Investigation of the effects of diisobutyl phthalate on rat testicular tissue: a histopathological and morphometric evaluation

Yücel BAŞIMOĞLU KOCA

doi:10.15625/2615-9023/21119

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Authors

Yücel Başımoğlu Koca Zoology Section, Department of Biology, Faculty of Science, Aydın Adnan Menderes University, Aydın, Turkey

DOI:

https://doi.org/10.15625/2615-9023/21119

Keywords:

Diisobutyl phthalate, testis, infertility, histopathology, histomorphometry.

Abstract

Phthalates are a group of chemicals used to make plastics more durable, and they are often called plasticisers. Additionally, these chemicals are found in hundreds of products such as floor coverings, lubricating oils, and personal care products (soaps, shampoos, hair sprays). Consumer products containing phthalates can result in human exposure through direct contact and use, indirectly through leaching into the other products or general environmental contamination. In this study, the effects of Diisobutyl phthalate a commonly used phthalate, were investigated histopathologically and morphometrically to determine whether it is one of the causes of increased infertility in recent years. Two study groups of albino Wistar albino rats (total n: 40) were formed; the control group (untreated control group, solvent-corn oil the control group) and the experimental group. DiBP was administered by oral gavage to the experimental group in 3 different doses (0.25–0.5–1 mL/kg/day) mixed with corn oil every day for 28 days. At the end of the experiment, testicular tissue samples taken from all the experimental and control animals were evaluated histopathologically and morphometrically by light microscopy after routine preparation. Degeneration/atrophic tubules were quite prominent in the sections. Tubules containing degenerated germ cells and tubules devoid of germ cells were observed. It was determined that in most tubules, only tubules covered with Sertoli cells remained due to germ cell death. In addition, multinucleated giant cells were frequently encountered in such tubules. Dilatation and thickening in the basal lamina of the seminiferous tubule were accompanied by decreased PAS-positive reaction. The morphometric results supported the histopathological findings. Significant dose-related morphometrical changes (p<0.0001), including seminiferous tubule diameter, tubular lumen diameter, spermatogenic cell line height and basal lamina thickness were observed between the control and administration groups. According to the control, sham and G1, the number of these multinucleated cells (MGC) increased in G2 and G3 but these increases were statistically insignificant (p > 0.9999). In conclusion, it was observed that irreversible damage occurred in the testicular tissues of DiBP-exposed groups, and it was decided that this could be the cause of infertility. Therefore, we recommend the use of an alternative plasticiser with proven reliability.

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References

Alhasnani M. A., Loeb S., Hall S. J., Cauolo F. S., Solano A. E., Spade D. J., 2022. Interaction between mono-(2-ethylhexyl) phthalate and retinoic acid alters Sertoli cell development during fetal mouse testis cord morphogenesis. Current Res Toxicol (CRTOX): 100087. https://doi.org/ 10.1016/j.crtox.2022.100087

ATSDR, Agency for Toxic Substances and Disease Registry,1995.Toxicological profile for Diethylphytalate. Toxicological profile for Di-n-Octylphytalate. Atlanta, GA: U.S. Department of Health and Human Services, Public Health Service. Available from: https://www.atsdr.cdc.gov/ toxprofiles/tp73-p.pdf

ATSDR, Agency for Toxic Substances and Disease Registry, 1997. Toxicological profile for Di-n-Octylphytalate. Atlanta, GA: U.S. Department of Health and Human Services, Public Health Service. Available from: https://www.atsdr.cdc.gov/ toxprofiles/tp95.pdf

ATSDR, Agency for Toxic Substances and Disease Registry, 2001. Toxicological profile for Di-n-Butyl Phytalate. Atlanta, GA: U.S. Department of Health and Human Services, Public Health Service. Available from: https://www.atsdr.cdc.gov/ ToxProfiles/tp135-p.pdf

ATSDR, Agency for Toxic Substances and Disease Registry, 2022.Toxicological profile for Di(2-ethylhexyl) Phytalate (DEHP). Atlanta, GA: U.S. Department of Health and Human Services, Public Health Service. Available from: https://www.at-sdr.cdc.gov/ToxProfiles/tp9.pdf

Bancroft J. D. and Cook H. C., 1994. Manual of Histological Techniques and Their Diagnostic Application. 2nd edition. Edinburgh, Churchill Livingstone: 457.

