This study evaluated, in vitro, the loss of tooth substance after cavity preparation for direct and indirect restorations and its relationship with fracture strength of the prepared teeth. Sixty sound human maxillary first premolars were assigned to 6 groups (n=10). MOD direct composite cavities (Groups I, II and III) and indirect inlay cavities (Groups IV, V and VI) were prepared maintaining standardized dimensions: 2-mm deep pulpal floors, 1.5-mm wide gingival walls and 2-mm high axial walls. Buccolingual width of the occlusal box was established at 1/4 (Groups I and IV), 1/3 (Groups II and V) or 1/2 (Groups III and VI) of the intercuspal distance. Teeth were weighed (digital balance accurate to 0.001 g) before and after preparation to record tooth substance mass lost during cavity preparation. The prepared teeth were submitted to occlusal loading to determine their fracture strength using a universal testing machine at a crosshead speed of 0.5 mm/min. Data were analyzed by two-way ANOVA and Tukey test (alpha= 0.05). 1/4-inlay cavities had higher percent mean mass loss (9.71%) than composite resin cavities with the same width (7.07%). 1/3-inlay preparations also produced higher percent mean mass loss (13.91%) than composite resin preparations with the same width (10.02%). 1/2-inlay cavities had 21.34% of mass loss versus 16.19% for the 1/2-composite resin cavities. Fracture strength means (in kgf) were: GI = 187.65; GII = 143.62; GIII = 74.10; GIV = 164.22; GV = 101.92; GVI = 50.35. Statistically significant difference (p