The Hormetic Effect of Arsenic Trioxide on Rat Pulpal Cells: An In Vitro Preliminary Study

Mohannad Nassar; Ahmad Dargham; Ahmed Jamleh; Yukihiko Tamura; Noriko Hiraishi; Junji Tagami

doi:10.1055/s-0040-1718637

European Journal of Dentistry, Table of Contents

CC BY-NC-ND 4.0 · Eur J Dent 2021; 15(02): 222-227
DOI: 10.1055/s-0040-1718637

Original Article

The Hormetic Effect of Arsenic Trioxide on Rat Pulpal Cells: An In Vitro Preliminary Study

Mohannad Nassar

¹Department of Preventive and Restorative Dentistry, College of Dental Medicine, University of Sharjah, Sharjah, United Arab Emirates

,

Ahmad Dargham

²Ras Al Khaimah College of Dental Sciences, Ras Al Khaimah Medical and Health Sciences University, Ras Al Khaimah, United Arab Emirates

,

Ahmed Jamleh

³Restorative and Prosthetic Dental Sciences, College of Dentistry, King Saud bin Abdulaziz University for Health Sciences, National Guard Health Affairs, Riyadh, Saudi Arabia

⁴King Abdullah International Medical Research Centre, National Guard Health Affairs, Riyadh, Saudi Arabia

,

Yukihiko Tamura

⁵Department of Bio-Matrix (Dental Pharmacology), Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan

,

Noriko Hiraishi

⁶Department of Cariology and Operative Dentistry, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan

,

Junji Tagami

⁶Department of Cariology and Operative Dentistry, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan

› Author Affiliations

Abstract

Objectives Despite the agreement that there is no longer any indication for arsenic use in modern endodontics, some concerns are surfacing about the minute amount of arsenic trioxide (As₂O₃) released from Portland cement-based materials. The present study investigated the effect of different concentrations of As₂O₃ on rat pulpal cells and the efficacy of N-acetylcysteine (NAC) in preventing As₂O₃-mediated toxicity.

Materials and Methods Cytotoxicities of 50, 10, or 5 µm As₂O₃ and the effect of cells co-treatment with 50 µm As₂O₃ and 5,000 µm NAC or 500 µm NAC were tested at 24 hours or 3 days. Cell viability was assessed by means of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and cellular morphological changes were observed under phase contrast microscope.

Statistical Analysis Two-way analysis of variance with Tukey’s post-hoc test was used to evaluate differences between the groups (α = 0.05).

Results At both exposure times, 50 µm As₂O₃ resulted in lower optical density (OD) values when compared with 10 or 5 µm As₂O₃. At 24 hours, 10 µm As₂O₃ resulted in a higher OD value compared with the control; however, at 3 days the difference was statistically insignificant. At each exposure time, the OD value of 5 µm As₂O₃ group was comparable to the control and 10 µm As₂O₃ group. There were no significant differences between 50 µm As₂O₃ group and 500 μm NAC+50 μm As₂O₃ group; however, these two groups had lower OD values when compared with 5,000 μm NAC+50 μm As₂O₃ group at 24 hours and 3 days. The latter group showed significantly lower OD value in comparison with the control at 24 hours and 3 days. Control cells were polygonal-shaped while 50 µm As₂O₃-treated cells exhibited contracted and spherical morphology with increased intercellular spaces. At 24 hours, 10 μm and 5 µm As₂O₃-treated cells were slightly hypertrophic. Cells co-treated with NAC and As₂O₃ showed increased intercellular spaces and lower cellular density compared with the control.

Conclusions As₂O₃ displayed a hormetic effect on pulpal cells; however, the proliferative effect induced by low As₂O₃ concentrations should be interpreted with caution. NAC did not prevent As₂O₃-mediated toxicity; however, it demonstrated potential for ameliorating this toxicity.

