Tissue Matrices Research Articles

Analyzing 3D facial morphology: Insights from a comparative European and South African study on population affinity, sex, age, and allometry

Variable growth patterns and multifactorial mechanisms cause variation in facial shape. These differences in facial morphology pose challenges for craniofacial reconstruction. Three-dimensional (3D) imaging modalities are a valuable resource for examining these variations. In this study, we used geometric morphometric methods to evaluate the effects of population affinity, sex, age, and allometry on the variation and covariation of hard and soft tissue facial morphology matrices in a sample of French and white South African individuals. Seventy-six and 108 cone-beam computed tomography scans of white South African and French nationals, respectively, were retrospectively acquired. Three-dimensional anatomical structures (hard and soft tissue matrices) were extracted using MeVisLab© v. 2.7.1 software for dense landmarking of 43 craniometric, 50 capulometric, and 559 sliding landmarks. Geometric morphometric analyses were used to quantify shape differences attributed to population affinity, sex, age, and allometry and assess the covariation between hard tissue structures and soft facial tissues. Hard and soft tissue facial matrices were influenced by population differences, sexual dimorphism, and aging. Compared to sex and age, population affinity had the strongest influence on variation. In French individuals, all hard and soft tissue matrices were sexually dimorphic, except for the eyes and left external auditory meatus (EAM). In white South Africans, sexual dimorphism was observed for the mouth, midface, and left EAM. Significant shape differences were also observed for specific age groups. The underlying skull and overlying soft tissues were strongly associated with the nose and anterior nasal aperture (correlation, r2-PLS = 0.976), followed by the right ear and right EAM (r2-PLS = 0.875) and the left ear and left EAM (r2-PLS = 0.871) in white South Africans. For French individuals, relatively weak to moderate correlations were observed, and the covariation between matrices was nonsignificant, except for the association between the right ear and right EAM (r2-PLS = 0.499). The smallest covariation was observed between the mouth and midfacial matrix in both populations (South African: r2-PLS = 0.464; French: r2-PLS = 0.367), which was also nonsignificant. This study revealed that 3D imaging technology and geometric morphometric methods can accurately quantify and visualize facial morphology differences. These methods can also evaluate the association between skull structure and soft facial features.

Nanozyme-enhanced enzyme control: Computational strategies for precise and sustained enzymatic activity in tissue engineering

Nanomaterials are revolutionizing tissue engineering by significantly enhancing enzyme-mediated chemical processes and cellular activities, thereby advancing the development of more effective therapeutic strategies. This research delves into the intricate dynamics of enzyme diffusion within tissue matrices, providing a comprehensive computational modeling framework aimed at optimizing enzyme concentration control in tissue scaffolds. Through advanced numerical algorithms and computational analysis, the study simulates optimal control strategies for regulating nanozyme levels, ensuring precise and sustained enzymatic activity within the scaffolds. A novel aspect of this work is the integration of videographic records, which offers an enriched understanding of the complex interactions between nanomaterials and biological tissues, providing detailed insights into the system’s operational dynamics. The study bridges the gap between experimental methodologies and theoretical models, aligning with the cutting-edge research in chemical physics and physical chemistry by offering novel approaches to tissue engineering challenges. The findings underscore the critical role of nanomaterials in achieving precise enzymatic control, which is pivotal for the development of advanced biomimetic systems and improved therapeutic outcomes. This work contributes to the ongoing frontier research in physical phenomena within chemistry, biology, and materials science, by providing significant new insights into enzyme delivery mechanisms and their application in biomedical contexts. The research not only highlights the importance of innovative methodologies in tissue engineering but also sets the stage for future experimental validations and potential clinical applications

