Search

Including sensor information in medical interventions aims to support surgeons to decide on subsequent action steps by characterizing tissue intraoperatively. With bladder cancer, an important issue is tumor recurrence because of failure to remove the entire tumor. Impedance measurements can help to classify bladder tissue and give the surgeons an indication on how much tissue to remove. Over the years of research, it became obvious that electrical impedance spectroscopy is a very promising tool for tissue differentiation, but also a very sensitive one. While differentiation in preliminary studies shows great potential, challenges arise when transferring this concept to real, intraoperative conditions, mainly due to the influence of preoperative radiotherapy, possibly different tumor types, and mechanical tissue deformations due to peristalsis or unsteady contact force of the sensor. This work proposes a patient-based classification approach that evaluates the distance of an unknown measurement to a healthy reference of the same patient, essentially a relative classification of the difference in impedance that is robust against inter-individual differences and systematic errors. A diversified dataset covering multiple disturbance scenarios is recorded. Two alternatives to define features from the impedance data are investigated, namely using measurement points and model-based parameters. Based on the distance of the feature vector of a unknown measurement to a healthy reference, a Gaussian process classifier is trained. The approach achieves a high classification accuracy of up to 100% on noise-free impedance data recorded under controlled conditions. Even when the differentiation is more ambiguous due to external disturbances, the presented approach still achieves a classification accuracy of 80%. These results are a starting point to tackle intraoperative bladder tissue characterization and decrease the recurrence rate. [ABSTRACT FROM AUTHOR]

-2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside (THSG) is a bioactive component in the Chinese herb Polygonum multiflorum, recognized for its anti-inflammatory and lipid-lowering properties. Human dental pulp stem cells (hDPSCs) have excellent capabilities in tooth regeneration, wound healing, and neural repair. The exosomes (Exo) released by hDPSCs contain bioactive molecules that influence cell proliferation, differentiation, and immune responses. Therefore, we aimed to unveil the potential of THSG-Exo and evaluate its regenerative capabilities through the in vitro experiment and rat bone defect model. The effects of hDPSC-derived exosomes, with or without THSG treatment, on repair and bone regeneration were evaluated through in vitro and in vivo studies. Finally, we conducted a proteomic analysis to meticulously compare the compositional contents of the two types of exosomes. In vitro data showed that 10 and 100 μM THSG-Exo enhanced cell proliferation and osteogenic differentiation, reducing wound size to 40 % of its original size. In our maxillary bone defect rat model, THSG-Exo significantly increased bone volume, trabecular thickness, and bone density in the bone defect area. In addition, proteomic analysis of THSG-Exo revealed diverse proteins linked to bone differentiation and tissue repair, including bone morphogenetic protein-1 (BMP-1) and tumor necrosis factor (TNF)-α-stimulated gene 6 (TNFAIP6). Our searches in functional databases revealed that THSG-Exo is involved in numerous biological pathways. THSG-Exo enhanced cell proliferation, wound healing, and osteogenesis in vitro , while also expediting tissue repair and bone regeneration in vivo. The protein diversity of THSG-Exo contributes significant value in both basic and regenerative medicine. [ABSTRACT FROM AUTHOR]

Breast cancer remains one of the most challenging diseases to treat due to its heterogeneity, propensity to recur, capacity to spread to distant vital organs, and, ultimately, patient death. Estrogen receptor-positive illness comprises the most common breast cancer subtype. Preclinical progress is hampered by the scarcity of medication-naïve estrogen receptor-positive tumour models that recapitulate metastatic development and treatment resistance. It is becoming increasingly clear that loss of differentiation and increased cellular stemness and plasticity are important causes of cancer evolution, heterogeneity, recurrence, metastasis, and treatment failure. Therefore, it has been suggested that reprogramming cancer cell differentiation could offer an effective method of reversing cancer through terminal differentiation and maturation. In this context, the hormone prolactin is well recognized for its pivotal involvement in the development of the mammary glands lobuloalveolar tissue and the terminal differentiation that drives the production of the milk protein gene and lactation. Additionally, numerous studies have examined the engagement of prolactin in breast cancer as a differentiation player that resulted in the ablation of tumour growth and progression. Here, we showed that a pre-treatment of the estrogen-positive breast cancer cell line with prolactin led to a considerable improvement in the sensitivity of this cancer cell to Tamoxifen endocrine therapy. We also showed a favourable prognostic value of prolactin receptors/estrogen receptors 1 (or alpha) co-expression on breast cancer patients outcomes, and this co-expression is highly correlated with the well-differentiated breast tumour type. Our results revealed a fruitful aspect of the effects of prolactin in improving the responses of breast cancer cells to conventional endocrine therapy. Moreover, these findings further validated the ability of prolactin as a persuader of a more differentiated and less aggressive breast cancer phenotype. Hence, it suggested a potential implication of prolactin as a therapeutic candidate. [ABSTRACT FROM AUTHOR]

The androgen receptor (AR) is central in prostate tissue identity and differentiation, and controls normal growth-suppressive, prostate-specific gene expression. It also drives prostate tumorigenesis when hijacked for oncogenic transcription. The execution of growth-suppressive AR transcriptional programs in prostate cancer (PCa) and the potential for reactivation remain unclear. Here, we use a genome-wide approach to modulate canonical androgen response element (ARE) motifs—the classic DNA binding elements for AR—to delineate distinct AR transcriptional programs. We find that activating these AREs promotes differentiation and growth-suppressive transcription, potentially leading to AR+ PCa cell death, while ARE repression is tolerated by PCa cells but deleterious to normal prostate cells. Gene signatures driven by ARE activity correlate with improved prognosis and luminal phenotypes in PCa patients. Canonical AREs maintain a normal, lineage-specific transcriptional program that can be reengaged in PCa cells, offering therapeutic potential and clinical relevance. Androgen response elements (AREs) regulation produce opposite effects in normal and cancer prostate cells. Here, authors engineer a modifier of ARE-containing chromatin (MACC) to define the elements responsible for a normal growth-suppressive program, which can be reengaged in prostate cancer cells. [ABSTRACT FROM AUTHOR]

