Osteoporosis and other metabolic bone diseases are prevalent in the aging population. While bone has the capacity to regenerate throughout life, bone formation rates decline with age and contribute to reduced bone density and strength. Identifying mechanisms and pathways that increase bone accrual in adults could prevent fractures and accelerate healing. G protein-gated inwardly rectifying K+ (GIRK) channels are key effectors of G protein-coupled receptor signaling. Girk3 was recently shown to regulate endochondral ossification. Here, we demonstrate that deletion of Girk3 increases bone mass after 18 weeks of age. Male 24-week-old Girk3 -/- mice have greater trabecular bone mineral density and bone volume fraction than wildtype (WT) mice. Osteoblast activity is moderately increased in 24-week-old Girk3 -/- mice compared to WT mice. In vitro, Girk3-/- bone marrow stromal cells (BMSCs) are more proliferative than WT BMSCs. Calvarial osteoblasts and BMSCs from Girk3 -/- mice are also more osteogenic than WT cells, with altered expression of genes that regulate the wingless-related integration site (Wnt) family. Wnt inhibition via Dickkopf-1 (Dkk1) or β-catenin inhibition via XAV939 prevents enhanced mineralization, but not proliferation, in Girk3 -/- BMSCs and slows these processes in WT cells. Finally, selective ablation of Girk3 from cells expressing Cre recombinase from the 2.3 kb-Col1a1 promoter, including osteoblasts and osteocytes, is sufficient to increase bone mass and bone strength in male mice at 24 weeks of age. Taken together, these data demonstrate that Girk3 regulates progenitor cell proliferation, osteoblast differentiation, and bone mass accrual in adult male mice.

Zika virus (ZIKV) impacts the developing brain. Here, a technique was applied to define, in 3D, developmental changes in the brains of ZIKV-infected mice. Postnatal day 1 mice were uninfected or ZIKV-infected, then analysed by iodine staining and micro-CT scanning (diffusible iodine contrast-enhanced micro-CT; diceCT) at 3-, 6-, and 10-days post-infection (dpi). Multiple brain regions were visualised using diceCT: the olfactory bulb, cerebrum, hippocampus, midbrain, interbrain, and cerebellum, along with the lens and retina of the eye. Brain regions were computationally segmented and quantitated, with increased brain volumes and developmental time in uninfected mice. Conversely, in ZIKV-infected mice, no quantitative differences were seen at 3 or 6 dpi when there were no clinical signs, but qualitatively, diverse visual defects were identified at 6-10 dpi. By 10 dpi, ZIKV-infected mice had significantly lower body weight and reduced volume of brain regions compared to 10 dpi-uninfected or 6 dpi ZIKV-infected mice. Nissl and immunofluorescent Iba1 staining on post-diceCT tissue were successful, but RNA extraction was not. Thus, diceCT shows utility for detecting both 3D qualitative and quantitative changes in the developing brain of ZIKV-infected mice, with the benefit, post-diceCT, of retaining the ability to apply traditional histology and immunofluorescent analysis to tissue.

Imprecision in breast conserving surgery results in high rates of take back to theatre for reexcision of margins. This paper reviews the various approaches to improving the precision of oncological margin control in breast conserving surgery. The review describes the rationale for improved tissue characterization over tumor localization and explores technology-free approaches, as well as progress being made to develop and test innovative technological solutions.

Purpose: To evaluate the efficacy of vertical guided bone regeneration (GBR) in the mandible utilizing a non-resorbable membrane and a bone graft combination of autogenous bone chips, and high-temperature processed (HTP) xenograft, through CT scans and microCT analysis. Materials and Methods: Patients underwent vertical ridge augmentation procedures prior to implant placement. The surgical procedure included flap elevation and placement of a bone graft comprising a 1:1 combination of autogenous posterior mandible-derived bone chips, and HTP xenograft graft particles covered with a d-PTFE membrane trimmed to suit the 3D shape of the bone defect. This was fastened securely with titanium screws and pins, and a layer of native collagen membrane. Post-operative complications and ridge measurements were assessed. Pre bone augmentation and pre implant placement bone parameters were obtained from CT scans. Biopsy specimens collected during implantation were examined by microCT. Results: All 13 study procedures were successful without any complications. The results revealed average vertical and horizontal bone gains of 3.35 mm and 5.15 mm respectively. A total of 33 implants were successfully placed in the augmented areas, without the need for further bone augmentation. MicroCT analysis revealed 48% bone, 15% filler material, and 37% non-calcified tissue in the augmented region compared to 65% bone, 3% filler material, and 32% non-calcified tissue in the pristine bone. Conclusions: A mixture of autogenous bone and HTP xenograft, covered with a d-PTFE membrane and a layer of native collagen membrane is effective for vertical GBR.