New perspectives on the role of platelet factors in enhancing wound regeneration

Aim. To analyze the use of biological factors in the stimulation of the wound healing process. In the course of the study, we analysed relevant domestic and foreign literature sources on the given topic.Methods. The literature was reviewed using the key query ‘the role of biological factors in wound healing stimulation’ through the eLIBRARY and PubMed databases.Results. Currently, the range of therapeutic approaches is broad and diverse, incorporating both traditional and experimental methods such as advanced dressings, tissue matrices, growth factors (GFs), cell therapy, and nanotechnology. The wound healing process is regulated by a complex interplay of intercellular, intracellular, and extracellular signalling mechanisms across various phases of healing.Conclusion. The application of platelet-based therapies in different medical fields has shown promising outcomes in certain conditions, such as acute and chronic injuries of bone and cartilage. However, platelet-based preparations have yet to gain widespread clinical use. Several studies have demonstrated the involvement of platelets and related products, such as platelet microparticles (PMPs) and exosomes, in multiple phases of wound healing. The presence of a substantial number of biologically active molecules within platelet granules—exhibiting anti-inflammatory, angiogenic, proliferative, and other properties—renders platelets particularly attractive for use in regenerative medicine, including the stimulation of wound healin

Comparative analysis of high-throughput RNA extraction kits in Naïve Non-Human Primate (NHP) tissues for downstream applications utilizing Xeno Internal Positive Control (IPC)

Ribonucleic acid (RNA) extraction and purification play pivotal roles in molecular biology and cell and gene therapy, where the quality and integrity of RNA are critical for downstream applications. Automated high-throughput systems have gained interest due to their potential for scalability and reduced labor requirements compared to manual methods. However, ensuring high-throughput capabilities, reproducibility, and reliability while maintaining RNA yield and purity remains challenging.This study evaluated and compared the performance of four commercially available high-throughput magnetic bead-based RNA extraction kits across six types of naïve non-human primate (NHP) tissue matrices: brain, heart, kidney, liver, lung, and spleen. The assessment focused on RNA purity, yield, and extraction efficiency (EE) using Xeno Internal Positive Control (IPC) spiking.Samples (∼50 mg) were homogenized via bead-beating and processed according to the manufacturer's protocol on the KingFisher Flex platform in eight replicates. RNA purity and yield were measured using a NanoDrop® spectrophotometer, while EE was evaluated via real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR).The findings indicate consistent high RNA purity across all tested extraction kits, yet substantial variation in RNA yield. Extraction efficiency exhibited variations across tissue types, with decreasing trends observed from brain to lung tissues. These results underscore the importance of careful kit selection and method optimization for achieving reliable downstream applications. The MagMAX™ mirVana™ Total RNA Isolation Kit stands out as the most accurate and reproducible, making it the preferred choice for applications requiring high RNA quality and consistency. Other kits, such as the Maxwell® HT simplyRNA Kit, offer a good balance between cost and performance, though with some trade-offs in precision. These findings highlight the importance of selecting the appropriate RNA isolation method based on the specific needs of the research, underscoring the critical role of accurate nucleic acid extraction in gene and cell therapy research.In conclusion, this study highlights the critical factors influencing RNA extraction performance, emphasizing the need for researchers and practitioners to consider both kit performance and tissue characteristics when designing experimental protocols. These insights contribute to the ongoing efforts to enhance the reproducibility and reliability of RNA extraction methods in molecular biology and cell/gene therapy applications.

Advancing Synthetic Hydrogels through Nature-Inspired Materials Chemistry.

Synthetic extracellular matrix (ECM) mimics that can recapitulate the complex biochemical and mechanical nature of native tissues are needed for advanced models of development and disease. Biomedical research has heavily relied on the use of animal-derived biomaterials, which is now impeding their translational potential and convoluting the biological insights gleaned from in vitro tissue models. Natural hydrogels have long served as a convenient and effective cell culture tool, but advances in materials chemistry and fabrication techniques now present promising new avenues for creating xenogenic-free ECM substitutes appropriate for organotypic models and microphysiological systems. However, significant challenges remain in creating synthetic matrices that can approximate the structural sophistication, biochemical complexity, and dynamic functionality of native tissues. This review summarizes key properties of the native ECM, and discusses recent approaches used to systematically decouple and tune these properties in synthetic matrices. The importance of dynamic ECM mechanics, such as viscoelasticity and matrix plasticity, is also discussed, particularly within the context of organoid and engineered tissue matrices. Emerging design strategies to mimic these dynamic mechanical properties are reviewed, such as multi-network hydrogels, supramolecular chemistry, and hydrogels assembled from biological monomers.

