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Cytotechnology

, Volume 68, Issue 2, pp 213–222 | Cite as

Cytotoxicity and genotoxicity of butyl cyclohexyl phthalate

  • Çinel Köksal
  • Ayse Nalbantsoy
  • N. Ülkü Karabay Yavaşoğlu
Original Research

Abstract

Butyl cyclohexyl phthalate (BCP) is frequently used in personal care products, medical and household applications. The aim of this study is therefore to evaluate possible cytotoxicity and genotoxicity of BCP using in vitro and in vivo assays. The in vitro cytotoxic effect of BCP was investigated on mouse fibroblastic cell line (L929 cells) by MTT assay. The result showed that BCP inhibits cell proliferation in a concentration-dependent manner (IC50 value = 0.29 µg/mL). For genotoxicity assessment, tested concentrations of BCP demonstrated mutagenic activity in the presence of S9 mix with the Salmonella strain TA100 in the Ames test. Results showed that BCP is a secondary mutagenic substance even in low concentrations. The data obtained from 28-days repeated toxicity tests on mice revealed that BCP caused abnormalities of chromosome number, in a dose-dependent manner. Additionally, DNA damage, particularly DNA strand breaks, was assessed by Comet assay. The test result shows that BCP seemed to have genotoxic potential at a high level of exposure.

Keywords

Butyl cyclohexyl phthalate Cytotoxicity Genotoxicity Mutagenicity Chromosome aberration 

Notes

Acknowledgments

The study is supported by Ege University, Faculty of Science (Project Number is 2010/FEN/018).

Conflict of interest

The authors have declared that they have is no conflict of interest.

