micro-CT Analysis Research Articles

Mll4 regulates postnatal palate growth and midpalatal suture development.

MLL4, also known as KMT2D, is a histone methyltransferase that acts as an important epigenetic regulator during various organogenesis programs. Mutations in the MLL4 gene are the major cause for Kabuki syndrome, a human developmental disorder that involves craniofacial birth defects, including anomalies in the palate. The purpose of this study was to investigate the role of Mll4 and the underlying mechanisms in the development and growth of the palate. We generated a novel conditional knockout (cKO) mouse model with tissue-specific deletion of Mll4 in the palatal mesenchyme. By using micro-computed tomography (CT), histology, cell mechanism assays, and gene expression analysis approaches, we examined the development and growth of the palate in the Mll4 -cKO mice. Gross intra-oral examination at adult stages showed that Mll4 -cKO mice had defects along the midline of the palate, which included disrupted rugae pattern and widened midpalatal suture. Micro-CT-based skeletal analysis in the adult mice revealed that the overall palate width was decreased in the Mll4 -cKO mice. By using whole-mount and histological staining approaches at perinatal stages, we identified that the midline defects started to appear as early as 1 day prior to birth, manifesting initially as a widened midpalatal suture, accompanied by increased cell apoptosis in the suture mesenchyme cells. Genome-wide analysis of mRNA expression in the midpalatal suture tissue showed that Mll4 is essential for timely expression of major genes for cartilage development, such as Col2a1 and Acan , at birth. These results were validated through immunofluorescence staining, confirming that the expression of chondrogenic markers Sox9 and Col2a1 were markedly decreased, whereas that of the osteogenic marker Runx2 remained unchanged, in the midpalatal suture of the Mll4 -cKO mice. Indeed, time-course histological analysis during postnatal palate growth revealed retardation in the development of the suture cartilage in the Mll4 -cKO mice. In parallel, time-course micro-CT analysis during postnatal palatogenesis confirmed a transverse growth deficit in the palate of the Mll4 -cKO mice. Taken together, our results show that Mll4 is essential for timely occurrence of key cellular and molecular events that lead to proper midpalatal suture development and palate growth.

Vasorin-deficient mice display disturbed vitamin D and mineral homeostasis in combination with a low bone mass phenotype

Vasorin (Vasn) is a pleiotropic molecule involved in various physiological and pathological conditions, including cancer. Vasn has also been detected in bone cells of developing skeletal tissues but no function for Vasn in bone metabolism has been implicated yet. Therefore, this study aimed to investigate if Vasn plays a significant role in bone biology. First, we investigated tissue distribution of Vasn expression, using lacZ knock-in reporter mice. We detected clear Vasn expression in skeletal elements of postnatal mice. In particular, osteocytes and bone forming osteoblasts showed high expression of Vasn, while the bone marrow was devoid of signal. Vasn knockout mice (Vasn−/−) displayed postnatal growth retardation and died after four weeks. MicroCT analysis of femurs from 22- to 25-day-old Vasn−/−mice demonstrated reduced trabecular and cortical bone volume corresponding to a low bone mass phenotype. Ex vivo bone marrow cultures demonstrated that osteoclast differentiation and activity were not affected by Vasn deficiency. However, osteogenesis of Vasn−/− bone marrow cultures was disturbed, resulting in lower numbers of alkaline phosphate positive colonies, impaired mineralization and lower expression of osteoblast marker genes. In addition to the bone phenotype, these mice developed a vitamin D3-related phenotype with a strongly reduced circulating 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 and urinary loss of vitamin D binding protein. In conclusion, Vasn-deficient mice suffer from severe disturbances in bone metabolism and mineral homeostasis.

