The changes in quality of mandibular bone mineral in otherwise totally immobilized rhesus monkeys.

The status of bone mineral and osteocalcin in the young adult Rhesus monkey mandible was assessed following a 14-day period of postcranial immobilization, and after 7- and 28-day recovery periods. Specimens of cortical bone taken from the compact bone at the inferior border of the jaws were ground in liquid nitrogen and sieved to a particular size below 20 micron. The bone powder was then fractionated in a bromoform-toluene density gradient to determine its mineralization profile (Ca, P, CO3, and osteocalcin), and X-ray diffraction was used to determine apatite crystal size in some fractions. There was no change in the chemistry of the mandibular bone from the immobilized animals. However, the mineralization profile in that group showed a significant shift toward the higher density fractions, indicating the presence of a greater than normal content of mature well-mineralized bone. While this trend was accentuated in the jaws following a 7-day postimmobilization recovery period, partial recovery of the normal profile was observed after a 28-day recovery period. The osteocalcin profile shifted like the mineralization profile during the immobilization and recovery periods. X-ray diffraction analyses showed that the shift in the mineralization profile during the immobilization period was associated with a decrease in apatite crystal size.

Calcergy and calciphylaxis: timed appearance of gamma-carboxyglutamic acid and osteocalcin in mineral deposits.

gamma-Carboxyglutamate (Gla), a calcium binding amino acid whose synthesis depends on vitamin K, has been found in association with pathologic calcifications. It is of interest therefore to examine the role of Gla-containing proteins in the formation of nonskeletal mineralized tissues. Calcergy and calciphylaxis, experimentally induced models of pathologic calcification, offer the opportunity to study the formation of mineral deposits in the absence of an endochondral sequence of bone formation. Before induction of subcutaneous calcinosis by topical treatment with the direct calcergen, KMnO4, or by challenging dihydrotachysterol-sensitized animals with FeCl2, control specimens contain no gamma-carboxyglutamic acid. With the initial formation of cytoplasmic vesicles, calcium content of the tissues increases and Gla is detected. Gla levels are further elevated with the appearance of poorly crystalline apatite-like crystallites. Origin of protein bound Gla was established by positive identification of osteocalcin by radioimmunoassay. Gla and osteocalcin appear concomitant with the earliest mineral deposits observed by electron microscopy and micro X-ray analysis. The formation of organized extracellular pattern by X-ray diffraction allowed sufficient mineral accumulation for detection with calcium, Gla, and osteocalcin increasing allowed sufficient mineral accumulation for detection of an apatite-like pattern by X-ray diffraction with calcium, Gla, and osteocalcin increasing proportionately as mineral is deposited.