References

  1. Adler ID (1984) Cytogenetic tests in mammals. In: Venitt S, Parry JM (eds) Mutagenicity testing, a practical approach. IRL Press, Oxford, pp 275–306Google Scholar
  2. Ahbab MA, Undeger U, Barlas N, Basaran N (2014) In utero exposure to dicyclohexyl and di-n-hexyl phthalate possess genotoxic effects on testicular cells of male rats after birth in the comet and TUNEL assays. Hum Exp Toxicol 33:230–239Google Scholar
  3. Anderson D, Yu TW, Hıncal F (1999) Effect of some phthalate esters in human cells in the comet assay. Teratog Carcinog Mutagen 19:275–280CrossRefGoogle Scholar
  4. Azqueta A, Lorenzo Y, Collins AR (2009) In vitro comet assay for DNA repair: a warning concerning application to cultured cells. Mutagenesis 24:379–381CrossRefGoogle Scholar
  5. HSDB Hazardous Substance Data Bank (2011) http://toxnet.nlm.nih.gov/cgi-bin/sis/htmlgen?HSDB
  6. Brendler-Schwaab S, Hartmann A, Pfuhler S, Speit G (2005) The in vivo comet assay: use and status in genotoxicity testing. Mutagenesis 20:245–254CrossRefGoogle Scholar
  7. Carrano AV, Natarajan AT (1988) International considerations for population monitoring using cytogenetic techniques. Commission for Protection against Environmental Mutagens and Carcinogens. Mutat Res 204:379–406CrossRefGoogle Scholar
  8. Collins AR (2004) Comet assay for DNA damage and repair: principles, applications and limitations. Mol Biotechnol 26:249–261CrossRefGoogle Scholar
  9. Collins AR (2009) Investigating oxidative DNA damage and its repair using the comet assay. Mutat Res 681:24–32CrossRefGoogle Scholar
  10. Collins AR, Dusinska A (2009) Applications of the comet assay in human biomonitoring, chap 9. In: Dhawan A, Anderson D (eds) The comet assay in toxicology. The Royal Society of Chemistry Press, Cambridge, pp 201–219Google Scholar
  11. Collins AR, Dusinska A, Franklin M, Somorovska M, Petrovska H, Duthie S, Fillion L, Panayoitidis M, Raslova K, Vaughan N (1997) Comet assay in human biomonitoring studies: reliability, validation and applications. Environ Mol Mutagen 30:139–146CrossRefGoogle Scholar
  12. Committee on the Health Risks of Phthalates, National Research Council (2008) Phthalates and cumulative risk assessment: the task ahead. National Academies Press, WashingtonGoogle Scholar
  13. Cotelle S, Ferard JF (1999) Comet assay in genetic ecotoxicology: a review. Environ Mol Mutagen 34:246–255CrossRefGoogle Scholar
  14. Erkekoglu P, Rachidi W, Rosa DV, Giray B, Favier A, Hincal F (2010) Protective effect of selenium supplementation on the genotoxicity of DEP and MEP treatment in LNCaP cells. Free Radic Biol Med 49:559–566CrossRefGoogle Scholar
  15. Foster PMD, Mylchreest E, Gaido KW, Sar M (2001) Effects of phthalate esters on the developing reproductive tract of male rats. Hum Reprod Update 7:231–235CrossRefGoogle Scholar
  16. Fujii J, Luchi Y, Matsuki S, Ishii T (2003) Cooperative function of antioxidant and redox systems against oxidative stress in male reproductive tissue. Asian J Androl 5:231–242Google Scholar
  17. Harris CA, Henttu P, Parker MG, Sumpter JP (1997) The estrogenic activity of phthalate esters in vitro. Environ Health Perspect 105:802–811CrossRefGoogle Scholar
  18. Hauser R, Meeker JD, Singh NP, Silva MJ, Ryan L, Duty S, Calafat AM (2007) DNA damage in human sperm is related to urinary levels of phthalate monoester and oxidative metabolites. Hum Reprod 22:688–695Google Scholar
  19. Heudorfa U, Sundermann V, Angerer J (2007) Phthalates: toxicology and exposure. Int J Hyg Environ Health 210:623–634CrossRefGoogle Scholar
  20. IARC International Agency for Research on Cancer (2011) http://monographs.iarc.fr/ENG/Preamble/CurrentPreamble.pdf
  21. Karabay Yavasoglu NU, Köksal C, Dagdeviren M, Aktug H, Yavasoglu A (2014) Induction of oxidative stress and histological changes in liver by subacute doses of butyl cyclohexyl phthalate. Environ Toxicol 29:345–353Google Scholar
  22. Koch HM, Drexler H, Angerer J (2003) An estimation of the daily intake of di (2-ethylhexyl) phthalate (DEHP) and other phthalates in the general population. Int J Hyg Environ Health 206:77–83CrossRefGoogle Scholar
  23. Koo HJ, Lee BM (2005) Human monitoring of phthalates and risk assessment. J Toxicol Environ Health A 68:1379–1392CrossRefGoogle Scholar
  24. Latini G, Del Vecchio A, Massaro M, Verrotti A, De Felice C (2006) Phthalate exposure and male infertility. Toxicology 226:90–98CrossRefGoogle Scholar
  25. Lee KH, Lee BM (2007) Study of mutagenicities of phthalic acid and terephthalic acid using in vitro and in vivo genotoxicity tests. J Toxicol Environ Health A 70:1329–1335CrossRefGoogle Scholar
  26. Mladenov E, Iliakis G (2011) Induction and repair of DNA double strand breaks: the increasing spectrum of non-homologous end joining pathways. Mutat Res 711:61–72CrossRefGoogle Scholar
  27. Møller P (2006) The alkaline comet assay: towards validation in biomonitoring of DNA damaging exposures. Basic Clin Pharmacol 98:336–345CrossRefGoogle Scholar
  28. Mossman T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63CrossRefGoogle Scholar
  29. MSDS (2010) Material safety data sheet for butyl cyclohexyl phthalate. Chemical Service Inc., West ChesterGoogle Scholar
  30. Muniz JF, McCauley L, Scherer J, Lasarev M, Koshy M, Kow YW, Nazar-Stewart V, Kisby GE (2008) Biomarkers of oxidative stress and DNA damage in agricultural workers: a pilot study. Toxicol Appl Pharm 227:97–107CrossRefGoogle Scholar
  31. Norppa H, Bonassi S, Hansteen IL, Hagmar L, Strömberg U, Rössner P, Boffetta P, Lindholm C, Gundy S, Lazutka J, Cebulska-Wasilewska A, Fabiánová E, Srám RJ, Knudsen LE, Barale R, Fucic A (2006) Chromosomal aberrations and SCEs as biomarkers of cancer risk. Mutat Res 600:37–45CrossRefGoogle Scholar
  32. OECD 407 (1995) Repeated dose 28-day oral toxicity study in rodents. Organization for Economic Cooperation and Development, ParisGoogle Scholar
  33. OECD 425 (2001) Acute oral toxicity: up-and-down procedure. Organization for Economic Cooperation and Development, ParisGoogle Scholar
  34. OECD 471 (1997) Bacterial reverse mutation test. Organization for Economic Cooperation and Development, ParisCrossRefGoogle Scholar
  35. Park SY, Choi J (2007) Cytotoxicity, genotoxicity and ecotoxicity assay using human cell and environmental species for the screening of the risk from pollutant exposure. Environ Int 33:817–822CrossRefGoogle Scholar
  36. Saillenfait AM, Gallissot F, Sabaté JP (2009) Differential developmental toxicities of di-n-hexyl phthalate and dicyclohexyl phthalate administered orally to rats. J Appl Toxicol 29:510–521CrossRefGoogle Scholar
  37. Schmid W (1976) The micronucleus test for cytogenetic analysis. In: Hollaender A (ed) Chemical mutagens: principles and methods for their detection, vol 4. Plenum Press, New York, pp 31–53CrossRefGoogle Scholar
  38. Sharpe RM (2001) Hormones and testis development and the possible adverse effects of environmental chemicals. Toxicol Lett 120:221–232CrossRefGoogle Scholar
  39. Silva MJ, Barr DB, Reidy JA, Malek NA, Hodge CC, Caudill SP, Brock JW, Needham LL, Calafat AM (2004) Urinary levels of seven phthalate metabolites in the US population from the National Health and Nutrition Examination Survey (NHANES) 1999–2000. Environ Health Perspect 112:331–338CrossRefGoogle Scholar
  40. 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 Mutagen 35:206–221Google Scholar
  41. Zimmermann M (1983) Ethical guidelines for investigations of experimental pain in conscious animals. Pain 16:109–110CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Çinel Köksal
    • 1
  • Ayse Nalbantsoy
    • 2
  • N. Ülkü Karabay Yavaşoğlu
    • 3
  1. 1.Center for Drug Research & Development and Pharmacokinetic ApplicationsEge UniversityBornova, IzmirTurkey
  2. 2.Department of Bioengineering, Faculty of EngineeringEge UniversityBornova, IzmirTurkey
  3. 3.Department of Biology, Faculty of ScienceEge UniversityBornova, IzmirTurkey

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