Mammalian placentas exhibit unparalleled structural diversity, despite sharing a common ancestor and principal functions. The bulk of structural studies in placental research has used two-dimensional (2D) histology sectioning, allowing significant advances in our understanding of mammalian placental structure. However, 2D histology sectioning may be limited if it does not provide accurate information of three-dimensional (3D) tissue architecture. Here, we propose correlative 3D X-ray histology (3D-XRH) as a tool with great potential for resolving mammalian placental structures. 3D-XRH involves scanning a formaldehyde-fixed, paraffin embedded (FFPE) tissue block with 3D X-ray microscopy (microCT) prior to histological sectioning to generate a 3D image volume of the embedded tissue piece. The subsequent 2D histology sections can then be correlated back into the microCT image volume to couple histology staining (or immunolabelling) with 3D tissue architecture. 3D-XRH is non-destructive and requires no additional sample preparation than standard FFPE histology sectioning, however the image volume provides 3D morphometric data and can be used to guide microtomy. As such, 3D-XRH introduces additional information to standard histological workflows with minimal effort or disruption. Using primary examples from porcine, bovine, equine, and canine placental samples, we demonstrate the application of 3D-XRH to quantifying placental structure as well as discussing the limitations and future directions of the methodology. The wealth of information derived from 2D histological sectioning in the biomedical, veterinary, and comparative reproductive sciences provides a rich foundation from which 3D-XRH can build on to advance the study of placental structure and function.

Placental structure is linked to function across morphological scales. In the placenta, changes to gross anatomy, such as surface area, volume, or blood vessel arrangement, are associated with suboptimal physiological outcomes. However, quantifying each of these metrics requires different laborious semi-quantitative methods. Here, we demonstrate how, with minimal sample preparation, whole-organ computed microtomography (microCT) can be used to calculate gross morphometry of the equine placenta and a range of additional metrics, including branching morphometry of placental vasculature, non-destructively from a single dataset. Our approach can be applied to quantify the gross structure of any large mammalian placenta.

OBJECTIVE: This ex vivo study aimed to evaluate the influence of different filling techniques on the filling removal from oval root canals filled with bioceramic sealer.
METHODS: Thirty-six mandibular incisors with single oval canals were filled with bioceramic sealer following the techniques (n = 12): single-cone, modified McSpadden, or continuous wave of condensation, and scanned by micro-computed tomography (micro-CT). The filling was removed using the Clearsonic ultrasound tip and Reciproc system up to the R40 instrument, and the working time was recorded. The teeth were scanned again by micro-CT. Percentage of remnant volume (mm³) of the filling material, dentin thickness, and root canal transportation were measured. The data was analyzed using parametric and non-parametric tests with a significance level of 5%.
RESULTS: It took more time to remove the filling material using the continuous wave of condensation technique (p < 0.05), followed by the modified McSpadden and single-cone techniques. There was no difference (p > 0.05) when comparing the percentage of remaining filling material among the three groups, nor did it among the segments of each tooth. There was also no difference in the analysis of dentin thickness and transportation when comparing the groups (p > 0.05).
CONCLUSIONS: The filling technique did not influence the amount of remaining filling material, dentin thickness, and transportation. The working time was longer with thermoplastic filling techniques.
CONCLUSIONS: Endodontic retreatment in teeth filled with bioceramic sealers increases with their use. Several techniques are used to fill the root canals, thus, it is important to know whether the filling technique influences the non-surgical endodontic retreatment.

