Assessment of chemical mixtures using biomarkers of combined biological activity: A screening study in human placentas

Reprod Toxicol. 2021 Mar;100:143-154. doi: 10.1016/j.reprotox.2021.01.002. Epub 2021 Jan 11.


Humans are simultaneously exposed to complex mixtures of chemicals with limited knowledge on potential health effects, therefore improved tools for assessing these mixtures are needed. As part of the Human Biomonitoring for Europe (HBM4EU) Project, we aimed to examine the combined biological activity of chemical mixtures extracted from human placentas using one in vivo and four in vitro bioassays, also known as biomarkers of combined effect. Relevant endocrine activities (proliferative and/or reporter gene assays) and four endpoints were tested: the estrogen receptor (ER), androgen receptor (AR), and aryl hydrocarbon receptor (AhR) activities, as well as thyroid hormone (TH) signaling. Correlations among bioassays and their functional shapes were evaluated. Results showed that all placental extracts agonized or antagonized at least three of the abovementioned endpoints. Most placentas induced ER-mediated transactivation and ER-dependent cell proliferation, together with a strong inhibition of TH signaling and the AR transactivity; while the induction of the AhR was found in only one placental extract. The effects in the two estrogenic bioassays were positively and significantly correlated and the AR-antagonism activity showed a positive borderline-significant correlation with both estrogenic bioassay activities. However, the in vivo anti-thyroid activities of placental extracts were not correlated with any of the tested in vitro assays. Findings highlight the importance of comprehensively mapping the biological effects of "real-world" chemical mixtures present in human samples, through a battery of in vitro and in vivo bioassays. This approach should be a complementary tool for epidemiological studies to further elucidate the combined biological fingerprint triggered by chemical mixtures.

Keywords: Bioassay; Biomarkers; Chemical mixtures; Combined effect; Endocrine disruption; HBM4EU; Placenta; Reproduction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Androgen Receptor Antagonists
  • Animals
  • Antithyroid Agents / analysis
  • Biological Assay
  • Biological Monitoring
  • Biomarkers / analysis*
  • Endocrine Disruptors / analysis
  • Environmental Exposure*
  • Environmental Pollutants / adverse effects*
  • Europe
  • Female
  • Genes, Reporter
  • Humans
  • MCF-7 Cells
  • Male
  • Placenta / chemistry*
  • Pregnancy
  • Receptors, Androgen / analysis
  • Receptors, Androgen / genetics
  • Receptors, Aryl Hydrocarbon / genetics
  • Receptors, Estrogen / genetics
  • Signal Transduction
  • Thyroid Hormones / metabolism
  • Xenopus laevis


  • Androgen Receptor Antagonists
  • Antithyroid Agents
  • Biomarkers
  • Endocrine Disruptors
  • Environmental Pollutants
  • Receptors, Androgen
  • Receptors, Aryl Hydrocarbon
  • Receptors, Estrogen
  • Thyroid Hormones