Localisation of bone-forming cells during fracture healing by osteocalcin immunocytochemistry: an experimental study of the rabbit tibia.

An immunocytochemical method was used to localise osteocalcin-producing cells during fracture healing in a rabbit model. In preliminary studies, tibial growth plates from young rabbits were used as a source of new bone formation, in order to determine the optimal tissue preparatory techniques. In the present study, a tibial shaft fracture was created in adult rabbits to study closed fracture healing. An indirect peroxidase method was used to stain paraffin-embedded tissue sections for osteocalcin. Osteocalcin-producing cells were positively identified at the periosteal and endosteal surfaces near the fracture site. Osteocalcin staining was not demonstrated in the surrounding soft tissues. At the interface between newly formed bone trabeculae and the cartilage layer within the callus, chondrocytic cells consistently showed localisation of osteocalcin. Within cartilaginous areas of the callus, some chondrocytes showed positive staining for osteocalcin. These cells were often seen in the proximity of blood vessels. The findings suggest that during fracture healing, under certain conditions, chondrocytes are capable of producing osteocalcin and thus could be considered capable of possible transformation into osteoblasts.