Introduction and Purpose: Mouse mesenchymal stem cells (MSCs) provide a resourceful tool to study physiological and pathological aspects of adipogenesis. Bone marrow-derived MSCs (BM-MSCs) and adipose tissue-derived MSCs (ASCs) are widely used for these studies. Since there is a wide spectrum of methods available, the purpose is to provide a focused hands-on procedural guide for isolation and characterization of murine BM-MSCs and ASCs and to effectively differentiate them into adipocytes. Methods and Results: Optimized harvesting procedures for murine BM-MSCs and ASCs are described and graphically documented. Since macrophages reside in bone-marrow and fat tissues and regulate the biological behaviour of BM-MSCs and ASCs, we included a procedure to deplete macrophages from the MSC preparations. The identity and stemness of BM-MSCs and ASCs were confirmed by flow cytometry using established markers. Since the composition and concentrations of adipogenic differentiation cocktails differ widely, we present a standardized four-component adipogenic cocktail, consisting of insulin, dexamethasone, 3-isobutyl-1-methylxanthine, and indomethacin to efficiently differentiate freshly isolated or frozen/thawed BM-MSCs and ASCs into adipocytes. We further included visualization and quantification protocols of the differentiated adipocytes. Conclusion: This laboratory protocol was designed as a step-by-step procedure for harvesting murine BM-MSCs and ASCs and differentiating them into adipocytes. [ABSTRACT FROM AUTHOR]

Simple Summary: Induced pluripotent stem cells have revolutionized biological research. These are adult cells that can be reprogrammed to differentiate into any cell type in the body. Although iPSCs have been widely studied in mammals, recent progress has extended their use to avian species, particularly chickens. Chicken iPSCs provide a unique model for investigating avian development, pathology, and genetics. Through the manipulation of these cells, researchers may get insights into cellular differentiation, disease progression, and possible tissue healing mechanisms. The development of efficient reprogramming methods, both viral and non-viral techniques, has significantly improved the generation of chicken iPSCs. In addition, a better understanding of the underlying molecular mechanisms, such as key signaling pathways and transcription factors, has driven the science forward. There are many uses of chicken-induced pluripotent stem cells. They may be used in the modeling of human diseases, the formulation of new drugs, and the engineering of genetically modified birds for agricultural purposes. As research proceeds, we are expecting more revolutionary discoveries and new uses of chicken iPSCs. The advent of iPSCs has greatly advanced the fields of degenerative biology, disease capabilities, and regenerative medicine. iPSCs are a new and compelling model system for studying biological processes, as well as evaluating novel therapeutic interventions. The concept of iPSCs was created in 2006 by Takahashi and Yamanaka, reprogramming mouse fibroblasts with the transcription factors Oct4, Sox2, Klf4, and c-Myc OSKM. Similar to ESCs, iPSCs can self-renew and differentiate into all types of cells. This approach has since been applied to the avian species (e.g., chickens), allowing new opportunities for scientific investigation. Thus, the chicken model organism fills the important gap between mammalian and non-mammalian vertebrates. The relatively short life cycle of the chicken model organism, alongside well-characterized genetics, makes it a perfect candidate for stem cell studies. With advancements in the technology behind avian iPSCs, it is now possible to generate pluripotent cells with high production rates. This allows new perspectives regarding germline transmission, tissue differentiation, and genetic trait improvement. This review summarizes the major advances made in the generation of chicken iPSCs using novel reprogramming techniques and the identification of species-specific transcription factors important for maintaining pluripotency. The applications of avian iPSCs in genetic engineering studies, disease resistance, and avian conservation are also discussed. This review will also indicate how the use of iPSCs in poultry biotechnology can further improve meat and egg production. Finally, the review discusses current challenges, such as incomplete understanding of the avian reprogramming environment and epigenetic modifications, and suggests future directions for refining the iPSC technology in avian mode. [ABSTRACT FROM AUTHOR]

In head and neck surgery, continuous intraoperative tissue differentiation is of great importance to avoid injury to sensitive structures such as nerves and vessels. Hyperspectral imaging (HSI) with neural network analysis could support the surgeon in tissue differentiation. A 3D Convolutional Neural Network with hyperspectral data in the range of 400 − 1000 nm is used in this work. The acquisition system consisted of two multispectral snapshot cameras creating a stereo-HSI-system. For the analysis, 27 images with annotations of glandular tissue, nerve, muscle, skin and vein in 18 patients undergoing parotidectomy are included. Three patients are removed for evaluation following the leave-one-subjectout principle. The remaining images are used for training, with the data randomly divided into a training group and a validation group. In the validation, an overall accuracy of 98.7% is achieved, indicating robust training. In the evaluation on the excluded patients, an overall accuracy of 83.4% has been achieved showing good detection and identification abilities. The results clearly show that it is possible to achieve robust intraoperative tissue differentiation using hyperspectral imaging. Especially the high sensitivity in parotid or nerve tissue is of clinical importance. It is interesting to note that vein was often confused with muscle. This requires further analysis and shows that a very good and comprehensive data basis is essential. This is a major challenge, especially in surgery. [ABSTRACT FROM AUTHOR]

Tactile information acquired through palpation plays a crucial role in relation to surface characterisation and tissue differentiation - an essential clinical requirement during surgery. In the case of Minimally Invasive Surgery, access is restricted, and tactile feedback available to surgeons is therefore reduced. This paper presents a novel stiffness controllable, dynamic force range sensor that can provide remote haptic feedback. The sensor has an abraded optical fibre integrated into a silicone dome. Forces applied to the dome change the curvature of the optical fibres, resulting in light attenuation. By changing the pressure within the dome and thereby adjusting the sensor's stiffness, we are able to modify the force measurement range. Results from our experimental study demonstrate that increasing the pressure inside the dome increases the force range whilst decreasing force sensitivity. We show that the maximum force measured by our sensor prototype at 20 mm/min was 5.02 N, 6.70 N and 8.83 N for the applied pressures of 0 psi (0 kPa), 0.5 psi (3.45 kPa) and 1 psi (6.9 kPa), respectively. The sensor has also been tested to estimate the stiffness of 13 phantoms of different elastic moduli. Results show the elastic modulus sensing range of the proposed sensor to be from 8.58 to 165.32 kPa. [ABSTRACT FROM AUTHOR]

The generation of a self-formed, ectodermal, autonomous multi-zone (SEAM) from human induced pluripotent stem cells (hiPSCs) offers a unique perspective to study the dynamics of ocular cell differentiation over time. Here, by utilising single-cell transcriptomics, we have (i) identified, (ii) molecularly characterised and (iii) ascertained the developmental trajectories of ectodermally-derived ocular cell populations which emerge within SEAMs as they form. Our analysis reveals interdependency between tissues of the early eye and delineates the sequential formation and maturation of distinct cell types over a 12-week period. We demonstrate a progression from pluripotency through to tissue specification and differentiation which encompasses both surface ectodermal and neuroectodermal ocular lineages and the generation of iPSC-derived components of the developing cornea, conjunctiva, lens, and retina. Our findings not only advance the understanding of ocular development in a stem cell-based system of human origin, but also establish a robust methodological paradigm for exploring cellular and molecular dynamics during SEAM formation at single-cell resolution and highlight the potential of hiPSC-derived systems as powerful platforms for modelling human eye development and disease. Single-cell transcriptomic analyses delineate the differentiation of human iPSCs into ectodermal ocular lineages, highlighting defined developmental trajectories and the sequential formation and maturation of distinct ocular cell types in vitro. [ABSTRACT FROM AUTHOR]

