Figure 1

Exposure to high glucose levels causes deficiencies in cranial osteogenesis in chick embryos.

(A–C) Alcian Blue/Alizarin Red S double staining was performed in control (A), 50 mM L-glucose treated (B) and 50 mM D-glucose treated (C) 12-day chick embryo skulls, respectively. Alcian Blue stains cartilage (blue) and Alizarin Red stains ossified bone (red). We focused on the region of FR (indicated by the black arrows) and PA developmental defects (indicated by the white arrows). (D–F) Alizarin Red S staining was performed in control (D), 50 mM L-glucose treated (E) and 50 mM D-glucose treated (F) 12-day chick embryo skulls, respectively. (D’–F’) The high magnification images from the sites indicated by the black dotted squares in (D–F), respectively. Impaired PA was indicated by the black arrows. (G) The bar chart showing the frequency comparison of PA developmental defects following exposure to high levels of glucose. (H) The bar chart showing the area of PA following glucose treatment. *P < 0.05 indicates a significant difference between the experimental and control embryos. Abbreviations: Con, control; L-Glc, L-Glucose; D-Glc, D-Glucose; PA, parietal bone; FR, frontal bone and SQ, squamous bone. Scale bars = 5 mm in (A–C), 2 mm in (D–F) and 1 mm in (D’–F’).