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Glucocorticoid-induced osteoporosis (GIOP) represents the most prevalent form of secondary osteoporosis. Aucubin (AU), a principal active component found in traditional herbal medicines such as Eucommia ulmoides, has been demonstrated to enhance osteoblast differentiation. Nonetheless, the precise therapeutic effects of AU on GIOP and the complex underlying regulatory mechanisms warrant further investigation. We first established a GIOP model in female mice and then assessed the therapeutic effects of AU using micro-CT analysis, biomechanical testing, measurements of serum calcium (Ca) and phosphorus (P) levels, and histological analyses using Hematoxylin and Eosin (HE) and Masson staining. Subsequently, non-targeted metabolomics was employed in order to study the effects of AU on serum metabolites in GIOP mice. The levels of the factors related to these metabolites were quantified using real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and western blot analyses. Finally, the effects of AU on osteoblastic and osteoclastic differentiation were examined. We found that AU significantly ameliorated bone microarchitecture and strength in GIOP mice. It mitigated pathological damages such as impairment of trabecular bone structure and reduction in collagen fibers, while concurrently elevating serum levels of Ca and P. Non-targeted metabolomics revealed that Arachidonic acid (AA) metabolism serves as a common pathway between the control and GIOP groups, as well as between the high-dose AU (AUH) and GIOP groups. AU notably upregulates prostaglandin-endoperoxide synthase 2 (PTGS2) and microsomal prostaglandin-E synthase 1 (PTGES) expression and downregulates prostaglandin-H2 D-isomerase (PTGDS) expression. Furthermore, AU treatment increased the expression of runt-related transcription factor 2 (Runx2) and transcription factor Sp7 (Osterix), enhanced serum alkaline phosphatase (ALP) activity, and reduced osteoclast expression. These results indicate that AU is a potential drug for treating GIOP, and its mechanism is related to regulating AA metabolism and promoting osteoblast differentiation. However, the key targets of AU in treating GIOP still need further exploration., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of competing interest We declare we have no competing interests., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Microcomputed tomography (µCT) is a nondestructive X-ray imaging method used in plant physiology to visualize in situ plant tissues that enables assessments of embolized xylem vessels. Whereas evidence for X-ray-induced cellular damage has been reported, the impact on plant physiological processes such as carbon (C) uptake, transport, and use is unknown. Yet, these damages could be particularly relevant for studies that track embolism and C fluxes over time. We examined the physiological consequences of µCT scanning for xylem embolism over 3 mo by monitoring net photosynthesis (Anet), diameter growth, chlorophyll (Chl) concentration, and foliar nonstructural carbohydrate (NSC) content in 4 deciduous tree species: hedge maple (Acer campestre), ash (Fraxinus excelsior), European hornbeam (Carpinus betulus), and sessile oak (Quercus petraea). C transport from the canopy to the roots was also assessed through 13C labeling. Our results show that monthly X-ray application did not impact foliar Anet, Chl, NSC content, and C transport. Although X-ray effects did not vary between species, the most pronounced impact was observed in sessile oak, marked by stopped growth and stem deformations around the irradiated area. The absence of adverse impacts on plant physiology for all the tested treatments indicates that laboratory-based µCT systems can be used with different beam energy levels and doses without threatening the integrity of plant physiology within the range of tested parameters. However, the impacts of repetitive µCT on the stem radial growth at the irradiated zone leading to deformations in sessile oak might have lasting implications for studies tracking plant embolism in the longer-term., Competing Interests: Conflict of interest statement. None declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of American Society of Plant Biologists.)

Competing Interests: Disclosure The authors report no conflicts of interest in this work.

Two-directional beam-tracking (2DBT) is a method for phase-contrast imaging and tomography that uses an intensity modulator to structure the X-ray beam into an array of independent circular beamlets that are resolved by a high-resolution detector. It features isotropic spatial resolution, provides two-dimensional phase sensitivity, and enables the three-dimensional reconstructions of the refractive index decrement, δ, and the attenuation coefficient, μ. In this work, the angular sensitivity and the spatial resolution of 2DBT images in a synchrotron-based implementation is reported. In its best configuration, angular sensitivities of ∼20 nrad and spatial resolution of at least 6.25 µm in phase-contrast images were obtained. Exemplar application to the three-dimensional imaging of soft tissue samples, including a mouse liver and a decellularized porcine dermis, is also demonstrated., (open access.)

Background: Visualizing (micro)vascular structures remains challenging for researchers and clinicians due to limitations in traditional radiological imaging methods. Exploring the role of vascular development in craniofacial malformations in experimental settings can enhance understanding of these processes, with the effectiveness of high-resolution imaging techniques being crucial for successful research in this field. Micro-CT imaging offers 3D microstructural insights, but requires contrast-enhancing staining agents (CESAs) for visualizing (micro)-vascular tissues, known as contrast-enhanced micro-CT (CECT). As effective contrast agents are crucial for optimal visualization, this review focuses on comparative studies investigating such agents for micro-vascular tissue imaging using micro-CT. Furthermore, we demonstrate the utilization of B-Lugol solution as a promising contrast agent for acquiring high-quality micro-CT images of (micro)vascular structures in human embryonic samples., Method: This scoping review followed Preferred Reporting Items for Systematic Reviews and Meta-analysis Protocols. PubMed database provided relevant articles, screened initially by title and abstract. Inclusion and exclusion criteria defined outcomes of interest., Results: From an initial search, 273 records were identified, narrowed down to 9 articles after applying our criteria. Additionally, two articles were added through citation searching. This, a total of 11 articles were incorporated in this study., Conclusion: This micro-CT contrast agent review underscores the need for tailored choices based on research goals. Both Barium sulfate and Iodine-based agents showing excellent results, providing high resolution (micro) vascular content, especially in ex-vivo specimens. However, careful consideration of protocols and tissue characteristics remains imperative for optimizing the effectiveness of micro-CT imaging for the study of cranio-facial vascular development., (© 2024. The Author(s).)

Study Design: Diagnostic test study., Objective: To determine the reliability and validity or diagnostic accuracy of two previously described endplate structural defect (EPSD) assessment methods., Summary of Background Data: Studies of EPSD may further the understanding of pathoanatomic mechanisms underlying back pain. However, clinical imaging methods used to document EPSD have not been validated, leaving uncertainty about what the observations represent., Materials and Methods: Using an evaluation manual, 418 endplates on CT sagittal slices obtained from 19 embalmed cadavers (9 men and 10 women, aged 62-91 yr) were independently assessed by two experienced radiologists and a novice for EPSD using the two methods. The corresponding micro-CT (µCT) from the harvested T7-S1 spines were assessed by another independent rater with excellent intra-rater reliability (k=0.96)., Results: Inter-rater reliability was good for the presence (k=0.60-0.69) and fair for specific phenotypes (k=0.43-0.58) of EPSD. Erosion, for which the Brayda-Bruno classification lacked a category, was mainly (82.8%) classified as wavy/irregular, while many notched defects (n=15, 46.9%) and Schmorl's nodes (n=45, 79%) were recorded as focal defects using Feng's classification. When compared to µCT, endplate fractures (n=53) and corner defects (n=28) were routinely missed on CT. Endplates classified as wavy/irregular on CT corresponded to erosion (n=29, 21.2%), jagged defects (n=21, 15.3%), calcification (n=19, 13.9%), and other phenotypes on µCT. Some focal defects on CT represented endplate fractures (n=21, 27.6%) on µCT. Overall, with respect to the presence of an EPSD, there was a sensitivity of 70.9% and a specificity of 79.1% using Feng's method, and 79.5% and 57.5% using Brayda-Bruno's method. Poor to fair inter-rater reliability (k=0.26-0.47) was observed for defect dimensions., Conclusion: There was good inter-rater reliability and evidence of criterion validity supporting assessments of EPSD presence using both methods. However, neither method contained all needed EPSD phenotypes for optimal sensitivity, and specific phenotypes were often misclassified., Competing Interests: The authors report no conflicts of interest., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Background: Trigeminal ganglion (TG) plays an important role in the process of orthodontic pain. It's necessary to design an accurate, precise and minimally invasive trigeminal ganglion injection guide plate to study TG., Methods: Micro-CT was used to obtain the Dicom format data, and three-dimensional (3D) software (mimics and magics23.03) was used to reconstruct 3D head models. Design and modifications of the TG injection guide plate were performed in Magic 23.03 software, and the guide plate was produced by a 3D stereolithography printer. X-ray, micro-CT, Evans blue, and virus transduction were used to demonstrate the accuracy of the guide-assisted injection. Pain levels were evaluated after using the injection guide by a bite force test and Von Frey test., Results: X-ray and micro-CT tests confirmed that the injection needle reached the bilateral trigeminal ganglia fossa. The Evans blue test and virus transduction proved that the injected drug could be accurately injected into the bilateral trigeminal ganglion and the lentivirus could be successfully transfected. The percentage of accurate injection was 10/10 (bilateral trigeminal ganglia). Orofacial pain induced by the trigeminal ganglion injection was mild and returned to baseline within seven days., Conclusion: The injection guide described in this study is viable and reliable for the delivery of drugs and virus transduction into the trigeminal ganglia., Competing Interests: Declaration of Competing Interest All authors declare that they have no conflicts of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: The human peripheral nervous system embodies anatomical, physiologic, and diagnostic perplexities that remain unexplained. Yet in the course of human history, there are no mechanisms, such as computed tomography (CT) or radiography, by which to image the peripheral nervous system in vivo using a contrast agent that is identified by ionizing radiation, which would aid in surgical navigation, diagnostic radiology, and basic science thereof., Methods: A novel class of contrast was created by linking iodine to lidocaine. The radiodensity of 0.5% experimental contrast molecule was compared with a control of 1% lidocaine by placing 1.5-mL aliquots of each liquid into centrifuge tubes and performing micro-computed tomography (micro-CT) synchronously under identical settings. Physiologic binding to the sciatic nerve was evaluated by injecting 10 mg of the experimental contrast and 10 mg of the control into the contralateral sciatic nerve, and documenting loss of hindlimb function and recovery. In vivo visualization of the sciatic nerve was evaluated by injecting 10 mg of experimental contrast or control into either sciatic nerve and imaging the hindlimbs under identical conditions using micro-CT., Results: The mean Hounsfield unit of the contrast was 56.09 compared with -0.48 for control (116-fold increase, P = .0001). Hindlimb paresis revealed similar degree of paresis, baseline recovery, and time to recovery. In vivo enhancement between the contralateral sciatic nerves was similar., Conclusion: Iodinated lidocaine offers a viable mechanism for in vivo peripheral nerve imaging using CT; however, it requires modification to improve in vivo radiodensity., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

