Fractal analysis is a powerful method for the morphological study of complex systems that is increasingly applied to biomedical images. Spatial resolution and image segmentation are crucial for the discrimination of tissue structures at the multiscale level. In this work, we have applied fractal analysis to high-resolution X-ray phase contrast micro-tomography (XrPCμT) images in both uninjured and injured tissue of a mouse spinal cord. We estimated the fractal dimension (FD) using the box-counting method on tomographic slices segmented at different threshold levels. We observed an increased FD in the ipsilateral injured hemicord compared with the contralateral uninjured tissue, which was almost independent of the chosen threshold. Moreover, we found that images exhibited the highest fractality close to the global histogram threshold level. Finally, we showed that the FD estimate largely depends on the image histogram regardless of tissue appearance. Our results demonstrate that the pre-processing of XrPCμT images is critical to fractal analysis and the estimation of FD.