OBJECTIVE: Cortical bone geometry is commonly used to investigate biomechanical properties of primate mandibles. However, the ontogeny of these properties is less understood. Here we investigate changes in cortical bone cross-sectional properties throughout capuchin ontogeny and compare captive versus wild, semi-provisioned groups. Tufted capuchins (Sapajus spp.) are known to consume relatively hard/tough foods, while untufted capuchins (Cebus spp.) exploit less mechanically challenging foods. Previous research indicates dietary differences are present early in development and adult Sapajus mandibles can resist higher bending/shear/torsional loads.
METHODS: This study utilized microCT scans of 22 Cebus and 45 Sapajus from early infancy to adulthood from three sample populations: one captive Cebus, one captive Sapajus, and one semi-provisioned, free-ranging Sapajus. Mandibular cross-sectional properties were calculated at the symphysis, P3, and M1. If the tooth had not erupted, its position within the crypt was used. A series of one-way ANOVAs were performed to assess differences between and within the sample populations.
RESULTS: Mandible robusticity increases across ontogeny for all three sample populations. Sapajus were better able to withstand bending and torsional loading even early in ontogeny, but no difference in shear resistance was found. Semi-provisioned, free-ranging Sapajus tend to show increased abilities to resist bending and torsional loading but not shear loading compared to captive Sapajus.
CONCLUSIONS: This study helps advance our understanding of the primate masticatory system development and opens the door for further studies into adaptive plasticity in shaping the masticatory apparatus of capuchins and differences in captive versus free-ranging sample populations.

Detailed osteological descriptions of the craniomandibular complex of passerine birds are lacking for most species, limiting our understanding of their diversity and evolution. Cowbirds (genus Molothrus) are a small but widespread group of New World nine-primaried songbirds, well-known for their unique brooding parasitic behavior. However, detailed osteological data for cowbirds and other Icteridae are currently scarce and several features of their skulls remain undescribed or poorly known. To address this issue, a detailed comparative osteology of cowbird skulls is presented here for the first time based on data from x-ray microcomputed tomography, dry skeletal data, and multivariate analyses of linear morphometric data. Cowbird skulls offer some functional insights, with many finch-like features probably related to a seed-rich diet that distinguishes them from most other icterids. In addition, features previously overlooked in earlier studies might provide valuable phylogenetic information at different levels of passerine phylogeny (Passerida, Emberizoidea, Icteridae, and Agelaiinae), including some of the otic region and nasal septum. Comparisons among cowbirds show that there is substantial cranial variation within the genus, with M. oryzivorus being the most divergent cowbird species. Within the genus, distantly related species share similar overall skull morphology and proportions, but detailed osteological data allow species identification even in cases of strong convergence. Further efforts are warranted to furnish baseline data for future studies of this iconic group of Neotropical birds and to fully integrate it into phylogenetic comparative frameworks.

3D printing technology is becoming a widely adopted alternative to traditional polymer manufacturing methods. The most important advantage of 3D printing over traditional manufacturing methods, such as injection molding or extrusion, is the short time from the creation of a new design to the finished product. Nevertheless, 3D-printed parts generally have lower strength and lower durability compared to the same parts manufactured using traditional methods. Resistance to the environmental conditions in which a 3D-printed part operates is important to its durability. One of the most important factors that reduces durability and degrades the mechanical properties of 3D-printed parts is temperature, especially rapid temperature changes. In the case of inhomogeneous internal geometry and heterogeneous material properties, rapid temperature changes can have a significant impact on the degradation of 3D-printed parts. This degradation is more severe in high-humidity environments. Under these complex service conditions, information on the strength and fatigue behavior of 3D-printed polymers is limited. In this study, we evaluated the effects of high humidity and temperature changes on the durability and strength properties of 3D-printed parts. Samples made of commonly available materials such as ABS (Acrylonitrile Butadiene Styrene), ASA (Acrylonitrile-Styrene-Acrylate), HIPS (High-Impact Polystyrene), and PLA (Poly(lactic acid)) were subjected to temperature cycling, from an ambient temperature to -20 °C, and then were heated to 70 °C. After thermal treatment, the samples were subjected to cyclic loading to determine changes in their fatigue life relative to non-thermally treated reference samples. The results of cyclic testing showed a decrease in durability for samples made of ASA and HIPS. The ABS material proved to be resistant to the environmental effects of shocks, while the PLA material exhibited an increase in durability. Changes in the internal structure and porosity of the specimens under temperature changes were also evaluated using microcomputed tomography (microCT). Temperature changes also affected the porosity of the samples, which varied depending on the material used.