Metabolic and Skeletal Characterization of the KK/Ay Mouse Model-A Polygenic Mutation Model of Obese Type 2 Diabetes.

Type 2 diabetes (T2D) increases fracture incidence and fracture-related mortality rates (KK.Cg-Ay/J. The Jackson Laboratory; Available from: https://www.jax.org/strain/002468 ). While numerous mouse models for T2D exist, few effectively stimulate persistent hyperglycemia in both sexes, and even fewer are suitable for bone studies. Commonly used models like db/db and ob/ob have altered leptin pathways, confounding bone-related findings since leptin regulates bone properties (Fajardo et al. in Journal of Bone and Mineral Research 29(5): 1025-1040, 2014). The Yellow Kuo Kondo (KK/Ay) mouse, a polygenic mutation model of T2D, is able to produce a consistent diabetic state in both sexes and addresses the lack of a suitable model of T2D for bone studies. The diabetic state of KK/Ay stems from a mutation in the agouti gene, responsible for coat color in mice. This mutation induces ectopic gene expression across various tissue types, resulting in diabetic mice with yellow fur coats (Moussa and Claycombe in Obesity Research 7(5): 506-514, 1999). Male and female KK/Ay mice exhibited persistent hyperglycemia, defining them as diabetic with blood glucose (BG) levels consistently exceeding 300 mg/dL. Notably, male control mice in this study were also diabetic, presenting a significant limitation. Nevertheless, male and female KK/Ay mice showed significantly elevated BG levels, HbA1c, and serum insulin concentration when compared to the non-diabetic female control mice. Early stages of T2D are characterized by hyperglycemia and hyperinsulinemia resulting from cellular insulin resistance, whereas later stages may feature hypoinsulinemia due to ß-cell apoptosis (Banday et al. Avicenna Journal of Medicine 10(04): 174-188, 2020 and Klein et al. Cell Metabolism 34(1): 11-20, 2022). The observed hyperglycemia, hyperinsulinemia, and the absence of differences in ß-cell mass suggest that KK/Ay mice in this study are modeling the earlier stages of T2D. While compromised bone microarchitecture was observed in this study, older KK/Ay mice, representing more advanced stages of T2D, might exhibit more pronounced skeletal manifestations. Compared to the control group, the femora of KK/Ay mice had higher cortical area and cortical thickness, and improved trabecular properties which would typically be indicative of greater bone strength. However, KK/Ay mice displayed lower cortical tissue mineral density in both sexes and increased cortical porosity in females. Fracture instability toughness of the femora was lower in KK/Ay mice overall compared to controls. These findings indicate that decreased mechanical integrity noted in the femora of KK/Ay mice was likely due to overall bone quality being compromised.

Evaluation of the Ejection Pressure for Tracking Internal Cracks during Compaction in Bilayer Tablet Formulations Using Experimental and Finite Element Methods.

This study aimed to evaluate the ejection pressure and the correlation of the findings with the occurrence of internal cracks within bilayer tablets (BLTs) consisting of metformin HCl (MF) and evogliptin tartrate (EG). Then, the mechanism of tablet failure was provided by the finite element method (FEM). The ejection pressure and the difference in diameter depending on MAIN-P were evaluated to understand the correlation between ejection pressure and change in the BLT internal structure. The ejection pressure and the difference in diameter increased as the MAIN-P increased, then steeply decreased from 350 MPa to 375 MPa of MAIN-P, despite there being no pattern in compaction breaking force and porosity. The mechanical integrity at the BLT interface was weakened by internal cracks, reducing ejection pressure. The stress distribution analysis during the compression revealed that crack formation caused by entrapped air located at the center of the BLT interface may not propagate due to concentrated stress, which promotes a tight bond at the edge of the BLT. Furthermore, complete delamination can occur in the ejection process due to localized and intensive shear stresses at the BLT interface. These findings indicate that the mechanisms of internal cracking and delamination were successfully confirmed by FEM simulation. Moreover, measuring ejection pressure before BLT manufacturing can prevent invisible tablet cracks without damaging the tablets.

Pulmonary vasculature development in congenital diaphragmatic hernia: a novel automated quantitative imaging analysis.

