micro-CT Data Research Articles

On the application of the finite cell method to static analysis of trabecular bone tissue specimen using high‐resolution microCT data

On the application of the finite cell method to static analysis of trabecular bone tissue specimen using high‐resolution microCT data

MICP treated sand: insights into the impact of particle size on mechanical parameters and pore network after biocementation

Microbiologically Induced Calcium Carbonate Precipitation (MICP) is a technology for improving soil characteristics, especially strength, that has been gaining increasing interest in literature during the last few years. Although a lot of influencing factors on the result of MICP are known, particle size and shape of the particles remain poorly understood. While destructive measuring of compressive strength or calcium carbonate content are important for the characterization of samples these methods give no insight into the internal structures and pore networks of the samples. X-ray microcomputed tomography (micro-CT) is a technique that is used to characterize the internals of rocks and to a certain degree MICP-treated soils. However, the impact of filtering and image processing of micro-CT Data depending on the type of MICP sample is poorly described in the literature. In this study, single fractions of local quarry were treated with MICP through the ureolytic microorganism Sporosarcina pasteurii to investigate the influence of particle size distribution on calcium carbonate content, unconfined compressive strength and the reduction of water permeability. Additionally, micro-CT was conducted to obtain insights into the resulting pore system. The impact of the Gauss filter und Non-local means filter on the resulting images and data on the pore network are discussed. The results show that particle size has a significant impact on the result of all tested parameters of biosandstone with lower particle size leading to higher strength and generally higher calcium carbonate content. Micro-CT data showed that the technology is feasible to gain valuable insights into the internal structures of biosandstone but the resolution and signal-to-noise ratio remain challenging, especially for samples with particle sizes smaller than 125 µm.

Lost & Found - Rediscovery of H. Hauffen’s Carychium material in the Dominik Bilimek Collection, BOKU University, including a contemporary assessment within the genus Zospeum (Gastropoda, Ellobioidea, Carychiinae)

The syntypes of Carychium reticulatum Hauffen, 1856 and Carychium bidentatum Hauffen, 1856, thought to be lost to science, have been rediscovered in a recent inventory of the shell collection of Domink Bilimek (1813–1884) at the University of Natural Resources and Life Sciences in Vienna, Austria (BOKU). In this work, we present (1) the historical context of the Dominik Bilimek and Heinrich (also: Henrik) Hauffen collections and (2) provide a contemporary image dataset of Hauffen’s original material while considering today’s taxonomic understanding of the genus Zospeum. We clarify the taxonomic status of the syntypes by using light microscopy, Scanning Electron Microcopy (SEM) and 3D X-ray Micro-CT data in conjunction with Hauffen’s original illustrations and compare them to contemporary investigations of their closest congeners. Analysis of Zospeum reticulatum (Hauffen, 1856) is supported by historical documentation from Heinrich Hauffen’s original annotations and Bilimek’s diary entries, as well as from individual collection labels in Hauffen’s own script (verified by archive material from the Archive of the National Museum of Slovenia, NMS). Hauffen’s descriptions are based on apertural morphology and shell microstructure. Our re-investigation of Zospeum reticulatum provides corroborating evidence supporting its assignment to Zospeum spelaeum (Rossmaessler, 1839) while that of Carychium bidentatum corroborates its status as junior synonym of Zospeum costatum (Freyer, 1855).

Mechanical properties of Acropora cervicornis aragonite skeleton by using multiscale models based on micro-CT data

Mechanical properties of <i>Acropora cervicornis</i> aragonite skeleton by using multiscale models based on micro-CT data

Effect of 3D printed teeth and virtual simulation system on the pre-clinical access cavity preparation training of senior dental undergraduates