The transcription factor Runx2 plays a crucial role in regulating osteogenic differentiation and skeletal development. This factor not only controls the expression of genes involved in bone formation, but also interacts with signaling pathways such as the Notch pathway, which are essential for body development. However, studies have produced conflicting results regarding the relationship between Runx2 and the Notch pathway. Some studies suggest a synergistic interaction between these molecules, while others suggest an inhibitory one, for example, the interplay between Notch signaling, Runx2, and vitamin D3 in osteogenic differentiation and bone remodeling. The findings suggest a complex relationship between Notch signaling and osteogenic differentiation, with ongoing research needed to clarify the mechanisms involved and resolve existing contradictions regarding role of Notch in this process. Additionally, there is increasing evidence of contradictory roles for Runx2 in various tissues and organs, both under normal conditions and in pathological states. This diversity of roles makes Runx2 a potential therapeutic target, offering new directions for research. In this review, we have discussed the mechanisms of osteogenic differentiation and the important role of Runx2 in this process. We have also examined its relationship with different signaling pathways. However, there are still many uncertainties and inconsistencies in our current understanding of these interactions. Additionally, given that Runx2 is also involved in numerous other events in various tissues, we have tried to comprehensively examine its functions outside the skeletal system. [ABSTRACT FROM AUTHOR]

The physiological functions of specific areas of the nervous system, such as the spinal cord, are directional dependent, relying on the ordered arrangement of cells and extracellular matrix. Here, an automated electrospinning method is used to make fibers with different diameters by changing the concentrations of polycaprolactone/gelatine solutions. The impact of fiber diameter, orientation, and the inclusion of superparamagnetic iron oxide nanoparticles on neural stem cell (NSC) behavior and differentiation is investigated. Fibers with an average diameter of 1189 ± 274 nm greatly improve neuronal differentiation. The highest relative expression of the neuronal marker Tuj‐1 and the formation of interconnected neuronal networks with neurites aligned along the fiber axis demonstrate this. Incorporating SPIONS into the fibers do not damage the cells, and aligned fibers do much better than random fibers at cell proliferation, migration, and neurite alignment. Further, magnetoresponsive fiber‐based hydrogels are fabricated by embedding SPIONS‐loaded fibers within a collagen matrix, which enables remote alignment of the fibers via a magnetic field. Postcrosslinking maintains this alignment and induces significant neurite orientation within the hydrogels. The potential use of automated, magnetically responsive electrospun fibers as flexible supports for NSC differentiation and neural tissue engineering guidance is demonstrated. [ABSTRACT FROM AUTHOR]

Poplars are essential tree species with critical roles in wood production, ecological conservation, environmental protection, and climate change mitigation. Plants of different poplar populations exhibit varying responses to biotic and abiotic stresses, largely driven by phytohormones—key regulators of plant growth and stress responses. Phytohormones control fundamental processes such as cell division, growth, tissue differentiation, organ development, and resistance to environmental challenges such as temperature extremes and drought. In this study, we conducted a quantitative analysis of phytohormone levels in the leaves of two populations of poplar species from the same geographical region over a one-year period, covering the early, mid, and late growth stages. Our results revealed that salicylic acid (SA) and jasmonic acid (JA) concentrations in the stress-resistant poplar (RP) increased significantly during the late growth stage (September), particularly in response to elevated pathogen pressure and fluctuating climatic conditions. In contrast, the susceptible poplar (SP) consistently exhibited lower levels of SA and JA across all growth stages, particularly during the mid-growth stage (characterized by high temperatures and low precipitation) and the late growth stage, when pathogen stress typically intensifies. The limited increase in phytohormone levels in the SP suggests a weaker defense response compared with the RP. These findings demonstrate that climatic factors significantly influence the phytohormonal dynamics in poplars, with the RP exhibiting more robust regulation of SA and JA to mitigate both biotic and abiotic stresses. This study provides a theoretical framework for better understanding the resistance mechanisms in Populus and offers insights for improving stress tolerance in future breeding programs. [ABSTRACT FROM AUTHOR]

Near-infrared (NIR) spectroscopy is a powerful non-invasive technique for assessing the optical properties of human tissues, capturing spectral signatures that reflect their biochemical and structural characteristics. In this study, we investigated the use of NIR reflectance spectroscopy combined with chemometric analysis to distinguish between patients with Essential Tremor (ET) and healthy individuals. ET is a common movement disorder characterized by involuntary tremors, often making it difficult to clinically differentiate from other neurological conditions. We hypothesized that NIR spectroscopy could reveal unique optical fingerprints that differentiate ET patients from healthy controls, potentially providing an additional diagnostic tool for ET. We collected NIR reflectance spectra from both extracranial (biceps and triceps) and cranial (cerebral cortex and brainstem) sites in ET patients and healthy subjects. Using Partial Least Squares Discriminant Analysis (PLS-DA) and Partial Least Squares (PLS) regression models, we analyzed the optical properties of the tissues and identified significant wavelength peaks associated with spectral differences between the two groups. The chemometric analysis successfully classified subjects based on their spectral profiles, revealing distinct differences in optical properties between cranial and extracranial sites in ET patients compared to healthy controls. Our results suggest that NIR spectroscopy, combined with machine learning algorithms, offers a promising non-invasive method for the in vivo characterization and differentiation of tissues in ET patients. [ABSTRACT FROM AUTHOR]

Abstract Including sensor information in medical interventions aims to support surgeons to decide on subsequent action steps by characterizing tissue intraoperatively. With bladder cancer, an important issue is tumor recurrence because of failure to remove the entire tumor. Impedance measurements can help to classify bladder tissue and give the surgeons an indication on how much tissue to remove. Over the years of research, it became obvious that electrical impedance spectroscopy is a very promising tool for tissue differentiation, but also a very sensitive one. While differentiation in preliminary studies shows great potential, challenges arise when transferring this concept to real, intraoperative conditions, mainly due to the influence of preoperative radiotherapy, possibly different tumor types, and mechanical tissue deformations due to peristalsis or unsteady contact force of the sensor. This work proposes a patient-based classification approach that evaluates the distance of an unknown measurement to a healthy reference of the same patient, essentially a relative classification of the difference in impedance that is robust against inter-individual differences and systematic errors. A diversified dataset covering multiple disturbance scenarios is recorded. Two alternatives to define features from the impedance data are investigated, namely using measurement points and model-based parameters. Based on the distance of the feature vector of a unknown measurement to a healthy reference, a Gaussian process classifier is trained. The approach achieves a high classification accuracy of up to 100% on noise-free impedance data recorded under controlled conditions. Even when the differentiation is more ambiguous due to external disturbances, the presented approach still achieves a classification accuracy of 80%. These results are a starting point to tackle intraoperative bladder tissue characterization and decrease the recurrence rate.