PURPOSE: Impaired fetal lung vasculature determines the degree of pulmonary hypertension in the congenital diaphragmatic hernia (CDH). This study aims to demonstrate the morphometric measurements that differ in pulmonary vessels of fetuses with CDH. METHODS: Nitrofen-induced CDH Sprague-Dawley rat fetuses were scanned with microcomputed tomography. The analysis of the pulmonary vascular tree was performed with artificial intelligence. RESULTS: The number of segments in CDH was significantly lower than that in the control group on the left (U = 2.5, p = 0.004) and right (U = 0, p = 0.001) sides for order 1(O1), whereas there was a significant difference only on the right side for O2 and O3. The pooled element numbers in the control group obeyed Horton's law (R2 = 0.996 left and R2 = 0.811 right lungs), while the CDH group broke it. Connectivity matrices showed that the average number of elements of O1 springing from elements of O1 on the left side and the number of elements of O1 springing from elements of O3 on the right side were significantly lower in CDH samples. CONCLUSION: According to these findings, CDH not only reduced the amount of small order elements, but also destroyed the fractal structure of the pulmonary arterial trees.

Shaping Efficiency of Rotary and Reciprocating Kinematics of Engine-driven Nickel-Titanium Instruments in Moderate and Severely curved Root Canals Using Microcomputed Tomography: A Systematic Review of Ex Vivo Studies.

INTRODUCTION: This systematic review aimed to compare reciprocating with continuous rotary instrumentation kinematics, by means of microcomputed tomography evaluations, in extracted human permanent teeth with moderate and severe canal curvatures. METHODS: The research protocol was registered in the International Prospective Register of Systematic Reviews and given the reference number CRD42023404035. An electronic search was undertaken in MEDLINE (PubMed), EBSCO, Scopus, Web of Science databases until December 2021. Manual screening of issues in endodontic journals and references of relevant articles were assessed individually. The risk of bias (RoB) of the included articles was evaluated with the QUIN tool (Quality Assessment Tool for In Vitro Studies). RESULTS: Among 1640 retrieved articles, 49 were included in the qualitative synthesis. Fifteen articles had low RoB, 33 articles had medium RoB, and only 1 study was at high RoB. Continuous rotary systems had better centering ability in both moderate and severe canal curvatures and resulted in less apical transportation in severely curved root canals. None of the kinematic systems was capable of instrumenting the entire canal surface area. Reciprocating kinematics systems tended to provide higher increase in surface area of severely curved canals and produced fewer dentinal microcracks in moderately curved canals. CONCLUSIONS: The evidence presented in this review suggests that continuous rotary system seems to be better than reciprocating system in solving the major issues encountered during root canal instrumentation of extracted teeth with moderate and severe curvatures.

Bone Marrow Adipose Tissue Is Not Required for Reconstitution of the Immune System Following Irradiation in Male Mice.

Bone marrow adipose tissue (BMAT) is hypothesized to serve as an expandable/contractible fat depot which functions, in part, to minimize energy requirements for sustaining optimal hematopoiesis. We investigated whether BMAT is required for immune reconstitution following injury. Male wild type (WBB6F1, WT) and BMAT-deficient WBB6F1/J-KitW/KitW-v/J (KitW/W-v) mice were lethally irradiated. Irradiation was followed by adoptive transfer of 1000 purified WT hematopoietic stem cells (HSCs). The extent of immune reconstitution in blood, bone marrow, and lymph nodes in the irradiated mice was determined using HSCs from green fluorescent protein (GFP)-expressing mice. We also evaluated skeletal response to treatment. Detection of GFP-positive B and T cells in peripheral blood at 4 and 9 weeks following adoptive transfer and in bone marrow and lymph nodes following necropsy revealed excellent immune reconstitution in both WT and BMAT-deficient mice. Adipocytes were numerous in the distal femur of WT mice but absent or rare in KitW/W-v mice. Bone parameters, including length, mass, density, bone volume, microarchitecture, and turnover balance, exhibited few differences between WT and BMAT-deficient mice. The minimal differences suggest that BMAT is not required for reconstitution of the immune system following lethal radiation and is not a major contributor to the skeletal phenotypes of kit signaling-deficient mice.

Xylem-dwelling pathogen unaffected by local xylem vessel network properties in grapevines (Vitis spp.).