Effects of the Irrigation Needle Design on Root Canal Disinfection and Cleaning

IntroductionThis ex vivo study evaluated the disinfecting and cleaning effects of root canal preparation using sodium hypochlorite irrigation with 3 different needle designs. MethodsMesial roots from extracted mandibular molars with Vertucci class II configuration were anatomically matched based on micro-computed tomography (micro-CT) analyses and distributed into 3 groups (n = 18/group). The canals were contaminated with a mixed bacterial culture for 30 days and then subjected to preparation using 2.5% sodium hypochlorite irrigation with open-ended flat needle (3 mm short of the working length - WL), closed-ended side-vented irrigation needle (1 mm short of the WL), or a closed-ended plastic needle with 2 lateral outlets (TruNatomy) (1 mm short of the WL). Bacteriological samples were taken from the canals before (S1) and after preparation (S2). After another micro-CT scan, the roots were sectioned and samples were also taken from the apical canal segment (S3). Bacterial reduction was assessed by quantitative real-time polymerase chain reaction. The amount of accumulated hard tissue debris was evaluated by micro-CT. ResultsA substantial reduction in bacterial counts was observed in all 3 groups when comparing S1 with S2 (99.95%, 98.93%, and 98.90% in the open-ended, closed-ended, and TruNatomy needle groups, respectively) (P < .01). No significant differences were observed between groups for bacterial reduction in the full and apical canal (P > .05). There were no intergroup differences in the amount of accumulated hard tissue debris either (P > .05). The open-ended needle group showed significantly more specimens with quantitative real-time polymerase chain reaction negative results for bacteria in S3 than TruNatomy (P < .05). ConclusionWhen used up to an appropriate insertion depth, the 3 needle types result in similar antibacterial and cleaning performance, provided variables such as needle size and irrigant type, volume and flow rate are controlled. Exclusive analysis of the apical segment, including the isthmus, revealed that the open-ended needle yielded more cases negative for bacteria.

Differential effects of clopidogrel and/or aspirin on the healing of tooth extraction wound bone tissue.

A multitude of variables influence the healing of tooth extraction wounds, and delayed or non-healing extraction wounds might complicate later prosthodontic therapy. In this research, we analyzed the effects of systemic clopidogrel and aspirin alone or in combination on the healing of tooth extraction wounds in mice in order to provide experimental evidence for the healing of extraction wounds in patients who are clinically treated with the two medicines. 7-week-old ICR mice were randomly divided into four groups: control group (CON), clopidogrel group (CLOP), aspirin group (ASP), and clopidogrel combined with aspirin group (CLOP + ASP); left upper first molar was extracted, after which mice in 1week of adaptive feeding, CLOP/ASP/CLOP + ASP groups were respectively administered with clopidogrel (10mg/kg/d), aspirin (15mg/kg/d), clopidogrel (10mg/kg/d)+aspirin (15mg/kg/d), and the control group was given an equal amount of 0.9% saline by gavage. Mice in each group were euthanized at 14 and 28days postoperatively, and the maxilla was extracted. The tissues in the extraction sockets were examined using MicroCT and sectioned for HE staining, Masson staining, and TRAP staining, and immunohistochemistry staining (for TRAP, RANKL and osteoprotegerin). MicroCT analysis showed that at day 14, BS/BV was significantly lower in CLOP and CLOP + ASP groups compared to control and ASP groups, while BV/TV, Tb.Th was significantly higher. At day 28, BV/TV was significantly higher in the CLOP + ASP group compared to the CLOP group, with p < 0.05 for all results. HE staining and Masson trichrome staining findings revealed that at day 28, the mesenchyme in the bone was further decreased compared to that at day 14, accompanied with tightly arranged and interconnected bone trabeculae. In the quantitative analysis of Masson, the fraction of newly formed collagen was significantly higher in the CLOP group in comparison with that in the CON group (p < 0.05). At day 14, the ASP group had substantially more TRAP-positive cells than the CLOP and CLOP + ASP groups (p < 0.05). In immunohistochemical staining, RANKL expression was found to be significantly higher in the ASP group than those in the other three groups at day 28 (p < 0.05); OPG expression was significantly higher in the CLOP group and the CLOP + ASP group compared with that at day 14, and was higher than that in the ASP group at day 14 and day 28. OPG/RANKL was significantly higher in the CLOP and the CLOP + ASP groups than in the ASP group (p < 0.05). Clopidogrel alone promotes osteogenesis in the extraction wound, whereas aspirin alone inhibits alveolar bone healing. When the two drugs were combined, the healing effect of the extraction wound was more similar to that of the clopidogrel alone group. These results indicated that clopidogrel could promote the healing of the tooth extraction wound, and neutralize the adverse effect of ASP on osteogenesis when the two drugs were used in combination.