The interaction between the surgical instrument and tissue is a yet little-exploited source of information. Vibroacoustic signals resulting from an interaction can be analyzed to extract relevant information related to the tissue. This work presents a summary of feasibility studies with synthetic-, animal-, and ex-vivo tissue specimen. A standard palpation probe is equipped with a vibration measurement system. A manual and robot-assisted setup is used for data acquisition. Simple features derived from the signals are used in a subsequent classification step using kNN and SVM. The obtained results show the capability to differentiate various tissues, such as subchondral bone with an accuracy of 98.44%. This demonstrates the potential of the approach to provide tissue related information based on instrument interactions.

Background: Cancer is the leading cause of disease-related mortality in children. Causes of leukemia, the most common form, are largely unknown. Growing evidence points to an origin in-utero, when global redistribution of DNA methylation occurs driving tissue differentiation. Methods: Epigenome-wide DNA methylation was profiled in surrogate (blood) and target (bone marrow) tissues at birth, diagnosis, remission and relapse of pediatric pre-B acute lymphoblastic leukemia (pre-B ALL) patients. Double-blinded analyses was performed between prospective cohorts extending from birth to diagnosis and retrospective studies backtracking from clinical disease to birth. Validation was carried out using independent technologies and populations. Results: The imprinted and immuno-modulating VTRNA2-1 was hypermethylated (FDR<0.05) at birth in nested cases relative to controls in all tested populations (totaling 317 cases and 483 controls), including European and Hispanic ancestries. VTRNA2-1 methylation was stable over follow-up years after birth and across surrogate, target and other tissues (n=5,023 tissues; 30 types). When profiled in leukemic tissues from two clinical cohorts (totaling 644 cases), VTRNA2-1 methylation exhibited higher levels at diagnosis relative to controls, it reset back to normal levels at remission, and then re-increased to above control levels at relapse. Hypermethylation was significantly associated with worse pre-B ALL patient survival and with reduced VTRNA2-1 expression (n=2,294 tissues; 26 types), supporting a functional and translational role for VTRNA2-1 methylation. Conclusion: This study provides proof-of-concept to detect at birth epigenetic precursors of pediatric pre-B ALL. These alterations were reproducible with different technologies, in three continents and in two ethnicities, and can offer biomarkers for early detection and prognosis as well as actionable targets for therapy. Key points: • Precursors of pediatric acute lymphoblastic leukemia may be of epigenetic origin, detectable since birth and affecting patient prognosis. • These epigenetic precursors can be robust over several years and across several populations, ethnicities and surrogate and target tissues. [ABSTRACT FROM AUTHOR]

Our research explores the detrimental effects of microplastic (MP) exposure on adipose tissue aging and function, emphasizing the potential health risks associated with environmental pollutants. Utilizing both in vivo and in vitro models, we discovered that MPs accumulate in adipose tissues, leading to cellular senescence, inflammation, and hindered adipogenic differentiation. Notably, our findings demonstrate that MPs prompt an aging response in both epididymal and inguinal white adipose tissue, increase senescence-associated β-galactosidase activity, and upregulate key senescence and inflammatory markers. Furthermore, we show that MPs disrupt normal adipogenic differentiation by reducing lipid droplet formation and downregulating critical adipogenic markers. These insights highlight the urgent need for further investigation into the long-term consequences of MP pollution on biological aging and underscore the importance of developing public health strategies to mitigate these effects. [ABSTRACT FROM AUTHOR]

Primary cilia on granule cell neuron progenitors in the developing cerebellum detect sonic hedgehog to facilitate proliferation. Following differentiation, cerebellar granule cells become the most abundant neuronal cell type in the brain. While granule cell cilia are essential during early developmental stages, they become infrequent upon maturation. Here, we provide nanoscopic resolution of cilia in situ using large-scale electron microscopy volumes and immunostaining of mouse cerebella. In many granule cells, we found intracellular cilia, concealed from the external environment. Cilia were disassembled in differentiating granule cell neurons—in a process we call cilia deconstruction—distinct from premitotic cilia resorption in proliferating progenitors. In differentiating granule cells, cilia deconstruction involved unique disassembly intermediates, and, as maturation progressed, mother centriolar docking at the plasma membrane. Unlike ciliated neurons in other brain regions, our results show the deconstruction of concealed cilia in differentiating granule cells, which might prevent mitogenic hedgehog responsiveness. Ciliary deconstruction could be paradigmatic of cilia removal during differentiation in other tissues. [ABSTRACT FROM AUTHOR]

