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Effects of hydrogen peroxide (H2O2) on alkaline phosphatase activity and matrix mineralization of odontoblast and osteoblast cell lines

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Abstract

Hydrogen peroxide (H2O2), an oxidizing agent, has been widely used as a disinfectant. Recently, because of its reactive properties, H2O2 has also been used as a tooth bleaching agent in dental care. This is a cause for concern because of adverse biological effects on the soft and hard tissues of the oral environment. To investigate the influence of H2O2 on odontoblasts, the cells producing dentin in the pulp, we assessed cellular viability, generation of reactive oxygen species (ROS), alkaline phosphatase (ALP) activity, and nodule formation of an odontoblastic cell line (MDPC-23) after treatment with H2O2, and compared those with the effects on preosteoblastic MC3T3-E1 cells. Cytotoxic effects of H2O2 began to appear at 0.3 mmol/L in both MDPC-23 and MC3T3-E1 cells. At that concentration, the accumulation of intracellular ROS was confirmed by a fluorescent probe, DCFH-DA. Although more ROS were detected in MDPC-23, the increasing pattern and rate are similar between the two cells. When the cells were treated with H2O2 at concentrations below 0.3 mmol/L, MDPC-23 displayed a significant increase in ALP activity and mineralized bone matrix, while MC3T3-E1 cells showed adverse effects of H2O2. It is known that ROS are generally harmful by-products of aerobic life and represent the primary cause of aging and numerous diseases. These data, however, suggest that ROS can induce in vitro cell differentiation, and that they play a more complex role in cell physiology than simply causing oxidative damage.

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Abbreviations

ROS:

reactive oxygen species

ALP:

alkaline phosphatase

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

DCFH-DA:

2′,7′-dichlorodihydrofluorescein diacetate

DCF:

dichlorofluorescin

References

  • Allen RG, Tresini M. Oxidative stress and gene regulation. Free Rad Biol Med. 2000;28:463–99.

    Article  CAS  PubMed  Google Scholar 

  • Ames BN, Shigenaga MK. Oxidants are a major contributor to aging. Ann NY Acad Sci. 1992;663:85–96.

    CAS  PubMed  Google Scholar 

  • Arana-Chavez VE, Massa LF. Odontoblast: the cells forming and maintaining dentin. Int J Biochem Cell Biol. 2004;36:1367–73.

    CAS  PubMed  Google Scholar 

  • Bai XC, Lu D, Bai J, et al. Oxidative stress inhibits osteoblastic differentiation on bone cells by ERK and NF-κB. Biochem Biophys Res Commun. 2004;314:197–207.

    Google Scholar 

  • Bai XC, Lu D, Liu AL, et al. Reactive oxygen species stimulates receptor activator of NF-κB ligand expression in osteoblast. J Biol Chem. 2005;280:17497–506.

    Google Scholar 

  • Benetti AR, Valera MC, Mancini MNG, et al. In vitro penetration of beaching agents into the pulp chamber. Int Endod J. 2004;37:120–4.

    Article  CAS  PubMed  Google Scholar 

  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem. 1976;72:248–54.

    Article  CAS  PubMed  Google Scholar 

  • Dahl JE, Pallesen U. Tooth bleaching—a crictical review of the biological aspects. Crit Rev Oral Biol Med. 2003;14:292–304.

    CAS  PubMed  Google Scholar 

  • Dandona P, Thusu K, Cook S, et al. Oxidative damage to DNA in diabetes mellitus. Lancet. 1996;347:444–5.

    Article  CAS  PubMed  Google Scholar 

  • Floyd RA. Role of oxygen free radicals in carcinogenesis and brain ischemia. FASEB J. 1990;4:2587–97.

    CAS  PubMed  Google Scholar 

  • Garrett IR, Boyce BF, Oreffo ROC, et al. Oxygen-derived free radicals stimulate osteoclastic bone resorption in rodent bone in vitro and in vivo. J Clin Invest. 1990;85:632–9.

    CAS  PubMed  Google Scholar 

  • Gökay O, Müjdeci A, Algın E. Peroxide penetration into the pulp from whitening strips. J Endod. 2004;30:887–9.

    PubMed  Google Scholar 

  • Goldberg M, Septier D. Phospholipids in amelogenesis and dentinogenesis. Crit Rev Oral Biol Med. 2002;13:276–90.

    CAS  PubMed  Google Scholar 

  • Hanks CT, Sun ZL, Fang DN, et al. Cloned 3T6 cell line from CD-1 mouse fetal molar dental papillae. Connect Tissue Res. 1998;37:233–49.

    CAS  PubMed  Google Scholar 

  • Haywood VB. Nightguard vital bleaching, a history and product update. Part I. Esthet Dent Update. 1991;2:63–6.

    Google Scholar 

  • Jenner P. Oxidative damage in neurodegenerative disease. Lancet. 1994;344:796–8.

    Article  CAS  PubMed  Google Scholar 

  • Johnson K, Hashimoto S, Lotz M, et al. Up-regulated expression of the phosphodiesterase nucleotide pyrophosphatase family member PC-1 is a marker and pathogenic factor for knee meniscal cartilage matrix calcification. Arthritis Rheum.2001;44:1071–81.

    CAS  PubMed  Google Scholar 

  • Kirsch T, Nah HD, Shapiro IM, Pacifici M. Regulated production of mineralization-competent matrix vesicles in hypertrophic chondrocytes. J Cell Biol.1997;137:1149–60.

    Article  CAS  PubMed  Google Scholar 

  • LeBel CP, Bondy SC. Sensitive and rapid quantification of oxygen reactive species formation in rat synaptosomes. Neurochem Int. 1990;17:435–41.

    Article  CAS  Google Scholar 

  • Martindale JL, Holbrook NJ. Cellular response to oxidative stress: signaling for suicide and survival. J Cell Physiol. 2002;192:1–15.

    Article  CAS  PubMed  Google Scholar 

  • Mody N, Parhami F, Sarafian TA, et al. Oxidative stress modulates osteoblastic differentiation of vascular and bone cells. Free Rad Biol Med. 2001;31:509–19.

    Article  CAS  PubMed  Google Scholar 

  • Ren J-G, Zheng R-L, Shi Y-M, et al. Apoptosis, redifferentiation and arresting proliferation simultaneously triggered by oxidative stress in human hepatoma cell. Cell Biol Int. 1998;22:41–9.

    Google Scholar 

  • Shackelford RE, Kaufmann WK, Paules RS. Oxidative stress and cell cycle checkpoint function. Free Rad Biol Med. 2000;28:1387–404.

    Article  CAS  PubMed  Google Scholar 

  • Slater AF, Stefan C, Nobel I, et al. Signalling mechanisms and oxidative stress in apoptosis. Toxicol Lett. 1995;82:149–53.

    Article  PubMed  Google Scholar 

  • Sulieman M, Addy M, Macdonald E, et al., The bleaching depth of a 35% hydrogen peroxide based in-office product: a study in vitro. J Dent. 2005;33:33–40.

    Article  CAS  PubMed  Google Scholar 

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Correspondence to H.-C. Yang.

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Lee, D.H., Lim, BS., Lee, YK. et al. Effects of hydrogen peroxide (H2O2) on alkaline phosphatase activity and matrix mineralization of odontoblast and osteoblast cell lines. Cell Biol Toxicol 22, 39–46 (2006). https://doi.org/10.1007/s10565-006-0018-z

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  • DOI: https://doi.org/10.1007/s10565-006-0018-z

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