d3000.2022.146

Palabras 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

2022-03-21

Número

Sección

Mechanisms of Oral Disease