The aim of this study was to explore the feasibility of a time-resolved reflectance imaging system employing a single photodetector to assess the activity of caries lesions that exploits the differential absorption of water at 1300 and 1950 nm. The time-resolved reflectivity of 10 active and 10 arrested lesions on the proximal surfaces and 5 active and 5 arrested lesions on the occlusal surfaces of extracted teeth were monitored simultaneously at 1300 and 1950 nm during forced air drying for 60 s. The presence of a highly mineralized surface zone measured with microcomputed tomography (microCT) was used to indicate lesion activity. Multiple kinetic parameters were extracted from the acquired short wavelength infrared (SWIR) intensity versus time dehydration curves and used to assess lesion activity. Differences in the reflectivity between curves acquired at 1300 and 1950 nm due to differential absorption of water provided improved discrimination between active and arrested lesions over the use of 1950 nm alone. This study demonstrates that it is feasible to use a device with a single photodetector operating at 1950 nm to collect dehydration curves for the assessment of lesion activity and that a system employing two SWIR wavelengths with differential water absorption can improve the performance of lesion activity assessment., (© 2024. The Author(s), under exclusive licence to Springer-Verlag London Ltd., part of Springer Nature.)

The purpose of this study was to evaluate the efficacy of different techniques in removing calcium silicate intracanal medicament (Bio-C Temp). Forty human single-canaled premolars were randomly distributed into five groups (n = 8). All root canals were instrumented, then filled with Bio-C Temp. Following 1-week incubation, the intra-canal medicament was removed using one of five techniques according to tooth group: conventional syringe irrigation, Endo Activator, passive ultrasonic irrigation (PUI), ProTaper Universal F3 and XP-endo Finisher (XPF). Micro-CT scanning was performed before and after removal of Bio-C Temp. All techniques significantly reduced the volume of Bio-C Temp (p < 0.001) without reaching complete elimination. The percentage of Bio-C Temp removed was significantly higher in the XPF group (98.2%) compared to conventional syringe irrigation (70.6%), the Endo Activator (75.7%), and the ProTaper Universal (76.6%). There was no significant difference between the XPF and PUI (95.1%) groups. None of the removal techniques were able to completely remove Bio-C Temp from the root canal. However, XPF was the most effective method, but was not statistically significant when compared to PUI. Clinical Relevance: This study demonstrated that both XPF and PUI outperform conventional irrigation in removing Bio-C Temp intracanal medicament., (© 2024. The Author(s).)

Background: The existence of voids within the mineral trioxide aggregate (MTA) composition is one of the factors that can influence the treatment outcome. The primary objective of this study was to quantitatively assess and compare the MTA orthograde obturation quality in severe curved root canals using two different MTA compaction techniques: manual compaction with K-file, or Auger technique using micro-computed tomography (micro-CT) imaging., Methods: For this study, 26 mandibular first molar teeth with severely curved mesiobuccal root canals were selected. These samples were randomly divided into two groups. All root canals were instrumented using ProTaper Gold rotary files up to the F3 file at the working length. In one group, OrthoMTA was compacted using a stainless steel K-file, while in the other group, the Auger technique was employed for compaction into the root canals. Once the MTA had completely set, the filled root canals were subjected to scanning using a high-resolution micro-CT scanner. The porosity volume was determined as a percentage in relation to the overal volume of the canal, and the collected data were subjected to analysis using SPSS software, with the significance level set at P < 0.05., Results: The two techniques had no significant difference in open, closed, and total mean porosity. In both groups, the mean of open porosity was significantly more than closed porosity., Conclusions: According to the results of the present study, neither of these two techniques is preferred to the other, and factors such as working time, etc., can be considered to choose the more appropriate clinical technique., (© 2024. The Author(s).)

We developed a nondestructive three-dimensional microbial visualization method utilizing synchrotron radiation X-ray microscale computed tomography to better understand the relationship between microorganisms and their surrounding habitats. The method was tested and optimized using a mixture of axenic Escherichia coli and Comamonas testosteroni . The osmium-thiocarbohydrazide-osmium method was used to stain all the microbial cells, and gold in situ hybridization was used to detect specific phylogenetic microbial groups. The stained samples were embedded in epoxy resin for microtomographic analysis. Differences in X-ray absorbances were calculated by subtracting the pre-L 3 -edge images from the post-L 3 -edge images to visualize the osmium and gold signals. Although we successfully detected cells stained with osmium, those labeled with gold were not detected, probably because of the insufficient density of gold atoms in the microbial cells. We then applied the developed technique to anaerobic granules and visualized the distribution of microbial cells and extracellular polymeric substances. Empty spaces were highlighted to determine the cavity distribution in granules. Numerous independent cavities of different sizes were identified in the granules. The developed method can be applied to various environmental samples for deeper insights into microbial life in their habitats., Importance: Microorganisms inhabit diverse environments and often form biofilms. One factor that affects their community structure is the surrounding physical environment. The arrangement of residential space within the formed biofilm plays a crucial role in the supply and transportation of substances, as well as the discharge of metabolites. Conventional approaches, such as scanning electron microscopy and confocal laser scanning microscopy combined with fluorescence in situ hybridization, have limitations as they provide information primarily from the biofilm surface and cross-sections. In this study, we developed a method for detecting microorganisms in biofilms using synchrotron radiation X-ray microscale computer tomography. The developed method allows nondestructive three-dimensional observation of biofilms at a single-cell resolution (voxel size of approximately 200 nm), facilitating an understanding of the relationship between microorganisms and their physical habitats., Competing Interests: The authors declare no conflict of interest.

Osteoporosis (OP) is a metabolic bone disease that can lead to major health challenges. The theory of Traditional Chinese medicine believes that kidney-Yin deficiency (KYD) is the main cause of postmenopausal osteoporosis. This study was aimed to investigate the effect of EZW on anti-osteoporosis with KYD, and explore potential mechanisms from the perspective of the kidney, bone and bone marrow through analysis of metabolomics and proteomics. The model of OP with KYD was established by rats treated with bilateral ovariectomy (OVX), and then given intragastric administration of thyroid and reserpine to induce. Micro-CT was applied to determine the microstructures of bone. Serum levels associated with bone turnover markers and kidney-Yin deficiency were detected by enzyme-linked immunosorbent (ELISA) assay. The differential metabolites in the kidney, bone and bone marrow were analyzed by metabolomics. The differentially expressed proteins in these three tissues were detected via proteomics. The findings suggested that EZW could alleviate a variety of metabolites and proteins among the kidney, bone and bone marrow, primarily in amino acid metabolism, carbohydrate metabolism, nucleotide metabolism and lipid metabolism, thus leading to improvements of OP with KYD, which provided theoretical basis for clinical treatment of EZW on OP with KYD., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The auditory ossicles amplify and transmit sound from the environment to the inner ear. The distribution of bone mineral density is crucial for the proper functioning of sound transmission as the ossicles are suspended in an air-filled chamber. However, little is known about the distribution of bone mineral density along the human ossicular chain and within individual ossicles. To investigate this, we analyzed fresh-frozen human specimens using synchrotron-based phase-contrast microtomography. In addition, we analyzed the volume and porosity of the ossicles. The porosity for the auditory ossicles lies, on average, between 1.92% and 9.85%. The average volume for the mallei is 13.85 ± 2.15 mm 3 , for the incudes 17.62 ± 4.05 mm 3 and 1.24 ± 0.29 mm 3 for the stapedes. The bone density distribution showed a similar pattern through all samples. In particular, we found high bone mineralization spots on the anterior crus of the stapes, its footplate, and along areas that are crucial for the transmission of sound. We could also see a correlation between low bone mineral density and holey areas where the bone is only very thin or missing. Our study identified a similar pattern of bone density distribution within all samples: regions exposed to lower forces generally show higher bone density. Further, we observed that the stapes shows high bone mineral density along the anterior crus and its footplate, which may indicate its importance in transmitting sound waves to the inner ear., (© 2024. The Author(s).)

