Skip to main content

Advertisement

SpringerLink
Log in

Mercuric dichloride induces DNA damage in human salivary gland tissue cells and lymphocytes

  • Inorganic Compounds
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

Amalgam is still one of the most frequently used dental filling materials. However, the possible adverse effects especially that of the mercuric component have led to continued controversy. Considering that mercury may be released from amalgam fillings into the oral cavity and also reach the circulating blood after absorption and resorption, it eventually may contribute to tumorigenesis in a variety of target cells. The present investigation focuses on genotoxic effects below a cytotoxic dose level of mercuric dichloride (HgCl2) in human samples of salivary glands and lymphocytes to elucidate a possible role in tumor initiation. DNA migration due to single strand breaks, alkali labile sites and incomplete excision repair was quantified with the aid of the single cell microgel electrophoresis (Comet) assay. The concepts of Olive Tail Moment, percentage of DNA in the Tail and Tail Length were used as measures of DNA damage. To control for cytotoxic effects, the trypan blue exclusion test was applied. Human samples of the parotid salivary gland and lymphocytes of ten donors were exposed to HgCl2 concentrations from 1 to 50 μM. N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and dimethyl sulfoxide (DMSO) served as controls. Increasing dose-dependent DNA migration could be demonstrated after exposure to HgCl2 in cells of the salivary glands and lymphocytes. In both cell types a significant increase in DNA migration could be shown starting from HgCl2 concentrations of 5 μM in comparison to the negative control. The viability of the cell systems was not affected except at the highest concentration (50 μM) tested. These data indicate genotoxic effects of mercuric dichloride in human salivary glands and lymphocytes at concentrations not leading to cytotoxic effects or cell death. Consequently, a contributory role in oral salivary gland tumor initiation warrants further investigation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Albertini RJ, Anderson D, Douglas GR, Hagmar L, Hemminki K, Merlo F, Natarajan AT, Norppa H, Shuker DEG, Tice R, Waters MD, Aitio A (2000) IPCS guidelines for the monitoring of genotoxic effects of carcinogens in humans. Mutat Res 463:111–172

    Article  PubMed  CAS  Google Scholar 

  • Ascani G, Pieramici T, Messi M, Lupi E, Rubini C, Balercia P (2006) Salivary gland tumors: a retrospective study of 454 patients. Minerva Stomatol 55:209–214

    PubMed  CAS  Google Scholar 

  • Bahia O, De Amorim MI, Burbano RR, Vincent S, Dubeau H (1999) Genotoxic effects of mercury on in vitro cultures of human cells. An Acad Bras Cienc 71:437–443

    Google Scholar 

  • Bloching M, Grummt T, Reich W (2005) Genotoxikologische Prüfverfahren zum Nachweis des Einflusses der Mundgesundheit auf die Entstehung von Plattenepithelkarzinomen im oberen Aerodigestivtrakt. HNO-Informationen 84:146

    Google Scholar 

  • Clarkson TW (2002) The three modern faces of mercury. Environ Health Perspect 110:11–23

    PubMed  CAS  Google Scholar 

  • Clarkson TW, Magos L, Myers GJ (2003) The toxicology of mercury-current exposures and clinical manifestations. N Engl J Med 349:1731–1737

    Article  PubMed  CAS  Google Scholar 

  • Collins AR (2004) The Comet Assay for DNA Damage and Repair: Principles, Applications and Limitations. Mol Biotechnol 26:249–261

    Article  PubMed  CAS  Google Scholar 

  • Cotelle S, Ferard JF (1999) Comet assay in genetic ecogenotoxicology: a review. Environ Mol Mutagen 34:246–255

    Article  PubMed  CAS  Google Scholar 

  • Davies L, Welch HG (2006) Epidemiology of head and neck cancer in the United States. Otolaryngol Head Neck Surg 135:451–457

    Article  PubMed  Google Scholar 

  • De Flora S, Bennicelli C, Bagnasco M (1994) Genotoxicity of mercury compounds. A review. Mutat Res 317:57–79

    Google Scholar 

  • Dunsche A, Kastel I, Terheyden H, Springer IN, Christophers E, Brasch J (2003) Oral lichenoid reactions associated with amalgam: improvement after amalgam removal. Br J Dermatol 148:70–76

