TY - JOUR
T1 - Calcification in rat developing mandibular bone demonstrated by whole mount staining, micro-computed tomography and scanning electron microscopy with energy dispersive X-ray spectroscopy
AU - Henmi, Akiko
AU - Okata, Hiroshi
AU - Mikami, Yasuto
AU - Sasano, Yasuyuki
N1 - Funding Information:
advice and comments on the study. We thank Dr. Miyuki Mayanagi for her excellent technical assistance. We also thank Drs. Rika Hayashi, Masahiro Kozuka, Shunichi Shishido, and Yusuke Kakiuchi for their help. This work was supported in part by grants-in-aid (23592694, 24792170, 15K11002, 15K11003, 17K17081) from the Japan Society for the Promotion of Science.
Publisher Copyright:
© Biomedical Research Foundation. All rights reserved.
PY - 2017
Y1 - 2017
N2 - The study was designed to investigate calcification in developing rat mandibular bone using whole mount staining, micro-computed tomography (micro-CT) and scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX). Wistar rats at embryonic days 16, 18, and 20 and postnatal weeks 1 and 6 were used. Rats were fixed with 4% paraformaldehyde and heads were resected, frozen and sectioned for histology, then analysed with SEM-EDX. Some of the specimens were observed with micro-CT. Other rats were fixed and stained with alcian blue and alizarin red for whole mount staining. Histology and whole mount staining showed that osteoid was deposited around Meckel’s cartilage at day 15 and developed into bone at day 16. Accumulation of Ca and P was identified in the bone matrix with SEM-EDX. The area of bone expanded until week 6. The Ca/P ratio increased, whereas the C/Ca and C/P ratios decreased during development. Micro-CT demonstrated an increase in radio-opacity with bone development. The results suggest that rat mandibular bone formation is initiated around Meckel’s cartilage at day 15. Deposition and maturation of the calcium phosphate mineral increase gradually with decrease in the organic component as the rat mandible develops.
AB - The study was designed to investigate calcification in developing rat mandibular bone using whole mount staining, micro-computed tomography (micro-CT) and scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX). Wistar rats at embryonic days 16, 18, and 20 and postnatal weeks 1 and 6 were used. Rats were fixed with 4% paraformaldehyde and heads were resected, frozen and sectioned for histology, then analysed with SEM-EDX. Some of the specimens were observed with micro-CT. Other rats were fixed and stained with alcian blue and alizarin red for whole mount staining. Histology and whole mount staining showed that osteoid was deposited around Meckel’s cartilage at day 15 and developed into bone at day 16. Accumulation of Ca and P was identified in the bone matrix with SEM-EDX. The area of bone expanded until week 6. The Ca/P ratio increased, whereas the C/Ca and C/P ratios decreased during development. Micro-CT demonstrated an increase in radio-opacity with bone development. The results suggest that rat mandibular bone formation is initiated around Meckel’s cartilage at day 15. Deposition and maturation of the calcium phosphate mineral increase gradually with decrease in the organic component as the rat mandible develops.
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U2 - 10.2220/biomedres.38.277
DO - 10.2220/biomedres.38.277
M3 - Article
C2 - 29070777
AN - SCOPUS:85032337781
VL - 38
SP - 277
EP - 284
JO - Biomedical Research (Japan)
JF - Biomedical Research (Japan)
SN - 0388-6107
IS - 5
ER -