Simple Summary: Skeletal muscle is an endocrine organ capable of responding to and releasing hormones and hormonally active molecules. In particular, sex hormones (e.g., testosterone) specifically regulate metabolic activity, growth, and tissue differentiation. However, the biological and clinical responses to the same type of hormone treatment differ between subjects with male and female karyotypes. Characterization of the biomolecular mechanisms underlying the sexual dimorphism in response to sex hormones is crucial for the design of new methodological approaches aimed at highlighting sex differences in the patho/physiological aspects caused by the use (and abuse) of sex hormones. To date, in vivo and ex vivo studies are very limited, mainly for ethical reasons; it is therefore of fundamental importance to use experimental approaches based on cellular models at the preclinical level. On the other hand, most in vitro studies do not take into account the chromosomal structure of the cell models used, with the result that 'sex' is often a 'confounding factor'. This highlights the need for basic research to develop preclinical models that take into account chromosomal sex differences to ensure the transferability of the data obtained in vivo and to adequately design relevant experimental studies. Therefore, given the role of skeletal muscle in biological responses to testosterone, the aim of this project is to assess the presence of sex differences in endocrine–metabolic cellular responses to increasing doses of testosterone by simulating functional modifications of skeletal muscle cells in vitro. Muscle tissue is an important target of sex steroids, and particularly, testosterone plays essential roles in muscle cell metabolism. Wide ranges of studies have reported sex differences in basal muscle steroidogenesis, and recently several genes have been identified to be regulated by androgen response elements that show innate sex differences in muscle. However, studies accounting for and demonstrating cell sexual dimorphism in vitro are still scarce and not well characterized. Here, we demonstrated the ability of 46XX and 46XY human primary skeletal muscle cells to differently activate steroidogenesis in vitro, likely related to sex-chromosome onset, and to differently induce hormone release after increasing doses of testosterone exposure. Cells were treated with testosterone at concentrations of 0.5, 2, 5, 10, 32, and 100 nmol/L for 24 h. Variations in 17β-HSD, 5α-R2, CYP-19 expression, DHT, estradiol, and androstenedione release, as well as IL6 and IL8 release, were analyzed, respectively, by RT-PCR, ELISA, and luminex-assay. Following testosterone treatments, and potentially at any concentration level, an increase in the expression of 17β-HSD, 5α-R2, and CYP-19 was observed in 46XY cells, accompanied by elevated levels of DHT, androstenedione, and IL6/IL8 release. Following the same treatment, 46XX cells exhibited an increase in 5α-R2 and CYP-19 expression, a conversion of androgens to estrogens, and a reduction in IL6 and IL8 release. In conclusion, this study demonstrated that sex-chromosome differences may influence in vitro muscle cell steroidogenesis and hormone homeostasis, which are pivotal for skeletal muscle metabolism. [ABSTRACT FROM AUTHOR]

Various materials have been used to promote human dental pulp stem cells (hDPSCs) differentiation to produce dentin bridge formation with less-than-optimal results. Polydatin (PD), a naturally present material with osteogenic properties can be a promising material in the pulp regeneration/repair process. The aim of this study was to evaluate the effect of (PD) on the viability and differentiation of human dental pulp stem cells. PD effect on hDPSCs in terms of cellular viability, alkaline phosphatase (ALP) production, and messenger RNAs (mRNA) of odontogenic markers production using quantitative reverse transcription polymerase chain reaction (RT-qPCR) were evaluated. In addition, mineral deposits were detected with Alizarin red stain. The viable hDPSCs in the presence of 0.01 μM and 0.1 μM PD were significantly higher than the control on days 3 and 7, respectively. In addition, ALP activity of hDPSCs was significantly increased with 0.01, 0.1, and 1 μM of PD. In addition, increased expression mRNAs of ALP, osteocalcin (OC), osteonectin (ON), osteopontin (OP), Runt-related transcription factor-2 (RUNX-2), dentin sialophosphoprotein (DSPP), and dentin matrix protein-1 (DMP-1) was observed after PD treatment, however, the difference was not statistically significant. Furthermore, increased size of mineral deposits was observed with PD. PD promoted the expression of markers associated with odontogenic differentiation and mineralized tissue deposition in hDPSCs. [ABSTRACT FROM AUTHOR]

Human periodontal ligament stem cells (hPDLSCs) differentiate into periodontal ligament (PDL) fibroblasts, osteoblasts, and cementoblasts. To identify inducers of PDL fibroblastic differentiation, monoclonal antibody series were developed a series of against membrane/extracellular matrix (ECM) molecules through decoy immunization. The anti-PDL13 antibody targets ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1), renowned for regulating skeletal and soft tissue mineralization. ENPP1 accumulates in the periodontal ligament region of tooth roots, and specifically localizes to the cell boundaries and elongated processes of the fibroblastic cells. As ENPP1 expression increases during fibroblastic differentiation, mineralization induced by tissue-nonspecific alkaline phosphatase (TNAP), a pyrophosphate-degrading enzyme, is completely inhibited. This is consistent with ENPP1 and TNAP acting in opposition, and TGF-ß1-induced ENPP1 expression creates an essential environment for PDL fibroblast differentiation. Representative fibroblastic differentiation markers decrease with endogenous ENPP1 inhibition by siRNA and antibody blocking. ENPP2 generates lipid signaling molecules. In contrast to ENPP1, ENPP2 disappears in TGF-ß1-induced PDL fibroblasts. Ectopic expression of ENPP2 hinders TGF-ß1-induced PDL fibroblastic differentiation. Suppression of ENPP1 and ENPP2 leads to severe defects in undifferentiated and differentiated cells, demonstrating that these two factors play opposing roles in soft and hard tissue differentiation but can complement each other for cell survival. In conclusion, increased ENPP1 is crucial for TGF-ß1-induced PDL differentiation, while ENPP2 and TNAP can inhibit ENPP1. ENPP1 and ENPP2 exhibit complementary functions in the cell survival. [ABSTRACT FROM AUTHOR]

In recent years, with the advent of a super-aged society, lifelong dental care has gained increasing emphasis, and implant therapy for patients with an edentulous jaw has become a significant option. However, for implant therapy to be suitable for elderly patients with reduced regenerative and immunological capabilities, higher osteoconductive and antimicrobial properties are required on the implant surfaces. Silicon nitride, a non-oxide ceramic known for its excellent mechanical properties and biocompatibility, has demonstrated high potential for inducing hard tissue differentiation and exhibiting antibacterial properties. In this study, silicon nitride was deposited on pure titanium metal surfaces and evaluated for its biocompatibility and antibacterial properties. The findings indicate that silicon nitride improves the hydrophilicity of the material surface, enhancing the initial adhesion of rat bone marrow cells and promoting hard tissue differentiation. Additionally, the antibacterial properties were assessed using Staphylococcus aureus, revealing that the silicon nitride-coated surfaces exhibited significant antibacterial activity. Importantly, no cytotoxicity was observed, suggesting that silicon nitride-coated titanium could serve as a novel implant material. [ABSTRACT FROM AUTHOR]