BackgroundThe objective of the present study was to evaluate the effect of 3D printed teeth and virtual simulation system on the pre-clinical access cavity preparation training of senior dental undergraduates.MethodsThe 3D printed teeth were manufactured based on the micro-CT data of an extracted lower first molar. Ninety-eight senior dental undergraduate students were required to finish the access cavity preparation of lower first molar within 20 min on plastic and 3D printed teeth on the manikin system as well as on a virtual simulation machine respectively with randomly selected sequences. Expert dentists scored the operated teeth. The scores from the virtual simulation system were also recorded. All the scores were analyzed and compared. Following the procedure, two questionnaires were sent to students to further evaluate the feelings and optimal training sequence.ResultsNo significant differences were found between plastic and 3D printed teeth scores, while virtual simulation achieved a valid/invalid area removal ratio of 96.86% ± 5.08% and 3.97% ± 1.85%, respectively. Most students found plastic teeth training the easiest and favored the three-training combination (36.36%). 71.42% of the students thought the virtual simulation training should be put at the first place of the three trainings. Over 80% of students agreed with incorporating 3D printed teeth and virtual simulation into their routine training courses. In addition, the general advantages and disadvantages of the virtual simulation system and 3D printed teeth training received almost equal recognition by students.ConclusionsVirtual simulation system training + plastic teeth training + 3D printed teeth training might be the optimal training sequence. Virtual simulation system training could not completely replace the traditional training methods on the manikin system at the moment. With further modifications, 3D printed teeth could be expected to replace the plastic teeth for the pre-clinical access cavity preparation training.

Crebanine mitigates glucocorticoid-induced osteonecrosis of the femoral head by restoring bone remodelling homeostasis via attenuating oxidative stress.

The onset of osteonecrosis of the femoral head (ONFH) is intimately associated with the extensive administration of glucocorticoids (GCs). Long-term stimulation of GCs can induce oxidative stress in both osteoclasts (OCs) and osteoblasts (OBs), resulting in the disturbance of bone remodelling. An alkaloid named crebanine (CN) demonstrates pharmacological properties including anti-inflammation and reactive oxygen species (ROS) modulation. Our objective is to assess the therapeutic potential of CN in treating ONFH and elucidate the associated underlying mechanisms. The network pharmacology analysis uncovered that CN played a role in regulating ROS metabolism. Invitro, CN demonstrated its ability to reduce the dexamethasone (DEX)-stimulated generation of OCs and suppress their resorptive function by downregulating the level of osteoclast marker genes. Concurrently, CN also mitigated DEX-induced damage to OBs, facilitating the restoration of osteoblast marker gene expression, cellular differentiation and function. These effects were achieved by CN augmenting the antioxidant system to reduce intracellular ROS levels. Furthermore, invitro results were corroborated by micro-CT and histological data, which also showed that CN attenuated MPS-induced ONFH in mice. This study highlights the therapeutic potential of CN in counteracting GCs-induced ONFH.

Sound reception and hearing capabilities in the Little Penguin (Eudyptula minor): first predicted in-air and underwater audiograms.

Despite increasing concern about the effects of anthropogenic noise on marine fauna, relevant research is limited, particularly in those inaccessible species, such as the Little Penguin (Eudyptula minor). In this study, we collected freshly deceased Little Penguins for dissection and micro-computed tomography (microCT) scans. The head structures, including the ear apparatus, were reconstructed based on high-resolution imaging data for the species. Moreover, three-dimensional finite-element models were built based on microCT data to simulate the sound reception processes and ear responses to the incident planar waves at the selected frequencies. The received sound pressure fields and motion (i.e. displacement and velocity) of the internal ear-related structures were modelled. The synergistic response of ear components to incident aerial and underwater sounds was computed to predict the hearing capabilities of the Little Penguins across a broad frequency range (100 Hz-10 kHz), both in air and under water. Our predicted data showed good agreement with other diving birds in both the form and range of auditory sensitivity. This study demonstrates a promising method to study hearing in other inaccessible animals. The outputs from this study can inform noise impact mitigation and conservation management.

Getting Nosy: Olfactory Rosette Morphology and Lamellar Microstructure of Two Chondrichthyan Species.

