Micro-computed Tomography Imaging Research Articles

Phenotypic convergence is more frequent and stronger in herbivorous freshwater fishes.

Constraints on phenotypic evolution can lead to patterns of convergent evolution, by limiting the 'pool' of potential phenotypes in the face of endogenous (functional, developmental) or exogenous (competition, predation) selective pressures. Evaluation of convergence depends on integrating ecological and morphological data within a robust, comparative phylogenetic context. The staggering diversity of teleost fishes offers a multitude of lineages adapted for similar ecological roles, and therefore, offers numerous replicated evolutionary experiments for exploring phenotypic convergence. However, our understanding of fish feeding systems has been primarily shaped by marine species, with the monolithic exception of freshwater cichlids. Here we use piranhas and pacus (Serrasalmidae) to explore the evolution of different feeding ecologies and their morphological proxies in Neotropical freshwater environments. Specifically, we explore whether convergence is more widespread among plant-eating fishes, arising from strong constraints on phenotypic evolution in herbivores. Using osteological micro-computed tomographic imaging (μCT), we describe the major axes of morphological variation in pacus and piranhas, regarding their diet and feeding behaviors. Next, we evaluated whether herbivorous niches are less labile than other dietary guilds and whether herbivorous species' phenotypes evolve at a slower evolutionary rate than other taxa. We then assess how convergent herbivorous taxa are, using three different suites of morphological characters (dental, jaw, and abdominal morphometrics). Ecologically, herbivory is not a dead end, exhibiting similar observed transition rates as those between carnivores and omnivores. However, we documented widespread convergence in herbivores and that herbivores have slower rates of phenotypic evolution than carnivores. Most instances of convergence are found in herbivorous taxa, specifically in frugivores and folivores. Moreover, instances of 'complete' convergence, indicated by positive convergence metrics observed in more than one morphometric dataset, were only found in herbivores. Herbivores do appear to evolve under constrained circumstances, but this has not limited their ecological lability.

An efficient finite element mesh generation methodology based on μCT images of multi-layer woven composites

High-fidelity models are essential for accurate finite element (FE) simulations of composite material behavior. This paper proposes an efficient meshing methodology based on micro-Computed Tomography (μCT) images. U-Net convolutional neural network was used for image segmentation. Connected yarns were then separated using an improved procedure based on watershed algorithm and geometric transformations. The proposed Constrained Delaunay-Advancing Front Technique (CD-AFT) surface reconstruction algorithm extracts point cloud of yarns from segmented images and outputs high-quality and smooth orientable manifold watertight triangulated surface. Intersecting meshes of yarns are separated through node position detection and Laplacian moving. Experimental results show that proposed methodology is capable of accomplishing mesh generation for different mesh sizes. Compared with commercial software, it has obvious advantages in mesh quality and size control. Since the proposed method operates independently of commercial software and manual operation, it facilitates the automated generation of numerous high-fidelity models from μCT images for FE simulations.

Multiscale and multimodal imaging for three-dimensional vascular and histomorphological organ structure analysis of the pancreas

Exocrine and endocrine pancreas are interconnected anatomically and functionally, with vasculature facilitating bidirectional communication. Our understanding of this network remains limited, largely due to two-dimensional histology and missing combination with three-dimensional imaging. In this study, a multiscale 3D-imaging process was used to analyze a porcine pancreas. Clinical computed tomography, digital volume tomography, micro-computed tomography and Synchrotron-based propagation-based imaging were applied consecutively. Fields of view correlated inversely with attainable resolution from a whole organism level down to capillary structures with a voxel edge length of 2.0 µm. Segmented vascular networks from 3D-imaging data were correlated with tissue sections stained by immunohistochemistry and revealed highly vascularized regions to be intra-islet capillaries of islets of Langerhans. Generated 3D-datasets allowed for three-dimensional qualitative and quantitative organ and vessel structure analysis. Beyond this study, the method shows potential for application across a wide range of patho-morphology analyses and might possibly provide microstructural blueprints for biotissue engineering.

A reliable deep-learning-based method for alveolar bone quantification using a murine model of periodontitis and micro-computed tomography imaging

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 mm3 to 0.50 mm3 on day 9 and then increased to 0.63 mm3 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.

