Analysis of Chemotactic Property of CCN2/CTGF in Intramembranous Osteogenesis

Chemotaxis is a directed migration of cells in response to a gradient of extracellular molecules called chemoattractants. Development, growth, remodeling, and fracture healing of bones are advanced through intramembranous osteogenesis. Chemotaxis of preosteoblasts toward future bone formation sites observed in the early stage of intramembranous osteogenesis is a critical cellular process for normal bone formation. However, molecular biological mechanisms of the chemotaxis of preosteoblasts are not fully understood. We have recently clarified, for the first time, the critical role of the cellular communication network factor 2 (CCN2)/connective tissue growth factor (CTGF)-integrin α5-Ras axis for chemotaxis of preosteoblasts during new bone formation through intramembranous osteogenesis. In this chapter, we describe in detail the procedures of the in vivo and in vitro assays to investigate the chemotactic property of CCN2/CTGF and its underlying molecular biological mechanisms during intramembranous osteogenesis.