Folic acid rescues corticosteroid‐induced vertebral malformations in chick embryos through targeting TGF‐β signaling. Issue 11 (23rd April 2020)

• Record Type: Journal Article
• Authors: Cai, Hongmei; Lin, Li; Wang, Guang; Berman, Zachary; Yang, Xuesong; Cheng, Xin
• Is Part Of: Journal of cellular physiology. Volume 235:Issue 11(2020:Nov.)
• Page Start: 8626
• Page End: 8639
• Publication Date: 2020-04-23
• Subjects: dexamethasone (Dex) -- folic acid (FA) -- growth plate -- osteogenesis -- vertebral development; Physiology -- Periodicals; Cell physiology -- Periodicals; 571.6
• DOI: 10.1002/jcp.29707
• Languages: English
• ISSNs: 0021-9541

Abstract

Abstract: Folic acid (FA) is routinely supplemented in the food of pregnant women or women planning a pregnancy, but whether FA exerts a positive effect on preventing fetal bone malformation remains obscure. In this study, we first exposed chick embryos with different concentrations of FA (1–10, 000 pmol/egg) and studied vertebral mineralization and ossification through alcian blue and alizarin red as well as hematoxylin and eosin staining. Morphological measurements of the thoracic vertebral bodies demonstrated that 100 pmol/egg FA exhibited the tendency of shortening the growth plate, extended the ossification center, and increased the amount of Type I collagen. Second, we suggested that FA treatment promotes osteogenesis by demonstrating increased RUNX family transcription factor 2 (Runx2) and Osterix expressions in MC3T3‐E1 and ATDC5 cells. Transforming growth factor‐β (TGF‐β) signaling was also upregulated by FA exposure, and addition of smad2/3 small interfering RNA knocks down FA‐induced increased p‐smad2/3, Runx2, and Osterix expression in vitro during chondrogenesis induction. Third, we employed dexamethasone (Dex), exposed chick embryos as an animal model of skeletal developmental retardation, to explore whether FA could rescue the loss of embryonic bone mass. Micro‐computed tomography imaging showed that the addition of FA improved the reduction of bone mass in our model. Histological analysis of the vertebral bodies revealed that FA dramatically improved the delayed turnover of the zones of growth plate caused by Dex exposure. Immunofluorescence on the chick embryonic vertebrae and chondrocytes showed that FA supplementation upregulated the expression of TGF‐β1, p‐smad2/3, and improved Runx2 as well as Osterix expression in the Dex + FA group compared with the Dex group. Lastly, we found that supplementation with TGF‐β1 (1 ng/egg) rescued bone mass loss caused by Dex as was also seen in FA exposure. Taken together these results, our data revealed that FA supplementation was able to rescue Dex exposure‐induced inhibitive osteogenesis through targeting on the TGF‐β signaling pathway. Abstract : In summary, dexamethasone (Dex) exposure downregulates transforming growth factor‐β (TGF‐β) signaling while folic acid (FA) treatment rescues it. TGF‐β signaling triggers the transcription of differentiative markers such as Runx2 and Osterix. Subsequently, the process of growth plate cartilage differentiation and osteogenic differentiation are accelerated. FA treatment rescues Dex‐induced vertebral malformation in chick embryos.