BACKGROUND AND AIMS: Xylella fastidiosa (Xf) is the xylem-dwelling bacterium associated with Pierce's Disease (PD), which causes mortality in agriculturally important species, such as grapevine (Vitis vinifera L.). Development of PD symptoms in grapevines depends on the ability of Xf to produce cell wall-degrading enzymes to break up intervessel pit membranes and systematically spread through xylem vessel network. Our objective was to investigate whether PD resistance could be mechanistically linked to xylem vessel network local connectivity. METHODS: We used high-resolution X-ray micro-computed tomography (microCT) imaging to identify and describe the type, area, and spatial distribution of intervessel connections for six different grapevine genotypes from three genetic backgrounds, with varying resistance to PD (four PD resistant and two PD susceptible). KEY RESULTS: Our results suggest that PD resistance is unlikely to derive from local xylem network connectivity. The intervessel pit area (Ai) varied from 0.07 ±0.01 SD mm2 mm-3 in Lenoir to 0.17 ±0.03 SD mm2 mm-3 in Blanc do Bois, both PD resistant. Intervessel contact fraction (Cp) was not statically significant, but the two PD susceptible genotypes, Syrah (0.056 ±0.015 SD) and Chardonnay (0.041 ±0.013 SD), were among the most highly connected vessel networks. Neither Ai nor Cp explained differences in PD resistance among the six genotypes. Bayesian re-analysis of our data shows moderate evidence against the effects of the traits analyzed, Ai (BF01=4.88), mean vessel density (4.86) and relay diameter (4.30), relay density (3.31), and solitary vessel proportion (3.19). CONCLUSIONS: Our results show that radial and tangential xylem network connectivity is highly conserved within the six different Vitis genotypes we sampled. The way that Xf traverses the vessel network may limit the importance of local network properties to its spread and may confer greater importance on host biochemical responses.

Impact of drug load and polymer molecular weight on the 3D microstructure of printed tablets.

This study investigates the influence of drug load and polymer molecular weight on the structure of tablets three-dimensionally (3D) printed from the binary mixture of prednisolone and hydroxypropyl methylcellulose (HPMC). Three different HPMC grades, (AFFINISOLTM HPMC HME 15LV, 90 Da (HPMC 15LV); 100LV, 180 Da (HPMC 100LV); 4M, 500 Da (HPMC 4M)), which are suitable for hot-melt extrusion (HME), were used in this study. HME was used to fabricate feedstock material, i.e., filaments, at the lowest possible extrusion temperature. Filaments of the three HPMC grades were prepared to contain 2.5, 5, 10 and 20 % (w/w) prednisolone. The thermal degradation of the filaments was studied with thermogravimetric analysis, while solid-state properties of the drug-loaded filaments were assessed with the use of X-ray powder diffraction. Prednisolone in the freshly extruded filaments was determined to be amorphous for drug loads up to 10%. It remained physically stable for at least 6 months of storage, except for the filament containing 10% drug with HPMC 15LV, where recrystallization of prednisolone was detected. Fused deposition modeling was utilized to print honeycomb-shaped tablets from the HME filaments of HPMC 15LV and 100LV. The structural characteristics of the tablets were evaluated using X-ray microcomputed tomography, specifically porosity and size of structural elements were investigated. The tablets printed from HPMC 15LV possessed in general lower total porosity and pores of smaller size than tablets printed from the HPMC 100LV. The studied drug loads were shown to have minor effect on the total porosity of the tablets, though the lower the drug load was, the higher the variance of porosity along the height of the tablet was observed. It was found that tablets printed with HPMC 15LV showed higher structural similarity with the virtually designed model than tablets printed from HPMC 100LV. These findings highlight the relevance of the drug load and polymer molecular weight on the microstructure and structural properties of 3D printed tablets.

Composite of bacterial cellulose and gelatin: A versatile biocompatible scaffold for tissue engineering.

