In dental research, restorative materials have been regularly subjected to alternating in vitro thermal stress in investigations since the 1950s, in order to simulate in vivo alternating temperature stress and to artificially stress them in vitro. The provocation temperature is mostly 5 degrees C for cold provocation, and 55 degrees C for hot provocation. These temperatures are determined quite arbitrarily based on very few examinations in vivo. Extensive temperature data for the approximal space of teeth, which is decisive for the success of fillings adhesively attached to dentin, has so far not been addressed. The objective of this study was to examine the interproximal temperature characteristics created in the space of all teeth in vivo with thermal alternating stress, and therefore to validate the in vitro standardized thermal alternating stress of 5-55 degrees C. Fifteen study participants with healthy teeth were used to determine the temperature in each inter-dental space, resulting from hot/cold provocation in the upper and lower jaw, from the central incisor to the second molars. This was performed by a thermal element (cable sensor GTF 300, Greisinger Electronic GmbH, Regenstauf, Germany). The temperature sensor was attached with dental floss into the interproximal space and the temperature was recorded by the computer. The participants in the pilot test had to state when they were able to sip an 85 degrees C hot drink. That particular temperature value was taken for hot provocation as maximum temperature reference. Cold ice water (0 degrees C) was used for cold provocation as minimum temperature reference. The respective recordings with a total of 14 measurements for each individual were performed simultaneously in the upper and lower jaw. The study participants were to start with hot provocation, followed by cold provocation. This cycle was repeated at least once with an individual dwell time. The highest recorded approximal space temperature was 52.8 degrees C in the lower jaw, between the first and the second premolar. The lowest temperature of 13.7 degrees C was recorded in two participants in the upper jaw, between the 1st and 2nd incisor, and between the two central incisors. The mean of the maximum temperatures was 43.8+/-3.7 degrees C, and the mean of the minimum temperatures 24.2+/-4.6 degrees C. The mean initial temperature was 35.2+/-1.3 degrees C. None of the recordings reached either the upper threshold (55 degrees C) or the lower threshold (5 degrees C). This study showed that the actual thermal stress in the interproximal space of teeth is slightly lower than the one used in in vitro examinations. For class II cavities, most of the alternating temperature stress limits selected at 5-55 degrees C cover the actually occurring temperature interval quite well.