Comprehensive three-dimensional microCT and signaling analysis reveal the teratogenic effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin on craniofacial bone development in mice.

Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in utero can result in osteogenic defect during palatogenesis, but the effects on other craniofacial bones and underlying mechanisms remain to be characterized. By treating pregnant mice with TCDD (40 μg/kg) at the vital craniofacial patterning stages (embryonic day 8.5, 10.5 and 12.5), and scanning and reconstructing the skulls at embryonic day 18.5 using microCT, we found that TCDD exposure at the earlier and later patterning stages induced variable craniofacial malformations, including premature fusion of metopic and coronal sutures, truncated palatal processes of maxillary and palatine bones, as well as opening oriented pterygoid processes. Further in vitro determination of the underlying mechanisms using human fetal palatal mesenchymal cells (hFPMCs) revealed that TCDD suppressed a wide variety of osteogenic genes responsible for osteoblast commitment and bone matrix synthesis and mineralization, through activating aryl hydrocarbon receptor (AhR) signaling and subsequently inhibiting estrogen signaling. The attenuation of AhR signaling significantly blocked the osteogenic toxicity, and partly restored the expressing level of estrogen receptor α (ERα). Additional treatment with ERα agonist (PPT) significantly relieved the activation of AhR and rescued the impairment of osteogenesis caused by TCDD. Together, our findings demonstrated that TCDD was teratogenic in numerous cranial neural crest cell-derived craniofacial bone development, and disrupted multiple genes for osteogenic differentiation via the TCDD-mediated AhR/ ERα signaling cross-talk.