Deletion of vitamin D receptor exacerbated temporomandibular joint pathological changes under abnormal mechanical stimulation

AimsTemporomandibular disorder can cause degenerative pathological changes by aseptic inflammation in the temporomandibular joint (TMJ). Vitamin D (VD) is known for maintaining calcium homeostasis, and recent studies indicated that VD and the vitamin D receptor (VDR) are important in inflammatory responses. In this study, we explored the anti-inflammatory effect of VD-VDR signaling axis in TMJ pathological degeneration. Main methodsMice ablated for Vdr (Vdr−/−res) were fed with a rescue diet to avoid hypocalcemia. With abnormal mechanical stimulation, unilateral anterior crossbite (UAC) induced temporomandibular disorders in mice. Histological staining, immunohistochemistry staining, and micro-CT analysis were performed to evaluate TMJ pathological changes. To identify the mechanisms in the aseptic inflammatory process, in vitro experiments were conducted on wild-type (WT) and Vdr−/− chondrocytes with compressive mechanical stress loading, and the related inflammatory markers were examined. Key findingsVdr−/−res mice did not develop rickets with a high calcium rescue diet. The TMJ cartilage thickness in Vdr−/−res mice was significantly decreased with mechanical stress stimulation compared to WT mice. UAC-induced bone resorption was obvious, and the number of osteoclasts significantly increased in Vdr−/−res mice. The proliferation was inhibited and the gene expression of Il1b, Mmp3, and Mmp13 was significantly increased in Vdr−/− chondrocytes. However, WT chondrocytes showed significantly increased Tnfa gene expression as a response to mechanical stress but not in Vdr−/− chondrocytes. SignificanceVD-VDR is crucial in TMJ pathological changes under abnormal mechanical stimulation. Deletion of Vdr exacerbated inflammatory response excluding TNFα, inhibited chondrocyte proliferation, and promoted bone resorption in TMJ.

In vivo Assessment of AMP2, a Novel Ceramic-Binding BMP-2, in Ovine Lumbar Interbody Fusion.

Assessment of bone formation in an ovine interbody fusion study. To compare OsteoAdapt SP, which consists of AMP-2, a modified variant of recombinant human bone morphogenetic protein (rhBMP-2) bound to a tricalcium phosphate-containing carrier, to autologous iliac crest bone graft (ICBG) in a lumbar interbody fusion model. Treatment of lumbar disk degeneration often involves spinal fusion to reduce pain and motion at the affected spinal segment by insertion of a cage containing bone graft material. Three graft materials were compared in this study-ICBG and OsteoAdapt SP (low or high dose). The sheep underwent lateral lumbar fusion surgery with PEEK or Titanium interbody cages packed with OsteoAdapt SP (low or high dose) or ICBG. Outcomes were evaluated at 8-, 16- and 26- weeks. Newly formed bone quality, bone mineralization, and fusion were assessed by manual palpation, qualitative and semi-quantitative histopathology, histomorphometry, computed tomography (CT), and micro-CT (mCT) analysis. OsteoAdapt SP was implanted into 43 animals and ICBG into 21 animals (L3-L4). No group showed evidence of systemic toxicity by multiple assessments. All levels were fused by manual palpation at 26 weeks. Serial CT scans showed increasing fusion scores over time. Both doses of OsteoAdapt SP resulted in robust new bone formation and progression of fusion in the interbody cage. Range of motion tests for treatment groups was lower compared with ICBG at 8- and 16 weeks. Similarly, histology at eight weeks demonstrated more robust new bone formation for both OsteoAdapt SP groups compared to autograft. We have demonstrated the preclinical safety and efficacy of OsteoAdapt SP in a clinically relevant large animal model, supporting faster and more robust new bone formation within the interbody cage, comparable to or better than the gold standard, ICBG, in all measures.

Enhanced antibiofilm potential of low-intensity pulsed ultrasound combined with 0.35% povidone-iodine in a rat model of periprosthetic joint infection.

Although low-intensity pulsed ultrasound (LIPUS) combined with disinfectants has been shown to effectively eliminate portions of biofilm in vitro, its efficacy in vivo remains uncertain. Our objective was to assess the antibiofilm potential and safety of LIPUS combined with 0.35% povidone-iodine (PI) in a rat debridement, antibiotics, and implant retention (DAIR) model of periprosthetic joint infection (PJI). A total of 56 male Sprague-Dawley rats were established in acute PJI models by intra-articular injection of bacteria. The rats were divided into four groups: a Control group, a 0.35% PI group, a LIPUS and saline group, and a LIPUS and 0.35% PI group. All rats underwent DAIR, except for Control, which underwent a sham procedure. General status, serum biochemical markers, weightbearing analysis, radiographs, micro-CT analysis, scanning electron microscopy of the prostheses, microbiological analysis, macroscope, and histopathology evaluation were performed 14 days after DAIR. The group with LIPUS and 0.35% PI exhibited decreased levels of serum biochemical markers, improved weightbearing scores, reduced reactive bone changes, absence of viable bacteria, and decreased inflammation compared to the Control group. Despite the greater antibiofilm activity observed in the PI group compared to the LIPUS and saline group, none of the monotherapies were successful in preventing reactive bone changes or eliminating the infection. In the rat model of PJI treated with DAIR, LIPUS combined with 0.35% PI demonstrated stronger antibiofilm potential than monotherapy, without impairing any local soft-tissue.