Texturally-enhanced 55S0P and 45S10P Bioactive Glass ceramic particles: Sol-gel fabrication, nano-characterization and comprehensive Bio-evaluation for applications in Bone tissue engineering

Bioactive glass ceramics (BGC) play a pivotal role in bone tissue engineering, specifically addressing challenges associated with bone repair and regeneration. In this study, we developed silica-based 55S0P and 45S10P spherical BGC nanoparticles (NPs), without and with P2O5 content, using a modified Stöber sol-gel process with base hydrolysis. The fabricated spherical NPs with pronounced textural features effectively contribute to enhanced surface roughness, a desirable parameter for improved biological activity in physiologically relevant solutions and tissue matrices. A comprehensive evaluation of the structural properties of these novel spherical BGC-NPs using diverse characterization techniques revealed distinct nano-textural features in the form of cracks and pores. The HR-TEM confirmed the nanoscale dimensions (170–190 nm) of particles with spherical surface morphology and also systematically analysed via DLS, FE-SEM studies. An augmented surface area with mesopores when phosphate added to the silica glass network was revealed by BET analysis. On the biological front, the study records the formation of an HCA layer on the surfaces of the BGC-NPs, the thickness of which increased with an increase in both phosphate content and immersion time (0, 3, 14 & 28 days) in the SBF solution. The zeta potential values attain a maximum from −22 mV to −23.6 mV for 55S0P and −21.5 mV to −26.4 mV for 45S10P BGC-NPs, and improved with increased immersion time, thereby favouring cellular adhesion for desired biological responses. In vitro, cell culture studies on MG-63 cells, after 24 and 72 h of incubation, demonstrated higher cell adhesion and proliferation, with pronounced biocompatibility. In addition, these BGC-NPs exhibited good hemocompatibility even at the highest test concentration (15 mg/ml) and demonstrated effective antibacterial activity against K. pneumoniae and E. coli. A combinatorial analysis of structural and biological properties revealed that the 45S10P BGC-NPs are more potent than 55S0P BGC-NPs for desired biological performances. In conclusion, the results suggest that the sol-gel fabricated BGC-NPs with enhanced nano-textural features are evidently suitable for applications in bone tissue engineering and regenerative medicine.

Reinforced tissue matrix to strengthen the abdominal wall following reversal of temporary ostomies or to treat incisional hernias.

Abdominal wall deficiencies or weakness are a common complication of temporary ostomies, and incisional hernias frequently develop after colostomy or ileostomy takedown. The use of synthetic meshes to reinforce the abdominal wall has reduced hernia occurrence. Biologic meshes have also been used to enhance healing, particularly in contaminated conditions. Reinforced tissue matrices (RTMs), which include a biologic scaffold of native extracellular matrix and a synthetic component for added strength/durability, are designed to take advantage of aspects of both synthetic and biologic materials. To date, RTMs have not been reported to reinforce the abdominal wall following stoma reversal. To evaluate the effectiveness of using an RTM to reinforce the abdominal wall at stoma takedown sites. Twenty-eight patients were selected with a parastomal and/or incisional hernia who had received a temporary ileostomy or colostomy for fecal diversion after rectal cancer treatment or trauma. Following hernia repair and proximal stoma closure, RTM (OviTex® 1S permanent or OviTex® LPR) was placed to reinforce the abdominal wall using a laparoscopic, robotic, or open surgical approach. Post-operative follow-up was performed at 1 month and 1 year. Hernia recurrence was determined by physical examination and, when necessary, via computed tomography scan. Secondary endpoints included length of hospital stay, time to return to work, and hospital readmissions. Evaluated complications of the wound/repair site included presence of surgical site infection, seroma, hematoma, wound dehiscence, or fistula formation. The observational study cohort included 16 male and 12 female patients with average age of 58.5 years ± 16.3 years and average body mass index of 26.2 kg/m2 ± 4.1 kg/m2. Patients presented with a parastomal hernia (75.0%), incisional hernia (14.3%), or combined parastomal/incisional hernia (10.7%). Using a laparoscopic (53.6%), robotic (35.7%), or open (10.7%) technique, RTMs (OviTex® LPR: 82.1%, OviTex® 1S: 17.9%) were placed using sublay (82.1%) or intraperitoneal onlay (IPOM; 17.9%) mesh positioning. At 1-month and 1-year follow-ups, there were no hernia recurrences (0%). Average hospital stays were 2.1 d ± 1.2 d and return to work occurred at 8.3 post-operative days ± 3.0 post-operative days. Three patients (10.7%) were readmitted before the 1-month follow up due to mesh infection and/or gastrointestinal issues. Fistula and mesh infection were observed in two patients each (7.1%), leading to partial mesh removal in one patient (3.6%). There were no complications between 1 month and 1 year (0%). RTMs were used successfully to treat parastomal and incisional hernias at ileostomy reversal, with no hernia recurrences and favorable outcomes after 1-month and 1-year.