To smell, fish rely on passive water flow into their olfactory chambers and through their olfactory rosettes to detect chemical signals in their aquatic environment. The olfactory rosette is made up of secondarily folded tissues called olfactory lamellae. The olfactory morphology of cartilaginous fishes varies widely in both rosette gross morphology and lamellar microstructure. Previous research has shown differences in lamellar sensory morphology depending on the position along the rosette in hammerheads (family Sphyrnidae). Here, we investigate if this pattern continues in members of two other chondrichthyan families: Squalidae and Chimaeridae. Using contrast-enhanced microCT and scanning electron microscopy, we investigated patterns in lamellar morphology based on lamellar position along the olfactory rosette in Pacific spiny dogfish (Squalus suckleyi) and spotted ratfish (Hydrolagus colliei). We describe the gross olfactory rosette anatomy and lamellar microstructure of both species. We also put forth a new method, combining 3D morphological microCT data with 2D SEM microstructure data to better approximate lamellar sensory surface area. We found that in both species, lamellae in the center of the rosette were larger with more secondary folds. However, we found no significant differences in lamellar sensory surface area among lamellar positions. Previously, differences in lamellar sensory morphology have been tied to the internal fluid dynamics of the olfactory chamber. It is possible that the internal flow dynamics of these species are like other chondrichthyan models, where water flow patterns differ in the lateral vs the medial part of the organ, and the consistent distribution of sensory tissue does not correspond to this flow. Alternatively, the olfactory morphology of these species may result in uniform flow patterns throughout the olfactory chamber, correlating with the consistent distribution of sensory tissue throughout the organ. This study emphasizes that further investigations into chondrichthyan fluid dynamics is paramount to any future studies on the correlations between distribution of sensory tissues and water flow.

Exploring single-shot propagation and speckle based phase recovery techniques for object thickness estimation by using a polychromatic X-ray laboratory source.

Propagation and speckle-based techniques allow reconstruction of the phase of an X-ray beam with a simple experimental setup. Furthermore, their implementation is feasible using low-coherence laboratory X-ray sources. We investigate different approaches to include X-ray polychromaticity for sample thickness recovery using such techniques. Single-shot Paganin (PT) and Arhatari (AT) propagation-based and speckle-based (ST) techniques are considered. The radiation beam polychromaticity is addressed using three different averaging approaches. The emission-detection process is considered for modulating the X-ray beam spectrum. Reconstructed thickness of three nylon-6 fibers with diameters in the millimeter-range, placed at various object-detector distances are analyzed. In addition, the thickness of an in-house made breast phantom is recovered by using multi-material Paganin's technique (MPT) and compared with micro-CT data. The best quantitative result is obtained for the PT and ST combined with sample thickness averaging (TA) approach that involves individual thickness recovery for each X-ray spectral component and the smallest considered object-detector distance. The error in the recovered fiber diameters for both techniques is , despite the higher noise level in ST images. All cases provide estimates of fiber diameter ratios with an error of 3% with respect to the nominal diameter ratios. The breast phantom thickness difference between MPT-TA and micro-CT is about 10%. We demonstrate the single-shot PT-TA and ST-TA techniques feasibility for thickness recovery of millimeter-sized samples using polychromatic microfocus X-ray sources. The application of MPT-TA for thicker and multi-material samples is promising.

Monte Carlo multiscale modelling of photon and proton distribution in heterogenous tissue

Enhancements in radiotherapy treatment outcomes can be realised by minimising uncertainties in dose distribution. Current treatment planning struggles to accurately calculate dose distribution in complex heterogeneous areas, thereby increasing the uncertainty associated with dose distribution. This research focuses on studying microscopic dose distribution in the temporal bone and cochlea. This study utilised an open-access DICOM format dataset for resected temporal bone and cochlea tissue, employing the FLUKA MC code to simulate potential high-dose scenarios in volume-modulated arc therapy using the FLOOD option. Simulations were conducted at 23 photon and proton energy levels, ranging from 0.055 to 5.5 MeV for photons and 37.59–124.83 MeV for protons, to calculate dose distributions. The largest proportion of the dose (48.8%) was deposited in high-density bone at photon energies between 0.055 and 0.09 MeV. Above 0.125 MeV, a notable shift in dose distribution to low-density tissues occurred, reaching a deposition of 53%. In intermediate-density soft bone, dose distribution was 26.4% at 0.07 MeV and decreased to 19.7% at 2.5 MeV, reflecting a 29% difference in dose distribution across the energy spectrum. In proton simulations, dose distribution at low energy (37.59 MeV) revealed no significant changes across low (54.86%), intermediate (19.75%), and high-density (25.39%) areas. Similar patterns were observed at high energy (124.83 MeV), with dose distributions of 54.21% in low-, 19.79% in intermediate-, and 26% in high-density areas. The simulations demonstrated that proton dose distribution was not significantly influenced by tissue heterogeneity in micro-CT data. The photoelectric effect at low energy levels contributed minimally to the dose in soft bone, favouring higher deposition in high-density bone, despite a lower weighting factor at low energies compared with high energies.

