Mutagenicity Testing and DNA Damage Analysis of Malaysian White Portland Cement Using Ames Test and Comet Assay

Siti Suraya Zulkifli, KANNAN THIRUMULU PONNURAJ, Norhayati Luddin

Abstract


The aim of the current study was to determine the mutagenic effect and DNA damage of Malaysian white Portland cement (MWPC) and White mineral trioxide aggregate (WMTA) using Ames test and Comet assay. The cytotoxicity of WMTA and MWPC was evaluated using MTT assay, mutagenicity using Ames test in the absence and presence of metabolic activation system (S9 mix) on Salmonella strains (TA98, TA102, TA1535, TA1537 and TA1538) and DNA damage using Comet assay on human periodontal ligament fibroblast (HPLF) cell line. Concurrent negative and positive controls were included. Based on the MTT assay, the IC50for WMTA and MWPC was 18.71 and 19.91 mg/ml, IC25 was 3.33 and 3.55 mg/ml and IC10was 0.59 and 0.63 mg/ml respectively. These values were further employed in Comet assay. Ames test revealed that WMTA and MWPC did not cause any mutagenic effect as the number of revertant colonies was less than that of the negative control. In the Comet assay, no significant comet formation was found in HPLFs treated with WMTA, MWPC and negative control except the positive control. Hence, it can be concluded that MWPC is non-mutagenic and does not cause any DNA damage under the present test conditions.


Keywords


Cytotoxicity; DNA damage; Malaysian white Portland cement; mineral trioxide aggregate; mutagenicity

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References


. Gutmann JL, Harrison JW. Surgical endodontics. Boston: Blackwell Scientific Publications. 1991;7-36.

. Torabinejad M, Watson TF, Pitt Ford TR. Sealing ability of a mineral trioxide aggregate when used as a root end filling material. J Endod. 1993;19(12):591-595.

. Torabinejad M, Chivian N. Clinical applications of mineral trioxide aggregate. J Endod. 1999;25(3):197-205.

. Holland R, Souza Vd, Nery MJ, Faraco Júnior IM, Bernabé PFE, et al. Reaction of rat connective tissue to implanted dentin tubes filled with a white mineral trioxide aggregate. Braz Dent J. 2002;13(1):23-26.

. Camilleri J, Montesin FE, Papaioannou S, McDonald F, Pitt Ford TR. Biocompatibility of two commercial forms of mineral trioxide aggregate. Int Endod J. 2004;37(10):699-704.

. Ferris DM, Baumgartner JC. Perforation repair comparing two types of mineral trioxide aggregate. J Endod. 2004;30(6):422-424.

. Menezes R, Bramante CM, Letra A, Carvalho VGG, Garcia RB. Histologic evaluation of pulpotomies in dog using two types of mineral trioxide aggregate and regular and white Portland cements as wound dressings. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2004;98(3):376-379.

. Asgary S, Parirokh M, Eghbal MJ, Brink F. Chemical differences between white and gray mineral trioxide aggregate. J Endod. 2005;31(2):101-103.

. Saidon J, He J, Zhu Q, Safavi K, Spångberg LSW. Cell and tissue reactions to mineral trioxide aggregate and Portland cement. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2003;95(4):483-489.

. Parirokh M, Torabinejad M. Mineral trioxide aggregate: A comprehensive literature review-Part III: Clinical applications, drawbacks, and mechanism of action. J Endod. 2010;36(3):400-413.

. Torabinejad M, White DJ. Loma Linda University, assignee. Tooth filling material and method of use. 1995. US patent 5769638. June 23, 1998.

. Wucherpfennig AL, Green DB. PR 40 Mineral trioxide vs. Portland cement: Two biocompatible filling materials. J Endod. 1999;25(4):308.

. Estrela C, Bammann LL, Estrela C, Silva RS, Pécora JD. Antimicrobial and chemical study of MTA, Portland cement, calcium hydroxide paste, Sealapex and Dycal. Braz Dent J. 2000;11(1):3-9.

. Asgary S, Parirokh M, Eghbal MJ, Brink F. A comparative study of white mineral trioxide aggregate and white Portland cements using X‐ray microanalysis. Aust Endod J. 2004;30(3):89-92.

. De-Deus G, Coutinho-Filho T. The use of white Portland cement as an apical plug in a tooth with a necrotic pulp and wide-open apex: a case report. Int Endod J. 2007;40(8):653-660.

. Hwang YC, Lee SH, Hwang IN, Kang IC, Kim MS, et al. Chemical composition, radiopacity, and biocompatibility of Portland cement with bismuth oxide. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2009;107(3):e96-e102.

. Camilleri J, Montesin FE, Di Silvio L, Pitt Ford TR. The chemical constitution and biocompatibility of accelerated Portland cement for endodontic use. Int Endod J. 2005;38(11):834-842.

. De Deus G, Ximenes R, Gurgel-Filho ED, Plotkowski MC, Coutinho-Filho T. Cytotoxicity of MTA and Portland cement on human ECV 304 endothelial cells. Int Endod J. 2005;38(9):604-609.