Abstract 3801: Spatially resolved laser-activated cell sorting (SLACS) for high throughput spatial cell sorting in tumor research

Abstract Understanding of tumor heterogeneity and the microenvironment is a cornerstone in advancing cancer research and therapy. The Spatially resolved Laser-Activated Cell Sorter (SLACS) represents a novel approach in cancer research, allowing for high-throughput, precise cell isolation while preserving spatial information, which is crucial for studying tumor heterogeneity and the tumor microenvironment. The SLACS instrument utilizes laser-activated technology for selective isolation of cells from heterogeneous tissue samples, optimized for accuracy, speed, and minimal cell damage. This instrument has demonstrated effective isolation of specific cell types from complex tissue matrices, maintaining cell integrity and spatial information. Its high-throughput capacity significantly reduces sample processing time. This groundbreaking tool offers detailed exploration of tumor heterogeneity and the tumor microenvironment, providing valuable insights into cancer biology and potential therapeutic targets. Citation Format: Sumin Lee, Amos Chungwon Lee. Spatially resolved laser-activated cell sorting (SLACS) for high throughput spatial cell sorting in tumor research [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3801.

Visualization of PFOA accumulation and its effects on phospholipid in zebrafish liver by MALDI Imaging.

Exposure to poly- and perfluoroalkyl substances (PFASs) can result in bioaccumulation. Initial findings suggested that PFASs could accumulate in tissues rich in both phospholipids and proteins. However, our current understanding is limited to the average concentration of PFASs or phospholipid content across entire tissue matrices, leaving unresolved the spatial variations of lipid metabolism associated with PFOA in zebrafish tissue. To address gap, we developed a novel methodology for concurrent spatial profiling of perfluorooctanoic acid (PFOA) and individual phospholipids within zebrafish hepatic tissue sections, utilizing matrix-assisted laser desorption/ionization time of flight imaging mass spectrometry (MALDI-TOF-MSI). 5-diaminonapthalene (DAN) matrix and laser sensitivity of 50.0 were optimized for PFOA detection in MALDI-TOF-MSI analysis with high spatial resolution (25μm). PFOA was observed to accumulate within zebrafish liver tissue. H&E staining results corroborating the damage inflicted by PFOA accumulation, consistent with MALDI MSI results. Significant up-regulation of 15 phospholipid species was observed in zebrafish groups exposed to PFOA, with these phospholipid demonstrating varied spatial distribution within the same tissue. Furthermore, co-localized imaging of distinct phospholipids and PFOA within identical tissue sections suggested there could be two distinct potential interactions between PFOA and phospholipids, which required further investigation. The MALDI-TOF-IMS provides a new tool to explore in situ spatial distributions and variations of the endogenous metabolites for the health risk assessment and ecotoxicology of emerging environmental pollutants.