Synthesis of 0.4 ± 0.03 g/L per day of pure and porous bacterial cellulose (BC) scaffolds (scaffBC) and BC scaffolds modified with gelatin (scaffBC/Gel) was carried out using the Medusomyces gisevii Sa-28 bacterial strain. FT-IR spectroscopy and X-ray diffraction analysis showed that the scaffolds largely consist of crystalline cellulose I (Iα, Iß). Heating of BC with gelatin to 60 °C with subsequent lyophilization led to its modification by adsorption and binding of low-molecular fractions of gelatin and the formation of small pores between the fibers, which increased the biocompatibility and solubility of BC. The solubility of scaffBC and scaffBC/Gel was 20.8 % and 44.4 %, respectively, which enhances degradation in vivo. Light microscopy, scanning electron microscopy, and microcomputed tomography showed a uniform distribution of pores with a diameter of 100-500 µm. The chicken chorioallantoic membrane (CAM) model and subcutaneous implantation in rats confirmed low immunogenicity and intense formation of collagen fibers in both scaffolds and active germination of new blood vessels in scaffBC and scaffBC/Gel. The proliferative cellular activity of fibroblasts confirmed the safety of scaffolds. Taken together, the results obtained show that scaffBC/Gel can be used for the engineering of hard and soft tissues, which opens opportunities for further research.

Natural canal deviation and dentin thickness of mesial root canals of mandibular first molars assessed by microcomputed tomography

Abstract The aim of this study was to assess the centralization and dentin thickness of mesial root canals of the first mandibular molars by microcomputed tomography (micro-CT). Material and methods: Ninety-nine mandibular molars of Vertucci's type IV canals were scanned by micro-CT. The mesiodistal deviation and centroid were assessed, in both mesiobuccal (MB) and mesiolingual (ML) canals, for the apical 4mm and the full canal length. Results: The dentin thickness was similar for both MB and ML canals. The narrowest thickness was in the distal wall of an MB canal (0.07mm), while the widest was found in the mesial wall of an MB canal (2.46mm). In centroid analysis, both the MB and ML canals exhibited deviations when compared to the root centroid, along the full canal length and the apical 4mm. For the MB canal, the mean deviation was 0.83mm (0.02 mm-2.30 mm) for the full canal and 0.18mm (0.01 mm-1.01 mm) for apical 4mm. Similarly, for the ML canal, the mean deviation measured 0.83 mm (0.05mm-3.99mm) for the full canal and 0.21 mm (0.01mm-1.01mm) for the apical 4 mm. Overall, deviations were observed towards the mesial of the roots, with 69% for MB and 57% for ML canals for the full canal, and 51% for MB canals within the 4 mm. The exception was the ML canal, which exhibited a higher deviation towards distal in the apical 4mm, accounting for 52% of cases. The dentin thickness was consistent between the mesial canals of mandibular molars. However, there is no centrality of mesial canals in their roots, with frequent deviation to mesial.

Resumo O objetivo deste trabalho foi avaliar a centralização e a espessura da dentina dos canais radiculares mesiais de primeiros molares inferiores por meio de microtomografia computadorizada (micro-CT). Material e métodos: Noventa e nove molares inferiores com canais tipo IV de Vertucci foram escaneados por micro-TC. O desvio mesiodistal e o centroide foram avaliados para os canais mesiovestibular (MB) e mesiolingual (ML), nos 4mm apicais e em todo o comprimento do canal. Resultados: A espessura da dentina foi semelhante para os canais MB e ML. A espessura mais estreita foi encontrada na parede distal de um canal MB (0,07mm), enquanto a mais larga foi encontrada na parede mesial de um canal MB (2,46mm). Na análise centroide, tanto o canal MB quanto o ML exibiram desvios quando comparados ao centroide da raiz, ao longo de todo o comprimento do canal e nos 4 mm apicais. Para o canal MB, o desvio médio foi de 0,83mm (0,02mm-2,30mm) para canal inteiro e 0,18mm (0,01mm-1,01mm) para o apical de 4mm. Da mesma forma, para o canal ML, o desvio médio mediu 0,83 mm (0,05 mm-3,99 mm) para o canal inteiro e 0,21 mm (0,01 mm-1,01 mm) para os 4 mm apicais. No geral, foram observados desvios em direção mesial das raízes, sendo 69% para canais MB e 57% para canais ML para canal inteiro, e 51% para canais MB dentro dos 4 mm. A exceção foi o canal ML, que apresentou maior desvio para distal nos 4mm apicais, representando 52% dos casos. A espessura da dentina foi consistente entre os canais mesiais dos molares inferiores. Entretanto, não há centralidade dos canais mesiais em suas raízes, com frequente desvio para mesial.