Objective . Auto-segmentation in mouse micro-CT enhances the efficiency and consistency of preclinical experiments but often struggles with low-native-contrast and morphologically complex organs, such as the spleen, resulting in poor segmentation performance. While CT contrast agents can improve organ conspicuity, their use complicates experimental protocols and reduces feasibility. We developed a 3D Cycle Generative Adversarial Network (CycleGAN) incorporating anatomy-constrained U-Net models to leverage contrast-enhanced CT (CECT) insights to improve unenhanced native CT (NACT) segmentation. Approach. We employed a standard CycleGAN with an anatomical loss function to synthesize virtual CECT images from unpaired NACT scans at two different resolutions. Prior to training, two U-Nets were trained to automatically segment six major organs in NACT and CECT datasets, respectively. These pretrained 3D U-Nets were integrated during the CycleGAN training, segmenting synthetic images, and comparing them against ground truth annotations. The compound loss within the CycleGAN maintained anatomical fidelity. Full image processing was achieved for low-resolution datasets, while high-resolution datasets employed a patch-based method due to GPU memory constraints. Automated segmentation was applied to original NACT and synthetic CECT scans to evaluate CycleGAN performance using the Dice Similarity Coefficient (DSC) and the 95th percentile Hausdorff Distance (HD 95p ). Main results. High-resolution scans showed improved auto-segmentation, with an average DSC increase from 0.728 to 0.773 and a reduced HD95p from 1.19 mm to 0.94 mm. Low-resolution scans benefited more from synthetic contrast, showing a DSC increase from 0.586 to 0.682 and an HD 95p reduction from 3.46 mm to 1.24 mm. Significance. Implementing CycleGAN to synthesize CECT scans substantially improved the visibility of the mouse spleen, leading to more precise auto-segmentation. This approach shows the potential in preclinical imaging studies where contrast agent use is impractical., (© 2024 IOP Publishing Ltd.)

Orthodontic tooth movement (OTM) represents a dynamic process in which the alveolar bone undergoes resorption at compression sites and deposition at tension sites, orchestrated by osteoclasts and osteoblasts, respectively. This mechanism serves as a valuable model for studying various aspects of bone adaptation, including root resorption and the cellular response to mechanical force stimuli. The protocol outlined here offers a straightforward approach to investigate OTM, establishing 0.35 N as the optimal force in a mouse model employing a nickel-titanium (NiTi) coil spring. Utilizing micro-computed tomography analysis, we quantified OTM by assessing the discrepancy in the linear distance at the cement-enamel junction. The evaluation also included an analysis of orthodontic-induced inflammatory root resorption, assessing parameters such as root mineral density and the percentage of root volume per total volume. This comprehensive protocol contributes to advancing our understanding of bone remodeling processes and enhancing the ability to develop effective orthodontic treatment strategies.

The purpose of this protocol is to guide researchers in performing a palpation-guided technique of intra-articular knee injection in guinea pigs and assessment using micro-computed tomography. Dunkin-Hartley guinea pigs are robust models for osteoarthritis research as they spontaneously develop osteoarthritis in their knees. Intra-articular drug delivery is a common method to study the effects of an investigational drug in vivo. In humans, therapeutic agents administered via intra-articular injection can offer pain relief and delay further progression of osteoarthritis. As with any species, the introduction of a needle into a joint space has the potential to cause injury, which can result in pain, lameness, or infection. Such adverse events can compromise animal welfare, confound study results, and necessitate additional animals to achieve study objectives. As such, it is imperative to develop proper injection techniques to prevent complications, especially in longitudinal studies that require multiple, repeated intra-articular injections. Using the presented methodology, five guinea pigs received bilateral knee injections under general anesthesia. Seven days after injection, animals were humanely euthanized for analysis of osteoarthritis severity. No adverse events occurred following anesthesia or knee injections, including limping, pain, or infection. X-ray micro-computed tomography analysis of the knee can detect pathologic changes associated with osteoarthritis. Micro-computed tomography data indicates osteoarthritis is more severe in older animals, as indicated by increased bone mineral density and trabecular thickness with age. These results are consistent with histologic changes and Modified Mankin scores, an established and widely used scoring system to assess arthritis severity in these same animals. This protocol can be utilized to refine intra-articular injections in guinea pigs.

This study evaluated the impact of canal irrigation using surfactants associated to NaOCl on the percentage of voids of root canal filling through micro-computed tomographic (micro-CT) analysis. 27 mandibular mesial roots with Vertucci type II canal configuration with isthmus were selected and scanned in a micro-CT device. The specimens were assigned to three groups (n = 9), according to the irrigation solution during instrumentation: 2.5% NaOCl without additives (control), 2.5% NaOCl with 0.1% Benzalkonium Chloride and 2.5% NaOCl with 0.1% Tween 80. After obturation, the specimens were rescanned, and the percentage of voids were calculated. Data were analysed using one-way ANOVA with a significance level of 5%. The percentage of voids were similar among the groups (p > 0.05). None of the specimens showed completely voids-free areas. The addition of surfactants to NaOCl irrigation did not improve the quality of root canal filling of mandibular mesial root canals., (© 2024 Australian Society of Endodontology Inc.)

Molar-incisor hypomineralization (MIH) is a qualitative defect of dental enamel characterized by demarcated opacities present in permanent first molars and other teeth. It is considered a major clinical challenge in dentistry because it makes affected teeth more susceptible to fractures and dental caries. Its diagnosis is mainly clinical and there are few technological resources that allow for a more accurate diagnosis, especially with respect to the depth of the defect in the dental enamel. In this context, optical coherence tomography (OCT), which is routinely used in ophthalmology, can produce images of the depth of the dental enamel, making it a promising method. In this study, 33 teeth with different MIH severities were evaluated using OCT and microcomputed tomography (microCT). Semi-quantitative methods of grayscale pattern analysis were used to compare images obtained from different severities of MIH with the mineral density obtained through microCT. MicroCT evaluation revealed that hypomineralized enamel had a significantly lower mineral density than intact enamel. However, this difference was not observed between the mild and severe MIH lesions. In the OCT evaluation, significant differences were observed between the intact and hypomineralized enamel, and the gray value comparison provided a method for quantitative differentiation between the two. This study suggests that OCT could be a useful adjunct to traditional diagnostic methods for MIH, offering a noninvasive approach to evaluate enamel defects. RESEARCH HIGHLIGHTS: Combining optical coherence tomography with grayscale digital analysis shows potential as a promising method for diagnosing molar-incisor hypomineralization and assessing its level of severity., (© 2024 Wiley Periodicals LLC.)

Background: This study aimed to investigate the contribution of myeloid differentiation primary-response gene 88 (MyD88) on the differentiation of T helper type 17 (Th17) and regulatory T (Treg) cells and the emerging subgingival microbiota dysbiosis in Porphyromonas gingivalis-induced experimental periodontitis., Methods: Alveolar bone loss, infiltrated inflammatory cells, immunostained cells for tartrate-resistant acid phosphatase (TRAP), the receptor activator of nuclear factor-kB ligand (RANKL), and osteoprotegerin (OPG) were quantified by microcomputerized tomography and histological staining between age- and sex-matched homozygous littermates (wild-type [WT, Myd88 +/+ ] and Myd88 -/- on C57BL/6 background). The frequencies of Th17 and Treg cells in cervical lymph nodes (CLNs) and spleen were determined by flow cytometry. Cytokine expression in gingival tissues, CLNs, and spleens were studied by quantitative polymerase chain reaction (qPCR). Analysis of the composition of the subgingival microbiome and functional annotation of prokaryotic taxa (FAPROTAX) analysis were performed., Results: P. gingivalis-infected Myd88 -/- mice showed alleviated bone loss, TRAP + osteoclasts, and RANKL/OPG ratio compared to WT mice. A significantly higher percentage of Foxp3 + CD4 + T cells in infected Myd88 -/- CLNs and a higher frequency of RORγt + CD4 + T cells in infected WT mice was noted. Increased IL-10 and IL-17a expressions in gingival tissue at D14-D28 then declined in WT mice, whereas an opposite pattern was observed in Myd88 -/- mice. The Myd88 -/- mice exhibited characteristic increases in gram-positive species and species having probiotic properties, while gram-negative, anaerobic species were noted in WT mice. FAPROTAX analysis revealed increased aerobic chemoheterotrophy in Myd88 -/- mice, whereas anaerobic chemoheterotrophy was noted in WT mice after P. gingivalis infection., Conclusions: MyD88 plays an important role in inflammation-induced bone loss by modulating the dynamic equilibrium between Th17/Treg cells and dysbiosis in P. gingivalis-induced experimental periodontitis., (© 2024 American Academy of Periodontology.)