Swimming exercise alleviates pathological bone features in curdlan-injected SKG mice by inducing irisin expression

This study assessed the therapeutic potential of swimming exercise in the curdlan-injected SKG mouse model and investigated the modulatory effects of irisin on inflammation. Curdlan-injected SKG were randomly assigned to either a home-cage group or a swimming group for 6 weeks. Changes in clinical arthritis scores and ankle thickness were measured weekly. Post-swimming program, mice were anesthetized for collection of vastus lateralis muscle and blood, which was followed by histological analysis, micro-CT imaging of the ankle joints, and the measurement of pro-inflammatory cytokines and irisin levels. Additionally, curdlan-injected SKG mice were intravenously injected with recombinant irisin protein and observed. Finally, serum levels of irisin in healthy control and ankylosing spondylitis (AS) patient groups were measured by ELISA. The swimming group of curdlan-injected SKG mice exhibited significant improvements in arthritis and enthesitis compared to the home-cage group. In particular, micro-CT and histological analyses revealed a notable reduction in pathological bone features in the swimming group compared to the home-cage group. Muscle endurance was also enhanced in the swimming group compared to the home-cage group, as determined by the wire-hanging test. Intriguingly, irisin levels not only were statistically increased in the swimming group but, also, TNF-α, IL-1β, and IL-6 levels were decreased. Additionally, injection of irisin protein slightly attenuated both arthritis and enthesitis in curdlan-injected SKG mice. Meanwhile, irisin serum levels were declined in AS patients. Overall, we found that swimming exercise attenuated pathological bone features in an AS animal model, potentially mediated by increased irisin serum levels with associated anti-inflammatory effects.

Exploration of the miR-187-3p/CNR2 pathway in modulating osteoblast differentiation and treating postmenopausal osteoporosis through mechanical stress.

This study aimed to explore how mechanical stress affects osteogenic differentiation via the miR-187-3p/CNR2 pathway. To conduct this study, 24 female C57BL/6 mice, aged 8 weeks, were used and divided into four groups. The Sham and OVX groups did not undergo treadmill exercise, while the Sham + EX and OVX + EX groups received a 8-week treadmill exercise. Post-training, bone marrow and fresh femur samples were collected for further analysis. Molecular biology analysis, histomorphology analysis, and micro-CT analysis were conducted on these samples. Moreover, primary osteoblasts were cultured under osteogenic conditions and divided into GM group and CTS group. The cells in the CTS group underwent a sinusoidal stretching regimen for either 3 or 7 days. The expression of early osteoblast markers (Runx2, OPN, and ALP) was measured to assess differentiation. The study findings revealed that mechanical stress has a regulatory impact on osteoblast differentiation. The expression of miR-187-3p was observed to decrease, facilitating osteogenic differentiation, while the expression of CNR2 increased significantly. These observations suggest that mechanical stress, miR-187-3p, and CNR2 play crucial roles in regulating osteogenic differentiation. Both invivo and invitro experiments have confirmed that mechanical stress downregulates miR-187-3p and upregulates CNR2, which leads to the restoration of distal femoral bone mass and enhancement of osteoblast differentiation. Therefore, mechanical stress promotes osteoblasts, resulting in improved osteoporosis through the miR-187-3p/CNR2 signaling pathway. These findings have broad prospect and provide molecular biology guidance for the basic research and clinical application of exercise in the prevention and treatment of PMOP.