Augmenting X-ray micro-CT data with MICP data for high resolution pore-scale simulations of flow properties of carbonate rocks

Pore-scale modeling is essential in understanding and predicting flow and transport properties of rocks. Generally, pore-scale modeling is dependent on imaging technologies such as Micro Computed Tomography (micro-CT), which provides visual confirmation into the pore microstructures of rocks at a representative scale. However, this technique is limited in the ability to provide high resolution images showing the pore-throats connecting pore bodies. Pore scale simulations of flow and transport properties of rocks are generally done on a single 3D pore microstructure image. As such, the simulated properties are only representative of the simulated pore-scale rock volume. These are the technological and computational limitations which we address here by using a stochastic pore-scale simulation approach. This approach consists of constructing hundreds of 3D pore microstructures of the same pore size distribution and overall porosity but different pore connectivity. The construction of the 3D pore microstructures incorporates the use of Mercury Injection Capillary Pressure (MICP) data to account for pore throat size distribution, and micro-CT images to account for pore body size distribution. The approach requires a small micro-CT image volume (7–19 mm3) to reveal key pore microstructural features that control flow and transport properties of highly heterogeneous rocks at the core-scale. Four carbonate rock samples were used to test the proposed approach. Permeability calculations from the introduced approach were validated by comparing laboratory measured permeability of rock cores and permeability estimated using five well-known core-scale empirical model equations. The results show that accounting for the stochastic connectivity of pores results in a probabilistic distribution of flow properties which can be used to upscale pore-scale simulated flow properties to the core-scale. The use of the introduced stochastic pore-scale simulation approach is more beneficial when there is a higher degree of heterogeneity in pore size distribution. This is shown to be the case with permeability and hydraulic tortuosity which are key controls of flow and transport processes in rocks.

The cranial and postcranial morphology of Hutchemys rememdium and its impact on the phylogenetic relationships of Plastomenidae (Testudinata, Trionychidae)

Hutchemys rememdium is a poorly understood softshell turtle (Trionychidae) from the mid Paleocene of the Williston Basin of North America previously known only from postcranial remains. A particularly rich collection of previously undescribed material from the Tiffanian 4 North American Land Mammal Age (NALMA) of North Dakota is here presented consisting of numerous shells that document new variation, some non-shell postcrania, and cranial remains, which are described based on 3D models extracted from micro-CT data. Although the observed shell variation weakens previously noted differences with the younger species Hutchemys arctochelys from the Clarkforkian NALMA, the two taxa are still recognized as distinct. Parsimony and Bayesian phylogenetic analyses reaffirm the previously challenged placement of Hutchemys rememdium within the clade Plastomenidae, mostly based on novel observations of cranial characters made possible by the new material and the micro-CT data. The new topology supports the notion that the well-ossified plastron of plastomenids originated twice in parallel near the Cretaceous/Paleogene boundary, once in the Hutchemys lineage and once in the Gilmoremys/Plastomenus lineage. Hutchemys rememdium is notable for being the only documented species of trionychid in the mid Paleocene of the Williston Basin. The presence of multiple individuals in a carbonaceous claystone indicates this taxon lived in swamps and lakes and its expanded triturating surface suggests it had a durophagous diet.

Absorption Correction for 3D Elemental Distributions of Dental Composite Materials Using Laboratory Confocal Micro-X-ray Fluorescence Spectroscopy.