Introduction: Intraoral scanners commonly used in orthodontic offices now offer near-infrared imaging (NIRI) technology, advertised as a screening tool to identify interproximal caries. This study aimed to evaluate the reliability and validity of NIRI detection of interproximal carious lesions in a common intraoral scanner (iTero Element 5D; Align Technology, San Jose, Calif) with and without bitewing radiograph complement, compared with a microcomputed tomography (micro-CT) reference standard., Methods: Extracted human posterior teeth (premolars and molars) were selected for early (noncavitated) interproximal carious lesions (n = 39) and sound control surfaces (n = 47). The teeth were scanned via micro-CT for evaluation by 2 blinded evaluators using consensus scoring. The teeth were mounted to simulate anatomic interproximal contacts and underwent a NIRI scan using iTero Element 5D and bitewing radiographs. Two trained, calibrated examiners independently evaluated (1) near-infrared images alone with clinical photograph, (2) bitewing radiograph alone with clinical photograph, and (3) near-infrared images with bitewing radiograph and clinical photograph in combination, after at least a 10-day washout period between each evaluation., Results: Interrater reliability was highest for NIRI alone (k = 0.533) compared with bitewing radiograph alone (k = 0.176) or in combination (k = 0.256). NIRI alone showed high specificity (0.83-0.96) and moderate sensitivity (0.42-0.63) compared with a micro-CT reference standard. Dentin lesions were significantly more reliably detected than enamel lesions., Conclusions: After rigorous training and calibration, NIRI can be used with moderate reliability, high specificity, and moderate sensitivity to detect noncavitated interproximal carious lesions., (Copyright © 2024 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

This study investigated the root canal morphology of fused-rooted mandibular second molars based on the pulp chamber floor (PCF) and analysed the correlation between the external morphology of the radicular groove, and the internal morphology of the PCF and root canal configuration. A total of 291 fused-rooted teeth collected from the Chinese population were scanned using micro-computed tomography and a dental operating microscope was used for observing the PCFs. The classification of the PCF and root canal configuration were identified according to modified Min et al.'s and Gao et al.'s classifications, respectively. Additionally, a new radicular groove classification was proposed. The correlation among these morphological characteristics was investigated using the chi-square test and Fisher's exact test (p < 0.05). The results showed that 74.2% of teeth had C-shaped PCFs, while 21.0% had non-C-shaped PCFs. As for the root canal configurations, 37.5% of teeth were merging type, 40.9% were symmetrical type, and 14.8% were asymmetrical type. Statistical analysis revealed a significant correlation between the PCF types and the root canal configurations (p < 0.001). The dominant root canal types for teeth with C-shaped PCFs were merging and symmetrical types, while the asymmetrical type was not identified in non-C-shaped PCFs. In addition, significant morphological association between the root canals and radicular grooves was also revealed (p < 0.001). Teeth with different PCF morphologies exhibit specific patterns of root canal category distribution. Understanding the morphological nuances of the root canal based on the PCF can assist clinicians in predicting and identifying the canal configuration beneath the visible orifice., (© 2024 Australian Society of Endodontology Inc.)

This study assessed the ability of Twisted File Adaptive (TFA), TruNatomy (TRN) and VDW.Rotate (VR) instruments activated by continuous rotation (CR) and adaptive motion (AM) to shape curved root canals. Thirty mandibular molars with two separate mesial canals 20°-40° curved were collected and scanned using micro-computed tomography (μCT). The canals were then randomly assigned into six groups (n = 10): TRN, VR or TFA instruments activated by CR or AM. TRN groups 17.02, 20.04 and 26.04; VR groups 15.04, 20.05 and 25.06; TFA groups were shaped consecutively using 15K-file, 20.04 and 25.06. After they were shaped, the canals were scanned again. The volume of removed dentin, canal transportation and centring ratio were calculated μCT images. All data were analysed using the Kruskal-Wallis test or one-way analysis of variance (p < 0.05). With both kinematics, the TRN instrument removed the least amount of dentin, the VR-CR and the TFA-AM removed the most (p < 0.05). The transportation and centring ratios among all groups were similar (p > 0.05). The volume of dentin removed, the diameter or design features of the instruments and different kinematics did not affect the centring ratio and the amount of transportation and remained within safe limits. All three instruments activated by either kinematics were found to have similar effectiveness in shaping curved root canals., (© 2024 Australian Society of Endodontology Inc.)

We examined the ultrastructure of the mammalian os penis at the high-resolution synchrotron level. Previously, bacular microanatomy had only been investigated histologically. We studied the baculum of the harp seal (Pagophilus groenlandicus), in which the baculum varies more in size and shape than does a mechanically constrained bone (humerus). We (1) investigated the microarchitecture of bacula and humeri from the same seal specimens, and (2) described changes in bone micro- and macro-morphology associated with age (n = 15, age range = 1-35 years) and bone type. We analyzed cross-sectional geometry non-destructively through laboratory micro-computed tomography. We suggest that the midshaft may resist axial compression while the proximal region may resist torsion, based on measurements of cross-sectional and cortical areas, perimeter, ratio of maximum and minimum moments of inertia, and polar moment of inertia. In addition, midshaft bacula may be less mechanosensitive than humeri, based on microstructural variables (e.g., volume, surface area, diameter associated with lacunae and cortical porosity) analyzed across age groupings. Our findings related to the microarchitecture of the pinniped baculum provide a basis for further studies on development, mechanical properties, functions, and adaptations in this and other pinniped species. Our use of a multi-modal imaging approach was minimally destructive for reproducible and accurate comparison of three-dimensional bone ultrastructure. Such methods, coupled with multidisciplinary analyses, enable diverse studies of bone biology, life history, and evolution using museum collections., (© 2024 American Association for Anatomy.)

Objectives: This study aimed to investigate the relationship between the aspect ratios of mandibular molar roots at the apical 3-mm level and their root canal complexity., Design: This study used micro-CT imaging to analyze 163 two-rooted mandibular molars. The aspect ratios of the roots at the apical 3-mm level were categorized as "< 2.75" or "≥ 2.75" (mesial) and "< 1.75" or "≥ 1.75" (distal). A two-dimensional (2D) analysis focused on four apical axial cross-section levels to determine the presence of main and accessory canals and isthmus. Additionally, a three-dimensional (3D) assessment of the apical 4-mm of both roots examined main and accessory canals, apical foramina, apical deltas, and middle mesial canals., Results: Mesial roots with aspect ratios ≥ 2.75 showed a higher number of main canals at all levels compared to those with aspect ratios < 2.75 at the 3-mm level. Additionally, the ≥ 2.75 group exhibited more accessory canals and a higher average number of accessory canals. The 3D assessment confirmed significantly more accessory canals and apical foramina in the ≥ 2.75 group. The prevalence of roots with apical deltas was nearly double in the ≥ 2.75 group, and middle mesial canals were exclusively found in this group. In the distal root, the ≥ 1.75 group showed a significantly higher number of main canals at all axial levels. No significant differences were observed between groups in terms of accessory canals, apical foramina, or deltas., Conclusions: A higher root aspect ratio is related to higher anatomical complexity., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objectives: The purpose of this study was to compare the effectiveness of three different instruments on cement loss, porosity and micro-crack formation, which was not evaluated before, following scaling and root planning (SRP) using micro-computed tomography (micro-CT)., Methods: In this experimental study, 30 single-rooted extracted human teeth were used and divided into three groups. All the teeth were scanned with micro-CT before and after SRP. Group 1: SRP was performed with Gracey curettes, Group 2: SRP was performed by using an ultrasonic device, and Group 3: SRP was performed by using diamond burs. Cement loss from the root surface, porosity, and micro-crack formation in the root dentine were analysed. Micro-CT is used for qualitative and quantitative analysis of samples. The obtained data were analysed statistically (p < 0.05)., Results: Minimum cement loss following SRP was detected with ultrasonic scaler (26.98 mm 3 ), whereas the highest was created by diamond burs (96.20 mm 3 ) (p < 0.05). The total porosity values after SRP were 0.278%, 0.334% and 0.252% for Groups 1, 2 and 3, respectively. Although Group 3 had the least porosity values, there was no statistically significant difference between the groups. The highest micro-crack formation was seen in Group 2 and the lowest was in Group 1 with a significant difference (p < 0.05)., Conclusions: More cement loss was observed with diamond burs. Ultrasonic devices appear to be a viable alternative to instrumentation with curettes. However, ultrasonic devices should be used carefully because of micro-crack formation since the micro-crack resulting from instrumentation with hand instruments is the least of all., (© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Apical periodontitis is an inflammatory condition resulting from microbial invasion in the root canal system, causing periapical tissue destruction and bone resorption. This study investigated melatonin's effects, known for its antioxidant and anti-inflammatory properties, on experimentally induced apical periodontitis in rats. Three groups of rats were studied: control, apical periodontitis and apical periodontitis with melatonin treatment. Proinflammatory cytokines and enzyme levels in blood serum were measured, and micro-CT analysis assessed bone resorption. Results showed significantly elevated cytokines and enzyme levels in the apical periodontitis group compared to the control. However, in the melatonin-treated group, these levels were significantly reduced (p < 0.01-0.001). Micro-CT analysis indicated decreased periapical resorption cavity volume and surface area with melatonin treatment. This suggests that systemic melatonin administration can mitigate inflammation and reduce bone resorption in experimentally induced apical periodontitis in rats, potentially holding promise for human endodontic disease treatment pending further research., (© 2024 Australian Society of Endodontology Inc.)