Acceleration of bone repairation by BMSCs overexpressing NGF combined with NSA and allograft bone scaffolds

BackgroundRepairation of bone defects remains a major clinical problem. Constructing bone tissue engineering containing growth factors, stem cells, and material scaffolds to repair bone defects has recently become a hot research topic. Nerve growth factor (NGF) can promote osteogenesis of bone marrow mesenchymal stem cells (BMSCs), but the low survival rate of the BMSCs during transplantation remains an unresolved issue. In this study, we investigated the therapeutic effect of BMSCs overexpression of NGF on bone defect by inhibiting pyroptosis.MethodsThe relationship between the low survival rate and pyroptosis of BMSCs overexpressing NGF in localized inflammation of fractures was explored by detecting pyroptosis protein levels. Then, the NGF+/BMSCs-NSA-Sca bone tissue engineering was constructed by seeding BMSCs overexpressing NGF on the allograft bone scaffold and adding the pyroptosis inhibitor necrosulfonamide(NSA). The femoral condylar defect model in the Sprague–Dawley (SD) rat was studied by micro-CT, histological, WB and PCR analyses in vitro and in vivo to evaluate the regenerative effect of bone repair.ResultsThe pyroptosis that occurs in BMSCs overexpressing NGF is associated with the nerve growth factor receptor (P75NTR) during osteogenic differentiation. Furthermore, NSA can block pyroptosis in BMSCs overexpression NGF. Notably, the analyses using the critical-size femoral condylar defect model indicated that the NGF+/BMSCs-NSA-Sca group inhibited pyroptosis significantly and had higher osteogenesis in defects.ConclusionNGF+/BMSCs-NSA had strong osteogenic properties in repairing bone defects. Moreover, NGF+/BMSCs-NSA-Sca mixture developed in this study opens new horizons for developing novel tissue engineering constructs.

Comparing Warm and Cold Gutta-Percha Techniques for Root Canal Filling: An In Vitro Study

ABSTRACT Background: Gutta-percha is commonly used in endodontic therapy for obturating root canals after cleaning and shaping procedures. Warm and cold gutta-percha techniques are two common methods employed for obturation, each with its own advantages and limitations. Materials and Methods: Twenty extracted human teeth with single-rooted canals were selected and divided into two groups: warm gutta-percha obturation and cold gutta-percha obturation. In the warm gutta-percha group, gutta-percha was heated to a predetermined temperature using a warm gutta-percha heating device and injected into the root canal using a heated carrier. In the cold gutta-percha group, gutta-percha was applied directly to the root canal without prior heating. Root canal obturation quality was assessed using radiographic imaging to evaluate the presence of voids, completeness of obturation, and adaptation to the canal walls. Micro-computed tomography (micro-CT) analysis was also performed to quantify the volume of obturating material and assess the three-dimensional distribution of gutta-percha within the root canal space. Results: Both warm and cold gutta-percha techniques demonstrated effective obturation of root canals, with comparable outcomes in terms of void formation, completeness of obturation, and adaptation to canal walls. However, micro-CT analysis revealed differences in the volume and distribution of gutta-percha within the root canal space. Warm gutta-percha obturation exhibited more uniform distribution of gutta-percha material and higher volumetric filling compared to cold gutta-percha obturation. Additionally, the warm gutta-percha technique demonstrated superior flowability and adaptation to irregularities within the root canal system. Conclusion: In this in vitro study, warm gutta-percha obturation demonstrated advantages over cold gutta-percha obturation in terms of volumetric filling, distribution within the root canal space, and adaptation to canal irregularities.

Concurrent Joint Contact in Anterior Cruciate Ligament Injury induces cartilage micro-injury and subchondral bone sclerosis, resulting in knee osteoarthritis.

Anterior Cruciate Ligament (ACL) injury initiates post-traumatic osteoarthritis (PTOA) via two distinct processes: initial direct contact injury of the cartilage surface during ACL injury, and secondary joint instability due to the ACL deficiency. Using the well-established Compression-induced ACL rupture method (ACL-R) and a novel Non-Compression ACL-R model, we aimed to reveal the individual effects of cartilage compression and joint instability on PTOA progression after ACL injury in mice. Twelve-week-old C57BL/6J male were randomly divided to three experimental groups: Compression ACL-R, Non-Compression ACL-R, and Intact. Following ACL injury, we performed joint laxity testing and microscopic analysis of the articular cartilage surface at 0 days, in vivo optical imaging of matrix-metalloproteinase (MMP) activity at 3 and 7 days, and histological and microCT analysis at 0, 7, 14, and 28 days. The Compression ACL-R group exhibited a significant increase of cartilage roughness immediately after injury compared with the Non-Compression group. At 7 days, the Compression group exhibited increased MMP-induced fluorescence intensity and MMP-13 positive cell ratio of chondrocytes. Moreover, histological cartilage degeneration was observable in the Compression group at the same time point. Sclerosis of tibial subchondral bone in the Compression group was more significantly developed than in the Non-Compression group at 28 days. Both Compression and Non-Compression ACL injury initiated PTOA progression due to joint instability. However, joint contact during ACL rupture also caused initial micro-damage on the cartilage surface and initiated early MMP activity, which could accelerate PTOA progression compared to ACL injury without concurrent joint contact.

