Sox9 is a transcription factor belonging to the SRY family of high-mobility box proteins, and plays a major role in endochondral ossification. Sox9 is a potent activator of the type-2 collagen pheno-type marker of articular cartilage. Regulation of osteogenic molecular signals in periosteal bone formation has not yet been elucidated yet. The purpose of the present study was to analyze histologically the bone formation in surgically released and repositioned periosteum, and to determine expression of Sox9 and type-2 collagen in periosteal bone formation of tibia and calvaria. After surgery, the released tibial periosteum formed ectopic cartilage. At 7 days, a combination of endochondral and intramembranous ossification was apparent. Some fibroblasts derived from the released periosteum showed Sox9 expression. Chondrocytes and cartilage matrix both displayed type-2 collagen expression. At 7 days, an additional new bone was formed on the calvaria. Osteoblasts and fibroblasts derived from released calvarial periosteum did not express Sox9 or type-2 collagen. Sox9 was not expressed throughout the process periosteal bone formation on the calvaria. It is concluded that we revealed Sox9 and type-2 collagen expression in periosteal cells after periosteum release and that the generative potential of periosteal cells of calvaria is different from that of tibia.
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