Embedding Biomimetic Vascular Networks via Coaxial Sacrificial Writing into Functional Tissue.

Printing human tissues and organs replete with biomimetic vascular networks is of growing interest. While it is possible to embed perfusable channels within acellular and densely cellular matrices, they do not currently possess the biomimetic architectures found in native vessels. Here, coaxial sacrificial writing into functional tissues (co-SWIFT) is developed, an embedded bioprinting method capable of generating hierarchically branching, multilayered vascular networks within both granular hydrogel and densely cellular matrices. Coaxial printheads are designed with an extended core-shell configuration to facilitate robust core-core and shell-shell interconnections between printed branching vessels during embedded bioprinting. Using optimized core-shell ink combinations, biomimetic vessels composed of a smooth muscle cell-laden shell that surrounds perfusable lumens are coaxially printed into granular matrices composed of: 1) transparent alginate microparticles, 2) sacrificial microparticle-laden collagen, or 3) cardiac spheroids derived from human induced pluripotent stem cells. Biomimetic blood vessels that exhibit good barrier function are produced by seeding these interconnected lumens with a confluent layer of endothelial cells. Importantly, it is found that co-SWIFT cardiac tissues mature under perfusion, beat synchronously, and exhibit a cardio-effective drug response in vitro. This advance opens new avenues for the scalable biomanufacturing of vascularized organ-specific tissues for drug testing, disease modeling, and therapeutic use.

Corrigendum: Applying new approach methodologies to assess next-generation tobacco and nicotine products.

[This corrects the article DOI: 10.3389/ftox.2024.1376118.].

Hibernating/Awakening Nanomotors Promote Highly Efficient Cryopreservation by Limiting Ice Crystals.

The disruptions caused by ice crystal formation during the cryopreservation of cells and tissues can cause cell and tissue damage. Thus, preventing such damage during cryopreservation is an important but challenging goal. Here, a hibernating/awakening nanomotor with magnesium/palladium covering one side of a silica platform (Mg@Pd@SiO2) is proposed. This nanomotor is used in the cultivation of live NCM460 cells to demonstrate a new method to actively limit ice crystal formation and enable highly efficient cryopreservation. Cooling Mg@Pd@SiO2 in solution releases Mg2+/H2 and promotes the adsorption of H2 at multiple Pd binding sites on the cell surface to inhibit ice crystal formation and cell/tissue damage; additionally, the Pd adsorbs and stores H2 to form a hibernating nanomotor. During laser-mediated heating, the hibernating nanomotor is activated (awakened) and releases H2, which further suppresses recrystallization and decreases cell/tissue damage. These hibernating/awakening nanomotors have great potential for promoting highly efficient cryopreservation by inhibiting ice crystal formation.

Macrophage Polarization during MRONJ Development in Mice.

Macrophages are important regulators of bone remodeling, and M1 polarization is observed in the setting of medication-related osteonecrosis of the jaws (MRONJ). Here, we characterize the phenotype of macrophages during early stages of MRONJ development in zoledronate (ZA)-treated mice with periodontal disease and explore the role of rosiglitazone, a drug that has been reported to lower the M1/M2 macrophage ratio, in MRONJ burden. Mice received ZA, and experimental periodontal disease (EPD) was induced around their second left maxillary molar. The mice were euthanized 1, 2, or 4 wk later. Micro-computed tomography and histologic and immunohistochemical analyses were carried out. In a separate experiment, mice were treated with ZA in the absence or presence of rosiglitazone, EPD was induced for 5 wk, and the MRONJ burden was assessed. An M1 predilection was noted in ZA versus vehicle (Veh) mice at 1, 2, or 4 wk after ligature placement. M1 cells were found to be positive for MMP-13, and their presence coincided with disruption of the surrounding collagen network in ZA mice. Rosiglitazone caused a reversal in the M1/M2 polarization in Veh and ZA mice. Rosiglitazone did not cause significant radiographic changes 5 wk after EPD in Veh or ZA animals. Importantly, percentage osteonecrosis and bone exposure were decreased in the rosiglitazone-treated versus nontreated ZA sites 5 wk after EPD. Our data point to an important role of M1 macrophage polarization with an overexpression of MMP-13 in the early phases of MRONJ development and provide insight into the use of interventional approaches promoting an M2 phenotype as a preventative means to alleviate MRONJ burden.