Confocal micro-X-ray fluorescence (micro-XRF) spectroscopy facilitates three-dimensional (3D) elemental imaging of heterogeneous samples in the micrometer range. Laboratory setups using X-ray tube excitation render the method accessible for diverse research fields but interpretation of results and quantification remain challenging. The attenuation of X-rays in composites depends on the photon energy as well as on the composition and density of the material. For confocal micro-XRF, attenuation severely impacts elemental distribution information, as the signal from deeper layers is distorted by superficial layers. Absorption correction and quantification of fluorescence measurements in heterogeneous composite samples have so far not been reported. Here, an absorption correction approach for confocal micro-XRF combining density information from microcomputed tomography (micro-CT) data with laboratory X-ray absorption spectroscopy (XAS) and synchrotron transmission measurements is presented. The energy dependency of the probing volume is considered during the correction. The methodology is demonstrated on a model composite sample consisting of a bovine tooth with a clinically used restoration material.

Chondrosarcoma evaluation using hematein-based x-ray staining and high-resolution 3D micro-CT: a feasibility study

BackgroundChondrosarcomas are rare malignant bone tumors diagnosed by analyzing radiological images and histology of tissue biopsies and evaluating features such as matrix calcification, cortical destruction, trabecular penetration, and tumor cell entrapment.MethodsWe retrospectively analyzed 16 cartilaginous tumor tissue samples from three patients (51-, 54-, and 70-year-old) diagnosed with a dedifferentiated chondrosarcoma at the femur, a moderately differentiated chondrosarcoma in the pelvis, and a predominantly moderately differentiated chondrosarcoma at the scapula, respectively. We combined a hematein-based x-ray staining with high-resolution three-dimensional (3D) microscopic x-ray computed tomography (micro-CT) for nondestructive 3D tumor assessment and tumor margin evaluation.ResultsWe detected trabecular entrapment on 3D micro-CT images and followed bone destruction throughout the volume. In addition to staining cell nuclei, hematein-based staining also improved the visualization of the tumor matrix, allowing for the distinction between the tumor and the bone marrow cavity. The hematein-based staining did not interfere with further conventional histology. There was a 5.97 ± 7.17% difference between the relative tumor area measured using micro-CT and histopathology (p = 0.806) (Pearson correlation coefficient r = 0.92, p = 0.009). Signal intensity in the tumor matrix (4.85 ± 2.94) was significantly higher in the stained samples compared to the unstained counterparts (1.92 ± 0.11, p = 0.002).ConclusionsUsing nondestructive 3D micro-CT, the simultaneous visualization of radiological and histopathological features is feasible.Relevance statement3D micro-CT data supports modern radiological and histopathological investigations of human bone tumor specimens. It has the potential for being an integrative part of clinical preoperative diagnostics.Key points• Matrix calcifications are a relevant diagnostic feature of bone tumors.• Micro-CT detects all clinically diagnostic relevant features of x-ray-stained chondrosarcoma.• Micro-CT has the potential to be an integrative part of clinical diagnostics.Graphical

Super-resolution of clinical CT: Revealing microarchitecture in whole bone clinical CT image data

Osteoporotic fractures, prevalent in the elderly, pose a significant health and economic burden. Current methods for predicting fracture risk, primarily relying on bone mineral density, provide only modest accuracy. If better spatial resolution of trabecular bone in a clinical scan were available, a more complete assessment of fracture risk would be obtained using microarchitectural measures of bone (i.e. trabecular thickness, trabecular spacing, bone volume fraction, etc.). However, increased resolution comes at the cost of increased radiation or can only be applied at small volumes of distal skeletal locations. This study explores super-resolution (SR) technology to enhance clinical CT scans of proximal femurs and better reveal the trabecular microarchitecture of bone. Using a deep-learning-based (i.e. subset of artificial intelligence) SR approach, low-resolution clinical CT images were upscaled to higher resolution and compared to corresponding MicroCT-derived images. SR-derived 2-dimensional microarchitectural measurements, such as degree of anisotropy, bone volume fraction, trabecular spacing, and trabecular thickness were within 16 % error compared to MicroCT data, whereas connectivity density exhibited larger error (as high as 1094 %). SR-derived 3-dimensional microarchitectural metrics exhibited errors <18 %. This work showcases the potential of SR technology to enhance clinical bone imaging and holds promise for improving fracture risk assessments and osteoporosis detection. Further research, including larger datasets and refined techniques, can advance SR's clinical utility, enabling comprehensive microstructural assessment across whole bones, thereby improving fracture risk predictions and patient-specific treatment strategies.