The Effects of Massage with Topical Agents (Mepiform Ultra Scar Gel or Scarnos Gel) on Scar Tissue Thickness and Fibroblast Proliferation in Rats

Background: Keloid and hypertrophic scars are prominent scars that are excessively repaired with upregulated synthesis, deposition, and accumulation of collagen. Topical agents are used to reduce inflammation and fibrotic changes via reduced transepithelial water loss. Increased mechanical pressure applied through scar massage can accelerate scar maturation by inducing fibroblast proliferation, which enhances the remodeling of connective tissue matrices and collagen degradation.Methods: This study comparatively analyzed the effectiveness of topical agents applied to post-burn wounds on the dorsal surface of Sprague-Dawley rats through twice-daily massaging. Postoperative histological analysis of the tissues was performed after surgical en bloc removal of the treatment area at 4, 10, and 16 weeks.Results: Histological analyses revealed larger amounts of fibroblasts in Mepiform and Scarnos gel-treated tissue than in Vaseline-treated tissue. Granulation was prevented in scars treated with the topical agents.Conclusion: Mepiform Ultra scar gel and Scarnos gel, accompanied by massaging, may be effective anti-scarring topical agents to alleviate contact burn scars in Sprague-Dawley rats.

Proteomic analysis of decellularized mice liver and kidney extracellular matrices

BackgroundThe extracellular matrix (ECM) is a three-dimensional network of proteins that encases and supports cells within a tissue and promotes physiological and pathological cellular differentiation and functionality. Understanding the complex composition of the ECM is essential to decrypt physiological processes as well as pathogenesis. In this context, the method of decellularization is a useful technique to eliminate cellular components from tissues while preserving the majority of the structural and functional integrity of the ECM.ResultsIn this study, we employed a bottom-up proteomic approach to elucidate the intricate network of proteins in the decellularized extracellular matrices of murine liver and kidney tissues. This approach involved the use of a novel, perfusion-based decellularization protocol to generate acellular whole organ scaffolds. Proteomic analysis of decellularized mice liver and kidney ECM scaffolds revealed tissue-specific differences in matrisome composition, while we found a predominantly stable composition of the core matrisome, consisting of collagens, glycoproteins, and proteoglycans. Liver matrisome analysis revealed unique proteins such as collagen type VI alpha-6, fibrillin-2 or biglycan. In the kidney, specific ECM-regulators such as cathepsin z were detected.ConclusionThe identification of distinct proteomic signatures provides insights into how different matrisome compositions might influence the biological properties of distinct tissues. This experimental workflow will help to further elucidate the proteomic landscape of decellularized extracellular matrix scaffolds of mice in order to decipher complex cell–matrix interactions and their contribution to a tissue-specific microenvironment.

Juvenile Primary Ossifying Fibroma of Maxillary Sinus: A Case Report and Review of the Literature

Fibro-osseous lesions (FOLs) are a group of benign bone disorders, defined by the replacement of functional bone by fibrous connective tissue matrices. Our case report delves into the presentation of a specific FOL, namely the juvenile ossifying fibroma (JOF). This disease’s rare occurrence in a clinical setup makes documentation of such presentations vital to otorhinolaryngologists across the globe. Our article highlights the presentation of such a case of JOF in a 15-year-old male child from Pakistan. We discuss differentials, our process towards reaching a diagnosis via biopsy, and the disease’s resolution via surgical excision. Due to JOF mimicking a plethora of similar otorhinolaryngology-adjacent diseases, our article wishes to provide insight with regards to approaching patients with relevant presenting complaints, eliminating differentials, and deciding on a particular treatment regimen.

Opto-Lipidomics of Tissues.

Lipid metabolism and signaling play pivotal functions in biology and disease development. Despite this, currently available optical techniques are limited in their ability to directly visualize the lipidome in tissues. In this study, opto-lipidomics, a new approach to optical molecular tissue imaging is introduced. The capability of vibrational Raman spectroscopy is expanded to identify individual lipids in complex tissue matrices through correlation with desorption electrospray ionization (DESI) - mass spectrometry (MS) imaging in an integrated instrument. A computational pipeline of inter-modality analysis is established to infer lipidomic information from optical vibrational spectra. Opto-lipidomic imaging of transient cerebral ischemia-reperfusion injury in a murine model of ischemic stroke demonstrates the visualization and identification of lipids in disease with high molecular specificity using Raman scattered light. Furthermore, opto-lipidomics in a handheld fiber-optic Raman probe is deployed and demonstrates real-time classification of bulk brain tissues based on specific lipid abundances. Opto-lipidomics opens a host of new opportunities to study lipid biomarkers for diagnostics, prognostics, and novel therapeutic targets.