Background: Chronic thromboembolic pulmonary hypertension (CTEPH) is a serious pulmonary vascular disease characterized by residual thrombi in the pulmonary arteries and distal pulmonary microvascular remodeling. The pathogenesis of CTEPH remains unclear, but many factors such as inflammation, immunity, coagulation and angiogenesis may be involved. Monocytes are important immune cells that can differentiate into macrophages and dendritic cells and play an important role in thrombus formation. However, the distribution, gene expression profile and differentiation trajectory of monocyte subsets in CTEPH patients have not been systematically studied. This study aims to reveal the characteristics and functions of monocytes in CTEPH patients using single-cell sequencing technology, and to provide new insights for the diagnosis and treatment of CTEPH. Methods: Single-cell RNA sequencing (scRNA-seq) were performed to analyze the transcriptomic features of peripheral blood mononuclear cells (PBMCs) from healthy controls, CTEPH patients and the tissues from CTEPH patients after the pulmonary endarterectomy (PEA). We established a CTEPH rat model with chronic pulmonary embolism caused by repeated injection of autologous thrombi through a central venous catheter, and used flow cytometry to detect the proportion changes of monocyte subsets in CTEPH patients and CTEPH rat model. We also observed the infiltration degree of macrophage subsets in thrombus tissue and their differentiation relationship with peripheral blood monocyte subsets by immunofluorescence staining. Results: The results showed that the monocyte subsets in peripheral blood of CTEPH patients changed significantly, especially the proportion of CD16+ monocyte subset increased. This monocyte subset had unique functional features at the transcriptomic level, involving processes such as cell adhesion, T cell activation, coagulation response and platelet activation, which may play an important role in pulmonary artery thrombus formation and pulmonary artery intimal remodeling. In addition, we also found that the macrophage subsets in pulmonary endarterectomy tissue of CTEPH patients showed pro-inflammatory and lipid metabolism reprogramming features, which may be related to the persistence and insolubility of pulmonary artery thrombi and the development of pulmonary hypertension. Finally, we also observed that CD16+ monocyte subset in peripheral blood of CTEPH patients may be recruited to pulmonary artery intimal tissue and differentiate into macrophage subset with high expression of IL-1β, participating in disease progression. Conclusion: CD16+ monocytes subset had significant gene expression changes in CTEPH patients, related to platelet activation, coagulation response and inflammatory response. And we also found that these cells could migrate to the thrombus and differentiate into macrophages with high expression of IL-1b involved in CTEPH disease progression. We believe that CD16+ monocytes are important participants in CTEPH and potential therapeutic targets. [ABSTRACT FROM AUTHOR]

Sugar beet (Beta vulgaris L.) is one of the most important sugar crops, accounting for nearly 30% of the world's annual sugar production. And it is mainly distributed in the northwestern, northern, and northeastern regions of China. However, Cercospora leaf spot (CLS) is the most serious and destructive foliar disease during the cultivation of sugar beet. In plants, the bZIP gene family is one of important family of transcription factors that regulate many biological processes, including cell and tissue differentiation, pathogen defense, light response, and abiotic stress signaling. Although the bZIP gene family has been mentioned in previous studies as playing a crucial role in plant defense against diseases, there has been no comprehensive study or functional analysis of the bZIP gene family in sugar beet with respect to biotic stresses. In this study, we performed a genomewide analysis of bZIP family genes (BvbZIPs) in sugar beet to investigate their phylogenetic relationships, gene structure and chromosomal localization. At the same time, we observed the stomatal and cell ultrastructure of sugar beet leaf surface during the period of infestation by Cercospora beticola Sacc (C. beticola). And identified the genes with significant differential expression in the bZIP gene family of sugar beet by qRT-PCR. Finally we determined the concentrations of SA and JA and verified the associated genes by qRT-PCR. The results showed that 48 genes were identified and gene expression analysis indicated that 6 BvbZIPs were significantly differential expressed in C. beticola infection. It is speculated that these BvbZIPs are candidate genes for regulating the response of sugar beet to CLS infection. Meanwhile, the observation stomata of sugar beet leaves infected with C. beticola revealed that there were also differences in the surface stomata of the leaves at different periods of infection. In addition, we further confirmed that the protein encoded by the SA signaling pathway-related gene BVRB_ 9g222570 in high-resistant varieties was PR1, which is closely related to systemic acquired resistance. One of the protein interaction modes of JA signal transduction pathway is the response of MYC2 transcription factor caused by JAZ protein degradation, and there is a molecular interaction between JA signal transduction pathway and auxin. Despite previous reports on abiotic stresses in sugar beet, this study provides very useful information for further research on the role of the sugar beet bZIP gene family in sugar beet through experiments. The above research findings can promote the development of sugar beet disease resistance breeding. [ABSTRACT FROM AUTHOR]

Objectives: To generate pseudo-CT (pCT) images of the pelvis from zero echo time (ZTE) MR sequences and compare them to conventional CT. Methods: Ninety-one patients were prospectively scanned with CT and MRI including ZTE sequences of the pelvis. Eleven ZTE image volumes were excluded due to implants and severe B1 field inhomogeneity. Out of the 80 data sets, 60 were used to train and update a deep learning (DL) model for pCT image synthesis from ZTE sequences while the remaining 20 cases were selected as an evaluation cohort. CT and pCT images were assessed qualitatively and quantitatively by two readers. Results: Mean pCT ratings of qualitative parameters were good to perfect (2–3 on a 4-point scale). Overall intermodality agreement between CT and pCT was good (ICC = 0.88 (95% CI: 0.85–0.90); p < 0.001) with excellent interreader agreements for pCT (ICC = 0.91 (95% CI: 0.88–0.93); p < 0.001). Most geometrical measurements did not show any significant difference between CT and pCT measurements (p > 0.05) with the exception of transverse pelvic diameter measurements and lateral center-edge angle measurements (p = 0.001 and p = 0.002, respectively). Image quality and tissue differentiation in CT and pCT were similar without significant differences between CT and pCT CNRs (all p > 0.05). Conclusions: Using a DL-based algorithm, it is possible to synthesize pCT images of the pelvis from ZTE sequences. The pCT images showed high bone depiction quality and accurate geometrical measurements compared to conventional CT. Critical relevance statement: pCT images generated from MR sequences allow for high accuracy in evaluating bone without the need for radiation exposure. Radiological applications are broad and include assessment of inflammatory and degenerative bone disease or preoperative planning studies. Key Points: pCT, based on DL-reconstructed ZTE MR images, may be comparable with true CT images. Overall, the intermodality agreement between CT and pCT was good with excellent interreader agreements for pCT. Geometrical measurements and tissue differentiation were similar in CT and pCT images. [ABSTRACT FROM AUTHOR]

