The in vitro behavior of fetal condylar cartilage in serum-free hormone-supplemented medium

Abstract

Mandibular condyles of fetal mice 19–20 days in utero were kept in a serum-free organ culture system for up to 14 days and were investigated for their capacity to develop osteoid and to mineralize in vitro. After 3 days in culture, the cartilage of the mandibular condyle appeared to have maintained all its inherent structural characteristics, including its various cell layers: chondroprogenitor, chondroblastic, and hypertrophic. After 7–9 days in culture, no chondroblasts could be seen; instead, most of the cartilage consisted of hypertrophic chondrocytes. In addition, various areas throughout the explant revealed the appearance of osteoidlike material. The process of matrix mineralization progressed with time, and by the 14th day new bonelike material was found to occupy a larger portion of the explant. The newly formed matrix reacted positively with antibodies against type I and type III collagens, as well as against bone alkaline phosphatase. Electron microscopic examination showed that the mineralization appeared to be associated with collagen fibers as well as the matrix vesicles. In composition, the in vitro-formed mineral deposits resembled hydroxyapatite crystals. Biochemical assays indicated that the newly formed tissue reacted strongly for alkaline phosphatase and incorporated 45Ca. The findings of the present study imply that fetal condylar cartilage possesses the potential to develop in vitro osseouslike tissue even in a system that is serum-free. Due to the fact that the newly formed extracellular matrix mineralized and reacted positively to bone markers as well as to cartilage macromolecules, it would seem justifiable to define the new tissue as chondroid bone.