Gold standard for determining bone density as a surrogate parameter of bone quality is measurement of bone mineral density (BMD) by dual energy X-ray absorptiometry (DXA), most commonly performed on the lumbar spine (L1-L4). Computed tomography (CT) data are often available for surgical planning prior to spine procedures, but currently this information is not standardized for bone quality assessment. Besides, measuring the Hounsfield-Units (HU) is also of great importance in the context of biomechanical studies. This in vitro study aims in comparing BMD from DXA and HU based on diagnostic CT scans. In addition, methods are presented to quantify local density variations within bones. One hundred and seventy-six vertebrae (L1-L4) from 44 body donors (age 84.0±8.7 years) were studied. DXA measurements were obtained on the complete vertebrae to determine BMD, as well as axial CT scans with a slice thickness of 1 mm. Using Mimics Innovation Suite image processing software (Materialise NV, Leuven, Belgium), two volumes (whole vertebra vs. spongious bone) were formed for each vertebra, which in turn were divided in their left and right sides. From these total of six volumes, the respective mean HU was determined. HU of the whole vertebra and just spongious HU were compared with the BMD of the corresponding vertebrae. Side specific differences were calculated as relative values. Whole bone and spongious HU correlated significantly (P>0.001; α=0.01) with BMD. A positive linear correlation was found, which was more pronounced for whole bone HU (R=0.72) than for spongious HU (R=0.62). When comparing the left and right sides within each vertebra, the HU was found to be 10% larger on average on one side compared to the opposite side. In some cases, the difference of left and right spongious bone can be up to 170%. There is a tendency for the side comparison to be larger for the spongious HU than for the whole vertebra. Determination of HU from clinical CT scans is an important tool for assessing bone quality, primarily by including the cortical portion in the calculation of HU. Unlike BMD, HU can be used to distinguish precisely between individual regions. Some of the very large side-specific gradients of the HU indicate an enormous application potential for preoperative patient-specific planning.