Transfer accuracy of 3D printed versus CAD/CAM milled surgical guides for temporary orthodontic implants: A preclinical micro CT study

ObjectivesTemporary 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. MethodsOverall, 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. ResultsIn 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. ConclusionsCAD/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 significanceDigital 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".

Surface Wetting Characterization in Pore-scale Multiphase Flow Simulations: A Ketton Carbonate Case Study

Direct numerical simulations are commonly applied to X-ray computed microtomography images of porous rocks to measure petrophysical properties, such as relative permeability. To complement and/or expand laboratory data with useful simulations, such simulations need to reflect the true wetting condition of reservoir rock. It is therefore important to investigate approaches for assigning surface wetting conditions in pore-scale multiphase flow simulations. Two-phase Lattice-Boltzmann simulations are conducted on a Ketton carbonate image using different effective wetting models and then compared to pore-scale experimental data on the same rock, which is published elsewhere in (Scanziani et al., 2018). Two different wetting models are established based on the aging of the Ketton sample with crude oil at connate water saturation. Model 1 provides uniform wetting on all oil treated surfaces. Model 2 attempts to capture the multiscale nature of carbonate by allowing for spatially varying wetting conditions on oil treated surfaces based on local microporosity. Assessment criteria are based on pore-scale morphological measures, which include oil surface area coverage, oil phase topology, fluid/fluid curvatures, and contact angles. Overall, Model 1 with an assigned contact angle of greater than +10° above the geometrically measured contact angle from the experimental data provided the best results based on our assessment criteria.

Polycaprolactone/β-Tricalcium Phosphate Composite Scaffolds with Advanced Pore Geometries Promote Human Mesenchymal Stromal Cells' Osteogenic Differentiation.

Critical-sized mandibular bone defects, arising from, for example, resections after tumor surgeries, are currently treated with autogenous bone grafts. This treatment is considered very invasive and is associated with limitations such as morbidity and graft resorption. Tissue engineering approaches propose to use 3D scaffolds that combine structural features, biomaterial properties, cells, and biomolecules to create biomimetic constructs. However, mimicking the complex anatomy and composition of the mandible poses a challenge in scaffold design. In our study, we evaluated the dual effect of complex pore geometry and material composition on the osteogenic potential of 3D printed scaffolds. The scaffolds were made of polycaprolactone (PCL) alone (TCP0), or with a high concentration of β-tricalcium phosphate (β-TCP) up to 40% w/w (TCP40), with two complex pore geometries, namely a star- (S) and a diamond-like (D) shape. Scanning electron microscopy and microcomputed tomography images confirmed high fidelity during the printing process. The D-scaffolds displayed higher compressive moduli than the corresponding S-scaffolds. TCP40 scaffolds in simulated body fluid showed deposition of minerals on the surface after 28 days. Subsequently, we assessed the differentiation of seeded bone marrow-derived human mesenchymal stromal cells (hMSCs) over 28 days. The early expression of RUNX2 in the cell nuclei confirmed the commitment toward an osteogenic phenotype. Moreover, alkaline phosphatase (ALP) activity and collagen deposition displayed an increasing trend in the D-scaffolds. Collagen type I was mainly present in the deposited extracellular matrix (ECM), confirming deposition of bone matrix. Finally, Alizarin Red staining showed successful mineralization on all the TCP40 samples, with higher values for the S-shaped scaffolds. Taken together, our study demonstrated that the complex pore architectures of scaffolds comprised TCP40 stimulated osteogenic differentiation and mineralization of hMSCs in vitro. Future research will aim to validate these findings in vivo.

Age and anatomical region-related differences in vascularization of the porcine meniscus using microcomputed tomography imaging.

Meniscal lesions in vascularized regions are known to regenerate while lack of vascular supply leads to poor healing. Here, we developed and validated a novel methodology for three-dimensional structural analysis of meniscal vascular structures with high-resolution microcomputed tomography (µCT). We collected porcine medial menisci from 10 neonatal (not-developed meniscus, n-) and 10 adults (fully developed meniscus, a-). The menisci were cut into anatomical regions (anterior horn (n-AH anda-AH), central body (n-CB anda-CB), and posterior horn (n-PH anda-PH). Specimens were cut in half, fixed, and one specimen underwent critical point drying and µCT imaging, while other specimen underwent immunohistochemistry and vascularity biomarker CD31 staining for validation of µCT. Parameters describing vascular structures were calculated from µCT. The vascular network in neonatal spread throughout meniscus, while in adult was limited to a few vessels in outer region, mostly on femoral side. n-AH, n-CB, and n-PH had 20, 17, and 11 times greater vascular volume fraction than adult, respectively. Moreover, thickness of blood vessels, in three regions, was six times higher in adults than in neonatal. a-PH appeared to have higher vascular fraction, longer and thicker blood vessels than both a-AH and a-CB. Overall, neonatal regions had a higher number of blood vessels, more branching, and higher tortuosity compared to adult regions. For the first time, critical point drying-based µCT imaging allowed detailed three-dimensional visualization and quantitative analysis of vascularized meniscal structures. We showed more vascularity in neonatal menisci, while adult menisci had fewer and thicker vascularity especially limited to the femoral surface.