Barlow N. J., McIntyre B. S., Foster P. M. D., 2004. Male reproductive tract lesions at 6, 12, and 18 months of age following in utero exposure to di(n-butyl) phthalate. Toxicologic Pathology 32: 79–90.

Başımoğlu Koca Y., 2019. Localization of Laminin and Fibronectin in Rat Testes after Diisobutyl Phthalate Exposure: Histopathologic and Immunohistochemical Study. Celal Bayar University Journal of Science, 15(3): 293–300. doi: 10.18466/ cbayarfbe.570613

Borch J., Axelstad M., Vinggaard A. M., Dalgaard M., 2006. Diisobutyl phthalate has comparable anti-androgenic effects to di-n-butyl phthalate in fetal rat testis. Toxicol letters, 163: 183–190.

CERHR, The National Toxicology Program (NTP) Center for the Evaluation of Risks to Human Reproduction., NTP-CERHR expert panel report on di(2-ethylhexyl) phthalate (DEHP), 2000. NTP-CERHR-DEHP-00. Sciences, International, Inc. 1800 Diagonal Road, Suite 500 Alexandria, VA 22314-2808. Available from: http://cerhr.niehs.nih.gov; accessed May 2022.

CERHR, The National Toxicology Program (NTP) Center for the Evaluation of Risks to Human Reproduction., 2003. Monograph on the potential human reproductive and developmental effects of di-isodecylphytalate (DIDP). NIH Publ No. 03-4485. Available from: https://ntp.niehs.nih.gov/sites/default/files/ntp/ohat/phthalates/didp/didp; accessed May 2022.

CERHR, The National Toxicology Program (NTP) Center for the Evaluation of Risks to Human Reproduction., 2005). Expert panel re-evaluation of DEHP, meeting summary. Available from: https://noharm-uscanada.org/sites/default/files/documents-files; accessed May 2022.

Cheng C. Y. and Mruk D. D., 2002. Cell junction dynamics in the testis: Sertoli–germ cell interactions and male contraceptive development. Physiol Rev, 82: 825–74.

Cheng C. Y. and Siu M. K. Y., 2008. Extracellular matrix and its role in spermatogenesis. Adv Exp Med Biol, 636: 74–91.

Creasy D. M., 2001. Pathogenesis of male reproductive toxicity. Toxicol Pathol, 29(1): 64–76.

Creasy D., Bube A., de Rijk E., Kandori H., Kuwahara M., Masson R., Nolte T., Reams R., Regan K., Rehm S., Rogerson P., Whitney K., 2012. Proliferative and nonproliferative lesions of the rat and mouse male reproductive system. Toxicol Pathol, 40: 40–121. https://doi.org/10.1177/0192623312454 337

Creasy D. M. and Maronpot R. R., 2020. National Toxicology Program (NTP) Nonneoplastic Lesion Atlas-A guide for standardizing terminology in toxicologic pathology for rodents. Testis, Seminiferous tubule-Giant cells. Available from: https://ntp.niehs.nih.gov/atlas/nnl/ reproductive-system-male/testis/semi-niferoustubule-giant cells; accessed March 2022.

Duty S. M., Calafat A. M., Silva M. J., Ryan L., Hauser R., 2005. Phthalate exposure and reproductive hormones in adult men. Human Reprod, 20(3): 604–610.

EC-European Commission, 2000. Substance ID:84-69-5. Diisobutyl phthalate. IUCLID Dataset. European Chemicals Breau. Available online at http://ecb.jrc.ec.euro-pa.eu/iuclid-datasheet/84695.pdf

EC-European Commission, 2004. Diisobutyl phthalate. Commission of the European Communities. European Breau. ECBI/116/04. Available online at http://ecb.jrc.it.classlab/agenta/_ag_Health_0305

Elsisi A. E, Carter D. E, Sipes I. G., 1989. Dermal absorption of phthalate diesters in rats. Fundam. Appl. Toxicol, 12: 70–77.

Fromme H., Gruber L., Seckin E., Raab U., Zimmermann S., Kiranoglu M, et al., 2011. Phthalates and their metabolites in breast milk–results from the Bavarian Monitoring of Breast Milk (BAMBI). Environ. Int, 37: 715–722.

Heudorf U., Mersch-Sundermann V., Angerer J., 2007. Phthalates Toxicology and Exposure. Int J Hyg Environ Health, 210: 623–634.

