Tooth acellular extrinsic fibre cementum incremental lines in humans are formed by parallel branched Sharpey's fibres and not by its mineral phase.

In humans, the growth pattern of the acellular extrinsic fibre cementum (AEFC) has been useful to estimate the age-at-death. However, the structural organization behind such a pattern remains poorly understood. In this study tooth cementum from seven individuals from a Mexican modern skeletal series were analyzed with the aim of unveiling the AEFC collagenous and mineral structure using multimodal imaging approaches. The organization of collagen fibres was first determined using: light microscopy, transmission electron microscopy (TEM), electron tomography, and plasma FIB scanning electron microscopy (PFIB-SEM) tomography. The mineral properties were then investigated using: synchrotron small-angle X-ray scattering (SAXS) for T-parameter (correlation length between mineral particles); synchrotron X-ray diffraction (XRD) for L-parameter (mineral crystalline domain size estimation), alignment parameter (crystals preferred orientation) and lattice parameters a and c; as well as synchrotron X-ray fluorescence for spatial distribution of calcium, phosphorus and zinc. Results show that Sharpey's fibres branched out fibres that cover and uncover other collagen bundles forming aligned arched structures that are joined by these same fibres but in a parallel fashion. The parallel fibres are not set as a continuum on the same plane and when they are superimposed project the AEFC incremental lines due to the collagen birefringence. The orientation of the apatite crystallites is subject to the arrangement of the collagen fibres, and the obtained parameter values along with the elemental distribution maps, revealed this mineral tissue as relatively homogeneous. Therefore, no intrinsic characteristics of the mineral phase could be associated with the alternating AEFC incremental pattern.

Mesoscale porosity at the dentin-enamel junction could affect the biomechanical properties of teeth.

In this paper, the 3D-morphology of the porosity in dentin is investigated within the first 350µm from the dentin-enamel junction (DEJ) by fluorescence confocal laser scanning microscopy (CLSM). We found that the porous microstructure exhibits a much more complex geometry than classically described, which may impact our fundamental understanding of the mechanical behavior of teeth and could have practical consequences for dental surgery. Our 3D observations reveal numerous fine branches stemming from the tubules which may play a role in cellular communication or mechanosensing during the early stages of dentinogenesis. The effect of this highly branched microstructure on the local mechanical properties is investigated by means of numerical simulations. Under simplified assumptions on the surrounding tissue characteristics, we find that the presence of fine branches negatively affects the mechanical properties by creating local stress concentrations. However, this effect is reduced by the presence of peritubular dentin surrounding the tubules. The porosity was also quantified using the CSLM data and compared to this derived from SEM imaging. A bimodal distribution of channel diameters was found near the DEJ with a mean value of 1.5-2µm for the tubules and 0.3-0.5µm for the fine branches which contribute to 30% of the total porosity (∼1.2%). A gradient in the branching density was observed from the DEJ towards the pulp, independently of the anatomical location. Our work constitutes an incentive towards more elaborate multiscale studies of dentin microstructure to better assess the effect of aging and for the design of biomaterials used in dentistry, e.g. to ensure more efficient bonding to dentin. Finally, our analysis of the tubular network structure provides valuable data to improve current numerical models.

Relation between the Macroscopic Pattern of Elephant Ivory and Its Three-Dimensional Micro-Tubular Network.

Macroscopic, periodic, dark and bright patterns are observed on sections of elephant tusk, in the dentin part (ivory). The motifs-also called Schreger pattern-vary depending on the orientation in the tusk: on sections perpendicular to the tusk axis, a checkerboard pattern is present whereas on sections longitudinal to it, alternating stripes are observed. This pattern has been used to identify elephant and mammoth ivory in archeological artifacts and informs on the continuous tissue growth mechanisms of tusk. However, its origin, assumed to be related to the 3D structure of empty microtubules surrounded by the ivory matrix has yet to be characterized unequivocally. Based on 2D observations of the ivory microtubules by means of a variety of imaging techniques of three different planes (transverse, longitudinal and tangential to the tusk axis), we show that the dark areas of the macroscopic pattern are due to tubules oblique to the surface whereas bright areas are related to tubules parallel to it. The different microstructures observed in the three planes as well as the 3D data obtained by SR-µCT analysis allow us to propose a 3D model of the microtubule network with helical tubules phase-shifted in the tangential direction. The phase shift is a combination of a continuous phase shift of π every 1 mm with a stepwise phase shift of π/2 every 500 µm. By using 3D modeling, we show how the 3D helical model better represents the experimental microstructure observed in 2D planes compared to previous models in the literature. This brings new information on the origin of the unique Schreger pattern of elephant ivory, crucial for better understanding how archaeological objects were processed and for opening new routes to rethink how biological materials are built.