Optimized extraction methodology for phenolic compounds in soil and plant tissues: Their implications in plant growth and gall formation.

Phenolic compounds, abundant secondary metabolites in plants, profoundly influence soil ecosystems, plant growth, and interactions with herbivores. Phenolic in soil microorganisms have the potential to impact a wide range of activities in plant-soil interactions. However, the existing methods for measuring microbial activity are typically time-consuming, intricate, and expensive. In this study, we propose modifications to the method used for the extraction and quantification of various types of phenolics in soil and plant tissues. There have been substantial advancements in research aimed at extracting, identifying, and quantifying phenolic compounds in the plant and soil samples. This study discusses the use of different methodologies in the analysis of phenolic compounds. In addition, we investigated the effect of phenolics on plant growth and cues in gall-forming under environmental disturbances.•This method is the optimum way to extract phenolic from soil and microbial activity in bulk and rhizosphere soil.•It can be used on any soil type and plant tissue, metabolites extracted from living organisms.

Condition of oral tissues in children with congenital cleft lip and palate.

OBJECTIVE: Aim: To study the condition of oral tissues in children with congenital complete cleft lip, alveolar process, hard and soft palate. PATIENTS AND METHODS: Materials and Methods: From the examined 470 children National Specialized Children's Hospital "OKHMATDYT" (Kyiv, Ukraine) with congenital cleft lip and palate was analyzed: 302 patients aged 8-18 years were subject to in-depth analysis for clinical and radiological - 192 with unilateral and 110 with bilateral complete cleft lip, alveolar process, hard and soft palate. RESULTS: Results: The average value of primary adention in patients with unilateral and bilateral complete combined cleft is 69.53%, but in females this indicator is higher and in unilateral cleft 92.18% for female against 53.17% for male. In females with unilateral cleft retention - 40.62% and overcomplete - 10,93%. The same high indicators in bilateral cleft: retention - 36.58% and overcomplete - 12.19%. Retention and overcomplete have higher values for men - 44.93% and 23.19%, respectively. Chewing efficiency in females with bilateral cleft as a result of primary dentition is below 80%. Affected by caries - 90.73% in both groups. Inflammatory processes in the periodontal tissues are revealed (80,75%): chronic catarrhal gingivitis, chronic hypertrophic gingivitis, chronic generalized periodontitis. Manifestations of atopic and angular cheilitis in 39.09% and 23.63% with bilateral cleft lip and palate, glossitis in 29.09%. CONCLUSION: Conclusions: Patients with congenital complete cleft lip, alveolar process, hard and soft palate have high rates of adentia, retention, overcomplete dentition and a wide range of diseases of the oral cavity, which negatively affects surgical and orthodontic rehabilitation.

Identifying Heterogeneous Micromechanical Properties of Biological Tissues via Physics-Informed Neural Networks.

The heterogeneous micromechanical properties of biological tissues have profound implications across diverse medical and engineering domains. However, identifying full-field heterogeneous elastic properties of soft materials using traditional engineering approaches is fundamentally challenging due to difficulties in estimating local stress fields. Recently, there has been a growing interest in data-driven models for learning full-field mechanical responses, such as displacement and strain, from experimental or synthetic data. However, research studies on inferring full-field elastic properties of materials, a more challenging problem, are scarce, particularly for large deformation, hyperelastic materials. Here, a physics-informed machine learning approach is proposed to identify the elasticity map in nonlinear, large deformation hyperelastic materials. This study reports the prediction accuracies and computational efficiency of physics-informed neural networks (PINNs) in inferring the heterogeneous elasticity maps across materials with structural complexity that closely resemble real tissue microstructure, such as brain, tricuspid valve, and breast cancer tissues. Further, the improved architecture is applied to three hyperelastic constitutive models: Neo-Hookean, Mooney Rivlin, and Gent. The improved network architecture consistently produces accurate estimations of heterogeneous elasticity maps, even when there is up to 10% noise present in the training data.