Exploration of material basis: Chemical composition profile, metabolic profile and pharmacokinetic characteristics in Xingbei Zhike granule

Headings ethnopharmacological relevanceXingbei Zhike granule (XBZK), a widely prescribed Chinese patent medicine, is known for its efficacy in clearing lung qi, relieving cough and reducing phlegm, as well as fever, dry and bitter taste, and irritability. Despite its clinical popularity, comprehensive investigations into its chemical composition, in vivo metabolism, and pharmacokinetic characteristics are limited. Aim of the studyThis study investigates the chemical composition, in vivo metabolism, and in vivo dynamics of XBZK to clarify its material basis and pharmacokinetic characteristics. Materials and methodsUltra-high performance liquid chromatography with Orbitrap tandem mass spectrometry (UPLC-Orbitrap-MS) was used to determine the chemical composition and in vivo metabolic profile of XBZK. Additionally, UPLC with triple quadrupole mass spectrometry (UPLC-TQ-MS/MS) was performed to quantify its main components and evaluate its in vivo dynamics in rat plasma. ResultsIn total, 57 components were identified in XBZK. Furthermore, 40 prototype components and 31 metabolites were detected in various biological matrices of rats, including plasma, tissues, bile, feces, and urine. After administration, the area under the curve (AUC) for ephedrine (Eph), pseudoephedrine (Peph), neotuberostemonine (Neo), amygdalin (Amy), and enoxolone (Eno) exhibited a strong linear relationship with the administered dose (r > 0.9) in all rats. And gender-related differences in the absorption of peiminine (Pmn), peimisine (Pms), and chrysin-7-O-glucuronide (Cog) were notable among rats, with male rats showing a dose-dependent pattern of absorption, while female rats exhibited minimal absorption. ConclusionsXBZK contains 57 components, primarily composed of flavonoids, alkaloids, and coumarins. The eight main components were rapidly absorbed and eliminated, with some, such as Eph, Peph, Neo, Amy and Eno, following a linear pharmacokinetic pattern. Furthermore, Pmn, Pms and Cog were well absorbed in male rats, showing a dose-dependent behavior.

Targeting Hyaluronan Synthesis in Cancer: A Road Less Travelled

Hyaluronan is one of the major components of the extracellular matrix and is involved in the regulation of multiple processes in both human physiology and disease. In human cancers, hyaluronan metabolism displays remarkable alterations, leading to the accumulation of large amounts of hyaluronan matrices in the tumoural tissues. The altered levels of hyaluronan in the tumours stem from the enhanced expression and activity of hyaluronan synthases in both tumour and stromal cells. Moreover, hyaluronidase activity is also upregulated in cancer, leading to the generation of lower molecular weight hyaluronan fragments that in turn assist tumour growth, neo-angiogenesis and the metastatic cascade. Hyaluronan accumulation in malignant tissues not only assists tumour growth and metastases but is also associated with worse outcomes in cancer patients. Therefore, targeting hyaluronan synthesis emerges as an interesting strategy that might be employed for cancer treatment. This review article summarises current evidence and discusses ways to move forward in the field of targeting hyaluronan synthesis for cancer therapy.

The Use of Acellular Tissue Matrices in Gender-Affirming Surgery: Review of the Literature and Institutional Experience.

Wound healing complications are not uncommon after genital gender-affirming surgery and can pose significant challenges for the reconstructive surgeon. Acellulartissue matrices are products that contain extracellular matrix compounds without living cells and are used to expedite and improve wound healing.Some of these products have been described for a variety of different indicationsin gender-affirming surgery. In this paper, the authors present a review of the current literature on the use of acellular tissue matrices in gender-affirming surgeryas well as the authors' institutionalexperience in using these products.

