Effect of nano-encapsulation of β-carotene on Xenopus laevis embryos development (FETAX)

Abstract

Vitamin A plays a vital role during embryo development as most precursor of embryonic retinoic acid, a key morphogen during embryogenesis. Carotenoids, including β-carotene, are important vegetal source of Vitamin A and in contrast to teratogenic potential of animal-derived retinoids, β‐carotene is usually considered freed from embryotoxic effects and supplements in pregnancy with β-carotene are suggested. The aim of the present work is to evaluate the effect of bulk and nano-encapsulated β-carotene on embryo development, by using the animal model Frog Embryo Teratogenesis Assay: Xenopus- FETAX. Xenopus laevis embryos were exposed from late gastrula till pharyngula (the phylotypic stage for vertebrates) to the concentrations of BULK β-carotene 150-3000 ng/mL and NANO β-carotene 0.75-30 ng/mL. At pharyngula stage, some embryos were processed for whole mount neural crest cell immunostaining, while others embryos were allowed to develop till tadpole for morphological and histological evaluation of neural crest cells-derived structures. In this model, the nano-encapsulated β-carotene induced specific malformations at craniofacial and eye level, while the bulk formulation only induced developmental delays. Finally, the applied alternative animal model resulted a rapid and sensitive screening method able to re-evaluate the teratogenic profile of nano-encapsulated micronutrients.