The Impact of Low-Power Therapeutic Lasers at 904†nm on the Healing Process of Wounds and the Relationships Between Extracellular Matrix Components and Myofibroblasts.

The goal of the study was to further our understanding of the functions that myofibroblasts do, how they interact with the matrix's extracellular elements, and how laser therapy at 904 nm works. Thirty Wistar rats with superficial wounds on the backs were used in this study. The laser therapy device used two energy levels of a 904 nm laser with an output power of 60 mW. The outcomes had been evaluated for one, three, five, seven, and fourteen days following laser therapy. Numerous methods, including as histology, immunohistochemistry, and scanning electron microscopy, have been used to evaluate the tissues.There was a statistically important (P ≤ .05) decrease in the number of inflammatory cells and the degree of edema after laser therapy. On the other hand, the amounts of elastic fiber as well as collagen slightly increased. On the third post-laser treatment day, there was a statistically significant distinction (P < .05) in the number of myofibroblasts with the desmin; smooth muscle alpha-actin phenotype between the laser-treated and control groups.These results suggest that, compared to a higher energy of 5.6 J, laser therapy at a lower energy of 3.6 J may be more effective in stimulating myofibroblast differentiation during the initial phases of wound healing. These findings show how low-power laser therapy promotes wound healing.

Comparison of cation and anion-mediated resolution enhancement of bioprinted hydrogels for membranous tissue fabrication.

Fabrication of engineered thin membranous tissues (TMTs) presents a significant challenge to researchers, as these structures are small in scale, but present complex anatomies containing multiple stratified cell layers. While numerous methodologies exist to fabricate such tissues, many are limited by poor mechanical properties, need for post-fabrication, or lack of cytocompatibility. Extrusion bioprinting can address these issues, but lacks the resolution necessary to generate biomimetic, microscale TMT structures. Therefore, our goal was to develop a strategy that enhances bioprinting resolution below its traditional limit of 150 µm and delivers a viable cell population. We have generated a system to effectively shrink printed gels via electrostatic interactions between anionic and cationic polymers. Base hydrogels are composed of gelatin methacrylate type A (cationic), or B (anionic) treated with anionic alginate, and cationic poly-L-lysine, respectively. Through a complex coacervation-like mechanism, the charges attract, causing compaction of the base GelMA network, leading to reduced sample dimensions. In this work, we evaluate the role of both base hydrogel and shrinking polymer charge on effective print resolution and cell viability. The alginate anion-mediated system demonstrated the ability to reach bioprinting resolutions of 70 µm, while maintaining a viable cell population. To our knowledge, this is the first study that has produced such significant enhancement in extrusion bioprinting capabilities, while also remaining cytocompatible.

How anatomy influences measurements of snakes.

Anatomy compromises the precision and accuracy of measurements made of the body length and head size of live snakes. Body measures (snout-vent length, SVL) incorporate many synovial intervertebral joints, each allowing flexion and limited extension and compression. Radiographs of the trunk in 14 phylogenetically diverse species in resting and stretched conditions combined with dissections and histological analysis of intervertebral joints show that the synovial nature of these joints underlies the variance in SVL measures. Similarly, the ubiquity and variety of viscoelastic tissues connecting mobile snout and jaw elements of alethinophidian snakes underlie variances in length and width measures of the head. For the overall size of the head and jaw apparatus, the part that can be most easily and relatively precisely measured for many snakes is the mandible because it has only one mobile joint. As to accuracy, the anatomy of intervertebral and cranial joints supports the hypothesis that in living snakes, the head and trunk have no exact size.

Therapeutic Application of Extracellular Vesicles Derived from Mesenchymal Stem Cells in Domestic Animals.