Paedomorphosis and retention of juvenile diet lead speciation in a group of Neotropical snakes (Colubroides-Philodryadini)

Dipsadidae is one of the largest clades of extant reptiles, showing an impressive morphological and ecological diversity. Despite this fact, the developmental processes behind its diversity are still largely unknown. In this study, we used 3D reconstructions based on micro-CT data and geometric morphometrics to evaluate the skull morphology of Philodryas agassizii, a small, surface-dwelling dipsadid that consume spiders. Adult individuals of P. agassizii exhibit a cranial morphology frequently observed in juveniles of other surface-dwelling colubroideans, represented in our analysis by its close relative Philodryas patagoniensis. Large orbits, gibbous neurocranial roof and a relatively short jaw complex are features present in juveniles of the latter species. Furthermore, we performed an extensive survey about diet of P. patagoniensis in which we detected an ontogenetic dietary shift, indicating that arthropods are more frequently consumed by juveniles of this dietary generalist. Thus, we infer that P. agassizzii retained not only the ancestral juvenile skull morphology but also dietary preferences. This study reveals that morphological changes driven by heterochronic changes, specifically paedomorphosis, influenced the retention of ancestral life history traits in P. agassizii, and therefore promoted cladogenesis. In this way, we obtained first evidence that heterochronic processes lead speciation in the snake megadiverse clade Dipsadidae.

Automated region growing-based segmentation for trabecular bone structure in fresh-frozen human wrist specimens

Bone strength depends on both mineral content and bone structure. Measurements of bone microstructure on specimens can be performed by micro-CT. In vivo measurements are reliably performed by high-resolution peripheral computed tomography (HR-pQCT) using dedicated software. In previous studies from our research group, trabecular bone properties on CT data of defatted specimens from many different CT devices have been analyzed using an Automated Region Growing (ARG) algorithm-based code, showing strong correlations to micro-CT.The aim of the study was to validate the possibility of segmenting and measuring trabecular bone structure from clinical CT data of fresh-frozen human wrist specimens. Data from micro-CT was used as reference. The hypothesis was that the ARG-based in-house built software could be used for such measurements.HR-pQCT image data at two resolutions (61 and 82 µm isotropic voxels) from 23 fresh-frozen human forearms were analyzed. Correlations to micro-CT were strong, varying from 0.72 to 0.99 for all parameters except trabecular termini and nodes. The bone volume fraction had correlations varying from 0.95 to 0.98 but was overestimated compared to micro-CT, especially at the lower resolution. Trabecular separation and spacing were the most stable parameters with correlations at 0.80-0.97 and mean values in the same range as micro-CT.Results from this in vitro study show that an ARG-based software could be used for segmenting and measuring 3D trabecular bone structure from clinical CT data of fresh-frozen human wrist specimens using micro-CT data as reference. Over-and underestimation of several of the bone structure parameters must however be taken into account.

Functional differences between the extraordinary eyes of deep-sea hyperiid amphipods.

The ocean's midwater is a uniquely challenging yet predictable and simple visual environment. The need to see without being seen in this dim, open habitat has led to extraordinary visual adaptations. To understand these adaptations, we compared the morphological and functional differences between the eyes of three hyperiid amphipods-Hyperia galba, Streetsia challengeri and Phronima sedentaria. Combining micro-CT data with computational modelling, we mapped visual field topography and predicted detection distances for visual targets viewed in different directions through mesopelagic depths. Hyperia's eyes provide a wide visual field optimized for spatial vision over short distances, while Phronima's and Streetsia's eyes have the potential to achieve greater sensitivity and longer detection distances using spatial summation. These improvements come at the cost of smaller visual fields, but this loss is compensated for by a second pair of eyes in Phronima and by behaviour in Streetsia. The need to improve sensitivity while minimizing visible eye size to maintain crypsis has likely driven the evolution of hyperiid eye diversity. Our results provide an integrative look at how these elusive animals have adapted to the unique visual challenges of the mesopelagic.