Slip and stress in block-in-matrix shear zones: 1. microstructure and mineralogy of a serpentine-filled dilational jog

We document the structural setting, microstructure, and mineralogy of a crack-seal-type dilational jog that developed in the stepover between two faults cutting through a phacoid of massive serpentinite, itself embedded within a serpentinite shear zone at the base of the Dun Mountain Ophiolite, New Zealand. Our outcrop and microstructural measurements allow us to constrain the boundary conditions for a numerical model (part 2) that quantitatively explores the relationships between stress, fault slip, and incremental cracking in block-in-matrix shear zones. The dilational jog is c. 3 cm wide and contains hundreds of crack-seal bands, each c. 20–30 μm wide. Internally, the jog comprises two mineralogically distinct crack-seal domains: serpentine-only domains and serpentine-andradite garnet domains. Additionally, individual crack-seal bands have a double-layer structure: in serpentine-only domains each band comprises a thin (<2 μm) layer of chrysotile and a thicker layer (c. 25 μm) of polygonal serpentine/lizardite, whereas each band in serpentine + andradite domains comprises a thinner (c. 5 μm) layer of microcrystalline andradite and a thicker layer (c. 15 μm) of polygonal serpentine/lizardite. Micro-CT analysis shows that the serpentine + andradite domains have conic or ellipsoidal shapes with long axes subparallel to the inferred jog opening direction, and that andradite is smeared along micro-transform surfaces inside the jog. Our conceptual microstructural model invokes jog formation during progressive serpentinization of the host rock. Incremental crack opening along the jog-wall rock interface promotes relatively rapid initial precipitation of chrysotile or andradite at high fluid:rock ratios. As cracks fill and pressure re-equilibrates, relatively slow growth of polygonal serpentine/lizardite is favoured until the cracks are sealed and the cycle repeats. Our observations suggest that the precipitation of andradite (instead of chrysotile) was controlled both by structural boundaries within the jog (e.g., micro-transform surfaces) and by local element transport (e.g., Ca from serpentinizing clinopyroxene grains in the host rocks) to patches of the crack wall.

Development of Foam Composites from Flax Gum-Filled Epoxy Resin

In the present work, an innovative range of foams based on flax gum-filled epoxy resin was developed, reinforced or not by flax fibers. Foams and composites with different gum and epoxy resin contents were produced and their mechanical and thermal performances were characterized. To enhance the organic flax gum filler’s cross-linking, we exploited the oxidized components’ reactivity with the amine hardener (isophorone diamine). We compared the materials obtained with those derived from the native components. The flax gum and fibers were primarily characterized by chemical analysis, NMR, and FTIR to evaluate the mild oxidation of the native materials. The formation of chemical bonds between the oxidized polymer chains, epoxy resin, and hardener was evidenced by FTIR, and the materials were then studied by SEM and X-ray computed micro-tomography (CT) and submitted to mechanical and thermal tests. The relevance of the oxidation treatment was highlighted through a significant increase in density and mechanical performance (+36% and +81%, respectively, for the 100% flax gum material). The positive effect of the flax fibers on homogeneity evidenced through micro-CT analysis was also clearly addressed. This set of promising results paves the way for the future development of fully flax-based insulation composite materials.

Dietary intervention rescues a bone porosity phenotype in a murine model of Neurofibromatosis Type 1 (NF1).

