Osteopontin mRNA expression during bone resorption: an in situ hybridization study of induced ectopic bone in the rat

To study the relationship between the expression of osteopontin mRNA and ectopic bone formation and resorption, in situ hybridization using 35S-labeled RNA probes was performed on ectopic bone that was induced in an experimental rat model. The expression of type I collagen and osteocalcin in this ectopic bone was also examined. After 6-week-old male Wistar rats were injected intravenously with colchicine at a dose of 1 mg/kg, trabecular bone-like ectopic calcified tissue had formed in the medial bone marrow cavity of the tibia on day 4, and then continued to increase progressing forward to the distal cavity. On day 8 peak growth was attained, after which it was resorbed within 4 days as a result of osteoclast recruitment. Subsequent in situ hybridization of sections of the ectopic bone at 4, 6, 8, and 10 days after this colchicine treatment revealed that high levels of the type I collagen mRNA were expressed in the osteoblasts of the mineralized ectopic bone surface, not only during the formation period but also during bone resorption. Although osteocalcin showed no specific signals throughout the experiments, osteopontin mRNA was expressed temporarily at day 10 during the initial phases of ectopic bone resorption, primarily in both osteoblasts and osteocytes and further in some osteoclasts. These results suggest that de novo synthesis of osteopontin is closely associated with bone resorption and could possibly be required to initiate and mediate this biological process.