Formational stages of natural fractures revealed by U-Pb dating and C-O-Sr-Nd isotopes of dolomites in the Ediacaran Dengying Formation, Sichuan Basin, southwest China

Deep core (>4.9 km) from Ediacaran Deng IV Member algal dolomites in the Gaoshiti-Moxi block in the Sichuan Basin, southwest China, reveals multiple generations of dolomite-lined and dolomite-filled opening-mode fractures. Three progressive stages of fracture formation are marked by crosscutting relations visible in the core, by acoustic emission experiments revealing evidence of past stress directions, and by fluid inclusions, U-Pb ages, C-O-Sr-Nd isotope patterns, and rare earth element data for dolomite cements in fractures, which document ages and differing thermal conditions and fluid compositions during fracture. In calcite-filled fractures, U-Pb ages and carbon and oxygen isotope signatures vary greatly, indicating that fractures developed with intensified tectonic activity marked by regional structures and with enhanced diagenetic alteration. In stage I, WNW-striking opening-mode fractures formed that contain dolomite deposits precipitated from basinal fluids between ca. 549 Ma and ca. 532 Ma. At this time, the Sichuan Basin experienced Xingkai taphrogenesis (rifting) from the late Neoproterozoic to early Cambrian. The central Sichuan paleo-uplift was undergoing ENE extension, and preexisting ESE- and nearly E-W−striking faults were oblique to the ENE principal stress orientation. This led to a local stress field favoring dextral shear near fault zones accommodated by the fractures. In stage II, ENE-striking fractures that are younger based on crosscutting relations contain dolomite deposits from basinal fluids with ages from ca. 423 Ma to ca. 411 Ma. Contemporaneous with Xuefeng thrusting, the central Sichuan paleo-uplift was in a NNE-striking transpressional stress field, which likely further generated ENE-striking fractures. In stage III, nearly N-S−striking fractures formed in the Gaoshiti-Moxi block. High-temperature fluids related to the Permian Emeishan large igneous province invaded these fractures from ca. 260 Ma to ca. 256 Ma. At this time, the Sichuan Basin was uplifted under the influence of the Emei taphrogenesis in the late Permian, and the central Sichuan paleo-uplift was subjected to E-W−striking extension. In fractures in these carbonate rocks, micro-computed tomography imaging reveals that macropores (>10 μm, 12.1%−21.8%) and small pores (2−10 μm, 76.6%−86.1%) dominate the dolomite mineral deposits, and that there are few (1.6%−1.8%) micropores or nanopores (<2 μm). Medium-sized throats (1−3 μm) are the main connecting channels. We infer that fractures served as conduits for fluid migration, leading to the dissolution of matrix pores adjacent to the fractures. This secondary porosity not only enhances reservoir storage capacity but also augments reservoir connectivity. Our study shows that in situ U-Pb dating and full-diameter rock acoustic emission data can effectively constrain the timing of fractures. By integrating this information with regional tectonic sequences and fracture diagenetic sequences from combined relative timing, geochemistry, and rock mechanics evidence, we clarify the factors controlling fracture formation.

