Cancellous centrum is a major component of the vertebral body and significantly contributes to its structural strength and fracture risk. We hypothesized that the variability of cancellous bone properties in the centrum is associated with vertebral strength. Microcomputed tomography (micro-CT)-based gray level density (GLD), bone volume fraction (BV/TV), and finite element modulus (E) were examined for different regions of the trabecular centrum and correlated with vertebral body strength determined experimentally. Two sets of images in the cancellous centrum were digitally prepared from micro-CT images of eight human vertebral bodies (T10–L5). One set included a cubic volume (1 per vertebral centrum, n = 8) in which the largest amount of cancellous material from the centrum was included but all the shell materials were excluded. The other set included cylindrical volumes (6 per vertebral centrum, n = 48) from the anterior (4 regions:​ front, center, left, and right of the midline of vertebra) and the posterior (2 regions:​ left and right) regions of the centrum. Significant positive correlations of vertebral strength with GLD (r² = 0.57, p = 0.03) and E (r² = 0.63, p = 0.02) of the whole centrum and with GLD (r² = 0.65, p = 0.02), BV/TV (r² = 0.72, p = 0.01) and E (r² = 0.85, p = 0.001) of the central region of the vertebral centrum were found. Vertebral strength decreased with increasing coefficient of variation of GLD, BV/TV, and E calculated from subregions of the vertebral centrum. The values of GLD, BV/TV, and E in centrum were significantly smaller for the anterior region than for the posterior region. Overall, these findings supported the significant role of regional variability of centrum properties in determining the whole vertebral strength.