Calcification and resorption of mouse Meckel’s cartilage analyzed by von Kossa and tartrate-resistant acid phosphatase histochemistry and scanning electron microscopy/energy-dispersive X-ray spectrometry
Calcification and resorption of mouse Meckel’s cartilage analyzed by von Kossa and tartrate-resistant acid phosphatase histochemistry and scanning electron microscopy/energy-dispersive X-ray spectrometry
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通过 von Kossa 和抗酒石酸酸性磷酸酶组织化学和扫描电子显微镜/能量色散 X 射线光谱法分析小鼠梅克尔软骨的钙化和吸收
DOI：
10.1007/s12565-021-00643-6
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发表时间：
2021
影响因子：
1.2
通讯作者：
Sasano Yasuyuki
中科院分区：
文献类型：
--
作者：
Nakamura Megumi;Yang Mu-Chen;Ashida Keijyu;Mayanagi Miyuki;Sasano Yasuyuki
梅克尔软骨对于下颌骨的正常发育至关重要。梅克尔软骨中间部分的命运是独特的，因为除了形成蝶下颌韧带的部分外，大部分软骨在出生后不久就消失了。梅克尔软骨消失的机制尚不清楚；因此，本研究旨在研究梅克尔软骨退化的过程，重点关注软骨基质钙化和破软骨细胞的出现。对胚胎第 15、16、17 和 18 天以及出生后第 2 天发育中的小鼠下颌骨进行处理，用阿新蓝和茜素红进行整体染色。将胚胎第15、16、17和18天的下颌骨固定并包埋在石蜡中。对相邻切片进行 von Kossa 和抗酒石酸酸性磷酸酶 (TRAP) 组织化学和扫描电子显微镜/能量色散 X 射线光谱 (SEM/EDS) 处理。用 von Kossa 组织化学和 SEM/EDS 检查钙化和钙、磷和碳的元素浓度。使用TRAP组织化学研究了破软骨细胞的参与情况。结果表明，软骨细胞肥大和软骨基质钙化后，梅克尔软骨的中间部分被软骨细胞吸收，钙化的梅克尔软骨的矿物质浓度与周围骨的矿物质浓度相当。这项研究有助于理解梅克尔软骨吸收的机制，并为下颌骨的发育提供有用的见解。
Meckel’s cartilage is essential for the normal development of the mandible. The fate of the intermediate portion of Meckel’s cartilage is unique as most of it disappears soon after birth except for the part that forms the sphenomandibular ligament. The mechanism of the disappearance of Meckel’s cartilage is unknown; therefore, this study was designed to investigate the process of Meckel’s cartilage degradation, focusing on cartilage matrix calcification and the appearance of chondroclasts. Developing mouse mandibles at embryonic days 15, 16, 17, and 18, and postnatal day 2 were processed for whole-mount staining with alcian blue and alizarin red. The mandibles on embryonic days 15, 16, 17, and 18 were fixed and embedded in paraffin. Adjacent sections were processed for von Kossa and tartrate-resistant acid phosphatase (TRAP) histochemistry and scanning electron microscopy/energy-dispersive X-ray spectrometry (SEM/EDS). Calcification and the element concentrations of calcium, phosphorus, and carbon were examined with von Kossa histochemistry and SEM/EDS. The involvement of chondroclasts was investigated using TRAP histochemistry. The results demonstrated that the intermediate portion of Meckel's cartilage is resorbed by chondroclasts after chondrocyte hypertrophy and cartilage matrix calcification and that the mineral concentration of calcified Meckel's cartilage is comparable to that of the surrounding bone. This study contributes to the understanding of the mechanism of Meckel's cartilage resorption and provides useful insights into the development of the mandible.