Monoliths for gas storage manufactured with precision pore engineering using 3D-printed templates

A fast, customizable, and systematic route for producing activated carbon monoliths for gas storage with low interparticle porosity (9 % to 24 %) containing tailored macro channels is disclosed. The production uses a three-dimensional (3D)-printed polymeric lattice, rich in oxygen, as a sacrificial template. Its interstitial voids are filled with a phenolic resin that is carbonized and activated at 1173 K under carbon dioxide (CO2). Sequential pyrolysis occurs via thermal treatment with the 3D-printed lattice removed first, rendering macro channels with the same shape as the template in accurately placed positions. Resulting in adsorbents with interparticle porosities ranging from 10 % to 23 %, calculated from X-ray micro-computed tomography (micro-CT) images. The axial resolution of these macro channels was similar to the SLA printing layer thickness used of 100 µm. Making it the best resolution attained to date in fabricating monoliths via a 3D-printed sacrificial templating method. Breakthrough experiments indicated healthy macro channel connectivity due to the reduced mass transfer zone. Moreover, excellent mechanical integrity was displayed, with a monolith withstanding 900 N before excessive fracturing occurred despite the presence of macro channels. Therefore, this method emphasizes its use for gas storage applications by enabling the adsorbent material to efficiently utilize the volumetric space while allowing a fast rate of adsorption over the entire structure.

3D reconstruction and multi-omics analysis reveal a unique pattern of embryogenesis in Ginkgo biloba.

Ginkgo (Ginkgo biloba L.) is one of the earliest extant species in seed plant phylogeny. Embryo development patterns can provide fundamental evidence for the origin, evolution, and adaptation of seeds. However, the architectural and morphological dynamics during embryogenesis in Ginkgo biloba (G. biloba) remain elusive. Herein, we obtained over 2200 visual slices from three stages of embryo development using micro-computed tomography imaging with improved staining methods. Based on 3D spatio-temporal pattern analysis, we found that a shoot apical meristem with seven highly differentiated leaf primordia, including apical and axillary leaf buds, is present in mature Ginkgo embryos. 3D rendering from the front, top, and side views showed two separate transport systems of tracheids located in the hypocotyl and cotyledon, representing a unique pattern of embryogenesis. Furthermore, the morphological dynamic analysis of secretory cavities indicated their strong association with cotyledons during development. In addition, we identified genes GbLBD25a (lateral organ boundaries domain 25a), GbCESA2a (cellulose synthase 2a), GbMYB74c (myeloblastosis 74c), GbPIN2 (PIN-FORMED 2) associated with vascular development regulation, and GbWRKY1 (WRKYGOK 1), GbbHLH12a (basic helix-loop-helix 12a), GbJAZ4 (jasmonate zim-domain 4) potentially involved in the formation of secretory cavities. Moreover, we found that flavonoid accumulation in mature embryos could enhance post-germinative growth and seedling establishment in harsh environments. Our 3D spatial reconstruction technique combined with multi-omics analysis opens avenues for investigating developmental architecture and molecular mechanisms during embryogenesis and lays the foundation for evolutionary studies of embryo development and maturation.

Shengxian decoction improves lung function in rats with bleomycin-induced idiopathic pulmonary fibrosis through the inhibition of PANoptosis

Ethnopharmacological relevanceShengxian decoction (SXD) is a classic Chinese medicinal formula that can effectively improve clinical symptoms and quality of life and delay disease progression in idiopathic pulmonary fibrosis (IPF) patients; however, the underlying mechanisms remain unclear. Aim of the studyThis study aimed to observe PANoptosis in bleomycin-induced IPF and to assess the efficacy and mechanism of action of SXD in the treatment of IPF. Materials and methodsFifty SD rats were randomly divided into the sham, IPF, IPF + pirfenidone (PFD), IPF + SXD-medium dose (SXD-M), and IPF + SXD-low dose (SXD-L) groups. Lung function analysis and microcomputed tomography imaging of the rats with IPF treated with oral pirfenidone or oral SXD for 28 days were performed. Hematoxylin and eosin (HE) staining and Masson's trichrome staining were used to observe pathological lung damage. Enzyme-linked immunosorbent assays (ELISAs) were used to determine the serum levels of IL-1β, IL-18, TNF-α, and IFN-γ. Pyroptosis, apoptosis, and necroptosis were assessed using TUNEL, TUNEL/caspase-1, and PI fluorescence staining, respectively. GSDMD, caspase-3, and MLKL were examined by immunohistochemistry. The expression of fibrin-, ZBP1-, pyroptosis-, apoptosis-, and necroptosis-related proteins in the lung tissue was determined by western blotting. ResultsSXD normalized lung function in rats with bleomycin-induced IPF and reduced serum inflammatory factor levels and lung tissue fibrosis. The underlying mechanism of action involves the inhibition of pyroptosis pathway proteins, such as NLRP3, caspase-1, cleaved caspase-1, and GSDMD; apoptotic pathway proteins, such as Bax, Bcl-2, cleaved caspase-3, and caspase-3; and necroptosis pathway proteins, such as RIPK1, RIPK3, p-MLKL and MLKL. These pathways are modulated by the PANoptosis initiator ZBP1. Notably, the efficacy of SXD is concentration dependent, with a medium dose exhibiting superior effectiveness compared to a low dose. ConclusionBleomycin induced PANoptosis in the lung tissue of rats with IPF. Additionally, SXD effectively delayed or reversed the early pathological changes in bleomycin-induced pulmonary fibrosis by inhibiting PANoptosis.

