Matrix constituents of early developing bone examined by freeze fracture

Specimens of aldehyde-fixed and glycerol-impregnated early developing bone matrix, obtained from rat calvaria, were examined by the freeze-fracture method. The developing bone matrix reveals collagen fibrils, numerous membranous structures and a granular background. The collagen fibrils, when viewed longitudinally, exhibit a substructure of thinner filaments (microfibrils) which appear to follow a twisted course along the fibril-axis. Some of the membranous structures are readily identified as osteoblast processes. Others, which are round, ovoid or irregular in shape, were found either with or without intramembrane particles (IMPs). It is concluded that the round or ovoid IMP-containing membranous structures correspond to matrix-vesicles. The nature of the IMP-free bodies, however, is uncertain. They may be artefacts or genuine matrix-vesicles deriving from unstable membrane domains which have a propensity for blebbing or budding off. Confirmation of the latter possibility might come from examination of directly frozen specimens.