3D renderings of ultrastructural bone features.

<p>[<b>a</b>] Rendering of osteocyte lacunae and canaliculi in the whole imaged volume overlayed over the bottom slice shown in grayscale. Colors correspond to connected components and grayscale to mass density. Note the difference in structure in the interstitia and osteon: the connected cells are all in the osteonal tissue, the others in the interstitial. The canaliculi are considerably reduced in the interstitia. [<b>b</b>] Zoom on the highlighted lacuna in A showing the interaction between the canaliculi [pink] and the cement line [green], and branching of the canaliculi.</p

Retrieved information in the reconstructed images.

<p>[<b>a</b>] Transverse, [<b>b</b>] frontal and [<b>c</b>] sagittal slices through the images reconstructed from phase data. Grayscale is proportional to local density. Osteocyte lacunae [Lc] and canaliculi [Ca] can clearly be seen. The heterogeneous organization of the matrix by mineralized collagen fibers can also be distinguished [box]. In this sample, a continuous change in collagen orientation can be seen between adjacent lamellae. The cement line [Cm], separating osteonal [On] and interstitial [It] tissue, can clearly be distinguished as more mineralized than the surrounding matrix. Tissue close to osteocyte lacunae is also hypermineralized. [<b>d</b>] Zoom on the boxed area in C. Matrix orientation is clearly visible and canaliculi are seen as black dots. [<b>e</b>] Mass density histograms in the three tissue types. [<b>f</b>] Samples were extracted from the mid diaphysis of a human femur. [<b>g</b>] The blue cylinder shows the imaged region inside the sample. [<b>h</b>] Schematic of a transverse section showing the organization of lamellar bone in osteons, interstitial tissue and cement lines. Blue circle shows the positioning of A. [<b>i</b>] Rendering of the electron density in the sample [blue] and porosity [yellow]. Structures such as osteocyte lacunae [Lc] and canaliculi [Ca], the cement line [Cm] and collagen fibers are revealed.</p

Experimental setup and image reconstruction.

<p>[<b>a</b>] Schematic of experimental setup. The X-ray beam [X] is monochromatized and focused into a focal spot [F] by X-ray reflective optics [KB]. The sample [S] is positioned on a translation-rotation stage downstream of the focus and imaged onto a stationary detector. Due to the resulting divergent beam, different spot-sample distances [D1] and different free space propagation distances [D2] imply different magnification factors on the detector. [<b>b</b>] Images were recorded at four focus-to-sample distances over a complete turn of the sample at 2999 projection angles. The images were used to reconstruct the phase shift at each angle [phase retrieval PR], which was used as input to a tomographic reconstruction algorithm to reconstruct the 3D local mass density.</p

MorphoJ script

MorphoJ script for all geometric morphometric analyses performed

TPS files

TPS files with digitized landmarks (of both lateral and dorsal views)

Thin plate spline

R script for thin plate spline

Histograms of the lacunar volumes for the three different sites are shown.

<p>Histograms are normalized to the area under the total number of lacunae for each site. Bin size is set to 50 µm³. The transparent areas indicate the standard error for each site based on the individual samples.</p

Histograms of all jaw lacunae grouped in either BRONJ or healthy bone.

<p>The shaded areas correspond to the standard error based on the different samples. Histograms are normalized to the absolute amount of lacunae, bin size is 50 µm³. It should be noted that even though the histograms of the two groups look very similar, there are differences between the histograms of individual donors.</p

Mean and standard deviation of the investigated properties for the different anatomical sites and pathology are summarized.

<p>The * indicates that those properties are statistically significantly different between the BRONJ and control jaw.</p

Investigated differences with respect to anatomical sites, assessed by analyses of variance (ANOVA), followed by post hoc multiple comparison Bonferroni tests are summarized.

<p>Significant differences between groups are indicated by a horizontal bar. If the significance level was reached, the p-Value and the F-Value are reported.</p