A New Method for Creating Impact-Induced Intra-Articular Fractures in a Rabbit Model Induces Severe Post-Traumatic Osteoarthritis.

The objective of this work was to develop a model of intra-articular fracture (IAF) in a rabbit and document the speed and severity of degenerative joint changes after fracture fixation. With Institutional Animal Care & Use Committee approval, impact-induced IAFs were created in the distal tibia of 16 New Zealand White rabbits. Fractures were fixed with a plate and screws. Pain and function were monitored at regular postoperative intervals with limb loading analysis. Twelve or 26 weeks after fracture, animals were euthanized for histological assessment of cartilage degeneration and micro-computed tomography analysis of bone histomorphometry. Eleven animals successfully completed the study. Maximum foot force in the fractured limb was 41% ± 21% lower than preoperative values ( P = 0.006) 12 weeks after fracture and remained 25% ± 13% lower ( P = 0.081) after 26 weeks. Cortical bone mineral density in micro-computed tomography images was 34% ± 13% lower 12 weeks after fracture ( P < 0.001) and remained (42% ± 8%) lower 26 weeks after fracture ( P < 0.001). Twelve weeks after fracture, Mankin scores of cartilage degeneration were significantly higher in the medial talus ( P = 0.007), lateral talus ( P < 0.001), medial tibia ( P = 0.017), and lateral tibia ( P = 0.002) of the fractured limb compared with the uninjured contralateral limb. Average Mankin scores in the talus increased from 12 to 26 weeks (5.9 ± 0.9 to 9.4 ± 0.4; P < 0.001 lateral; 5.4 ± 1.8 to 7.8 ± 2.0; P = 0.043 medial), indicating substantial and progressive joint degeneration. The ankle joint of the New Zealand White rabbit provides the smallest available model of impact-induced IAF that can be treated with clinically relevant techniques and replicates key features of healing and degeneration found in human patients.

Establishment of a Mouse Degenerative Model of Patellar Tendinopathy with Upregulation of Inflammation.

There is no mouse model of patellar tendinopathy. This study aimed to establish a mouse inflammatory and degenerative patellar tendon injury model, which will facilitate research on patellar tendinopathy using advanced molecular tools including transgenic models. Collagenase at different doses (low dose (LD), medium dose (MD), high dose (HD)) or saline was injected over the mouse patellar tendon. At weeks 1, 2, 4, and 8 post-injection, the tendons were harvested for histology and further examined by micro-computed tomography (microCT) imaging at week 8. The optimal dose group and the saline group were further evaluated by immunohistochemical staining, gait pattern, and biomechanical properties. The histopathological score increased dose-dependently post-collagenase injection. Ectopic mineralization was observed and increased with collagenase dose. The LD group was selected for further analysis. The expression of IL-10, TNF-α, and MMP-1 significantly increased post-injection. The changes of limb idleness index (ΔLII) compared to preinjury state were significantly higher, while the ultimate load, stiffness, ultimate stress, and maximum Young's modulus were significantly lower in the LD group compared to the saline group. A mouse inflammatory degenerative model of patellar tendon injury resembling tendinopathy was established as indicated by the dose-dependent increase in tendon histopathology, ectopic calcification, decrease in biomechanical properties, and pain-associated gait changes.

