A comparative study of osteoblasts: in situ versus isolated specimens.

Osteoblasts and osteoclasts are known to adhere tenaciously to bone surfaces even when the fibrous periosteal covering is removed. Brush-smear isolates and in situ, whole bone mounts of the osteogenic periosteum on the inner and outer calvarial tables of young adult rats were examined at the light microscopic level after exposing the cells by removing the fibrous periosteum. Based on staining, enzymatic activity, and overall morphology, most of the cells could be classified as osteogenic in nature. However, detailed analysis revealed that some cells exhibited certain unique morphologic appearances that suggested the existence of subpopulations or variant forms of the conventional or prototypic osteoblast. Three variant forms were the large nucleate, small nucleate, and multinucleate osteoblast-like cells, the latter with nuclear numbers ranging from 2-10. The finding of all such forms in smears, short-term cell cultures, and in situ specimens greatly lessened the probability that the unusual forms were artifactual. Phase contrast microscopic studies and sectioning of cellular isolates for study at both the light and electron microscopic levels provided additional support for the existence of the multinucleate osteoblast-like cell. The small size of these osteoblast-like subpopulations and the orientation of the cells to the commonly used plane of section of most histologic bone specimens, particularly with the multinucleate type, could explain why they had escaped earlier detection.