Introduction: In community settings with limited dental personnel and equipment, and in an era when the aerosol transmission of infectious agents is on the rise, a non-invasive approach to caries management is critical. To provide information on non-invasive material selection, the aim of this study was to compare the remineralization effect of commonly used fluoride-containing materials, adjunctive to the everyday use of fluoride toothpaste (F-toothpaste), on primary tooth natural dentin caries., Methods: Fifty-five specimens were randomly divided into five groups: 38% silver diamine fluoride (SDF), 5% fluoride varnish (F-varnish), glass-ionomer cement (GIC), deionized water (DW) with F-toothpaste slurry, and DW as a control group (n = 11). The lesion depth and mineral density were measured before and after bacterial pH-cycling using micro-computed tomography. The percentage of mineral density change (%MDchange) was quantified. The dependent t-test, Wilcoxon-Signed-Rank Test, and one way ANOVA with Bonferroni correction were used to analyse the data., Results: SDF application reduced lesion depth from 844.6 to 759.1 μm (p < 0.045) while increasing mineral density from 551.4 to 763.0 mgHA/cm 3 (p < 0.003). Only mineral density rose from 600.2 to 678.4 mgHA/cm 3 (p < 0.013) when GIC was used. The other groups showed no difference. The highest %MDchange was also found after SDF treatment (49.7%, p < 0.05), whereas GIC (17.2%, p < 0.05) presented a higher percentage than the F-varnish (2.0%), F-toothpaste (-1.1%) and no-treatment groups (-1.4%)., Conclusion: In this in vitro study, where the pH of cycling was almost neutral, using SDF as an adjunct to F-toothpaste resulted in the highest remineralization compared with other remineralizing materials., (© 2023 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

High cyclic strains induce formation of microcracks in bone, triggering targeted bone remodeling, which entails osteoclastic resorption. Racehorse bone is an ideal model for studying the effects of high-intensity loading, as it is subject to focal formation of microcracks and subsequent bone resorption. The volume of resorption in vitro is considered a direct indicator of osteoclast activity but indirect 2D measurements are used more often. Our objective was to develop an accurate, high-throughput method to quantify equine osteoclast resorption volume in μCT 3D images. Here, equine osteoclasts were cultured on equine bone slices and imaged with μCT pre- and postculture. Individual resorption events were then isolated and analyzed in 3D. Modal volume, maximum depth, and aspect ratio of resorption events were calculated. A convolutional neural network (CNN U-Net-like) was subsequently trained to identify resorption events on post-culture μCT images alone, without the need for pre-culture imaging, using archival bone slices with known resorption areas and paired CTX-I biomarker levels in culture media. 3D resorption volume measurements strongly correlated with both the CTX-I levels (p < 0.001) and area measurements (p < 0.001). Our 3D analysis shows that the shapes of resorption events form a continuous spectrum, rather than previously reported pit and trench categories. With more extensive resorption, shapes of increasing complexity appear, although simpler resorption cavity morphologies (small, rounded) remain most common, in acord with the left-hand limit paradigm. Finally, we show that 2D measurements of in vitro osteoclastic resorption are a robust and reliable proxy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Lance Frazer reports financial support was provided by Canon Medical Systems USA Inc. Jay Vaishnav reports a relationship with Canon Medical Systems USA Inc. that includes: employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: The purpose of this study was to assess the effects of systemically given krill oil (KO) on the development of new bone formation in the sutura palatina media following rapid maxillary expansion (RME)., Methods: 28 4-5 week-old male Wistar albino rats were randomly divided into 4 groups: Control (C), Only Expansion (OE) (no supplement but undergoing expansion and retention), KE (supplemented during both the expansion and retention phases), Krill Oil Nursery Group (KN) (supplemented during the 40-day nursery phase as well as during the expansion and retention phases). A 5-day RME was followed by a 12-day retention period. All rats were euthanized simultaneously. Micro-computerized tomography (Micro-CT), hemotoxylen-eosin (H&E) staining, and immunohistochemical analysis were conducted. Kruskal-Wallis and Dunn tests with Bonferonni corrrection were applied (p < 0.05)., Results: Expansion and KO supplementation did not cause a statistically significant change in bone mineral density (BMD), bone volume fraction (BV/TV), spesific bone surface (BS/BV) and trabecular thickness (Tb.Th). While the expansion prosedure increased the trabecular seperation (Tb.Sp), KO supplemantation mitigated this effect. The KE group exhibited a statistically significantly increase in trabecular number (Tb.N) compared to the OE group. Although receptor activator of nuclear factor-kappa-Β ligand (RANKL)/osteoprotegerin (OPG) ratios did not show significant differences between groups, the KE and OE groups demonstrated the lowest and highest value, respectively. KE showed a reduced amount of tartrate-resistant acid phosphatase (TRAP) compared to the OE., Conclusion: KO positively affected the architecture of the new bone formed in the mid-palatal suture. In this rat model of RME, results support the idea that administering of KO during the expansion period or beginning before the RME procedure may reduce relapse and enhance bone formation within the mid-palatal suture., (© 2024. The Author(s).)

Among all of the materials used in tissue engineering in order to develop bioequivalents, collagen shows to be the most promising due to its superb biocompatibility and biodegradability, thus becoming one of the most widely used materials for scaffold production. However, current imaging techniques of the cells within collagen scaffolds have several limitations, which lead to an urgent need for novel methods of visualization. In this work, we have obtained groups of collagen scaffolds and selected the contrasting agents in order to study pores and patterns of cell growth in a non-disruptive manner via X-ray computed microtomography (micro-CT). After the comparison of multiple contrast agents, a 3% aqueous phosphotungstic acid solution in distilled water was identified as the most effective amongst the media, requiring 24 h of incubation. The differences in intensity values between collagen fibers, pores, and masses of cells allow for the accurate segmentation needed for further analysis. Moreover, the presented protocol allows visualization of porous collagen scaffolds under aqueous conditions, which is crucial for the multimodal study of the native structure of samples.

This study evaluated the effect of ultrasonic agitation on the filling capacity of ready-to-use calcium silicate-based sealer Bio-C Sealer (BCS, Angelus, Paraná, Brazil) or powder-liquid BioRoot RCS (BR, Septodont, Saint-Maur-des-Fossés, France) using curved artificial canals by micro-computed tomography (micro-CT). Additionally, flow (mm) and flow area (mm2) were evaluated for both materials. Acrylic resin main canal (60° curvature and 5 mm radius, with 3 lateral canals in the cervical, middle, and apical thirds) were prepared up to size 40/.05 (Prodesign Logic, Brazil). The agitation method was used with ultrasonic tip (US, Irrisonic, Helse, Brazil): BCS, BCS/US, BR, and BR/US. All specimens were filled using the single-cone technique. The samples were scanned by micro-CT (8,74 µm) after obturation. The percentage of filling material and voids were calculated. Flow was evaluated based on ISO 6876/2012 standards (mm) and area (mm2). The data were statistically analyzed using ANOVA and Tukey tests (α = 0.05). BR/US showed lower percentage of filling material in the lateral canals than and, BCS/US (p<0.05). BR/US resulted in a higher percentage of voids than BR in the lateral apical third (p<0.05). BCS showed higher flow than BR (p<0.05). BCS and BR presented proper filling capacity in the simulated curved canals regardless of the use of ultrasonic agitation. However, BR/US showed more voids in the apical third. BCS demonstrates higher filling ability.