Microcomputed tomographic analysis of the ulnar trochlear notch in medium- and large-breed canine cadavers with and without medial coronoid disease.

Medial coronoid disease (MCD) is a common disease often associated with thoracic limb lameness in medium- and large-breed dogs. The term MCD includes subchondral changes of the medial coronoid process (MCP), as well as pathologies of both cartilaginous surfaces. As there are only a few comprehensive and detailed studies on the trabecular structure of the medial coronoid, the goal of this study was to compare the trabecular structure of the ulnar trochlear notch of canine cadavers with and without MCD using different micro-computed tomographic (micro-CT) parameters. Fifty-eight elbow joints from 29 canine cadavers of MCD-predisposed and non-predisposed breeds (control group) were examined radiographically, macroscopically and by microcomputed tomography. The study included elbow joints of eight Labrador Retrievers (21.6-37 kg), seven Golden Retrievers (26.3-42 kg), seven Bernese Mountain dogs (31-47 kg) and seven dogs of non-predisposed breeds (19.7-52 kg) (control group). The final diagnosis of MCD was based on necropsy and micro-computed tomographic examinations. Micro-CT examinations were performed using XtremeCT II (Scanco Medical, Zurich, Switzerland) and the following parameters were examined: bone volume fraction (BV/TV), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), trabecular number (Tb.N), connectivity density (Conn.D) and degree of anisotropy (DA). Twenty-four elbows of 44 elbows of the predisposed breeds (Labrador Retrievers, Golden Retrievers Bernese Mountain dogs) showed subchondral changes and lesions of the cartilage surfaces. The result of this study is a higher density (BV/TV) of the trabecular bone of the ulnar trochlear notch in elbows affected by MCD compared to the control group. The increased density due to trabecular reconstruction in the ulnar trochlear notch is likely the result of selectively increased loading during life.

Quantitative Analysis of Bone, Blood Vessels, and Metastases in Mice Tibiae Using Synchrotron Radiation Micro-Computed Tomography.

Bone metastases are one of the most dangerous consequences of breast cancer. Early diagnosis and treatment would slow down the development of the disease and increase the survival rates of patients. Bone micro-vasculature is believed to play a major role in the development of bone metastases. It could be used for both diagnosis and as a therapeutic target. Synchrotron radiation micro-computed tomography (SR-µCT) with a contrast agent of blood vessels has been used to analyze the bone vasculature both in healthy and in metastatic bone. However, few studies have investigated the local features of blood vessels around metastases so far. For this purpose, the metastases first need to be automatically segmented. This is a challenging task, however, since the metastases do not contribute a specific contrast to the three-dimensional (3D) SR-µCT images. Here, we propose a new method for the simultaneous segmentation of bone, blood vessels, and metastases from contrast enhanced 3D SR-µCT images based on the nnU-Net architecture. In this study, we showed that only minimal training data was required to achieve a high quality of segmentation. The proposed method allowed for the automatic segmentation of metastases and provided an improved segmentation of bone and blood vessels compared to previous methods while being much more efficient to apply once trained. Further, the automatic segmentation allowed for the measurement of vascular metastases interdistance and to restrict measurements to volumes of interest around the metastases. Finally, we quantitatively analyzed blood vessel parameters locally around metastases. This allowed for the demonstration that a combined anti-angiogenic treatment significantly decreased the volume and thickness of blood vessels close to metastases. The proposed method showed the capacity of the method to reveal new aspects of the blood vessel structure interaction with metastases. This could be further used to both define new targets for precocious detection of metastases as well as to study the kinetics of metastasis development in bone and the action of drugs on this process.

Study of Microstructural, Nutritional, and Biochemical Changes in Hulled and Hulless Barley during Storage Using X-ray and Infrared Techniques.

