Abstract
Background
Cleft palate (CP) constitutes the most frequently seen orofacial cleft and is often associated with low folate status. Folate plays an essential role in the human body as a major coenzyme in one-carbon metabolism, including DNA synthesis, repair, and methylation. Whether the administration of isolated folic acid (FA) supplements prevents the CP caused by genetic mutations is unknown, as is its effect on the mechanisms leading to palate fusion.
Methods
FA was administered to females from two different strains of transforming growth factor β3 heterozygous mice. Null mutant progeny of these mice exhibit CP in 100% of cases of varying severity. We measured cleft length, height of palatal shelf adhesion, and the number of proliferating mesenchymal cells. Immunohistochemistry was also carried for collagen IV, laminin, fibronectin, cytokeratin-17, and EGF.
Results
FA supplementation significantly reduced CP severity and improved palatal shelf adhesion in both strains both in vivo and in vitro. Medial edge epithelium proliferation increased, and its differentiation was normalized as indicated by the presence and disposition of collagen IV, laminin, fibronectin, and cytokeratin-17.
Conclusions
A maternal FA supplementation reduces the CP appearance by improving the mechanisms leading to palatal shelf adhesion.
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Acknowledgements
The authors wish to thank Ms. Alicia Cerro and Ms. Dolores Arroyo (Universidad Complutense) for their assistance with the histology. We also thank the technical team of the Centro de Microscopía y Citometría of the Universidad Complutense for their assistance with the assessment of cell proliferation and Professor Colin Robinson (University of Leeds), the anonymous referees, and Proof-Reading-Service UK for their helpful suggestions and English editing of this manuscript.
Funding
This study was supported by grants to the Complutense Research Group 920202 from the Spanish Ministerio de Economía y Competitividad (PS09/01762) and Banco Santander-Universidad Complutense Madrid (PR41/17-21009).
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Y.L.-G., E.M., L.N., E.M.-S., I.P.-L., M.I.A., and T.P. took care of the animals and/or acquired the data. Y.L.-G., A.d.R., M.C.B., J.M., and C.M.-Á. analyzed the results obtained. C.M.-Á. designed the experiments and wrote the manuscript. All authors approved the manuscript before submission.
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López-Gordillo, Y., Maldonado, E., Nogales, L. et al. Maternal folic acid supplementation reduces the severity of cleft palate in Tgf-β3 null mutant mice. Pediatr Res 85, 566–573 (2019). https://doi.org/10.1038/s41390-018-0267-6
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DOI: https://doi.org/10.1038/s41390-018-0267-6
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