Recently, the therapeutic potential of extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) has been extensively studied in both human and veterinary medicine. EVs are nano-sized particles containing biological components commonly found in other biological materials. For that reason, EV isolation and characterization are critical to draw precise conclusions during their investigation. Research on EVs within veterinary medicine is still considered in its early phases, yet numerous papers were published in recent years. The conventional adult tissues for deriving MSCs include adipose tissue and bone marrow. Nonetheless, alternative sources such as synovial fluid, endometrium, gingiva, and milk have also been intermittently used. Fetal adnexa are amniotic membrane/fluid, umbilical cord and Wharton's jelly. Cells derived from fetal adnexa exhibit an intermediate state between embryonic and adult cells, demonstrating higher proliferative and differentiative potential and longer telomeres compared to cells from adult tissues. Summarized here are the principal and recent preclinical and clinical studies performed in domestic animals such as horse, cattle, dog and cat. To minimize the use of antibiotics and address the serious issue of antibiotic resistance as a public health concern, they will undoubtedly also be utilized in the future to treat infections in domestic animals. A number of concerns, including large-scale production with standardization of EV separation and characterization techniques, must be resolved for clinical application.

Mapping of Human Polyomavirus in Renal Cell Carcinoma Tissues.

Worldwide, the incidence of renal cell carcinoma (RCC) is rising, accounting for approximately 2% of all cancer diagnoses and deaths. The etiology of RCC is still obscure. Here, we assessed the presence of HPyVs in paraffin-embedded tissue (FFPE) resected tissue from patients with RCC by using different molecular techniques. Fifty-five FFPE tissues from 11 RCC patients were included in this study. Consensus and HPyV-specific primers were used to screen for HPyVs. Both PCR approaches revealed that HPyV is frequently detected in the tissues of RCC kidney resections. A total of 78% (43/55) of the tissues tested were positive for at least one HPyV (i.e., MCPyV, HPyV6, HPyV7, BKPyV, JCPyV, or WUyV). Additionally, 25 tissues (45%) were positive for only one HPyV, 14 (25%) for two HPyVs, 3 (5%) for three HPyVs, and 1 one (1%) tissue specimen was positive for four HPyVs. Eleven (20%) RCC specimens were completely devoid of HPyV sequences. MCPyV was found in 24/55 RCC tissues, HPyV7 in 19, and HPyV6 in 8. The presence of MCPyV and HPyV6 was confirmed by specific FISH or RNA-ISH. In addition, we aimed to confirm HPyV gene expression by IHC. Our results strongly indicate that these HPyVs infect RCC and nontumor tissues, possibly indicating that kidney tissues serve as a reservoir for HPyV latency. Whether HPyVs possibly contribute to the etiopathogenesis of RCC remains to be elucidated.

RiboScreenTM Technology Delivers a Ribosomal Target and a Small-Molecule Ligand for Ribosome Editing to Boost the Production Levels of Tropoelastin, the Monomeric Unit of Elastin.

Elastin, a key structural protein essential for the elasticity of the skin and elastogenic tissues, degrades with age. Replenishing elastin holds promise for anti-aging cosmetics and the supplementation of elastic activities of the cardiovascular system. We employed RiboScreenTM, a technology for identifying molecules that enhance the production of specific proteins, to target the production of tropoelastin. We make use of RiboScreenTM in two crucial steps: first, to pinpoint a target ribosomal protein (TRP), which acts as a switch to increase the production of the protein of interest (POI), and second, to identify small molecules that activate this ribosomal protein switch. Using RiboScreenTM, we identified ribosomal protein L40, henceforth eL40, as a TRP switch to boost tropoelastin production. Drug discovery identified a small-molecule hit that binds to eL40. In-cell treatment demonstrated activity of the eL40 ligand and delivered increased tropoelastin production levels in a dose-dependent manner. Thus, we demonstrate that RiboScreenTM can successfully identify a small-molecule hit capable of selectively enhancing tropoelastin production. This compound has the potential to be developed for topical or systemic applications to promote skin rejuvenation and to supplement elastic functionality within the cardiovascular system.

Clinical efficacy of two vertical soft tissue augmentation techniques for peri-implant crestal bone level stability: A randomized clinical trial.

