Thorough pre-testing is critical in assessing the likely in vivo performance of spinal devices prior to clinical use. However, there is a lack of data available concerning the dynamic testing of lumbar (porcine model) total disc replacements in all six axes under preload conditions. The aim of this study was to provide new data comparing porcine lumbar spinal specimen stiffness between the intact state and after the implantation of an unconstrained total disc replacement, in 6 degrees of freedom. The dynamic, stiffness matrix testing of six porcine lumbar isolated disc specimens was completed using triangle waves at a test frequency of 0.1 Hz. An axial preload of 500 N was applied during all testing. Specimens were tested both in the intact condition and after the implantation of the total disc replacement. Sixteen key stiffness terms were identified for the comparison of the intact and total disc replacement specimens, comprising the 6 principal stiffness terms and 10 key off-axis stiffness terms. The total disc replacement specimens were significantly different to the intact specimens in 12 of these key terms including all six principal stiffness terms. The implantation of the total disc replacement resulted in a mean reduction in the principal stiffness terms of 100%, 91%, and 98% in lateral bending, flexion–extension, and axial rotation, respectively. The novel findings of this study have demonstrated that the unconstrained, low-friction total disc replacement does not replicate the stiffness of the intact specimens. It is likely that other low-friction total disc replacements would produce similar results due to stiffness being actively minimised as part of the design of low-friction devices, without the introduction of stiffening elements or mechanisms to more accurately replicate the mechanical properties of the natural intervertebral disc. This study has demonstrated, for the first time, a method for the quantitative comparative mechanical function testing of total disc replacements and provides baseline data for the development of future devices.