This study characterized the interactions of mineral trioxide aggregate with a synthetic tissue fluid composed of a neutral phosphate buffer saline solution and root canal dentin in extracted human teeth using inductively coupled plasma-atomic emission spectroscopy, scanning electron microscopy, energy dispersive X-ray analysis, and X-ray diffraction. Mineral trioxide aggregate exposed to synthetic tissue fluid at 37 degrees C released its metallic constituents and produced precipitates with a composition and structure similar to that of hydroxyapatite [Ca10(PO4)6(OH)2-HA]. Endodontically prepared teeth filled with mineral trioxide aggregate and stored in synthetic tissue fluid at 37 degrees C for 2 months produced at the dentin wall an adherent interfacial layer that resembled hydroxyapatite in composition. The authors conclude that Ca, the dominant ion released from mineral trioxide aggregate, reacts with phosphates in synthetic tissue fluid, yielding hydroxyapatite. The dentin-mineral trioxide aggregate interfacial layer results from a similar reaction. The sealing ability, biocompatibility, and dentinogenic activity of mineral trioxide aggregate is attributed to these physicochemical reactions.