Micro-CT data of early physiological cancellous bone formation in the lumbar spine of female C57BL/6 mice

Micro-CT provides critical data for musculoskeletal research, yielding three-dimensional datasets containing distributions of mineral density. Using high-resolution scans, we quantified changes in the fine architecture of bone in the spine of young mice. This data is made available as a reference to physiological cancellous bone growth. The scans (n = 19) depict the extensive structural changes typical for female C57BL/6 mice pups, aged 1-, 3-, 7-, 10- and 14-days post-partum, as they attain the mature geometry. We reveal the micro-morphology down to individual trabeculae in the spine that follow phases of mineral-tissue rearrangement in the growing lumbar vertebra on a micrometer length scale. Phantom data is provided to facilitate mineral density calibration. Conventional histomorphometry matched with our micro-CT data on selected samples confirms the validity and accuracy of our 3D scans. The data may thus serve as a reference for modeling normal bone growth and can be used to benchmark other experiments assessing the effects of biomaterials, tissue growth, healing, and regeneration

Gaps at the interface between dentine and self‐adhesive resin cement in post‐endodontic restorations quantified in 3D by phase contrast‐enhanced micro‐CT

Aim: To assess the extent of gaps between root dentine and titanium or fibreglass post restorations following cementation with a self-adhesive resin cement. Methodology: Fourteen root filled maxillary central incisors restored with prefabricated posts made of Fibreglass (n = 7) or Titanium (n = 7) and cemented with RelyX Unicem 2 were imaged by rapid, high-resolution phase contrast-enhanced micro-CT (PCE-CT) in a synchrotron X-ray imaging facility (ID19, ESRF, 34 KeV, 0.65 µm pixel resolution). Reconstructions were used to measure canal, cement and post perimeters and cross-sectional areas and interfacial gaps at 0.1 mm increments in the root canal space, along the cervical region of the tooth. Remnants of endodontic sealer (AH Plus), when present, were also quantified. Mann–Whitney and 2-way ANOVA tests were used to compare findings within slices and between the two post groups. Pearson correlation coefficients (r) were determined between the interfacial gaps and the other measured parameters. Results: Clearly detectable gaps were found in 45% (±14%) of the interfaces between dentine and cement, along the canal in the cervical area of the tooth beneath the core. The length of interfacial gaps was moderately correlated to the canal cross-sectional area, to the canal perimeter and to the canal area filled by cement (R = 0.52 ~ 0.55, P 0.01). Both post types had defect-free interfaces with cement. Endodontic sealer remnants were found on ~10% of the canal walls and were moderately correlated to the presence of gaps. Approximately 30% of the sealer-affected interfaces exhibited no detachment between dentine, sealer and cement. Conclusions: Self-adhesive cements had interfacial gaps along substantial regions of the root canal surface, which was not correlated with the amount of cement in the canal. PCE-CT proved to be an excellent non-destructive method to study root canal restorations of hydrated samples in 3D

Hard X-ray phase-contrast-enhanced micro-CT for quantifying interfaces within brittle dense root-filling-restored human teeth

Bonding of resin composite fillings, for example following root-canal treatment, is a challenge because remaining gaps grow and lead to failure. Here, phase-contrast-enhanced micro-computed tomography (PCE-CT) is used to explore methods of non-destructive quantification of the problem, so that countermeasures can be devised. Five human central incisors with damaged crowns were root-filled followed by restoration with a dental post. Thereafter, the crowns were rebuilt with a resin composite that was bonded conventionally to the tooth with a dental adhesive system (Futurabond U). Each sample was imaged by PCE-CT in a synchrotron facility (ID19, European Synchrotron Radiation Facility) with a pixel size of 650 nm. The reconstructed datasets from each sample were segmented and analysed in a semi-automated manner using ImageJ. PCE-CT at sub-micrometre resolution provided images with an impressive increased contrast and detail when compared with laboratory micro-computed tomography. The interface between the dental adhesive and the tooth was often strongly disrupted by the presence of large debonded gaps (on average 34% ± 15% on all surfaces). The thickness of the gaps spanned 2 µm to 16 µm. There was a large variability in the distribution of gaps within the bonding area in each sample, with some regions around the canal exhibiting up to 100% discontinuity. Although only several micrometres thick, the extensive wide gaps may serve as gateways to biofilm leakage, leading to failure of the restorations. They can also act as stress-raising `cracks' that are likely to expand over time in response to cyclic mechanical loading as a consequence of mastication. The observations here show how PCE-CT can be used as a non-destructive quantitative tool for understanding and improving the performance of clinically used bonded dental restorations

Accelerated Growth Plate Mineralization and Foreshortened Proximal Limb Bones in Fetuin-A Knockout Mice

PMCID: PMC3473050This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

Heterogeneity of the osteocyte lacuno-canalicular network architecture and material characteristics across different tissue types in healing bone

Various tissue types, including fibrous connective tissue, bone marrow, cartilage, woven and lamellar bone, coexist in healing bone. Similar to most bone tissue type, healing bone contains a lacuno-canalicular network (LCN) housing osteocytes. These cells are known to orchestrate bone remodeling in healthy bone by sensing mechanical strains and translating them into biochemical signals. The structure of the LCN is hypothesized to influence mineralization processes. Hence, the aim of the present study was to visualize and match spatial variations in the LCN topology with mineral characteristics, within and at the interfaces of the different tissue types that comprise healing bone. We applied a correlative multi-method approach to visualize the LCN architecture and quantify mineral particle size and orientation within healing femoral bone in a mouse osteotomy model (26 weeks old C57BL/6 mice). This approach revealed structural differences across several length scales during endochondral ossification within the following regions: calcified cartilage, bony callus, cortical bone and a transition zone between the cortical and callus region analyzed 21 days after the osteotomy. In this transition zone, we observed a continuous convergence of mineral characteristics and osteocyte lacunae shape as well as discontinuities in the lacunae volume and LCN connectivity. The bony callus exhibits a 34% higher lacunae number density and 40% larger lacunar volume compared to cortical bone. The presented correlations between LCN architecture and mineral characteristics improves our understanding of how bone develops during healing and may indicate a contribution of osteocytes to bone (re)modeling