Simple Summary: Tendon injuries are known to be difficult to heal, making effective treatments crucial. This study examines the potential of stem cells derived from sheep fat tissue for tendon repair, since little information is available on their tenogenic potential. The focus of this study was to understand how these stem cells can be induced to develop into tendon cells. Initially, the cells were grown in the lab to ensure that their essential properties were maintained. Two methods were then employed to promote the acquisition of tendon-like characteristics: exposing the cells to a tenogenic conditioned medium or seeding them on a scaffold that mimics a tendon structure. The results indicated that these stem cells could be expanded without aging or changes in their characteristics. When exposed to the tendon medium, the cells were able to start the tendon differentiation process but were not able to complete it. Conversely, when seeded on the scaffold, the cells changed their naïve genetic and protein profile and acquired that of the typical tendon cells. This suggests that these stem cells can be addressed towards tendon cells, being more effective when seeded on a tendon biomimetic scaffold. These findings highlight the potential use of these stem cell treatments for tendon injuries, particularly in veterinary medicine. Adipose-derived stem cells (ADSCs) hold promise for tendon repair, even if their tenogenic plasticity and underlying mechanisms remain only partially understood, particularly in cells derived from the ovine animal model. This study aimed to characterize oADSCs during in vitro expansion to validate their phenotypic properties pre-transplantation. Moreover, their tenogenic potential was assessed using two in vitro-validated approaches: (1) teno-inductive conditioned media (CM) derived from a co-culture between ovine amniotic stem cells and fetal tendon explants, and (2) short- (48 h) and long-term (14 days) seeding on highly aligned PLGA (ha-PLGA) electrospun scaffold. Our findings indicate that oADSCs can be expanded without senescence and can maintain the expression of stemness (Sox2, Oct4, Nanog) and mesenchymal (CD29, CD166, CD44, CD90) markers while remaining negative for hematopoietic (CD31, CD45) and MHC-II antigens. Of note, oADSCs' tendon differentiation potential greatly depended on the in vitro strategy. oADSCs exposed to CM significantly upregulated tendon-related genes (COL1, TNMD, THBS4) but failed to accumulate TNMD protein at 14 days of culture. Conversely, oADSCs seeded on ha-PLGA fleeces quickly upregulated the tendon-related genes (48 h) and in 14 days accumulated high levels of the TNMD protein into the cytoplasm of ADSCs, displaying a tenocyte-like morphology. This mechano-sensing cellular response involved a complete SOX9 downregulation accompanied by YAP activation, highlighting the efficacy of biophysical stimuli in promoting tenogenic differentiation. These findings underscore oADSCs' long-term self-renewal and tendon differentiative potential, thus opening their use in a preclinical setting to develop innovative stem cell-based and tissue engineering protocols for tendon regeneration, applied to the veterinary field. [ABSTRACT FROM AUTHOR]

The mortiño (Vaccinium floribundum Kunth), belonging to the Ericaceae family, is a native species of the Ecuadorian paramos. It has a shrub-like habit and produces edible fruits. In the Neotropics, it is primarily found in the Andes of Colombia and Ecuador, thriving in humid environments up to an elevation of 3700 m above sea level. In this review, a selection of studies was carried out that evaluated the taxonomy, reproduction biology, and nutraceutical, environmental properties, and industrial use of the mortiño (V. floribundum). The data gathered from various bioassays were essential in determining the appropriate techniques for tissue differentiation and assessing the quality of resulting plants. This work aims to generate a deep knowledge of the cultivation of mortiño, as well as the properties of its fruits and the benefits they provide for health. These are rich in compounds with antioxidant activity, so the consumption of V. floribundum fruits is related to health benefits. Besides, the environmental role of V. floribundum and its applications in various industries, especially in the development of nanoparticles contributes to the valorization of this plant. Overall, this research contributes to establishing sustainable methods for the propagation of Vaccinium floribundum, ensuring its successful cultivation and utilization for both commercial purposes and ecological preservation. [ABSTRACT FROM AUTHOR]

DNA methylation plays an important role in the development and tissue differentiation of eukaryotes. In this study, bisulfite sequencing (BS-seq) technology was used to analyze the DNA methylation profiles of liver tissues taken from Rongchang pigs at three postnatal feeding stages, including newborn, suckling, and adult. The DNA methylation pattern across the genomes or genic region showed little difference between the three stages. We observed 419 differentially methylated regions (DMRs) in promoters, corresponding to 323 genes between newborn and suckling stages, in addition to 288 DMRs, corresponding to 134 genes, between suckling and adult stages and 351 DMRs, corresponding to 293 genes, between newborn and adult stages. These genes with DMRs were mainly enriched in metabolic, immune-related functional processes. Correlation analysis showed that the methylation level of gene promoters was significantly negatively correlated with gene expression. Further, we found that genes related to nutritional metabolism, e.g., carbohydrate metabolism (FAHD1 and GUSB) or fatty acid metabolism (LPIN1 and ACOX2), lost DNA methylation in their promoter, with mRNA expression increased in newborn pigs compared with those in the suckling stage. A few fatty acid metabolism-related genes (SLC27A5, ACOX2) were hypomethylated and highly expressed in the newborn stage, which might satisfy the nutritional requirements of Rongchang pigs with high neonatal birth rates. In the adult stage, HMGCS2—which is related to fatty acid β-oxidation—was hypomethylated and highly expressed, which explains that the characteristics of high energy utilization in adult Rongchang pigs and their immune-related genes (CD68, STAT2) may be related to the establishment of liver immunity. This study provides a comprehensive analysis of genome-wide DNA methylation patterns in pig liver postnatal development and growth. Our findings will serve as a valuable resource in hepatic metabolic studies and the agricultural food industry. [ABSTRACT FROM AUTHOR]

Tissue engineering is based on combining cells with suitable scaffolds and growth factors. Recently, bone tissue engineering has been especially investigated deeply due to a large number of bone-related diseases. One approach to improve scaffolds is based on using piezoelectric materials as a way to influence the growing bone tissue by mechanical stress. Another method to stimulate tissue growth is by applying an external magnetic field to composites of magnetostrictive and piezoelectric materials, as well as the possibility to prepare oriented surfaces by orienting embedded magnetic fibers or nanoparticles. In addition, magnetic scaffolds without other special properties have also been reported to show improved properties for bone tissue and other tissue engineering. Here, we provide an overview of recent research on magnetic scaffolds for tissue engineering, differentiating between bone and other tissue engineering. We show the advantages of magnetic scaffolds, especially related to cell guidance and differentiation, and report recent progress in the production and application of such magnetic substrates for different areas of tissue engineering. [ABSTRACT FROM AUTHOR]

