Cytologic effects of primary tooth endodontic filling materials
Article Outline
Background/purpose
Primary tooth endodontic filling materials should be bio-compatible with periodontal tissue. The purpose of this study was to analyze the biologic effects of different endodontic filling materials for primary teeth on a human osteosarcoma cell line (U2OS).
Materials and methods
Experimental groups comprised different mixes of endodontic filling materials: zinc oxide-eugenol (ZnOE) + formocresol (FC); calcium hydroxide [Ca(OH)2] + FC; Ca(OH)2 + iodoform + deionized water; Ca(OH)2 + iodoform +camphorated parachlorophenol (CPC); Ca(OH)2 + CPC; and Vitapex. These were prepared and used to fill special glass rings, which were subsequently eluted in 10 mL of cell culture medium at 37ºC in a 5% carbon dioxide-in-air atmosphere for 24 hours. Cell culture medium alone was used as the control group. A DNA fragmentation assay was performed to determine the genotoxicity of each mix of materials. The level of cyclooxygenase (COX)-2 protein expression, the extent of dental material-elicited inflammation of U2OS cells, and the degree of mitogen-activated protein (MAP) kinase expression were determined using Western blot analysis.
Results
The results revealed that no DNA breakage was apparent after U2OS cells were treated with the various materials. COX-2 band expression dramatically declined in the ZnOE + FC group compared with the control group, although high levels of expression of the COX-2 band were noted for the Ca(OH)2 + FC and Ca(OH)2 + iodo-form + CPC groups. Band levels of extracellular signal-regulated kinase (ERK-1 and ERK-2) expression declined in the ZnOE + FC and Ca(OH)2 + CPC groups compared with the control group. p53 and caspase-3 protein bands appeared in all experimental groups.
Conclusion
The cytotoxic mechanism of endodontic filling materials on U2OS cells was induced by means of activation of the p53 and caspase-3 apoptosis signaling pathways.
Key Words: genotoxicity , inflammation , primary tooth , pulpectomy
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References
- . Antibacterial activity of dental restorative materials . Int Endod J . 1985;18:161–171
- . Evaluation of the antimicrobial potential of calcium hydroxide as an intracanal medicament . J Endod . 1983;9:372–374
- . Cytotoxicity of clove (Syzygium aromaticum) oil and its major components to human skin cells . Cell Prolif . 2006;39:241–248
- . Cytotoxicity of formocresol on cultured mammalian cells . Shigaku . 1988;75:1027–1038 [In Japanese]
- . Biocompatibility of various formula root filling materials for primary teeth . J Biomed Mater Res B Appl Biomater . 2007;80:486–490
- . A comparison of the inflammatory response produced by commercial eugenol and purified eugenol . J Dent Res . 1981;60:1724–1728
- . In vitro and in vivo effects of phenolic antioxidants against cisplatin-induced nephrotox-icity . J Biochem . 1999;125:383–390
- . Effects of isoeugenol on oxidative stress pathways in normal and streptozotocin-induced diabetic rats . J Biochem Mol Toxicol . 2001;15:159–164
- . Studies on anti-inflammatory activity of spice principles and dietary n-3 polyunsaturated fatty acids on carrageenan-induced inflammation in rats . Ann Nutr Metab . 1994;38:349–358
- Eugenol suppresses cyclo-oxygenase-2 expression in lipopolysaccharide-stimulated mouse macrophage RAW264.7 cells . Life Sci . 2003;73:337–348
- . Pulp response to ferric sulfate, diluted formocresol, and IRM in pulpotomized primary baboon teeth . ASDC J Dent Child . 1997;64:254–259
- . Pulpal tissue reaction to formocresol vs. ferric sulfate in pulpotomized rat teeth . J Clin Pediatr Dent . 1997;21:247–253
- . A review of the erroneously labeled “mummification” techniques of pulp therapy . Oral Surg Oral Med Oral Pathol . 1972;34:131–144