Background: This study evaluated the osseointegration potential of functionalised high-performance oxide ceramics (HPOC) in isolation or coated with BMP-2 or RGD peptides in 36 New Zeeland female rabbits using micro-computed tomography (micro CT). The primary outcomes of interest were to assess the amount of ossification evaluating the improvement in the bone volume/ total volume (BV/TV) ratio and trabecular thickness at 6 and 12 weeks. The second outcome of interest was to investigate possible differences in osteointegration between the functionalised silanised HPOC in isolation or coated with Bone Morphogenetic Protein 2 (BMP-2) or RGD peptides., Methods: 36 adult female New Zealand white rabbits with a minimum weight of three kg were used. One-third of HPOCs were functionalised with silicon suboxide (SiOx), a third with BMP-2 (sHPOC-BMP2), and another third with RGD (sHPOC-RGD). All samples were scanned with a high-resolution micro CT (U-CTHR, MILabs B.V., Houten, The Netherlands) with a reconstructed voxel resolution of 10 µm. MicroCT scans were reconstructed in three planes and processed using Imalytics Preclinical version 2.1 (Gremse-IT GmbH, Aachen, Germany) software. The total volume (TV), bone volume (BV) and ratio BV/TV were calculated within the coating area., Results: BV/TV increased significantly from 6 to 12 weeks in all HPOCs: silanised (P = 0.01), BMP-2 (P < 0.0001), and RGD (P < 0.0001) groups. At 12 weeks, the BMP-2 groups demonstrated greater ossification in the RGD (P < 0.0001) and silanised (P = 0.008) groups. Trabecular thickness increased significantly from 6 to 12 weeks (P < 0.0001). At 12 weeks, BMP-2 promoted greater trabecular thickness compared to the silanised group (P = 0.07), although no difference was found with the RGD (P = 0.1) group., Conclusion: Sinalised HPOC in isolation or functionalised with BMP-2 or RGD promotes in vivo osteointegration. The sinalised HOPC functionalised with BMP-2 demonstrated the greatest osseointegration., (© 2024. The Author(s).)

Background: This study aimed to assess the quality of various obturation techniques to fill perforation caused by internal root resorption using Micro-computed Tomography., Methods: Cone-beam computed tomography images of a maxillary central incisor tooth with perforating internal resorptive defect were used to create a 3D printed model of the affected tooth. The replicas were divided into four groups based on the obturation technique used. The techniques included Group 1: a polydimethylsiloxane-based sealer (GuttaFlow-2) with gutta-percha. Group 2: same as Group 1 except for using a pre-mixed Bioceramic-based sealer (NeoSEALER Flo). Group 3: the defect was filled entirely using the NeoSealer Flo Bioceramic-based sealer. Group 4: the samples were obturated using the warm vertical compaction technique with a resin-based sealer (ADSeal). The resin models were then scanned a micro-computed scanner to evaluate the percentage of voids in each group., Results: The results showed that NeoSEALER Flo groups had significantly the highest volume of voids while GuttaFlow-2 and warm vertical compaction groups had the lowest void volume., Conclusions: GuttaFlow-2 and warm vertical compaction techniques performed best in filling the internal resorptive defect., (© 2024. The Author(s).)

Aim: The aim of this study was to evaluate the removal efficiency of PRMTA and ECMPremixed applied to the coronal third according to the RET by UI and to examine the effect of different solutions on material removal., Materials and Methods: 40 permanent upper central teeth were used to simulate immature teeth. The samples were irrigated with 1.5% NaOCl and calcium hydroxide was placed. Samples were incubated in PBS. Then irrigation was done with 17% EDTA, the samples were randomly divided into 2 groups (n = 20):Group 1: PRMTA, Group 2: ECM Premixed. The materials were placed in the samples. Then the samples were scanned with micro-CT. Materials were removed by UI. Micro-CT scan of the samples was performed. Each material group was divided into 2 subgroups (n = 10): Group1 was MTAD, group2 was irrigated with 10% CA; then micro-CT was performed. Obtained images were positioned in DataViewer and analyzed with CTAn. The obtained data were statistically analyzed in IBM SPSS 25. The significance level was determined as 5%., Results: There was no significant difference between the initial volumes of the materials (p > 0.05). The amount of the remaining material after UI was significantly higher in the PRMTA (0.7471%) group compared to the ECM Premixed (0.0093%). There was no significant difference in terms of remaining material after irrigation with MTAD and CA in both groups (p > 0.05)., Conclusion: A great deal of the materials were removed by UI under the operation microscope. ECM Premixed was removed more effectively compared to the PRMTA. And, acidic solutions did not provide any additional benefit in material removal., (© 2024. The Author(s).)

Background: Development of reliable disease activity biomarkers is critical for diagnostics, prognostics, and novel drug development. Although computed tomography (CT) is the gold-standard for quantification of bone erosions, there are no consensus approaches or rationales for utilization of specific outcome measures of erosive arthritis in complex joints. In the case of preclinical models, such as sexually dimorphic tumor necrosis factor transgenic (TNF-Tg) mice, disease severity is routinely quantified in the ankle through manual segmentation of the talus or small regions of adjacent bones primarily due to the ease in measurement. Herein, we sought to determine the particular hindpaw bones that represent reliable biomarkers of sex-dependent disease progression to guide future investigation and analysis., Methods: Hindpaw micro-CT was performed on wild-type (n = 4 male, n = 4 female) and TNF-Tg (n = 4 male, n = 7 female) mice at monthly intervals from 2-5 (females) and 2-8-months (males) of age, since female TNF-Tg mice exhibit early mortality from cardiopulmonary disease at approximately 5-6-months. Further, 8-month-old WT (n = 4) and TNF-Tg males treated with anti-TNF monoclonal antibodies (n = 5) or IgG placebo isotype controls (n = 6) for 6-weeks were imaged with micro-CT every 3-weeks. For image analysis, we utilized our recently developed high-throughput and semi-automated segmentation strategy in Amira software. Synovial and osteoclast histology of ankle joints was quantified using Visiopharm., Results: First, we demonstrated that the accuracy of automated segmentation, determined through analysis of ~9000 individual bones by a single user, was comparable in wild-type and TNF-Tg hindpaws before correction (79.2±8.9% vs 80.1±5.1%, p = 0.52). Compared to other bone compartments, the tarsal region demonstrated a sudden, specific, and significant bone volume reduction in female TNF-Tg mice, but not in males, by 5-months (4-months 4.3± 0.22 vs 5-months 3.4± 0.62 mm3, p<0.05). Specifically, the cuboid showed significantly reduced bone volumes at early timepoints compared to other tarsals (i.e., 4-months: Cuboid -24.1±7.2% vs Talus -9.0±5.9% of 2-month baseline). Additional bones localized to the anterolateral region of the ankle also exhibited dramatic erosions in the tarsal region of females, coinciding with increased synovitis and osteoclasts. In TNF-Tg male mice with severe arthritis, the talus and calcaneus exhibited the most sensitive response to anti-TNF therapy measured by effect size of bone volume change over treatment period., Conclusions: We demonstrated that sexually dimorphic changes in arthritic hindpaws of TNF-Tg mice are bone-specific, where the cuboid serves as a reliable early biomarker of erosive arthritis in female mice. Adoption of automated segmentation approaches in pre-clinical or clinical models has potential to translate quantitative biomarkers to monitor bone erosions in disease and evaluate therapeutic efficacy., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Kenney et al. This 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.)

This study aimed to characterize the anatomical and physiological features of pits and fissures in primary and permanent molars by microtomographic (micro-CT) examination and three-dimensional (3D) printing. The occlusal surfaces of 84 primary molars and 60 permanent third molars were examined. The samples were scanned with micro-CT and the occlusal surface separated. The areas of the crown, its occlusal part, and fissures and pits were calculated. Digital impression of the occlusal surface was created and 3D printed. The frequency of different fissure types was determined by direct observation. Data were subjected to statistical analysis using Mann-Whitney U Test and chi-square test ( p < 0.05). There was statistically significant difference between the ratio of occlusal surface and the crown area for the molars in primary and permanent dentitions (24.78% and 28.85% respectively, p < 0.05). In terms of the percentage ratio of the fissure area to the occlusal surface (24.24% and 22.30%) and the fissure area to the crown (6.02% and 6.52%), no significant difference was observed ( p > 0.05). V-shaped fissures were predominant in both primary and permanent teeth, with a higher occurrence in primary dentition (59.48%, p < 0.05). Permanent molars exhibited a higher prevalence of I-type and U-type fissure configurations compared to primary molars ( p < 0.05), with I-type fissures being the least common in primary molars. In both dentitions there was no statistically significant difference in the prevalence of IK-configuration ( p > 0.05). The fissure depth was significantly greater in permanent molars than primary molars ( p < 0.05). In conclusion, this study revealed remarkable diversity in fissure morphology among primary and permanent molars., Competing Interests: The authors declare no conflict of interest., (©2024 The Author(s). Published by MRE Press.)