Neurofibromatosis type 1 (NF1) is a complex genetic disorder that affects a range of tissues including muscle and bone. Recent preclinical and clinical studies have shown that Nf1 deficiency in muscle causes metabolic changes resulting in intramyocellular lipid accumulation and muscle weakness. These can be subsequently rescued by dietary interventions aimed at modulating lipid availability and metabolism. It was speculated that the modified diet may rescue defects in cortical bone as NF1 deficiency has been reported to affect genes involved with lipid metabolism. Bone specimens were analyzed from wild type control mice as well as Nf1Prx1-/- (limb-targeted Nf1 knockout mice) fed standard chow versus a range of modified chows hypothesized to influence lipid metabolism. Mice were fed from 4 weeks to 12 weeks of age. MicroCT analysis was performed on the cortical bone to examine standard parameters (bone volume, tissue mineral density, cortical thickness) and specific porosity measures (closed pores corresponding to osteocyte lacunae, and larger open pores). Nf1Prx1-/- bones were found to have inferior bone properties to wild type bones, with a 4-fold increase in the porosity attributed to open pores. These measures were rescued by dietary interventions including a L-carnitine + medium-chain fatty acid supplemented chow previously shown to improve muscle histology function. Histological staining visualized these changes in bone porosity. These data support the concept that lipid metabolism may have a mechanistic impact on bone porosity and quality in NF1.

Micro-scale investigation of the pore network of sandstone in the Pannonian Basin to improve geothermal energy development

This study focuses on the Pannonian Basin, specifically in Szentes, Hungary a region of significant geothermal potential, with particular emphasis on the Dunántúl Group; a collective name for the Zagyva and Újfalu formations, which consists of slightly consolidated delta-front sandstone sediments. This research is pivotal in understanding the challenges associated with clogging in geothermal wells, a problem that has led to the premature shutdown of injection wells in the region. Our approach integrates classical petrophysical and mineralogical methods with advanced techniques such as micro-Computed X-Ray Tomography imaging, 3D image analysis, and digital rock simulations. Our findings indicate that the target geothermal rock formations within the Dunántúl Group exhibit high porosity (27–31 %) and variable permeability (60–400 mD), dependent on the location and specific characteristics of the formation. Our micro-CT analyses further identified that the presence of fine-grained materials in smaller pores and generally weak cementation of grains substantially contributes to these challenges.

Unraveling microstructural evolution in air-entrained mortar and paste: Insights from MIP and micro-CT tomography amid cyclic freezing-thawing damage

Understanding the microstructure evolution during the cyclic freezing-thawing process is crucial to studying the deterioration mechanisms of concrete under cyclic freezing-thawing. In this study, air voids were entrained into cement mortar and paste by AEA and polymeric microspheres. Mercury intrusion porosimetry (MIP) and micro-CT techniques were integrated to investigate the microstructure of the specimens with various freezing-thawing cycles. After subjecting the Ref. and AEA mixed mortar to 100 and 200 freezing-thaw cycles, an increase in porosity and pore entrapment was observed from the MIP results, suggesting the occurrence of damage induced by cyclic freezing-thawing. Conversely, in the microspheres mixed mortar, the presence of surface adsorbed moisture led to an initial increase in porosity and pore entrapment after 100 freezing-thaw cycles, followed by a decrease after 200 cycles, indicating a healing effect. Micro-CT analysis revealed that while the AEA mixed specimens exhibited higher air content than the microspheres mixed specimens, the latter showed a greater number of voids, which ultimately enhanced their resistance to freezing-thawing. The micro-CT results unveiled that air bubbles tended to merge in AEA mixed specimens, adversely affecting their resistance to freezing-thawing. Conversely, the microsphere-incorporated specimens displayed smooth void distribution curves, indicating a well-distributed air void system that enhances their freezing-thawing resistance. This work suggests that microspheres can be a great candidate for air-entraining materials. Combining the MIP and micro-CT analysis can better understand the microstructure change of concrete with cyclic freezing-thawing.

Insights in emerging Ti3C2Tx MXene-enriched polymeric coatings for metallic surface protection: Advancements in microstructure, anti-aging, and electrochemical performance