Miao medicine Gu Yan Xiao tincture inhibits mTOR to stimulate chondrocyte autophagy in a rabbit model of osteoarthritis

Ethnopharmacological relevanceThe Gu Yan Xiao tincture, a blend of traditional Chinese herbs, is traditionally used for osteoarthritis and related pain. This study investigated its mechanism of action in order to rationalize and validate its therapeutic use. Aim of the studyThis study analyzed, in a rabbit model of knee osteoarthritis, whether and how Gu Yan Xiao tincture exerts therapeutic benefits by modulating chondrocyte autophagy. Materials and methodsThe active constituents within the GYX tincture were identified using liquid chromatography-mass spectrometry. The rabbit model was established by injecting animals with type II collagenase intra-articularly, and the effects of topically applied tincture were examined on osteoarthritis lesions of the knee using histopathology, micro-computed tomography and x-ray imaging. Effects of the tincture were also evaluated on levels of inflammatory cytokines, matrix metalloproteases, and autophagy in chondrocytes. As a positive control, animals were treated with sodium diclofenac. ResultsThe tincture mitigated the reduction in joint space, hyperplasia of the synovium and matrix metalloproteases in serum that occurred after injection of type II collagenase in rabbits. These therapeutic effects were associated with inhibition of mTOR and activation of autophagy in articular chondrocytes. Inhibiting mTOR with rapamycin potentiated the therapeutic effects of the tincture, while inhibiting autophagy with 3-methyladenine antagonized them. ConclusionsGu Yan Xiao tincture mitigates tissue injury in a rabbit model of osteoarthritis, at least in part by inhibiting mTOR and thereby promoting autophagy in chondrocytes. These results rationalize the use of the tincture not only against osteoarthritis but also potentially other diseases involving inhibition of autophagy in bones and joints.

Atherosclerotic Calcifications Have a Local Effect on the Peel Behavior of Human Aortic Media.

Aortic dissections, characterized by the propagation of a tear through the layers of the vessel wall, are critical, life-threatening events. Aortic calcifications are a common comorbidity in both acute and chronic dissections, yet their impact on dissection mechanics remains unclear. Using micro-computed tomography (CT) imaging, peel testing, and finite element modeling, this study examines the interplay between atherosclerotic calcifications and dissection mechanics. Samples cut from cadaveric human thoracic aortas were micro-CT imaged and subsequently peel-tested to map peel tension curves to the location of aortic calcifications. Empirical mode decomposition separated peel tension curves into high and low-frequency components, with high-frequency effects corresponding to interlamellar bonding mechanics and low-frequency effects to peel tension fluctuations. Finally, we used an idealized finite element model to examine how stiff calcifications affect aortic failure mechanics. Results showed that atherosclerosis influences dissection behavior on multiple length scales. Experimentally, atherosclerotic samples exhibited higher peel tensions and greater variance in the axial direction. The variation was driven by increased amplitudes of low-frequency tension fluctuations in diseased samples, indicating that more catastrophic propagations occur near calcifications. The simulations corroborated this finding, suggesting that the low-frequency changes resulted from the presence of a stiff calcification in the vessel wall. There were also modifications to the high-frequency peel mechanics, a response likely attributable to alterations in the microstructure and interlamellar bonding within the media. Considered collectively, these findings demonstrate that dissection mechanics are modified in aortic media nearby and adjacent to aortic calcifications.

BBR affects macrophage polarization via inhibition of NF-κB pathway to protect against T2DM-associated periodontitis.

Periodontitis is intimately associated with the development of various systemic diseases, among which type 2 diabetes mellitus (T2DM) has a bidirectional relationship with the pathogenesis of periodontitis. The objective of the present work was to investigate the role of berberine (BBR) in periodontitis with T2DM and related mechanisms. The mRNA expression of macrophage polarization-related factors in the microenvironment of periodontal inflammation was detected using real-time quantitative PCR (RT-qPCR). The experimental periodontitis model was constructed in wild-type (WT) and T2DM (db/db) mice, which were administered BBR after 7 days of modeling. Alveolar bone loss (ABL) in each group of mice was measured utilizing micro-computed tomography images. RT-qPCR was performed to analyze the levels of macrophage polarization-related factors in mouse gingiva. Lastly, using western blotting and RT-qPCR, the signaling pathway of BBR affecting macrophage polarization in the microenvironment of periodontitis was explored. BBR inhibited M1 polarization and stimulated M2 polarization in the periodontitis microenvironment. BBR decreased ABL in the WT and T2DM periodontitis models. And BBR reduced the production of proinflammatory cytokines and increased anti-inflammatory cytokine expression in the gingiva of WT and T2DM model mice. Ultimately, BBR mediates its anti-inflammatory effects on periodontitis through inhibition of the NF-κB pathway. BBR had a therapeutic effect on T2DM-associated periodontitis via inhibiting the NF-κB pathway to affect macrophage polarization, which may have implications for the new pharmacological treatment of T2DM-associated periodontitis.