Objective: Evaluate the impact of adjustment procedures - cut-out-rescan (COR) and data exchange by over-scanning (DEOS) techniques - through CAD/CAM software on the marginal fit outcome of ceramic crowns., Methods and Materials: Twenty-eight de-identified teeth were adapted in a mandibular typodont set. Tooth #19 was prepared for a lithium disilicate crown and seven groups, G0 to G6 (n=10), were created based on the rescanned areas (mesial and/or buccal) on the typodont using an intraoral scanner through COR or DEOS techniques. A digital workflow was used to design and mill 70 crowns according to the groups. Each crown was temporarily cemented on tooth #19 and scanned with micro-computed tomography to measure the marginal fit. The data were analyzed statistically by the Kruskal-Wallis test followed by the Mann-Whitney test to compare the groups pairwise as a post-hoc (α=0.05)., Results: Statistically significant differences were found for vertical misfit (μm) between the groups for Marginal Gap Buccal (MGB) and Marginal Gap Mesial (MGM) (p=0.003 and p=0.029, respectively). No significant difference was found for Finish Line Buccal (FLB) and Finish Line Mesial (FLM) (p=0.062 and p=0.092, respectively). G3 (COR buccal and mesial) had the highest MGB (57.75 μm), statistically different from all other groups. G4 (DEOS buccal) (41.60 μm) was different from G6 (DEOS buccal and mesial) (44.21 μm) (p=0.023). For MGM, G0 (control) (53.96 μm) was different from G5 (DEOS mesial) (45.76 μm) and G6 (DEOS buccal and mesial) (48.56 vm) (p=0.013 and p=0.041, respectively) and G2 (COR mesial) (58.43 μm) was different from G5 (DEOS mesial) (45.76 μm) (p=0.016)., Conclusions: Despite a statistically significant difference in certain groups for both techniques, COR and DEOS techniques are viable options for image editing during acquisition. Lithium disilicate crowns can be produced with satisfactory marginal gap values utilizing a chairside CAD/CAM system., (©Operative Dentistry, 2024.)

Histological analysis is the core of follicular thyroid carcinoma (FTC) classification. The histopathological criteria of capsular and vascular invasion define malignancy and aggressiveness of FTC. Analysis of multiple sections is cumbersome and as only a minute tissue fraction is analyzed during histopathology, under-sampling remains a problem. Application of an efficient tool for complete tissue imaging in 3D would speed-up diagnosis and increase accuracy. We show that X-ray propagation-based imaging (XPBI) of paraffin-embedded tissue blocks is a valuable complementary method for follicular thyroid carcinoma diagnosis and assessment. It enables a fast, non-destructive and accurate 3D virtual histology of the FTC resection specimen. We demonstrate that XPBI virtual slices can reliably evaluate capsular invasions. Then we discuss the accessible morphological information from XPBI and their significance for vascular invasion diagnosis. We show 3D morphological information that allow to discern vascular invasions. The results are validated by comparing XPBI images with clinically accepted histology slides revised by and under supervision of two experienced endocrine pathologists.

Objectives: This study aimed to measure the distribution of silver ion (Ag + ), mineral recovery, and nanohardness in carious lesions and comprehensively evaluate the degree of dentin restoration., Methods: Sixty human teeth with root caries were randomly assigned to the control, silver diammine fluoride (SDF) [Safo], and SDF+Glass ionomer cement (GIC) treatment [Safo+Fuji] groups. Micro-computed tomography (micro-CT) was performed at five time points for each sample before/after treatment to evaluate mineral density within and around carious lesions. Three months following treatment, 12 samples were selected for synchrotron radiation X-ray fluorescence analysis to evaluate Ag + distribution, while 15 samples were selected for nanoindentation. Data were analyzed using Dunnett's T3 test for micro-CT and Wilcoxon rank sum test with Bonferroni correction (p = 0.017) for nanoindentation. The correlation between hardness and mineral change was analyzed using the Spearman rank correlation coefficient., Results: The Safo and Safo+Fuji groups showed significantly higher mineral recovery rates than did the control group (p < 0.001). In the Safo group, Ag + accumulated in the deeper layers rather than the superficial layer of caries. In the Safo+Fuji group, Ag + was found evenly distributed throughout caries, with only a few Ag + detected in the GIC layer. Hardness in the Safo+Fuji group was significantly higher compared with the Safo group at depths in the range of 10-50 µm., Conclusion: In the presence of GICs, SDF exhibited high remineralization capacity when diffusing throughout carious lesions over time. Combined treatment with SDF and GIC could strengthen root dentin even in the presence of caries., Clinical Significance: We found that combination treatment with SDF and GIC could increase mineral density in caries and improve the hardness of the tooth structure compared with fluoride-based agents alone. These findings might pave the way for future clinical trials to determine the therapeutic potential of nanotechnology-based restorative materials., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Objectives: This study focuses on artificial intelligence (AI)-assisted analysis of alveolar bone for periodontitis in a mouse model with the aim to create an automatic deep-learning segmentation model that enables researchers to easily examine alveolar bone from micro-computed tomography (µCT) data without needing prior machine learning knowledge., Methods: Ligature-induced experimental periodontitis was produced by placing a small-diameter silk sling ligature around the left maxillary second molar. At 4, 7, 9, or 14 days, the maxillary bone was harvested and processed with a µCT scanner (µCT-45, Scanco). Using Dragonfly (v2021.3), we developed a 3D deep learning model based on the U-Net AI deep learning engine for segmenting materials in complex images to measure alveolar bone volume (BV) and bone mineral density (BMD) while excluding the teeth from the measurements., Results: This model generates 3D segmentation output for a selected region of interest with over 98 % accuracy on different formats of µCT data. BV on the ligature side gradually decreased from 0.87 mm 3 to 0.50 mm 3 on day 9 and then increased to 0.63 mm 3 on day 14. The ligature side lost 4.6 % of BMD on day 4, 9.6 % on day 7, 17.7 % on day 9, and 21.1 % on day 14., Conclusions: This study developed an AI model that can be downloaded and easily applied, allowing researchers to assess metrics including BV, BMD, and trabecular bone thickness, while excluding teeth from the measurements of mouse alveolar bone., Clinical Significance: This work offers an innovative, user-friendly automatic segmentation model that is fast, accurate, and reliable, demonstrating new potential uses of artificial intelligence (AI) in dentistry with great potential in diagnosing, treating, and prognosis of oral diseases., Competing Interests: Declaration of competing interest The authors have no competing interests to declare., (Copyright © 2024. Published by Elsevier Ltd.)

Bone microdamage is common at subchondral bone (SCB) sites subjected to repeated high rate and magnitude of loading in the limbs of athletic animals and humans. Microdamage can affect the biomechanical behaviour of bone under physiological loading conditions. To understand the effects of microdamage on the mechanical properties of SCB, it is important to be able to quantify it. The extent of SCB microdamage had been previously estimated qualitatively using plain microcomputed tomography (μCT) and a radiocontrast quantification method has been used for trabecular bone but this method may not be directly applicable to SCB due to differences in bone structure. In the current study, SCB microdamage detection using lead uranyl acetate (LUA) and quantification by contrast-enhanced μCT and backscattered scanning electron microscopy (SEM) imaging techniques were assessed to determine the specificity of the labels to microdamage and the accuracy of damaged bone volume metrices. SCB specimens from the metacarpus of racehorses, with the hyaline articular cartilage (HAC) removed, were grouped into two with one group subjected to ex vivo uniaxial compression loading to create experimental bone damage. The other group was not loaded to preserve the pre-existing in vivo propagated bone microdamage. A subset of each group was stained with LUA using an established or a modified protocol to determine label penetration into SCB. The μCT and SEM images of stained specimens showed that penetration of LUA into the SCB was better using the modified protocol, and this protocol was repeated in SCB specimens with intact hyaline articular cartilage. The percentage of total label localised to bone microdamage was determined on SEM images, and the estimated labelled bone volume determined by μCT in SCB groups was compared. Label was present around diffuse and linear microdamage as well as oblique linear microcracks present at the articular surface, except in microcracks with high-density mineral infills. Bone surfaces lining pores with recent mineralisation were also labelled. Labelled bone volume fraction (LV/BV) estimated by μCT was higher in the absence of HAC. At least 50% of total labels were localised to bone microdamage when the bone area fraction (B.Ar/T.Ar) of the SCB was greater than 0.85 but less than 30% when B.Ar/T.Ar of the SCB was less than 0.85. To adjust for LUA labels on bone surfaces, a measure of the LV/BV corrected for bone surface area (LV/BV BS -1 ) was used to quantify damaged SCB. In conclusion, removal of HAC and using a modified labelling protocol effectively stained damaged SCB of the metacarpus of racehorses and represents a technique useful for quantifying microdamage in SCB. This method can facilitate future investigations of the effects of microdamage on joint physiology., (© 2024 The Authors. Journal of Anatomy published by John Wiley & Sons Ltd on behalf of Anatomical Society.)