Keywords

arsenic - heavy metal - hormesis - N-acetylcysteine - pulp cells - toxicity

Full Text

References

References
1 Agency for Toxic Substances and Disease Registry (ATSDR).. Toxicological Profile for Arsenic. Toxicological Profile for Arsenic U.S. Department of Health and Human Services. Public Health Service; 2007
2 Ratnaike RN. Acute and chronic arsenic toxicity. Postgrad Med J 2003; 79 (933) 391-396
3 Schmidt CW. Low-dose arsenic: in search of a risk threshold. Environ Health Perspect 2014; 122 (05) A130-A134
4 Hyson Jr JM. A history of arsenic in dentistry. J Calif Dent Assoc 2007; 35 (02) 135-139
5 Chen G, Sung PT. Gingival and localized alveolar bone necrosis related to the use of arsenic trioxide paste—two case reports. J Formos Med Assoc 2014; 113 (03) 187-190
6 Ali MS, Kano B. Endodontic materials: from old materials to recent advances. In: Khurshid Z, Najeeb S, Zafar MS, Sefat F. eds. Advanced Dental Biomaterials. Woodhead Publishing; Cambridge, UK:; 2019: 255-299
7 Oliveira MG, Xavier CB, Demarco FF, Pinheiro ALB, Costa AT, Pozza DH. Comparative chemical study of MTA and Portland cements. Braz Dent J 2007; 18 (01) 3-7
8 Guven Y, Tuna EB, Dincol ME, Aktoren O. X-ray diffraction analysis of MTA-Plus, MTA-Angelus and DiaRoot BioAggregate. Eur J Dent 2014; 8 (02) 211-215
9 Phair JW. Green chemistry for sustainable cement production and use. Green Chem 2006; 8 (09) 763-780
10 Nejatian T, Firouzmanesh P, Syed AU. Dental gypsum and investments. In: Khurshid Z, Najeeb S, Zafar MS, Sefat F. eds. Advanced Dental Biomaterials. Woodhead Publishing; 2019. Cambridge, UK: 37-54
11 Islam I, Chng HK, Yap AU. Comparison of the physical and mechanical properties of MTA and Portland cement. J Endod 2006; 32 (03) 193-197
12 Schembri M, Peplow G, Camilleri J. Analyses of heavy metals in mineral trioxide aggregate and Portland cement. J Endod 2010; 36 (07) 1210-1215
13 Minotti PG, Ordinola-Zapata R, Midena RZ. et al. Rat subcutaneous tissue response to calcium silicate containing different arsenic concentrations. J Appl Oral Sci 2015; 23 (01) 42-48
14 Monteiro Bramante C, Demarchi ACCO, de Moraes IG. et al. Presence of arsenic in different types of MTA and white and gray Portland cement. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2008; 106 (06) 909-913
15 Chang SW, Shon WJ, Lee W, Kum KY, Baek SH, Bae KS. Analysis of heavy metal contents in gray and white MTA and 2 kinds of Portland cement: a preliminary study. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010; 109 (04) 642-646
16 Dorileo MCGO, Bandeca MC, Pedro FLM. et al. Analysis of metal contents in Portland type V and MTA-based cements. ScientificWorldJournal 2014; 2014: 983728
17 International Organization for Standardization. Dentistry—Water-based cements—Part 1: Powder/liquid acid-base cements. Geneva, Switzerland; 2007. ISO 9917-1:2003.
18 De-Deus G, de Souza MCB, Sergio Fidel RA, Fidel SR, de Campos RC, Luna AS. Negligible expression of arsenic in some commercially available brands of Portland cement and mineral trioxide aggregate. J Endod 2009; 35 (06) 887-890
19 Duarte MAH, De Oliveira Demarchi ACC, Yamashita JC, Kuga MC, De Campos Fraga S. Arsenic release provided by MTA and Portland cement. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2005; 99 (05) 648-650
20 Sochman J. N-acetylcysteine in acute cardiology: 10 years later: what do we know and what would we like to know?!. J Am Coll Cardiol 2002; 39 (09) 1422-1428
