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The behavior of cultured rat bone cells growing on modified polyethylene terephthalate (mPET), glass, and machinable ceramic substrates containing enstatite (MgO, SiO2) and glass (CaO-P2O5-Al2O3) was studied. Cell attachment was measured directly on the substrates using an image analysis system. Electron microscopy observations and the MTT test revealed that cells are able to spread and proliferate on the material surface, keeping a healthy ultrastructure on all materials tested in the present study. After having colonized the surface of the materials, as shown by immunocytochemistry, the cells synthesize an osteoid-like matrix composed of osteocalcin, type I collagen, and fibronectin fibrils. The titration of alkaline phosphatase activity showed that the cells grown on the ceramic exhibit a greater osteogenic activity than those grown on controls (glass and mPET). This osteogenic activity results in a mineralization of the extracellular matrix in cultures on ceramic or plastic whereas only few calcium phosphate crystallite traces were revealed by Von Kossa staining on glass. Enstatite constitutes, therefore, an environment compatible with in vitrobone cell life. © 1998 John Wiley & Sons, Inc. J. Biomed Mater Res (Appl Biomater) 43: 215–225, 1998