The forgotten merits of GIC restorations: a systematic review

Objective: To reevaluate proven strengths and weakness of glass ionomer cements (GICs) and to identify agreement versus conflicting evidence in previous reports regarding the transition between GIC and the tooth, and the existence of an "interphase". Materials and methods: Relevant electronic databases (PubMed, Embase via Ovid and Medline via Web of science) were searched for publications of evidence relating to the transition zone at the GIC-tooth interphase. Studies were examined and grouped according to characteristics of GIC-tooth attachment area quantified by X-ray and optical microscopy techniques in 2D and 3D. Results: Inclusion criteria comprised of in vitro studies that showed images of the conventional GIC-tooth substrate attachments using at least one of the following techniques: SEM, CLSM, or μCT. The search identified 419 studies, from which 33 were included. Ten studies demonstrated the existence of an interphase layer and five studies quantified the layer thickness (1-15 μ). Twenty-nine publications studied different failure modes of the GIC-tooth interphase. Eleven studies described discontinuities inside the GIC bulk. Conclusion: The GIC-tooth interphase attributes evolve with time. Good attachment is evident even under compromised surface preparation. The GIC-tooth attachment area is resistant to acidic dissolution as compared to both tooth and GIC bulk. In general, studies revealed mostly intact GIC-tooth interphases with only some cracked interphases. Clinical significance: GIC bonds to the tooth structure and forms an acid resistant attachment zone that might enhance caries inhibition. Due to fluoride release and ease of use, GIC provides a cost effective treatment, ideal for low income or high caries populations

Artificial Intelligence as a Tool to Study the 3D Skeletal Architecture in Newly Settled Coral Recruits: Insights into the Effects of Ocean Acidification on Coral Biomineralization

Understanding the formation of the coral skeleton has been a common subject uniting various marine and materials study fields. Two main regions dominate coral skeleton growth: Rapid Accretion Deposits (RADs) and Thickening Deposits (TDs). These have been extensively characterized at the 2D level, but their 3D characteristics are still poorly described. Here, we present an innovative approach to combine synchrotron phase contrast-enhanced microCT (PCE-CT) with artificial intelligence (AI) to explore the 3D architecture of RADs and TDs within the coral skeleton. As a reference study system, we used recruits of the stony coral Stylophora pistillata from the Red Sea, grown under both natural and simulated ocean acidification conditions. We thus studied the recruit’s skeleton under both regular and morphologically-altered acidic conditions. By imaging the corals with PCE-CT, we revealed the interwoven morphologies of RADs and TDs. Deep-learning neural networks were invoked to explore AI segmentation of these regions, to overcome limitations of common segmentation techniques. This analysis yielded highly-detailed 3D information about the RAD’s and TD’s architecture. Our results demonstrate how AI can be used as a powerful tool to obtain 3D data essential for studying coral biomineralization and for exploring the effects of environmental change on coral growth

Carbonate substitution significantly affects the structure and mechanics of carbonated apatites

Bone mineral comprises nanoparticles of carbonate-substituted bioapatite similar to hydroxylapatite. Yet mechanical values of macroscopic-sized geological hydroxylapatite are often used to model bone properties due to a lack of experimental data for bioapatite. Here, we investigated the effects of carbonate substitution and hydration on biomimetic apatite response to load using in situ hydrostatic pressure loading and synchrotron x-ray diffraction. We find that increasing carbonate levels reduced the bulk modulus and elastic strain ratio. Elastic constants, determined using computational optimization techniques, revealed that compliance values and elastic moduli decreased with increasing carbonate content, likely a result of decreased bond strength due to CO32− substitution and Ca2+ loss. Hydration environment had no clear effects on the elastic properties likely due to dissolution and reprecipitation processes modifying the crystal structure organization. These results reinforce the need to consider carbonate composition when selecting mechanical properties and provide robust data for carbonate-substituted apatite stiffness

In situ compressibility of carbonated hydroxyapatite in tooth dentine measured under hydrostatic pressure by high energy X-ray diffraction

Tooth dentine and other bone-like materials contain carbonated hydroxyapatite nanoparticles within a network of collagen fibrils. It is widely assumed that the elastic properties of biogenic hydroxyapatites are identical to those of geological apatite. By applying hydrostatic pressure and by in situ measurements of the a- and c- lattice parameters using high energy X-ray diffraction, we characterize the anisotropic deformability of the mineral in the crowns and roots of teeth. The collected data allowed us to calculate the bulk modulus and to derive precise estimates of Young׳s moduli and Poisson׳s ratios of the biogenic mineral particles. The results show that the dentine apatite particles are about 20% less stiff than geological and synthetic apatites and that the mineral has an average bulk modulus K=82.7GPa. A 5% anisotropy is observed in the derived values of Young׳s moduli, with E11≈91GPa and E33≈96GPa, indicating that the nanoparticles are only slightly stiffer along their long axis. Poisson׳s ratio spans ν≈0.30-0.35, as expected. Our findings suggest that the carbonated nanoparticles of biogenic apatite are significantly softer than previously thought and that their elastic properties can be considered to be nearly isotropic