    Article  PubMed  CAS  Google Scholar 

  • Ganss C, Gottwald B, Traenckner I, Kupfer J, Eis D, Mönch J, Gieler U, Klimek J (2000) Relation between mercury concentrations in saliva, blood and urine in subjects with amalgam restaurations. Clin Oral Invest 4:206–211

    Article  CAS  Google Scholar 

  • Gopal KV (2003) Neurotoxic effects of mercury on auditory cortex networks growing on microelectrode arrays: a preliminary analysis. Neurotoxicol Teratol 25:69–76

    Article  PubMed  CAS  Google Scholar 

  • Grover P, Saleha Banu B, Dana Devi K, Begum S (2001) In vivo genotoxic effects of mercuric chloride in rat peripheral blood leucocytes using comet assay. Toxicology 167:191–197

    Article  PubMed  CAS  Google Scholar 

  • Halbach S, Kremers L, Willruth H, Mehl A, Welzl G, Wack FX, Hickel R, Greim H, (1998) Systemic transfer of mercury from amalgam fillings before and after cessation of emission. Environ Res 77:115–123

    Article  PubMed  CAS  Google Scholar 

  • Halbach S, Welzl G, Kremers L, Willruth H, Mehl A, Wack FX, Hickel R, Greim H (2000) Steady-state transfer and depletion kinetics of mercury from amalgam fillings. Sci Total Environ 259:13–21

    Article  PubMed  CAS  Google Scholar 

  • Harréus UA, Baumeister P, Wallner BC, Berghaus A, Kleinsasser NH (2005) Karzinogene und kokarzinogene Effekte von Metallen und Ethylalkohol in humanen Speicheldrüsenzellen. HNO 53:155–162

    Article  PubMed  Google Scholar 

  • Henderson L, Wolfreys A, Fedyk J, Bourner C, Windebank S (1998) The ability of the Comet assay to discriminate between genotoxins and cytotoxins. Mutagenesis 13:89–95

    Article  PubMed  CAS  Google Scholar 

  • Horn-Ross PL, Ljung BM, Morrow M (1997) Environmental factors and the risk of salivary gland cancer. Epidemiology 8:414–419

    Article  PubMed  CAS  Google Scholar 

  • IARC (1997) Monographs on the evaluation of carcinogenic risks to humans. Beryllium, cadmium, mercury, and exposures in the glass manufacturing industry. WHO 58

  • Ionescu JG, Novotny J, Stejskal VD, Latsch A, Blaurock-Busch E, Eisenmann-Klein M (2006) Increased levels of transition metals in breast cancer tissue. Neuro Endocrinol Lett 27, E-pub ahead of print

  • Kehe K, Reichl FX, Durner J, Walther U, Hickel R, Forth W (2001) Cytotoxicity of dental composite components and mercury compounds in pulmonary cells. Biomaterials 22:317–322

    Article  PubMed  CAS  Google Scholar 

  • Kim SH, Sharma RP (2004) Mercury induced apoptosis and necrosis in murine macrophages: role of calcium-induced reactive oxygen species and p38 mitogen-activated protein kinase signalling. Toxicol Appl Pharmacol 196:47–57

    Article  PubMed  CAS  Google Scholar 

  • Kleinsasser NH, Wallner BC, Kastenbauer ER, Muenzenrieder RK, Harréus UA (2000) Comparing genotoxic sensitivities of peripheral lymphocytes and mucosa of the upper aerodigestive tract of human donors using the Comet assay. Mutat Res 467:21–30

    PubMed  CAS  Google Scholar 

  • Kleinsasser NH, Juchhoff J, Wallner BC, Bergner A, Harréus UA, Gamarra F, Buehrlen M, Huber RM, Rettenmeier AW (2004) The use of mini-organ cultures of human upper aerodigestive tract epithelia in ecogenotoxicology. Mutat Res 561:63–73

    PubMed  CAS  Google Scholar 

  • Little MC, Watson REB, Pemberton MN, Griffiths CEM, Thornhill MH (2001) Activation of oral keratinocytes by mercuric chloride: relevance to dental amalgam-induced oral lichenoid reactions. Br J Dermatol 144:1024–1032

    Article  PubMed  CAS  Google Scholar 

  • Mackert JR, Berglund A (1997) Mercury exposure from dental amalgam fillings: absorbed dose and the potential for adverse health effects. Crit Rev Oral Biol Med 8:410–436