Four varieties of barley (Esma, AC Metacalf, Tradition, and AB Cattlelac), representing four Canadian barley classes, were stored at 17% moisture content (mc) for 8 week. Stored barely was characterized using synchrotron X-ray phase contrast microcomputed tomography, synchrotron X-ray fluorescence imaging, and mid-infrared spectroscopy at the Canadian Light Source, Saskatoon. The deterioration was observed in all the selected varieties of barley at the end of 8 week of storage. Changes due to spoilage over time were observed in the grain microstructure and its nutrient distribution and composition. This study underscores the critical importance of the initial condition of barley grain microstructure in determining its storage life, particularly under unfavorable conditions. The hulled barley varieties showed more deterioration in microstructure than the hulless varieties of barley, where a direct correlation between microstructural changes and alterations in nutritional content was found. All selected barley classes showed changes in the distribution of nutrients (Ca, Fe, K, Mn, Cu, and Zn), but the two-row AC Metcalf variety exhibited more substantial variations in their nutrient distribution (Zn and Mn) than the other three varieties during storage. The two-row class barley varieties showed more changes in biochemical components (protein, lipids, and carbohydrates) than the six-row class varieties.

Combining Raman Microspectroscopy and X-ray Microcomputed Tomography for the Study of Bone Quality in Apolipoprotein-Deficient Animal Models.

Raman microspectroscopy and X-ray microcomputed tomography (micro-CT) were used for assessment of the quality of the femur and tibia bones in apolipoprotein-deficient mice compared to control littermates. The cortical and trabecular bone was investigated separately. Raman spectra revealed no differences in the bioapatite-to-collagenous matrix ratio of the cortical bone. The quantities of calcium and collagen, which were measured using atomic absorption spectrometry and thermogravimetric analysis, respectively, were also found to be equal in the two groups. Density and morphometric parameters, which were measured using micro-CT, verified the cortical mineral stability. Bone quality indices were measured using Raman spectra. A decreased collagen crosslink (trivalent-to-divalent) ratio revealed delayed maturation of the collagen network. Such a decrease has been reported in the literature to be connected to decreased bone strength. For the trabecular bone, micro-CT revealed severe osteoporosis in the knock-out group, which was evident from a decreased mineral density, trabecular thickness and increased bone surface/volume ratio. The trabecular bone was not accessible for Raman spectroscopy. According to these results, the cortical and trabecular femur bone is expected to exhibit proneness to fracturing, each for a different reason. A combination of the two techniques was regarded as necessary for an overall assessment of bone quality.

Characterization of the Metal Fused Filament Fabrication Process for Manufacturing of Pure Copper Inductors.

This work presents a comprehensive investigation into the optimization of critical process parameters associated with metal fused filament fabrication (Metal-FFF) for the production of copper-based components. The study focused on three different commercial and one self-manufactured filament, each with unique chemical compositions. These filaments were systematically optimized and the density was characterized for all processing steps, as well as the electrical conductivity on the specimen scale. Remarkably, two of the studied filaments exhibited exceptional properties after sintering with forming gas (up to 94% density and 55.75 MS/m electrical conductivity), approaching the properties measured for established manufacturing methods like metal injection molding. Finally, the research was extended to component-scale applications, demonstrating the successful fabrication of inductors with integrated cooling channels. These components exhibited water tightness and were used in induction hardening experiments, validating the practical utility of the optimized Metal-FFF process. In summary, the results show great promise in advancing the utilization of Metal-FFF in industrial contexts, particularly in the production of high-performance copper components.

A fossil "Social Stomach": Micro-CT imaging unearths first fossil record of ant proventriculi and comparative analysis with recent species.

Eusociality is an extreme form of social behavior found in some invertebrates, especially in ants, which allows them to thrive and dominate the environment through their cooperative and organized behavior. Their complex colony structure and social behavior is provided, among other things, by morphological adaptations, such as the unique structure and a huge variety of proventriculi relative to other insect groups. Here, we report the first discovery of proventriculus by X-ray microcomputed tomography (µCT) in the fossil ants †Oecophylla brischkei Mayr, 1868 and †Oecophylla crassinoda Wheeler, 1922. This is a remarkable fossil find, as it opens up new perspectives for research on phylogeny, the evolution of ant sociality and feeding behavior. We present a 3D model of the proventriculus in Oecophylla smaragdina (Fabricius, 1775) and a 3D model of the proventriculus in †Oe. brischkei and compare them. Fossil proventriculi are not fundamentally different in structure from the recent. Apparently, already in the late Eocene, the lifestyle and sociality of ants of the genus Oecophylla were similar to the one we are currently observing.

