The fate of soft callus chondrocytes during long bone fracture repair.

Following fracture, the cartilaginous tissue of the soft callus is eventually replaced by bone. Removal of the cartilage is a critical part of the bone healing process but information concerning the changes in chondrocytes during this process is sparse. The aim of the study was to investigate the fate of chondrocytes in the soft callus during the bone repair process using a rabbit tibial fracture model. Fracture tissue was processed for collagen I-III and keratan sulphate immunohistochemistry to study changes in matrix composition and the TUNEL technique (terminal deoxynucleotidyl transferase medicated dUTP nick-end labelling) to identify death of soft callus chondrocytes. Transmission electron microscopy (TEM) was also carried out to investigate the ultrastructure of chondrocytes within the soft callus. Results showed that the size of the cartilage area decreased over time and that cartilage matrix was replaced with new matrix rich in collagen I and III. Chondrocytes became engulfed in the new matrix and appeared to stop producing cartilage matrix. Chondrocyte cell death was seen at the border of the soft callus, just within the newly produced matrix. TEM revealed that these dying/dead cells were not typically apoptotic in appearance. In conclusion, results indicate that chondrocytes of the soft callus die as a result of the progressive production of bone matrix which eventually engulfs them and leads to the remodelling of the area and eventual bone repair.