    Article  PubMed  CAS  Google Scholar 

  • McKelvey-Martin VJ, Green MH, Schmezer P, Pool-Zobel BL, De Meo MP, Collins A (1993) The single cell electrophoreses assay: a European review. Mutat Res 288:47–63

    PubMed  CAS  Google Scholar 

  • Olive PL, Durand RE, Le Riche J, Olivotto IA, Jackson SM (1993) Gel electrophoresis of individual cells to quantify hypoxic fraction in human breast cancer. Cancer Res 53:733–736

    PubMed  CAS  Google Scholar 

  • Phillips HJ (1973) Dye exclusion test for cell viability. In: Kruse PF, Patterson MK (Eds) Tissue culture methods and applications. Academic, New York, pp 406–408

    Google Scholar 

  • Rajaee-Behbahani N, Müller-Vogt U, Gliniorz R, Engelhardt E, Bartsch H, Schmezer P (1999) Effect of long-term cryopreservation on mutagen sensitivity and DNA repair capacity of peripheral human lymphocytes. J Cancer Res Clin Oncol 125:32

    Google Scholar 

  • Rao MV, Chinoy NJ, Suthar MB, Rajvanshi MI (2001) Role of ascorbic acid on mercuric chloride-induced genotoxicity in human blood cultures. Toxicol In Vitro 15:649–654

    Article  PubMed  CAS  Google Scholar 

  • Reichl FX, Walther UI, Durner J, Kehe K, Hickel R, Kunzelmann KH, Spahl W, Hume WR, Benschop H, Forth W (2001) Cytotoxicity of dental composite components and mercury compounds in lung cells. Dent Mater 17:95–101

    Article  PubMed  CAS  Google Scholar 

  • Rojas E, Lopez MC, Valverde M (1999) Single cell electrophoreses assay: methodology and applications. J Chromatogr B Biomed Sci Appl 722:225–254

    Article  PubMed  CAS  Google Scholar 

  • Schurz F, Sabater-Vilar M, Fink-Gremmele J (2000) Mutagenicity of mercury chloride and mechanisms of cellular defense: the role of metal-binding proteins. Mutagenesis 15:525–530

    Article  PubMed  CAS  Google Scholar 

  • Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, Miyamae Y, Rojas E, Ryu JC, Sasaki YF (2000) Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutag 35:206–221

    Article  CAS  Google Scholar 

  • Wallner BC, Harréus UA, Gamarra F, Sassen A, Kleinsasser NH (2005) Genotoxicity studies in mini-organ cultures of human nasal mucosa. HNO 53:1037–1046

    Article  PubMed  CAS  Google Scholar 

  • Walther UI, Walther SC, Liebl B, Reichl FX, Kehe K, Nilius M, Hickel R (2002) Cytotoxicity of ingredients of various dental materials and related compounds in L2- and A549 cells. J Biomed Mater Res 63:643–649

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. University of Regensburg, Regensburg, Germany 

    Katharina Schmid & Susanne Kroemer

  2. Department of Pathology, University of Regensburg, Regensburg, Germany

    Andrea Sassen

  3. Department of Otorhinolaryngology, Head and Neck Surgery, Technical University of Munich, Munich, Germany

    Rainer Staudenmaier

  4. Institute of Pharmacology and Toxicology, University of Munich, Munich, Germany

    Franz-Xaver Reichl

  5. Department of Otorhinolaryngology, Head and Neck Surgery, University of Munich, Munich, Germany

    Ulrich Harréus

  6. Department of Otorhinolaryngology, Head and Neck Surgery, University of Wuerzburg, Wuerzburg, Germany

    Rudolf Hagen & Norbert Kleinsasser

Authors
  1. Katharina Schmid
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Andrea Sassen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rainer Staudenmaier
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Susanne Kroemer
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Franz-Xaver Reichl
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ulrich Harréus
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Rudolf Hagen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Norbert Kleinsasser
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Norbert Kleinsasser.

Additional information

The authors declare that there is no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schmid, K., Sassen, A., Staudenmaier, R. et al. Mercuric dichloride induces DNA damage in human salivary gland tissue cells and lymphocytes. Arch Toxicol 81, 759–767 (2007). https://doi.org/10.1007/s00204-007-0208-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-007-0208-3

Keywords

Search

Navigation