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Differential scanning calorimetric (DSC) analyses were performed between −130° and 100°C on specimens prepared from nickel-titanium (NiTi) rotary endodontic instruments: ProFile (n = 5), Lightspeed (n = 4), and Quantec (n = 3). The ProFile and Lightspeed instruments were in the as-received condition, whereas the Quantec instruments were randomly selected from a dental clinic and had unknown history. The DSC plots showed that the ProFile and Lightspeed instruments analyzed had the superelastic NiTi property, with an austenite-finish (Af) temperature of approximately 25°C. Differences in DSC plots for the ProFile instruments and the starting wire blanks (n = 2) were attributed to the manufacturing process. The phase transformation behavior when the specimens were heated and cooled between −130° and 100°C, the temperature ranges for the phase transformations, and the resulting enthalpy changes were similar to those previously reported for nickel-titanium orthodontic wires having superelastic characteristics or shape memory behavior in the oral environment. The experiments demonstrated that DSC is a powerful tool for materials characterization of these rotary instruments, providing direct information not readily available from other analytical techniques about the NiTi phases present, which are fundamentally responsible for their clinical behavior.