21 Modi M, Kaul RK, Kannan GM, Flora SJ. Co-administration of zinc and n-acetylcysteine prevents arsenic-induced tissue oxidative stress in male rats. J Trace Elem Med Biol 2006; 20 (03) 197-204
22 Reddy PS, Rani GP, Sainath SB, Meena R, Supriya Ch. Protective effects of N-acetylcysteine against arsenic-induced oxidative stress and reprotoxicity in male mice. J Trace Elem Med Biol 2011; 25 (04) 247-253
23 Kasugai S, Adachi M, Ogura H. Establishment and characterization of a clonal cell line (RPC-C2A) from dental pulp of the rat incisor. Arch Oral Biol 1988; 33 (12) 887-891
24 Hashmi MZ, Naveedullah. Shen H, Zhu S, Yu C, Shen C. Growth, bioluminescence and shoal behavior hormetic responses to inorganic and/or organic chemicals: a review. Environ Int 2014; 64: 28-39
25 Kitchin KT. Recent advances in arsenic carcinogenesis: modes of action, animal model systems, and methylated arsenic metabolites. Toxicol Appl Pharmacol 2001; 172 (03) 249-261
26 Rossman TG. Mechanism of arsenic carcinogenesis: an integrated approach. Mutat Res 2003; 533 (1-2) 37-65
27 Pilger A, Rüdiger HW. 8-Hydroxy-2′-deoxyguanosine as a marker of oxidative DNA damage related to occupational and environmental exposures. Int Arch Occup Environ Health 2006; 80 (01) 1-15
28 Valavanidis A, Vlachogianni T, Fiotakis C. 8-hydroxy-2′-deoxyguanosine (8-OHdG): a critical biomarker of oxidative stress and carcinogenesis. J Environ Sci Health Part C Environ Carcinog Ecotoxicol Rev 2009; 27 (02) 120-139
29 Schmeisser S, Schmeisser K, Weimer S. et al. Mitochondrial hormesis links low-dose arsenite exposure to lifespan extension. Aging Cell 2013; 12 (03) 508-517
30 Lau ATY, Li M, Xie R, He QY, Chiu JF. Opposed arsenite-induced signaling pathways promote cell proliferation or apoptosis in cultured lung cells. Carcinogenesis 2004; 25 (01) 21-28
31 Liao WT, Lan CC, Lee CH, Yu HS. Concentration-dependent cellular responses of arsenic in keratinocytes. Kaohsiung J Med Sci 2011; 27 (09) 390-395
32 Dodson M, de la Vega MR, Harder B. et al. Low-level arsenic causes proteotoxic stress and not oxidative stress. Toxicol Appl Pharmacol 2018; 341: 106-113
33 Snow ET, Sykora P, Durham TR, Klein CB. Arsenic, mode of action at biologically plausible low doses: what are the implications for low dose cancer risk?. Toxicol Appl Pharmacol 2005; 207 (suppl 2) 557-564
34 Udensi UK, Graham-Evans BE, Rogers C, Isokpehi RD. Cytotoxicity patterns of arsenic trioxide exposure on HaCaT keratinocytes. Clin Cosmet Investig Dermatol 2011; 4: 183-190
35 Tokar EJ, Diwan BA, Waalkes MP. Arsenic exposure transforms human epithelial stem/progenitor cells into a cancer stem-like phenotype. Environ Health Perspect 2010; 118 (01) 108-115
36 Neuhaus KW. Teeth: malignant neoplasms in the dental pulp?. Lancet Oncol 2007; 8 (01) 75-78
37 Samanta J, Mondal A, Saha S, Chakraborty S, Sengupta A. Oleic acid protects from arsenic-induced cardiac hypertrophy via AMPK/FoxO/NFATc3 pathway. Cardiovasc Toxicol 2020; 20 (03) 261-280
38 Dash M, Maity M, Dey A. et al. The consequence of NAC on sodium arsenite-induced uterine oxidative stress. Toxicol Rep 2018; 5: 278-287
39 Kannan GM, Flora SJ. Combined administration of N-acetylcysteine and monoisoamyl DMSA on tissue oxidative stress during arsenic chelation therapy. Biol Trace Elem Res 2006; 110 (01) 43-59
40 Takahashi N, Yoshida T, Ohnuma A. et al. The enhancing effect of the antioxidant N-acetylcysteine on urinary bladder injury induced by dimethylarsinic acid. Toxicol Pathol 2011; 39 (07) 1107-1114