- . Pulp condition of successfully formocresol-treated primary molars . Scand J Dent Res . 1978;86:267–272
- . Distribution of 14C-formaldehyde after pulpot-omy with formocresol . J Am Dent Assoc . 1978;96:805–813
- . Formocresol concerns: a review . J Can Dent Assoc . 1987;53:401–404
- Lipopolysaccharide increases cyclooxygenase-2 expression in a colon carcinoma cell line through nuclear factor-kB activation . Oncogene . 2000;19:1225–1231
- . Involvement of NF-kB in the regulation of cyclooxygenase-2 protein expression in LPS-stimulated J774 macrophages . FEBS Lett . 1997;418:175–178
- . Recent advances towards understanding redox mechanisms in the activation of nuclear factor ?B . Free Radic Biol Med . 2001;28:1317–1327
- . Regulation of NF-?B activation by MAP kinase cascades . Immunobiology . 1997;198:35–49
- Atypical apoptotic death induced in L929 targets by exposure to tumor necrosis factor . J Interferon Cytokine Res . 1995;15:71–80
- . Effects of mineral triox-ide aggregate (MTA) extracts on mitogen-activated protein kinase activity in human osteosarcoma cell line (U2OS) . Biomaterials . 2003;24:3909–3913
- . Lack of genotoxicity of formocresol, paramonochlorophenol and calcium hydroxide on mammalian cells by comet assay . J Endod . 2004;30:593–596
- Scavenging property of three cresol isomers against H2O2, hypochlorite, superoxide and hydroxyl radicals . Food Chem Toxicol . 2002;40:1403–1413
- . Effect of eugenol on the genotoxic-ity of established mutagens in the liver . Food Chem Toxicol . 1996;34:33–42
- Genotoxicity and endoreduplication inducing activity of the food flavouring eugenol . Mutagenesis . 2006;21:199–204
- . Evaluation of the genotoxicity of zinc oxide eugenol-based, calcium hydroxide-based, and epoxy resin-based root canal sealers by comet assay . J Endod . 2001;27:744–748
- . In vitro evaluation of the cytotoxicity and genotoxicity of root canal medicines on human pulp fibroblasts . J Endod . 1998;24:604–606
- . Cell-transforming activity of fourteen chemical agents used in dental practice in Syrian hamster embryo cells . J Pharmacol Sci . 2003;93:497–500
- . Assessment of the genotoxicity of chemical substances used in dental practice: studies on the ability of chlorhexidine, glutaraldehyde, hydrogen peroxide, iodine, sodium arsenite, erythrosine B, fuchsin acid and fuchsin basic to induce unscheduled DNA synthesis in cultured Syrian hamster embryo cells . Odontology . 2000;87:587–596 [In Japanese, English abstract]
- . Connective tissue responses to calcium hydroxide-based root canal medicaments . Int Endod J . 1999;32:303–311
- . A histological comparison of calcium hydroxide plugs and dentin plugs used for the control of Gutta-percha root canal filling material . J Endod . 1984;10:283–293
- . Calcium hydroxide inhibits substrate adherence capacity of macrophages . J Endod . 1997;23:444–447
- . Fate of 45Ca-labeled calcium hydroxide in a root canal filling paste embedded in rat subcutaneous tissues . J Endod . 1987;13:220–223
- . Intracanal medication . In: Ingle JI , Backland LK editor. Endodontics . 4th ed.. Baltimore: Williams & Wilkins; 1994;p. 627–640
- . In vitro effect of parachlorophenol and camphorated para chlorophenol on macrophages . J Endod . 1997;23:728–730
- . Differential induction of apoptosis by capping agents during pulp wound healing . J Endod . 2003;29:41–43
- p53-mediated induction of Cox-2 counteracts p53- or genotoxic stress-induced apop-tosis . EMBO J . 2002;21:5635–5644
- COX-2 regulates p53 activity and inhibits DNA damage-induced apoptosis . Biochem Biophys Res Commun . 2005;328:1107–1112
PII: S1991-7902(09)60004-0
doi:10.1016/S1991-7902(09)60004-0
© 2009 Association for Dental Sciences of The Republic of China. Published by Elsevier Inc. All rights reserved.