This study provides new insights into the development of high-performance MXene-reinforced coatings to strengthen polymeric nanocomposites by enhancing microstructure, anti-aging properties, corrosion resistance, and robustness. MXene nanoparticles, labeled 25C and 80C, were synthesized using two different methods and incorporated at concentrations ranging from 0.1 to 2.0 wt% into epoxy composites. The results demonstrated that 80C MXene, characterized by its finer morphology and superior dispersion, significantly improved the composite's performance compared to 25C. Electrochemical Impedance Spectroscopy (EIS) tests, along with long-term exposure assessments, suggested that incorporating both types of MXene nanoparticles enhances the corrosion protection performance of epoxy coatings over time. Micro-CT analysis revealed that both types of MXene substantially reduced defects and voids in the polymeric matrix, resulting in enhanced protective performance. This void reduction confirms that the incorporation of both 25C and 80C MXene improves microstructural integrity by filling voids and creating a more continuous, uniform structure, particularly in samples with 0.1 % to 1.0 % MXene flakes. The findings also highlighted MXene's potential in modifying the anti-aging properties of epoxy by inhibiting free radical generation and enhancing the composite's resistance against corrosion. Both 25C and 80C MXene-epoxy groups exhibited a clear trend of diminishing free radical intensity with increasing MXene concentration up to 1.0 %, with free radical intensity reduced by over 40 % compared to neat epoxy. The relationship between MXene concentration and reinforcement was also investigated, revealing superior corrosion protection properties at concentrations of 0.5–1.0 wt%. This research offers a profound understanding of MXene's potential in polymer-based composites, laying a foundation for future investigations aimed at utilizing MXene to achieve superior material properties for a wide range of applications, particularly in the realm of metallic surface protection.

Titanium-doped phosphate glasses containing zinc and strontium applied in bone regeneration

Phosphate bioactive glass has been studied for its advanced biodegradability and active ion release capability. Our previous research found that phosphate glass containing (P2O5)–(Na2O)–(TiO2)–(CaO)–(SrO) or (ZnO) showed good biocompatibility with MG63 and hMSCs. This study further investigated the application of 5 mol% zinc oxide or 17.5 mol% strontium oxide in titanium-doped phosphate glass for bone tissue engineering. Ti-Ca-Na-Phosphate glasses, incorporating 5% zinc oxide or 17.5% strontium oxide, were made with melting quenching technology. The pre-osteoblast cell line MC3T3-E1 was cultured for indirect contact tests with graded diluted phosphate glass extractions and for direct contact tests by seeding cells on glass disks. The cell viability and cytotoxicity were analysed in vitro over 7 days. In vivo studies utilized the tibial defect model with or without glass implants. The micro-CT analysis was performed after surgery and then at 2, 6, and 12 weeks. Extractions from both zinc and strontium phosphate glasses showed no negative impact on MC3T3-E1 cell viability. Notably, non-diluted Zn-Ti-Ca-Na-phosphate glass extracts significantly increased cell viability by 116.8% (P < 0.01). Furthermore, MC3T3-E1 cells cultured with phosphate glass disks exhibited no increase in LDH release compared with the control group. Micro-CT images revealed that, over 12 weeks, both zinc-doped and strontium-doped phosphate glasses demonstrated good bone incorporation and longevity compared to the no-implant control. Titanium-doped phosphate glasses containing 5 mol% zinc oxide, or 17.5 mol% strontium oxide have promising application potential for bone regeneration research.Graphical

Protective effects of probiotics against menopausal symptoms in ovariectomized mice

With growing interest in the role of the gut microbiome in the pathogenesis of menopausal disorders, probiotic supplementation is emerging as a novel therapeutic strategy for postmenopausal women. In this study, we investigated the protective effects of several probiotic strains against menopausal symptoms using an ovariectomized (OVX) mouse model fed a high-fat diet, in comparison with the effects of 17β-estradiol (E2) replacement. Among the eight tested probiotic strains, we selected four (Limosilactobacillus fermentum MG901, Lactiplantibacillus plantarum MG4604, Ligilactobacillus salivarius MG4679, and Lacticaseibacillus reuteri MG5463) that showed superior estrogen-like activity in an uterotrophic assay. We then examined their effects in OVX mice. The probiotic MG4679 was tested in combination with MG5463. During the 12-week experimental period, oral administration of the four probiotics reduced blood glucose and body temperature and ameliorated depression-like behavior in OVX mice. In the Barnes maze test, MG901 and MG4604 improved the spatial memory deficits observed in OVX mice. Micro-CT analysis of mouse femurs revealed that MG5463 prevented the OVX-induced reduction in cortical bone mineral density and bone volume fraction. Importantly, similar to E2 replacement, most of the test probiotics increased Akt phosphorylation in skeletal muscle, estrogen receptor-α (ER-α) protein expression in the liver, and ERK phosphorylation in the brain of OVX mice. However, unlike E2 treatment, all probiotics did not affect uterine weight, uterine ER-α expression, and serum E2 concentration. These results indicated that the probiotics used in this study exhibited protective effects against menopausal symptoms, partly through estrogen-mimetic action, without unfavorable uterine stimulation in OVX mice.