Applications of hybrid structure light three‐dimensional scanning in neurodegenerative disease autopsy brain banking

AbstractBackgroundPostmortem magnetic resonance imaging (MRI) and three‐dimensional (3D) printing has been demonstrated to be a feasible approach for the creation of tissue matrices to aid in postmortem brain dissection. We aimed to extend this approach using hybrid structure light technology (visible and infrared) 3D scanning to generate brain models as a digital resource and operational tool for human neurodegenerative disease brain banks.MethodA consecutive series of 56 brains accessioned into the Glenn Biggs Institute Brain Bank were imaged using an EinScan H handheld 3D scanner. Five brains were selected for generation of tissue matrices, with infratentorial structures removed using 3D object post‐processing software tools. Matrices were printed using a Raise3D Pro2 printer and 1.75‐mm polylactic acid premium filament.ResultEach scan was approximately an hour to complete, including capture and alignment of a lateral and a medial surface scan for each brain. Raw scan files (DAT and BRAP‐PRJ) averaged 5.47 GB and 3D object files were 12,991 KB for 3MF and 18,067 KB for OBJ file types. For cases with tissue matrices, average print time was 42 hours with an average filament consumption of 338 g. Brain cutting with the tissue matrices was performed in a standardized manner, with superb control for section thickness and planar orientation compared to brains cut without a matrix.Conclusion3D scanning of postmortem autopsied brains is a simple and useful strategy to generate models with utility in guided tissue sectioning and storage (practice), surface area volumetric analysis (research) and virtual reality/augmented reality neuroanatomy training (education) for Alzheimer’s disease and related dementias.

Novel Humanized G6PD Deficient Mice Reveal Mechanisms of Increased Tolerance to Exercise As Gleaned By Multi-Omics in Models of Critical Speed

INTRODUCTION: Glucose-6-phosphate-dehydrogenase (G6PD) deficiency is the most common enzymatic deficiency in humans, affecting ~500 million people or ~6% of mankind. G6PD is the rate limiting enzyme of the pentose phosphate pathway (PPP), the main antioxidant pathway in mature red blood cells (RBCs). Activation of the PPP generates NADPH, an essential reducing equivalent for antioxidant metabolism that protects against intra- and extra-vascular hemolysis in response to oxidant stress. Impaired G6PD activity makes RBCs susceptible to hemolysis induced by oxidant challenges. Exercise-induced oxidant stress is a well-established concept in the field of physiology. Individuals with G6PD deficiency are recommended to refrain from strenuous exercise, as they are deemed to be at increased risk for hemolytic events in response to exercise-induced oxidant stress. While biochemically sound, this rationale has never been tested mechanistically, owing to ethical and technical limitations. As such, knowledge is limited regarding molecular impacts of exercise activity in G6PD deficient individuals. METHODS: Here we generated novel humanized mouse models carrying either (i) human canonical G6PD (normal activity), or two of the most common G6PD variants (ii) African (V68M - < 10% residual enzymatic activity) or (iii) Mediterranean (S188F - <1% residual enzymatic activity). We leveraged these mouse models to assess critical speed (CS), a functional measurement of the speed-to-duration relationship. To expand the molecular resolution of genetically associated adaptations to exercise, CS tests were combined with measurements of hemodynamics - including cardiac output, imaging (e.g., scanning electron microscopy of RBCs before and after exercise) and multi-omics (mass spectrometry-based metabolomics, lipidomics and proteomics) characterization of plasma, RBCs, and multiple tissue matrices including liver, heart, kidney, lung, spleen, and muscles (gastrocnemius and soleus). RESULTS: First of all, we confirmed a significant decrease in G6PD activity in RBCs from mice carrying the African and Mediterranean variants ( Figure 1). Unexpectedly, G6PD deficient mice showed ~10% faster CS compared to mice carrying the canonical human G6PD (p<0.0001 and p<0.02, Figure 1). While scanning electron microscopy showed differences before and after exercise, with accumulation of reversibly modified morphologies across all groups after the run, no significant differences were observed across groups. No significant changes in hemolysis markers were observed in plasma samples after exercise, neither at the metabolic (e.g., heme catabolites) or proteomics level (e.g., hemoglobin chains, RBC-derived proteins). Within hemodynamics, the hG6PD Med- mice are showing a higher efficiency in their cardiac function through cardiac function (p<0.005 - Figure 1). Pathway analysis of metabolomics data generated from RBC and plasma fractions showed significant decreases in arginine metabolism, increased glycolysis, and elevated Krebs cycle intermediates, in addition to expected differences in PPP. Specifically, in hG6PD A- RBC acylcarnitines accumulate after running. In hG6PD Med-, there are clear left ventricular differences highlighted in energy metabolism ( Figure 1). Multiomics analyses identified five major tissue clusters between heart, muscle, liver, kidney, and spleen, with G6PD function having the most significant metabolic impact on liver and soleus. RBC proteomics results highlighted deceased ferroportin (key iron transporter) in both the deficient groups, with markedly significant changes in the hG6PD Med- (p<0.0001). CONCLUSION: Novel humanized G6PD deficient mice maintain faster CS compared to mice. Omics approaches afford the opportunity to identify metabolic correlates to measurements of performance in relevant animal models, such as the CS model, which offer expanded mechanistic understanding of molecular bases of CS and the associated impact of G6PD deficiency. Our results not only challenge the assumption that exercise-induced oxidant stress predisposes the ~500M individuals with G6PD deficiency worldwide to increased hemolytic propensity, but rather indicate a metabolic advantage of functional polymorphisms to G6PD with respect to exercise performance, an observation that could contribute explaining the prevalence of such mutation in human populations.

