Combining High-Resolution Hard X-ray Tomography and Histology for Stem Cell-Mediated Distraction Osteogenesis

Distraction osteogenesis is a clinically established technique for lengthening, molding and shaping bone by new bone formation. The experimental evaluation of this expensive and time-consuming treatment is of high impact for better understanding of tissue engineering but mainly relies on a limited number of histological slices. These tissue slices contain two-dimensional information comprising only about one percent of the volume of interest. In order to analyze the soft and hard tissues of the entire jaw of a single rat in a multimodal assessment, we combined micro computed tomography (µCT) with histology. The µCT data acquired before and after decalcification were registered to determine the impact of decalcification on local tissue shrinkage. Identification of the location of the H&E-stained specimen within the synchrotron radiation-based µCT data collected after decalcification was achieved via non-rigid slice-to-volume registration. The resulting bi- and tri-variate histograms were divided into clusters related to anatomical features from bone and soft tissues, which allowed for a comparison of the approaches and resulted in the hypothesis that the combination of laboratory-based µCT before decalcification, synchrotron radiation-based µCT after decalcification and histology with hematoxylin-and-eosin staining could be used to discriminate between different types of collagen, key components of new bone formation.