Correlative Imaging of Three-Dimensional Cell Culture on Opaque Bioscaffolds for Tissue Engineering Applications.

Three-dimensional (3D) tissue engineering (TE) is a prospective treatment that can be used to restore or replace damaged musculoskeletal tissues, such as articular cartilage. However, current challenges in TE include identifying materials that are biocompatible and have properties that closely match the mechanical properties and cellular microenvironment of the target tissue. Visualization and analysis of potential 3D porous scaffolds as well as the associated cell growth and proliferation characteristics present additional problems. This is particularly challenging for opaque scaffolds using standard optical imaging techniques. Here, we use graphene foam (GF) as a 3D porous biocompatible substrate, which is scalable, reproducible, and a suitable environment for ATDC5 cell growth and chondrogenic differentiation. ATDC5 cells are cultured, maintained, and stained with a combination of fluorophores and gold nanoparticles to enable correlative microscopic characterization techniques, which elucidate the effect of GF properties on cell behavior in a 3D environment. Most importantly, the staining protocol allows for direct imaging of cell growth and proliferation on opaque scaffolds using X-ray MicroCT, including imaging growth of cells within the hollow GF branches, which is not possible with standard fluorescence and electron microscopy techniques.

Improvement of Plasterboard Properties by the Control of Polymethylhydrosiloxane Dosage, Stirring Time, and Drying Temperature Applied to the Calcium Sulfate Hemihydrate and Water Mixture.

Plasterboard is an important building material in the construction industry because it allows for quick installation of walls, partitions, and ceilings. Although a common material, knowledge about its performance related to modern polymers and fabrication conditions is still lacking. The present work analyzes how some manufacturing factors applied during the plaster board fabrication impact on some plasterboard properties, including water absorption, flexural strength, and thermal conductivity. The manufacturing variables evaluated are the dose (D) of polymethylhydrosiloxane (PMHS), the agitation time of the mixture (H), and the drying temperature of the plaster boards after setting (T). The results suggest that factors D, H, and T induce changes in the porosity and the morphological structure of the calcium sulfate dihydrate crystals formed. Performance is evaluated at two levels of each factor following a statistical method of factorial experimental design centered on a cube. Morphological changes in the crystals of the resulting boards were evaluated with scanning electron microscopy (SEM) and the IMAGEJ image analysis program. Porosity changes were evaluated with X-ray microcomputed tomography (XMT) and 3D image analysis tools. The length-to-width ratio of the crystals decreases as it goes from low PMHS dosage to high dosage, favoring a better compaction of the plasterboard under the right stirring time and drying temperature. In contrast, the porosity generated by the incorporation of PMHS increases when going from low-level to high-level conditions and affects the maximum size of the pores being generated, with a maximum value achieved at 0.6% dosage, 40 s, and 140 °C conditions. The presence of an optimal PMHS dosage value that is approximately 0.6-1.0% is evidenced. In fact, when comparing trails without and with PMHS addition, a 10% decrease in thermal conductivity is achieved at high H (60 s) and high T (150 °C) level conditions. Water absorption decreases by more than 90% when PMHS is added, mainly due to the hydrophobic action of the PMHS. Minimum water absorption levels can be obtained at high drying temperatures. Finally, the resistance to flexion is not affected by the addition of PMHS because apparently there are two opposing forces acting: on one hand is the decrease in the length-width ratio giving more compactness, and on the other hand is the generation of pores. The maximum resistance to flexion was found around a dosage of 0.6% PMHS. In conclusion, the results suggest that the addition of PMHS, the correct agitation time of the mixture, and the drying temperature reduce the water absorption and the thermal conductivity of the gypsum boards, with no significant changes in the flexural resistance.

Accumulated Hard Tissue Debris and Root Canal Shaping Profiles Following Instrumentation with Gentlefile, One Curve, and Reciproc Blue.

