The aim of the present study was to evaluate the effects of storage media on the longitudinal hardness changes of tissue conditioning materials. Four tissue-conditioning materials were used for fabrication of 80 disc-shaped specimens and divided in four groups, stored in four storage media. The specimens underwent artificial ageing corresponding to 30 nights of extra-oral storage. Hardness measurements were obtained at nine intervals between 8 and 240 hours after specimen fabrication. To test the effects of storage media on hardness we employed multivariate modelling (Bonferroni correction; α = 0.05). The materials exhibited varying hardness changes, most pronounced when stored in ambient air.