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.

Abstract

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.