Computed tomography (CT) and microcomputed tomography (μCT) require calibration against density phantoms scanned with specimens or during routine internal calibration for assessment of mineral concentration (MC) and density. In clinical studies involving bone, alternative calibration methods using bodily tissues and fluids ("phantomless" calibration) have been suggested. However, such tissues are seldom available in archeological and osteological research. This study investigates the potential of dental tissue as internal reference for calibration of μCT scans, facilitating the analysis of bone MC. We analyzed 70 molars from 24 extant primate species, including eight human teeth, each scanned with density phantoms for calibration. Our findings indicate that sampling specific regions of molars (lateral aspects of the mesial cusps) yields low variation in enamel and dentine MC values, averaging 1.27 g/cm 3 (±0.03) for dentine and 2.25 g/cm 3 (±0.03) for enamel. No significant differences were observed across molar types or among scanning procedures, including scanner model, resolution, and filters. An ad hoc test on 12 mandibles revealed low variance in MC between the conventional phantom and dental tissue calibration methods; all 36 measurements (low, medium, and high MC for each mandible) were within 0.05 g/cm 3 of each other -81% were within 0.03 g/cm 3 and 94% within 0.04 g/cm 3 . Based on these results, we propose a new "phantomless" calibration technique using these mean enamel and dentine MC values. The presented phantomless calibration method could aid in the assessment of bone pathology and enhance the scope of studies investigating bone structure and physical property variations in archeological, osteological, and laboratory-based research., (© 2024 The Authors. American Journal of Biological Anthropology published by Wiley Periodicals LLC.)

The vertebral column, a defining trait of all vertebrates, is organized as a concatenated chain of vertebrae, and therefore its support to the body depends on individual vertebral morphology. Consequently, studying the morphology of the vertebral centrum is of anatomical and clinical importance. Grass carp (GC) is a member of the infraclass Teleostei (teleost fish), which accounts for the majority of all vertebrate species; thus, its vertebral anatomical structure can help us understand vertebrate development and vertebral morphology. In this study, we have investigated the morphology and symmetry of the grass carp vertebral centrum using high-resolution micro-CT scans. To this end, three abdominal vertebrae (V9, V10, & V11) from eight grass carp were micro-CT scanned and then segmented using Dragonfly (ORS Inc.). Grass carp vertebral centrum conformed to the basic teleost pattern and demonstrated an amphicoelous shape (biconcave hourglass). The centrum's cranial endplate was smaller, less circular, and shallower compared to the caudal endplate. While the vertebral centrum demonstrated bilateral symmetry along the sagittal plane (left/right), the centrum focus was shifted dorsally and cranially, breaking dorsoventral and craniocaudal symmetry. The sum of these findings implies that the caudal aspect of grass carp vertebral centrum is bigger and more robust. Currently, we have no information whether this is due to nature, for example, differences in gene expression, or nurture, for example, environmental effect. As the vertebral parapophyses and spinous processes are slanted caudally, the direction of muscle action during swimming may create a gradient of stresses from cranial to caudal, resulting in a more robust caudal aspect of the vertebral centrum. Expanding our study to include additional quadrupedal and bipedal (i.e., human) vertebrae, as well as testing if these morphological aspects of the vertebrae are indeed plastic and can be affected by environmental factors (i.e., temperature or other stressors) may help answer this question., (© 2024 Anatomical Society.)

Subchondral bone remodeling, mediated by osteocytes within the lacuno-canalicular network, plays a crucial role in osteoarthritis (OA) progression. Following cell death, lacunae preserve integrity, offering insights into bone remodeling mechanisms. Limited and controversial data on osteocyte lacuna morphology in OA result from small sample sizes and two-dimensional (2D) techniques that have been used thus far. This study aimed to quantify three-dimensional (3D) osteocyte lacunar characteristics at well-defined tibial plateau locations, known to be differently affected by OA. Specifically, 11 tibial plateaus were obtained from end-stage knee-OA patients with varus deformity. Each plateau provided one sample from the less affected lateral compartment and two samples from the medial compartment, at minimum and maximum bone volume fraction (BV/TV) locations. High-resolution desktop micro-computed tomography (micro-CT) at 0.7 μm voxel resolution imaged the 33 samples. Lacuna number density (Lc.N/BV) and lacuna volume density (Lc.TV/BV) were significantly lower (p < 0.02) in samples from the medial side with maximum BV/TV compared to lateral side samples. In the medial compartment at maximum local BV/TV, mean lacuna volume (Lc.V), total lacuna volume (Lc.TV), and Lc.TV/BV were significantly (p < 0.001) lower than in the region with minimum BV/TV. Lc.N/BV was also significantly lower (p < 0.02) at the maximum local BV/TV location compared to the region with minimum BV/TV. Our findings suggest that subchondral bone lacunae adapt to the changing loads in end-stage OA., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Plants are able to naturally graft or inosculate their trunks, branches and roots together, this mechanism is used by humans to graft together different genotypes for a range of purposes. Grafts are considered successful if functional vascular connections between the two genotypes occur. Various techniques can evaluate xylem connections across the graft interface. However, these methods are generally unable to assess the heterogeneity and three-dimensional (3D) structure of xylem vessel connections. Here we present the use of X-ray micro-computed tomography to characterize the 3D morphology of grafts of grapevine. We show that xylem vessels form between the two plants of natural root and human-made stem grafts. The main novelty of this methodology is that we were able to visualize the 3D network of functional xylem vessels connecting the scion and rootstock in human-made stem grafts thanks to the addition of a contrast agent to the roots and improved image analysis pipelines. In addition, we reveal the presence of extensive diagonal xylem connections between the main axial xylem vessels in 2-year old grapevine stems. In conclusion, we present a method that has the potential to provide new insights into the structure and function of xylem vessels in large tissue samples., (© 2024 The Authors. Plant, Cell & Environment published by John Wiley & Sons Ltd.)

Objectives: Temporary anchorage devices (TADs) have become an integral part of comprehensive orthodontic treatments. This study evaluated the transfer accuracy of three-dimensional (3D) printed and computer-aided design/computer-aided manufacturing (CAD/CAM) milled surgical guides for orthodontic TADs using micro-computed tomography (CT) imaging in a preclinical trial., Methods: Overall, 30 surgical guides were used to place TADs into typodonts; 3D printing and CAD/CAM milling were used to produce the guides. The virtual target positions of the TADs were compared to the real positions in terms of spatial and angular deviations using digital superimposition. Micro-CT imaging was used to detect the positions. To evaluate reliability, two investigators collected the measurements twice. Intra-rater and inter-rater correlations were tested., Results: In total, 60 palatal TADs were evaluated. The mean coronal deviations in the print group ranged from 0.15 ± 0.20 mm to 0.71 ± 0.22 mm, whereas in the mill group, they ranged from 0.09 ± 0.15 mm to 0.83 ± 0.23 mm. At the apical tip, the overall deviations in the print group ranged from 0.14 ± 0.56 mm to 1.27 ± 0.66 mm, whereas in the mill group, they ranged from 0.15 ± 0.57 mm to 1.09 ± 0.44 mm. The mean intra-class and inter-class correlation coefficients ranged from 0.904 to 0.987. No statistically significant differences were found between the groups., Conclusions: CAD/CAM milled guides yielded spatial and angular accuracies comparable to those of 3D printed guides with notable deviations in the vertical positioning of TADs., Clinical Significance: Digital planning of orthodontic temporary implants combines clinical predictability and the safety of surrounding tissue. Therefore, the transfer accuracy of the guides is crucial. This preclinical study was the first to evaluate CAD/CAM milling for orthodontic guides and found its accuracy comparable to that of the current "gold standard"., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

This study aims to compare the shaping ability of three systems using micro-computed tomography (micro-CT). Moderately-curved mesial canals of 36 mandibular molars were assigned to three groups (n = 24); Protaper Next (PTN, 0.25, 0.06), WaveOne Gold (WOG, 0.25, 0.07), TruNatomy (TRN, 0.25, 0.04), and instrumentation was performed. Pre- and post-micro-CT scans were obtained. Canal volume and surface, structure model index (SMI), centroid shift, canal transportation, and untouched canal were analyzed. One-way ANOVA and Student's t-test were used for statistics. There was no difference in SMI, centroid shift, and centering ability between the study groups (p > .05). Removed dentin and canal surface changes were lower in TRN, while untouched dentin walls were higher (p < .05). Cross-sections became more rounded (p < .05), but not significant between groups (p > .05). Considering the removed dentin by TRN, it can be used in critical dentin thickness, such as the danger zone (DZ). PTN, WOG, and WOG kept the original canal course similarly. Untouched dentin by TRN (41%) was wider than PTN and WOG, consequently, meticulous irrigation is recommended. TRN, which provides a controlled increase in canal volume, can be used in thin dentin such as the DZ, however, its use should be supported by copious irrigation and brushing considering the rate of untouched dentin walls. RESEARCH HIGHLIGHTS: TRN presented a higher untouched dentin wall compared to PTN and WOG. Canal volume and surface changes were the lowest in the TRN group. The centering abilities were similar in PTN, WOG, and TRN., (© 2024 The Author(s). Microscopy Research and Technique published by Wiley Periodicals LLC.)