Reliability of Three-Dimensional Micro-Computed Tomography Root Canal Analysis Using Drishti Software: A Pilot Study

In recent years, micro-computed tomography (micro-CT) has become a promising tool for non-destructive imaging and the study of root canal morphology in endodontics. However, the reliability of such analyses heavily relies on the accuracy and consistency of the software used for image processing. Objective: This study aims to investigate the reliability of two different micro-CT voxel sizes using open-source three-dimensional (3D) software (Drishti v3.0) used to identify and quantify the Apical Foramen (AF) and Apical Constriction (AC) in mandibular premolars. Methods: Eight mandibular premolars were divided into two groups, with four samples in each group. Samples were scanned using a ZEISS X-Radia 520 Micro-CT system with two different voxel sizes of 23 µm (Group A) and 17 µm (Group B), respectively. The micro-CT data were reconstructed and transferred to Drishti Software, and individual image rendering was performed to visualize the images for data analysis. The position and measurement of AF and AC to the anatomical apex were measured individually and independently by single operators. Results: The reliability of the measurements assessed using Intraclass Correlation Coefficients (ICCs) between two voxel sizes was excellent and on par with previous research. This pilot study suggested that results for voxel sizes of 23 µm and 17 µm were comparable. Both voxel size 3D images demonstrated excellent and high-quality visualization of the root canal configuration, lateral and accessory canals, apical deltas, major AF, and AC. Conclusion: It is feasible and reliable to visualize and analyze 3D micro-CT images using open-source software (Drishti v3.0). Both voxel sizes of 23 µm and 17 µm are recommended for use in root canal study.

Lions & sea lions & bears, oh my: utilizing museum specimens to study the ossification sequence of carnivoran taxa

BackgroundMammalian skeletons are largely formed before birth. Heterochronic changes in skeletal formation can be investigated by comparing the order of ossification for different elements of the skeleton. Due to the challenge of collecting prenatal specimens in viviparous taxa, opportunistically collected museum specimens provide the best material for studying prenatal skeletal development across many mammalian species. Previous studies have investigated ossification sequence in a range of mammalian species, but little is known about the pattern of bone formation in Carnivora. Carnivorans have diverse ecologies, diets, and biomechanical specializations and are well-suited for investigating questions in evolutionary biology. Currently, developmental data on carnivorans is largely limited to domesticated species. To expand available data on carnivoran skeletal development, we used micro-computed tomography (micro-CT) to non-invasively evaluate the degree of ossification in all prenatal carnivoran specimens housed in the Harvard Museum of Comparative Zoology. By coding the presence or absence of bones in each specimen, we constructed ossification sequences for each species. Parsimov-based genetic inference (PGi) was then used to identify heterochronic shifts between carnivoran lineages and reconstruct the ancestral ossification sequence of Carnivora.ResultsWe used micro-CT to study prenatal ossification sequence in six carnivora species: Eumetopias jubatus (Steller sea lion, n = 6), Herpestes javanicus (small Indian mongoose, n = 1), Panthera leo (lion, n = 1), Urocyon cinereoargenteus (gray fox, n = 1), Ursus arctos arctos (Eurasian brown bear, n = 1), and Viverricula indica (small Indian civet, n = 5). Due to the relatively later stage of collection for the available specimens, few heterochronic shifts were identified. Ossification sequences of feliform species showed complete agreement with the domestic cat. In caniforms, the bear and fox ossification sequences largely matched the dog, but numerous heterochronic shifts were identified in the sea lion.ConclusionsWe use museum specimens to generate cranial and postcranial micro-CT data on six species split between the two major carnivoran clades: Caniformia and Feliformia. Our data suggest that the ossification sequence of domestic dogs and cats are likely good models for terrestrial caniforms and feliforms, respectively, but not pinnipeds.