Increased lifetime of a functional dentition demands renewed understanding of tooth and restorative wear processes, including development of improved assays for assessing tooth wear.  Recent studies have explored profilometry and microhardness indentation loss methods for evaluating wear related phenomena.  Objectives:  This study reports on a 3D profilometry method (3DP) developed to assess tooth wear phenomena and compares abrasivity assessment values with those obtained by microhardness indentation loss (IL) and standard radiotracer methodologies.  Methods:  A V-8 brushing machine was used to brush flattened enamel or dentin samples for 10,000 (3DP) or 1,500 (IL) strokes.  3DP samples had a masked control surface throughout brushing.  Combining pre- and post-brushing surface maps generated by a Taylor-Hobson Form Talysurf Series 2 profilometer and a computer algorithm designed to reposition the surfaces based on aligning demarcations on the protected surface permitted volume tooth substrate lost assessment from brushing which converted into an average delta z-axis mineral loss.  For IL studies, measuring the post-brushing Vickers Hardness indent change permitted average delta z-axis mineral loss evaluation.  Results:  Dentin wear for low (DCPD), mid (silica) and high (alumina-silica blend) RDA dentifrices measured:  11.7±4.2; 45.2±5.2; 55.5±5.8 microns respectively under standardized conditions after 10,000 strokes; while enamel wear measured:  0.32±0.16; 0.71±0.12; 1.13±0.16 microns after 1,500 strokes.  There were significant linear correlations between developed methodologies and RDA and REA assessments from historical accounts.  Conclusions:  Tested methodologies demonstrated enamel wear at <10% dentin values, following enamel ‘conditioning'.   Both 3DP and IL methods provide useful substitutes for standardized radiotracer techniques.  The 3DP method worked more reliably on dentin, while the IL method worked more reliably on enamel.  These methods may be ideal in applications to wear related testing including assessments of additive or synergistic contributions of dietary or chemical treatment effects to dentition or in in vivo applications of wear related testing.