Hild S. A., Reel J. R., Dykstra M. J., Mann P. C., Marshall G. R., 2007. Acute adverse effects of the indenopyridine CDB-4022 on the ultrastructure of Sertoli cells, spermatocytes, and spermatids in rat testis: Comparison to the known Sertoli cell toxicant di-n-pentylphthalate (DPP). J Androl, 28: 621–629. doi: 10.1016/ j.taap.2022.116262

HSDB, 2017. Diisobutyl Phthalate. National Library of Medicine, Bethesda, MD.

Koch H. M., Christensen K. L. Y., Harth V., Lorber M., Brüning T., 2012. Di-n-butyl phthalate (DnBP) and diisobutyl phthalate (DIBP) metabolism in a human volunteer after single oral doses. Arch. Toxicol, 86: 1829–1839.

Latini G., Wittassek M., Del Vecchio A., Presta G., De Felice C., Angerer J., 2009. Lactational exposure to phthalates in southern Italy. Environ. Int, 35: 236–239.

Li Q., Zhu Q., Tian F., Li J., Shi L., Yu Y., Zhu Y., Li H., Wang Y., Ge R. S., Li X., 2022. In utero di-(2-ethylhexyl) phthalate-induced testicular dysgenesis syndrome in male newborn rats is rescued by taxifolin through reducing oxidative stress. Toxicol Appl Pharmacol, 456: 11662. doi: 10.1016/ j.taap.2022.116262

Marsee K., Woodruff T. J., Axelrad D.A., Calafat A. M., Swan S. H., 2006. Estimated Daily phthalate exposures in a population of mothers of male infants exhibiting reduced anogenital distance. Environ Health Perspect, 114: 805–809.

Meistrich M. L., 1984. Stage-specific sensitivity of spermatogonia to different chemotherapeutic drugs. Biomed Pharmacother, 38: 137–142.

Meistrich M. M., 1986. Critical components of testicular function and sensitivity to disruption. Biol Reprod, 34: 17–28.

Mitchell R. T., Childs A. J., Anderson R. A., van den Driesche S., Saunders P. T. K., McKinnell C., Wallace W. H. B., Kelnar C. J. H., Sharpe R. M., 2012. Do Phthalates Affect Steroidogenesis by the Human Fetal Testis? Exposure of Human Fetal Testis Xenografts to Di-n-Butyl Phthalate. J Clin Endocrinol Metab, 97(3): 341–348. https://doi.org/10.1210/jc.2011-2411

OECD, Organisation for Economic Co-Operation and Development., 1995. Guidelines for Testing of Chemicals No: 407. Repeated Dose 28-day Oral Toxicity Study in Rodents, Organization for Economic Cooperation and Development, Paris, France.

Setchell B. P., 1990. Local control of testicular fluids. Reprod Fertil Dev, 2: 291–309.

Spade D. J., Hall S. J., Wilson S., Boekelheide K., 2015. Di-n-Butyl Phthalate Induces Multinucleated Germ Cells in the Rat Fetal Testis Through a Nonproliferative Mechanism. Biol Reprod, 93(5): 110.

Strucinski P., Goralczyk K., Ludwicki J.K., Czaja K., Hernik A., Korcz W., 2006. Levels of selected organochlorine insecticides, polychlorinated biphenyls, phthalates and perfluorinated aliphatic substances in blood. Rocz. Panstw. Zakl. Hig, 57: 99–112.

The chemical company, Diisobutyl Phthalate, 2021. Available from: https://thechemco.com/chemical/diisobutyl-phthalate; accessed, April 2021.

Wittassek M., Angerer J., Kolossa-Gehring M., Schafer S. D., Klockenbusch W., Dobler L..., 2009. Fetal exposure to phthalates-a pilot study. Int. J. Hyg. Environ. Health, 212: 492–498.

Wormuth M., Scheringer M., Vollenweider M., Hungerbuhler K., 2006. What are the sources of exposure to eight frequently used phthalic acid esters in Europeans? Risk Anal., 26: 803–824.

Yost E. E., Euling S. Y., Weaver J. A., Beverly B. E. J., Keshava N., Mudipalli A., Araga X., Blessinger T., Dishaw L., Hotchkiss A., Makris S. 2019. Environ Int., 125: 579–594. doi: 10.1016/j.envint. 2018.09.038