In situ three-dimensional visualization of Bursaphelenchus xylophilus inoculated in Pinus thunbergii using X-ray micro-computed tomography

Pine wilt disease, a devastating infectious disease of pine trees, is caused by the pinewood nematode, Bursaphelenchus xylophilus. It is important to understand the spatial dispersal processes of B. xylophilus within host-tree tissues to assess its pathogenic mechanism. However, this is not feasible with conventional microscopy-imaging techniques. In this study, we showed that X-ray micro-computed tomography (micro-CT) imaging can be a powerful tool for visualizing infected nematodes within host-tree tissues. We visualized 161 nematodes and 11 eggs within a Pinus thunbergii stem section, 47.3 mm3 in volume, using an appropriate segmentation of the micro-CT images. Quantitative measurements of the segmented region corresponding to the nematodes allowed for the calculation of the longitudinal length and a-value, which were similar to previous morphological descriptions of B. xylophilus. The technique adopted in this study can aid in understanding the behavior of and obtaining quantitative information on B. xylophilus within tree tissues.

Investigation of gradient properties for cement mortar interfacial transition zones and their effects on mechanical characteristics using micro-CT

The interfacial transition zone (ITZ) in cement-based composites with aggregates is widely recognized as the most vulnerable component, greatly affecting their physical and mechanical properties. In this study, the effect of the ITZ characteristics on the mechanical behavior of cement mortar using its actual microstructure was investigated with image-based numerical approaches in this study. The volumetric information of the ITZ of the mortar sample was obtained and characterized from high-resolution synchrotron micro-computed tomography (micro-CT) images. The mechanical properties of the mortar were evaluated using the finite element method, based on the components characteristics represented by grayscale information of the micro-CT images. It was found that the adhesion between the aggregates and binder was well replicated in the simulation with the multiple-phase model, based on the gradient characteristics and properties of the ITZ. Considering the ITZ thickness according to the aggregates size, a more realistic crack propagation trend can be observed. The obtained results confirmed the significant influence of the ITZ thickness and phase distribution characteristics on the mechanical properties of cement mortars, emphasizing the importance of considering ITZ phases for accurate material analysis.

A new animal model of lumbar disc degeneration in rabbits

BACKGROUND CONTEXTThere are many models of lumbar disc degeneration, but mechanical stress-induced lumbar disc degeneration is rare. Here we propose a mechanical stress-induced lumbar disc degeneration model to better understand the molecular mechanism of lumbar disc degeneration under stress stimulation. PURPOSETo design a new model of lumbar disc degeneration under mechanical stress. STUDY DESIGNThe anatomic approach of the oblique lateral approach to lumbar fusion surgery was used to design a longitudinal compression device across the vertebral body of the rabbit to impose longitudinal load on the lumbar disc. METHODSNew Zealand white rabbits (n=30) were used. Screws were used to cross the rabbits’ lumbar vertebral bodies, and both sides of the screws were pressurized. Continuous compression was then performed for 28 days. Adjacent unpressurized lumbar discs serve as controls for pressurized lumbar discs. At 28 days after surgery, micro-computed tomography (CT) and magnetic resonance imaging (MRI) were performed on the rabbits’ lumbar discs. After the imaging examination, lumbar disc samples were removed, Safranin-O fast green and immunofluorescence was performed to detect the expression level of intervertebral disc degeneration-related proteins. RESULTSThe CT results showed that the disc height did not decrease significantly after mechanical loading. The MRI results showed that the signals in the pressurized disc decreased 28 days after loading. The results of Safranin-O fast green showed that the cartilage component of the intervertebral disc after mechanical compression was significantly reduced. The immunofluorescence results showed that the expression of ADAMTS5 and MMP13 protein in the nucleus pulposus of the intervertebral disc after mechanical compression increased, while the expression of SOX9 decreased, and the difference was statistically significant. Aggrecan's protein expression decreased, but was not statistically significant. CONCLUSIONSThis study designed a reliable model of disc degeneration in rabbits. It is more likely to mimic disc compression in the human body. CLINICAL SIGNIFICANCEThis animal model can be used as a basic model to study the molecular physiological mechanisms of discogenic low back pain.