Ca-enriched amorphous mineral deposits associated with the plasma membranes of chondrocytes and matrix vesicles of rat epiphyseal cartilage

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Abstract
Electron microscopic study of tibial epiphyseal plates of young growing rats revealed amorphous-appearing electron dense deposits 5–35 nm in diameter, associated with the plasma membranes of more than 43% of the proliferative zone chondrocytes. Hypertrophic zone chondrocytes, however, revealed no plasma membrane-associated amorphous-appearing deposits. The membrane-associated densities were observable in unstained sections of tissues fixed in glutaraldehyde alone and in tissues double-fixed with glutaraldehyde and osmium tetroxide, and were extracted from ultrathin sections floated on neutral aqueous solutions of 4% ethyleneglycol bis-(β-aminoethyl ether) N,N′-tetraacetic acid (EGTA) for one-half hour. Energy dispersive X-ray analysis of the densities in scanning transmission electron microscope (STEM) mode revealed the presence of Ca, suggesting that the membrane-associated amorphous-appearing deposits are Ca-enriched. Similar deposits were observed in the membrane of matrix vesicles present in the longitudinal cartilaginous septae in the hypertrophic zone. Four types of matrix vesicles were encountered in the longitudinal cartilaginous septae; one type with amorphous-appearing deposits, another with crystallites, a third type with both amorphous-appearing and crystalline-like deposits, and a fourth that is empty. These observations are interpreted to indicate that chondrocytes of the reserve and proliferative zones play a direct role in mineralization by elaborating amorphous mineral deposits along their plasma membranes. These deposits are incorporated into budding matrix vesicles, which then play a role in the initiation of mineralization by supporting the spontaneous phase transformation of amorphous-appearing mineral to crystalline mineral.