OBJECTIVES: This study aimed to compare the efficacy of two techniques-acellular dermal matrix (ADM) grafting and tenting technique (TT)-for soft tissue height (STH) augmentation simultaneous to implant placement to minimize peri-implant crestal bone level (CBL) changes. METHODS: Forty patients with a healed single mandibular posterior edentulous site with a thin soft tissue phenotype were enrolled. Twenty patients received simultaneously to implant placement ADM grafting, while the others received submerged healing abutment (TT). Clinical peri-implant soft tissue height and radiographic CBL changes were measured at restoration delivery and 1-year follow-up. RESULTS: Both techniques effectively increased soft tissue thickness, resulting in a final average STH of 3.4 ± 0.5 mm after augmentation. On average, soft tissue increased by 1.6 ± 0.5 mm in group ADM and by 1.8 ± 0.4 mm in group TT after augmentation. In Group ADM, mesial CBL decreased from 0.4 ± 0.3 mm to 0.1 ± 0.2 mm, and distal CBL decreased from 0.5 ± 0.3 mm to 0.2 ± 0.3 mm over 1 year. In Group TT, mesial CBL remained stable at 0.3 ± 0.2 mm, while distal CBL reduced slightly from 0.5 ± 0.5 mm to 0.3 ± 0.2 mm. Both groups showed minimal changes in CBL, indicating great stability (pmesial = 0.003, pdistal = 0.004). TT was particularly effective in preventing mesial bone loss (pmesial = 0.019). The mesial CBL changes significantly differed between groups (p = 0.019), and not significantly at distal sites (p = 0.944). Neither treatment exhibited significant bone remodeling below the implant shoulder. CONCLUSION: This study suggests that both techniques were successful in STH augmentation, and they may effectively reduce peri-implant crestal bone level changes, with TT being slightly superior. TT was more prone to post-surgical complications. This RCT was not registered before participant recruitment and randomization.

Light in evaluation of molecular diffusion in tissues: Discrimination of pathologies.

The evaluation of the diffusion properties of different molecules in tissues is a subject of great interest in various fields, such as dermatology/cosmetology, clinical medicine, implantology and food preservation. In this review, a discussion of recent studies that used kinetic spectroscopy measurements to evaluate such diffusion properties in various tissues is made. By immersing ex vivo tissues in agents or by topical application of those agents in vivo, their diffusion properties can be evaluated by kinetic collimated transmittance or diffuse reflectance spectroscopy. Using this method, recent studies were able to discriminate the diffusion properties of agents between healthy and diseased tissues, especially in the cases of cancer and diabetes mellitus. In the case of cancer, it was also possible to evaluate an increase of 5% in the mobile water content from the healthy to the cancerous colorectal and kidney tissues. Considering the application of some agents to living organisms or food products to protect them from deterioration during low temperature preservation (cryopreservation), and knowing that such agent inclusion may be reversed, some studies in these fields are also discussed. Considering the broadband application of the optical spectroscopy evaluation of the diffusion properties of agents in tissues and the physiological diagnostic data that such method can acquire, further studies concerning the optimization of fruit sweetness or evaluation of poison diffusion in tissues or antidote application for treatment optimization purposes are indicated as future perspectives.

Attitudes of Oral Surgeons and Periodontists towards Immediate Dental Implant Placement.

Objectives: This cross-sectional study aimed to evaluate the factors that determine the choice of oral surgeons and periodontists to perform immediate dental implant placement. Material and Methods: An anonymous survey was carried out from January 6, 2024 to February 29, 2024. The questionnaire was distributed online to Lithuanian specialists - oral surgeons and periodontists, who perform implantation procedures. A total of 186 professionals were included in this survey. Chi-square test, its degrees of freedom was used for the analysis of variables. Results: The main reason for refusing immediate implant placement is a periapical lesion greater than 5 mm, reported by 91.7% of oral surgeons and 96.9% of periodontists. Good aesthetics and preservation of anatomical structures are identified as an advantage by 99.2% of oral surgeons and 92.3% of periodontists. In the aesthetic zone, for periodontists, the main criterion for choosing a method is the quantitative and qualitative indicators of the soft tissue of the extraction socket 96.9%, and for oral surgeons - the morphology of the bone walls of the socket 87.6%. Only 43.1% of periodontists and 33.9% of oral surgeons are familiar with and use extraction socket morphology assessment classifications for immediate dental implant placement. Conclusions: Taking into account study's results, it is recommended to adjust the teaching programs at Universities and to increase the knowledge of specialists performing dental implantation procedures, by carrying out continuous educational programs.