INTRODUCTION: This laboratory study aimed to evaluate the accumulated hard tissue debris (AHTD) and shaping performance following root canal instrumentation with a novel stainless steel rotary system (Gentlefile; MedicNRG, Kibbutz Afikim, Israel) compared with two single-file nickel-titanium instruments of different kinematics through micro-computed tomography (micro-CT) analysis. METHODS: Thirty isthmus-containing mesial roots of human mandibular molars with a curvature of 15°-35° were scanned with micro-CT at an isotropic resolution of 12 µm and randomly assigned to three groups (n = 10) according to the instruments used for canal preparation: Gentlefile (GF; #23/.04), One Curve (OC; #25/.06; Micro-Mega, Besancon, France), and Reciproc Blue R25 (RB; #25/.08; VDW, Munich, Germany). The AHTD, unprepared canal surface area, volume changes, surface area changes, and transportation were evaluated by comparing preoperative and postoperative micro-CT images. In addition, the time required for canal preparation was recorded. Data were statistically analyzed using the one-way analysis of variance (ANOVA) test or the Kruskal-Wallis test at a significance level of 5%. RESULTS: No significant differences were observed in the percentage of AHTD or in the unprepared canal surface area between the three groups (P > .05). The RB group demonstrated a significantly greater percentage increase in volume and surface area than the GF group (P < .05). The GF and OC groups showed significantly less transportation at 3 mm from the apex than the RB group (P < .001). The OC group required significantly less time for instrumentation than the RB and GF groups (P < .05). CONCLUSIONS: Instrumentation with GF (#23/.04), OC, and RB yielded similar levels of AHTD and unprepared surface area in isthmus-containing curved canals. GF achieved this with less dentin removal and apical transportation, at the expense of a longer preparation time.

Characterization of Channeling Effects Applied to Extended-Release Matrix Tablets Containing Pirfenidone.

Pirfenidone (PRF) is an anti-fibrotic agent that has been approved by the Food and Drug Administration (FDA) for the treatment of mild to moderate idiopathic pulmonary fibrosis. However, the current oral administration dosing regimen of PRF is complex and requires high doses. Patients are instructed to take PRF three times daily, with each dose consisting of up to three capsules or tablets (600 mg/d or 1.8 g/d of PRF) taken with food. To improve the dosing regimen, efforts are being made to develop an extended-release tablet with a zero-order release pattern. In this study, two types of extended-release matrix tablets were compared: non-channeled extended-release matrix tablets (NChMT) and channeled extended-release matrix tablets (ChMT). In vitro release tests, swelling and erosion index, rheology studies, and X-ray microcomputed tomography (XRCT), were conducted. The results indicated that ChMT maintained a zero-order release pattern with a constant release rate, while NChMT exhibited a decreased release rate in the latter half of the dissolution. ChMT exhibited accelerated swelling and erosion compared to other formulations, and this was made possible by the presence of channels within the tablet. These channels allowed for thorough wetting and swelling throughout the entire depth of the tablet. The formation of channels was confirmed through XRCT images. In conclusion, the presence of channels in ChMT tablets increased the rate of swelling and erosion, resulting in a zero-order release pattern. This development offers the potential to improve the dosage of PRF and reduce its associated side effects.

Location-specific pathology analysis of the monopodial pulmonary vasculature in a rabbit model of bronchopulmonary dysplasia-A pilot study.

The mammalian pulmonary vasculature consists of functionally and morphologically heterogeneous compartments. When comparing sets of lungs, for example, in disease models or therapeutic interventions, local changes may be masked by the overall heterogeneity of the organ structure. Therefore, alterations taking place only in a sub-compartment may not be detectable by global analysis. In the monopodial lung, the characterization of distinct vessel groups is difficult, due to the asymmetrical branching pattern. In this pilot study, a previously established method to classify segments of the monopodial pulmonary arterial tree into homogeneous groups was employed. To test its suitability for experimental settings, the method was applied to a hyperoxia (HYX, ≥95% oxygen) rabbit model of bronchopulmonary dysplasia and a normoxic control group (NOX, 21% oxygen). The method allowed the identification of morphological differences between the HYX and the NOX groups. Globally visible differences in lumen diameter were pinpointed to specific lung regions. Furthermore, local changes of wall dimension and cell layers in single compartments, that would not have been identifiable in an unfocused analysis of the whole dataset, were found. In conclusion, the described method achieves a higher precision in morphological studies of lung disease models, compared to a common, global analysis approach.