High-resolution multicontrast tomography with an X-ray microarray anode-structured target source.

Multicontrast X-ray imaging with high resolution and sensitivity using Talbot-Lau interferometry (TLI) offers unique imaging capabilities that are important to a wide range of applications, including the study of morphological features with different physical properties in biological specimens. The conventional X-ray TLI approach relies on an absorption grating to create an array of micrometer-sized X-ray sources, posing numerous limitations, including technical challenges associated with grating fabrication for high-energy operations. We overcome these limitations by developing a TLI system with a microarray anode-structured target (MAAST) source. The MAAST features an array of precisely controlled microstructured metal inserts embedded in a diamond substrate. Using this TLI system, tomography of a Drum fish tooth with high resolution and tri-contrast (absorption, phase, and scattering) reveals useful complementary structural information that is inaccessible otherwise. The results highlight the exceptional capability of high-resolution multicontrast X-ray tomography empowered by the MAAST-based TLI method in biomedical applications.

Nanoscale X-ray microscopic imaging of mammalian mineralized tissue.

A novel hard transmission X-ray microscope (TXM) at the Stanford Synchrotron Radiation Lightsource operating from 5 to 15 keV X-ray energy with 14 to 30 microm2 field of view has been used for high-resolution (30-40 nm) imaging and density quantification of mineralized tissue. TXM is uniquely suited for imaging of internal cellular structures and networks in mammalian mineralized tissues using relatively thick (50 microm), untreated samples that preserve tissue micro- and nanostructure. To test this method we performed Zernike phase contrast and absorption contrast imaging of mouse cancellous bone prepared under different conditions of in vivo loading, fixation, and contrast agents. In addition, the three-dimensional structure was examined using tomography. Individual osteocytic lacunae were observed embedded within trabeculae in cancellous bone. Extensive canalicular networks were evident and included processes with diameters near the 30-40 nm instrument resolution that have not been reported previously. Trabecular density was quantified relative to rod-like crystalline apatite, and rod-like trabecular struts were found to have 51-54% of pure crystal density and plate-like areas had 44-53% of crystal density. The nanometer resolution of TXM enables future studies for visualization and quantification of ultrastructural changes in bone tissue resulting from osteoporosis, dental disease, and other pathologies.

Mineral density volume gradients in normal and diseased human tissues.

Clinical computed tomography provides a single mineral density (MD) value for heterogeneous calcified tissues containing early and late stage pathologic formations. The novel aspect of this study is that, it extends current quantitative methods of mapping mineral density gradients to three dimensions, discretizes early and late mineralized stages, identifies elemental distribution in discretized volumes, and correlates measured MD with respective calcium (Ca) to phosphorus (P) and Ca to zinc (Zn) elemental ratios. To accomplish this, MD variations identified using polychromatic radiation from a high resolution micro-computed tomography (micro-CT) benchtop unit were correlated with elemental mapping obtained from a microprobe X-ray fluorescence (XRF) using synchrotron monochromatic radiation. Digital segmentation of tomograms from normal and diseased tissues (N=5 per group; 40-60 year old males) contained significant mineral density variations (enamel: 2820-3095 mg/cc, bone: 570-1415 mg/cc, cementum: 1240-1340 mg/cc, dentin: 1480-1590 mg/cc, cementum affected by periodontitis: 1100-1220 mg/cc, hypomineralized carious dentin: 345-1450 mg/cc, hypermineralized carious dentin: 1815-2740 mg/cc, and dental calculus: 1290-1770 mg/cc). A plausible linear correlation between segmented MD volumes and elemental ratios within these volumes was established, and Ca/P ratios for dentin (1.49), hypomineralized dentin (0.32-0.46), cementum (1.51), and bone (1.68) were observed. Furthermore, varying Ca/Zn ratios were distinguished in adapted compared to normal tissues, such as in bone (855-2765) and in cementum (595-990), highlighting Zn as an influential element in prompting observed adaptive properties. Hence, results provide insights on mineral density gradients with elemental concentrations and elemental footprints that in turn could aid in elucidating mechanistic processes for pathologic formations.

Structure, chemical composition and mechanical properties of coronal cementum in human deciduous molars.

OBJECTIVES: It was hypothesized that the coronal cementum containing collagen forms a weak junction with enamel unlike the well integrated DEJ and CDJ. METHODS: The hypothesis was investigated in two parts: (1) evaluate the structure, chemical composition and mechanical properties of coronal cementum and its junction with enamel using scanning electron microscopy, micro-X-ray computed tomography, and atomic force microscopy. The chemical composition and mechanical properties were determined by evaluating the spatial variations of inorganic (PO(4)(3-)nu(1) mode at 960 cm(-1)) and organic (C-H deformation at 1452 cm(-1); C-H stretch at 2940 cm(-1)) contents using Raman microspectroscopy and elastic modulus and hardness values using nanoindentation. (2) Estimate the strength and evaluate the microstructure of coronal cementum interface with enamel using SEM and MicroXCT. RESULTS AND CONCLUSIONS: Coronal cementum is heterogeneous because it is a combination of laminar acellular afibrillar cementum and acellular extrinsic fiber cementum with relatively higher organic content. It integrates micromechanically via a scallop-like weak interface with enamel unlike the biomechanically efficient DEJ and CDJ and is continuous with primary root cementum. A single tooth could exhibit all three types of cementum enamel junctions; an overlap, butt and a gap depending on the sectioning plane. The elastic modulus of coronal cementum (11.0+/-5.8 GPa) is significantly lower (p<0.05; Student's t-test with 95% confidence interval) than primary cementum (15.8+/-5.3 GPa).

Structure, chemical composition and mechanical properties of human and rat cementum and its interface with root dentin.

This work seeks to establish comparisons of the physical properties of rat and human cementum, root dentin and their interface, including the cementum-dentin junction (CDJ), as a basis for future studies of the entire periodontal complex using rats as animal models. In this study the structure, site-specific chemical composition and mechanical properties of cementum and its interface with root dentin taken from 9- to 12-month-old rats were compared to the physiologically equivalent 40- to 55-year-old human age group using qualitative and quantitative characterization techniques, including histology, atomic force microscopy (AFM), micro-X-ray computed tomography, Raman microspectroscopy and AFM-based nanoindentation. Based on results from this study, cementum taken from the apical third of the respective species can be represented as a woven fabric with radially and circumferentially oriented collagen fibers. In both species the attachment of cementum to root dentin is defined by a stiffness-graded interface (CDJ/cementum-dentin interface). However, it was concluded that cementum and the cementum-dentin interface from a 9- to 12-month-old rat could be more mineralized, resulting in noticeably decreased collagen fiber hydration and significantly higher modulus values under wet conditions for cementum and CDJ (E(rat-cementum)=12.7+/-2.6 GPa; E(rat-CDJ)=11.6+/-3.2 GPa) compared to a 40- to 55-year-old human (E(human-cementum)=3.73+/-1.8 GPa; E(human-CDJ)=1.5+/-0.7 GPa). The resulting data illustrated that the extensions of observations made from animal models to humans should be justified with substantial and equivalent comparison of data across age ranges (life spans) of mammalian species.