Pretreatment Effect of Folic Acid on 13-Cis-RA-Induced Cellular Damage of Developing Midfacial Processes in Cultured Rat Embryos

Rungarun Kriangkrai1, *, Suconta Chareonvit2, Sachiko Iseki3, Visaka Limwongse1
1 Department of Oral Biology, Faculty of Dentistry, Naresuan University, Phitsanulok, Thailand.
2 Department of Anatomy, Faculty of Dentistry, Chulalongkorn University, Bangkok, Thailand
3 Department of Molecular Craniofacial Embryology, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan

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Creative Commons License
© 2017 Kriangkrai et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: ( This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Department of Oral Biology, Faculty of Dentistry, Naresuan University, 99 Moo 9 Phitsanulok-Nakornsawan Road, Amphor Muang, Phitsanulok, Thailand; Tel: +6655966066; Fax: +6655966053; E-mail:



Excess treatment of 13-cis-RA (Accutane®) on pregnant women induces craniofacial malformation found in infants. However, the effect of folic acid on 13-cis-RA-induced cellular damages of developing midfacial processes is still unknown. The purpose of this study was to investigate the pretreatment effect of folic acid (FA) on 13-cis-RA-induced cellular damage in developing midfacial processes in rat embryos.

Materials and Methods:

The rat embryos at developing midfacial processes were performed by whole embryo culture in vitro, in the presence of 13-cis-RA (20 µM) with or without pre-treatment of FA (100 µM). The midfacial morphogenesis score, PCNA and TUNEL assay staining were evaluated for morphogenesis, cell proliferation and apoptosis of the midfacial processes, respectively.


The 13-cis-RA-treated embryos at 24h showed atrophy of midfacial processes with significantly decreased morphogenesis score and cell proliferation, and increased apoptotic cell death. In contrast, the embryos pre-treated with FA for 18h, followed by 13-cis-RA treatment for 24h (FA-RA) showed significantly greater morphogenesis score, increased cell proliferation and lower apoptotic cell death compared to those of the 13-cis-RA-treated embryos.


The results suggest that FA reduced the teratogenic effects of 13-cis-RA on midfacial process tissue. Future investigations regarding the anti-teratogenic mechanism of FA on the prevention of damages in midface processes induced by 13-cis-RA on pregnant woman are warranted.

Keywords: Apoptosis, Cell proliferation, Folic acid, Midfacial process, 13-Cis-RA, Rat embryo.