. Braz MG, Camargo EA, Salvadori DMF, Marques MEA, Ribeiro DA. Evaluation of genetic damage in human peripheral lymphocytes exposed to mineral trioxide aggregate and Portland cements. J Oral Rehab. 2006;33(3):234-239.

. Ribeiro DA, Sugui MM, Matsumoto MA, Duarte MAH, Marques MEA, et al. Genotoxicity and cytotoxicity of mineral trioxide aggregate and regular and white Portland cements on Chinese hamster ovary

(CHO) cells in vitro. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006;101(2):258-261.

. Ahmed HMA, Luddin N, Kannan TP, Mokhtar KI, Ahmad A. Chemical analysis and biological properties of two different formulations of white Portland cements. Scanning. 2015;38(4):303-316.

. Ahmed HMA, Luddin N, Kannan TP, Mokhtar KI, Ahmad A. Cell attachment properties of Portland Cement-based endodontic materials: Biological and methodological considerations. J Endod. 2014;40(10):1517-1523.

. Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65(1–2):55-63.

. Burton JD. The MTT assay to evaluate chemosensitivity. Methods Mol Med. 2005;110:69-78.

. Mortelmans K, Zeiger E. The Ames Salmonella/microsome mutagenicity assay. Mutat Res. 2000;455(1–2):29-60.

. Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, et al. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen. 2000;35(3):206-221.

. Ribeiro DA, Duarte MAH, Matsumoto MA, Marques MEA, Salvadori DMF. Biocompatibility in vitro tests of mineral trioxide aggregate and regular and white Portland cements. J Endod. 2005;31(8):605-607.

. Min KS, Kim HI, Park HJ, Pi SH, Hong CU, et al. Human pulp cells response to Portland cement in vitro. J Endod. 2007;33(2):163-166.

. Chang SW, Yoo HM, Park DS, Oh TS, Bae KS. Ingredients and cytotoxicity of MTA and 3 kinds of Portland cements. J Korean Acad Conser Dent. 2008;33(4):369-376.

. ISO 10993-3. International Organization for Standardization. Biological evaluation of medical devices – Tests for genotoxicity, carcinogenicity, and reproductive toxicity. 2014.

. Samiei M, Asgary S, Farajzadeh M, Bargahi N, Abdolrahimi M, et al. Investigating the mutagenic effects of three commonly used pulpotomy agents using the Ames test. Adv Pharm Bull. 2015;5(1):121-125.

. Ames BN, Lee FD, Durston WE. An improved bacterial test system for the detection and classification of mutagens and carcinogens. PNAS. 1973;70(3):782-786.

. Sahebgharani M, Partoazar A. Monitoring Ames assay on urine of clinical pathology laboratories technicians. J Pharmacol Toxicol. 2008;3(3):230-235.

. Mahon GAT, Green MHL, Middleton B, Mitchell I, de G, et al. Analysis of data from microbial colony assays. Statistical evaluation of mutagenicity test data. In:Kirkland, D.J. (Ed.), The Report of the

UKEMS Sub-committee on Guidelines for Mutagenicity Testing. Part III. Cambridge University Press, Cambridge. 1989; 26e65.

. Kaplan Ç, Diril N, Şahin S, Cehreli MC. Mutagenic potentials of dental cements as detected by the Salmonella/microsome test. Biomaterials. 2004;25(18):4019-4027.

. Noushad M, Kannan TP, Husein A, Abdullah H, Ismail AR. Genotoxicity evaluation of locally produced dental porcelain – An in vitro study using the Ames and Comet assays. Toxicol in Vitro. 2009;23(6):1145-1150.

. Catalano F, Mariano F, Maina G, Bianco C, Nuzzo J, et al. An unusual case of extensive self-inflicted cement burn. Ann Burns Fire Disasters. 2013;26(1):40-43.

. Kettering JD, Torabinejad M. Investigation of mutagenicity of mineral trioxide aggregate and other commonly used root-end filling materials. J Endod. 1995;21(11):537-539.

. Maron DM, Ames BM. Revised methods for the Salmonella mutagenicity test. Mutat Res. 1983;113(3-4):173-215.

. Hakura A, Suzuki S, Satoh T. Improvement of the Ames test

using human liver S9 preparation. In: Yan Z, Caldwell GW (eds). Optimization in Drug Discovery. Methods in Pharmacology and Toxicology. Humana Press, Totowa, New Jersey. 2004.

Singh NP, McCoy MT, Tice RR, Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 1988;175(1):184-191.

. Musa M, Kannan TP, Masudi SM, Ismail AR. Assessment of DNA damage caused by locally produced hydroxyapatite-silica nanocomposite using Comet assay on human lung fibroblast cell line. Mol Cell Toxicol. 2012;8(1):53-60.

. Frenzilli G, Bosco E, Barale R. Validation of single cell gel assay in human leukocytes with 18 reference compounds. Mutat Res Genet Toxicol Environ Mutagen. 2000;468(2):93-108.




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