Biomaterials for extrusion-based bioprinting and biomedical applications.

Amongst the range of bioprinting technologies currently available, bioprinting by material extrusion is gaining increasing popularity due to accessibility, low cost, and the absence of energy sources, such as lasers, which may significantly damage the cells. New applications of extrusion-based bioprinting are systematically emerging in the biomedical field in relation to tissue and organ fabrication. Extrusion-based bioprinting presents a series of specific challenges in relation to achievable resolutions, accuracy and speed. Resolution and accuracy in particular are of paramount importance for the realization of microstructures (for example, vascularization) within tissues and organs. Another major theme of research is cell survival and functional preservation, as extruded bioinks have cells subjected to considerable shear stresses as they travel through the extrusion apparatus. Here, an overview of the main available extrusion-based printing technologies and related families of bioprinting materials (bioinks) is provided. The main challenges related to achieving resolution and accuracy whilst assuring cell viability and function are discussed in relation to specific application contexts in the field of tissue and organ fabrication.

Variants in the Kallikrein Gene Family and Hypermobile Ehlers-Danlos Syndrome.

Hypermobile Ehlers-Danlos syndrome (hEDS) is a common heritable connective tissue disorder that lacks a known genetic etiology. To identify genetic contributions to hEDS, whole exome sequencing was performed on families and a cohort of sporadic hEDS patients. A missense variant in Kallikrein-15 (KLK15 p. Gly226Asp), segregated with disease in two families and genetic burden analyses of 197 sporadic hEDS patients revealed enrichment of variants within the Kallikrein gene family. To validate pathogenicity, the variant identified in familial studies was used to generate knock-in mice. Consistent with our clinical cohort, Klk15 G224D/+ mice displayed structural and functional connective tissue defects within multiple organ systems. These findings support Kallikrein gene variants in the pathogenesis of hEDS and represent an important step towards earlier diagnosis and better clinical outcomes.

Applications of extraembryonic tissue-derived cells in vascular tissue regeneration.

Vascular tissue engineering is a promising approach for regenerating damaged blood vessels and developing new therapeutic approaches for heart disease treatment. To date, different sources of cells have been recognized that offer assistance within the recovery of heart supply routes and veins with distinctive capacities and are compelling for heart regeneration. However, some challenges still remain that need to be overcome to establish the full potential application of these cells. In this paper, we review the different cell sources used for vascular tissue engineering, focusing on extraembryonic tissue-derived cells (ESCs), and elucidate their roles in cardiovascular disease. In addition, we highlight the intricate interplay between mechanical and biochemical factors in regulating mesenchymal stem cell (MSC) differentiation, offering insights into optimizing their application in vascular tissues.

Intelligent tumor tissue classification for Hybrid Health Care Units.

Introduction: In the evolving healthcare landscape, we aim to integrate hyperspectral imaging into Hybrid Health Care Units to advance the diagnosis of medical diseases through the effective fusion of cutting-edge technology. The scarcity of medical hyperspectral data limits the use of hyperspectral imaging in disease classification. Methods: Our study innovatively integrates hyperspectral imaging to characterize tumor tissues across diverse body locations, employing the Sharpened Cosine Similarity framework for tumor classification and subsequent healthcare recommendation. The efficiency of the proposed model is evaluated using Cohen's kappa, overall accuracy, and f1-score metrics. Results: The proposed model demonstrates remarkable efficiency, with kappa of 91.76%, an overall accuracy of 95.60%, and an f1-score of 96%. These metrics indicate superior performance of our proposed model over existing state-of-the-art methods, even in limited training data. Conclusion: This study marks a milestone in hybrid healthcare informatics, improving personalized care and advancing disease classification and recommendations.