Developmental roles of glomerular epithelial protein-1 in mice molar morphogenesis.

Glomerular epithelial protein-1 (Glepp1), a R3 subtype family of receptor-type protein tyrosine phosphatases, plays important role in the activation of Src family kinases and regulates cellular processes such as cell proliferation, differentiation, and apoptosis. In this study, we firstly examined the functional evaluation of Glepp1 in tooth development and morphogenesis. The precise expression level and developmental function of Glepp1 were examined by RT-qPCR, in situ hybridization, and loss and gain of functional study using a range of in vitro organ cultivation methods. Expression of Glepp1 was detected in the developing tooth germs in cap and bell stage of tooth development. Knocking down Glepp1 at E13 for 2days showed the altered expression levels of tooth development-related signaling molecules, including Bmps, Dspp, Fgf4, Lef1, and Shh. Moreover, transient knock down of Glepp1 revealed alterations in cellular physiology, examined by the localization patterns of Ki67 and E-cadherin. Similarly, knocking down of Glepp1 showed disrupted enamel rod and interrod formation in 3-week renal transplanted teeth. In addition, due to attrition of odontoblastic layers, the expression signals of Dspp and the localization of NESTIN were almost not detected after knock down of Glepp1; however, their expressions were increased after Glepp1 overexpression. Thus, our results suggested that Glepp1 plays modulating roles during odontogenesis by regulating the expression levels of signaling molecules and cellular events to achieve the proper structural formation of hard tissue matrices in mice molar development.

Immunoassay testing forbarbiturates using alternative matrices inpostmortem tissues from cats anddogs.

The barbiturate drug pentobarbital is commonly used by veterinarians for the euthanasia of domestic animals. During the veterinary forensic autopsy, it is sometimes necessary to determine whether the animal was chemically euthanized with pentobarbital. The use of a human immunochromatographic test for barbiturate screening utilizing dog or cat urine has been previously validated; however, the use of alternative matrices for this purpose is yet to be explored when urine is not available. Postmortem heart, liver, spleen, skeletal muscle, blood and/or urine samples from 20 dogs and 26 cats with a reported chemical euthanasia status were processed using two different methods, bead homogenization and sonication, and screened for barbiturates using a human immunochromatographic test. There was 100% agreement of the immunochromatographic test results using the sonication method with the reported euthanasia status of both dogs and cats. Using the bead homogenization method, agreement with the reported euthanasia status was 93.3% and 96.7% for dogs and cats, respectively, due to invalid test results from four dog and two cat samples. A subset of liver samples (10 canine and 10 feline) was analyzed via gas chromatography-mass spectrometry, and there was 100% agreement between the immunochromatographic test results and gas chromatography-mass spectrometry results for both cats and dogs. Overall, our results support the use of a variety of alternative matrices for barbiturate screening in cats and dogs.