Regeneration of articular cartilage remains a challenge for patients who have undergone cartilage injury, osteochondritis dissecans and osteoarthritis. Here, we describe a new recombinant silk fibroin with basic fibroblast growth factor (bFGF) binding peptide, which has a genetically introduced sequence PLLQATLGGGS, named P7. In this study, we cultured a human mesenchymal cell line derived from bone marrow, UE6E7-16, in wild-type fibroin sponge (FS) and recombinant silk fibroin sponge with P7 peptide (P7 FS). We compared cell proliferation, chondrogenic differentiation and cartilaginous tissue formation between the two types of sponge. After stimulation with bFGF at 3 ng/mL, P7 FS showed significantly higher cell growth (1.2-fold) and higher cellular DNA content (5.6-fold) than did wild-type FS. To promote chondrogenic differentiation, cells were cultured in the presence of TGF-β at 10 ng/mL for 28 days. Immunostaining of P7 FS showed SOX9-positive cells comparable to wild-type FS. Alcian-Blue staining of P7 FS also showed cartilaginous tissue formation equivalent to wild-type FS. A significant increase in cell proliferation in P7 FS implies future clinical application of this transgenic fibroin for regeneration of articular cartilage. To produce cartilaginous tissue efficiently, transgenic fibroin sponges and culture conditions must be improved. Such changes should include the selection of growth factors involved in chondrogenic differentiation and cartilage formation. [ABSTRACT FROM AUTHOR]

Challenges in directed differentiation and survival limit the clinical use of stem cells despite their promising therapeutic potential in regenerative medicine. Nanotechnology has emerged as a powerful tool to address these challenges and enable precise control over stem cell fate. In particular, nanomaterials can mimic an extracellular matrix and provide specific cues to guide stem cell differentiation and proliferation in the field of nanotechnology. For instance, recent studies have demonstrated that nanostructured surfaces and scaffolds can enhance stem cell lineage commitment modulated by intracellular regulation and external stimulation, such as reactive oxygen species (ROS) scavenging, autophagy, or electrical stimulation. Furthermore, nanoframework-based and upconversion nanoparticles can be used to deliver bioactive molecules, growth factors, and genetic materials to facilitate stem cell differentiation and tissue regeneration. The increasing use of nanostructures in stem cell research has led to the development of new therapeutic approaches. Therefore, this review provides an overview of recent advances in nanomaterials for modulating stem cell differentiation, including metal-, carbon-, and peptide-based strategies. In addition, we highlight the potential of these nano-enabled technologies for clinical applications of stem cell therapy by focusing on improving the differentiation efficiency and therapeutics. We believe that this review will inspire researchers to intensify their efforts and deepen their understanding, thereby accelerating the development of stem cell differentiation modulation, therapeutic applications in the pharmaceutical industry, and stem cell therapeutics. [ABSTRACT FROM AUTHOR]

Modern neurosurgery strives to maximize tumor removal while preserving healthy tissue integrity. Accurate intraoperative differentiation between tumor and healthy tissue is crucial yet challenging. Often neurosurgeons rely on their experience and haptic feedback during palpation to distinguish between tumor and healthy tissue. A commonly used hand-held tool for tissue removal during neurosurgery is the ultrasonic aspirator, which changes its electrical properties as it interacts with tissue. The goal is to equip the ultrasonic aspirator with the ability to differentiate between different types of tissue while at the same time not interfering with the surgical workflow and providing comprehensible outcomes. To this end, a hierarchical classification approach is employed as a proof of concept, enabling precise identification of tissue stiffness during resection.The hierarchical approach is compared with the standard flat classification, commonly used in machine learning. Within the hierarchical approach, two strategies are employed: mandatory leaf-node predictions (MLNP) and non-mandatory leaf-node predictions (NMLNP). The NMLNP allows prediction to revert to a parent node when certainty is low. Data are acquired on three artificial tissue models – differing in stiffness – with an ultrasonic aspirator in a hand-held manner. The dataset comprises 1,821 data points for training and 186 for testing after balancing.The results indicate a slight performance advantage for the hierarchical classification MLNP approach over the flat classification approach in the absence of confidence thresholds, with weighted F2-scores of 0.781 and 0.762, respectively. However, the application of confidence thresholds results in both approaches exhibiting comparable performance, with the hierarchical NMLNP approach achieving a weighted F1-score of 0.920, thereby demonstrating superior overall performance. The effects of enforcing these thresholds and excluding data with low certainty are thoroughly investigated. This work emphasizes the feasibility of tissue differentiation using a hand-held ultrasound aspirator while resecting tissue. Moreover, it highlights the capability of hierarchical classification in advancing tissue differentiation accuracy during neurosurgical procedures, which could ultimately aid surgeons and enhance the safety of intraoperative workflows.

ABSTRACT: Stem cells offer great promise in dentistry by regenerating critical oral tissues. Derived from sources such as dental pulp, periodontal ligament, and dental follicle, these cells can be pluripotent or multipotent and are capable of differentiating into various cell types, including odontoblasts and osteoblasts. This regenerative potential could transform treatments for dental conditions like periodontal disease and pulpitis. Advancing this field requires understanding stem cell differentiation and exploring tissue engineering and biomaterial scaffolds. Despite the potential, evolving legal and ethical issues shape the integration of these innovations. Research in dental stem cells represents a significant advance in regenerative medicine, promising to enhance dental treatments and patient care. [ABSTRACT FROM AUTHOR]

During animal embryogenesis, one of the earliest specification events distinguishes extraembryonic (EE) from embryonic tissue fates: the serosa in the case of the insects. While it is well established that the homeodomain transcription factor Zen1 is the critical determinant of the serosa, the subsequent realization of this tissue's identity has not been investigated. Here, we examine serosal differentiation in the beetle Tribolium castaneum based on the quantification of morphological and morphogenetic features, comparing embryos from a Tc-zen1 RNAi dilution series, where complete knockdown results in amnion-only EE tissue identity. We assess features including cell density, tissue boundary morphology, and nuclear size as dynamic readouts for progressive tissue maturation. While some features exhibit an all-or-nothing outcome, other key features show dose-dependent phenotypic responses with trait-specific thresholds. Collectively, these findings provide nuance beyond the known status of Tc-Zen1 as a selector gene for serosal tissue patterning. Overall, our approach illustrates how the analysis of tissue maturation dynamics from live imaging extends but also challenges interpretations based on gene expression data, refining our understanding of tissue identity and when it is achieved. [ABSTRACT FROM AUTHOR]

Background: Colorectal cancer is among the most common malignant tumors affecting the gastrointestinal tract. Liver metastases, a complication present in approximately 50% of colorectal cancer patients, are a considerable concern. Recently, studies have revealed the crucial role of miR-455 in tumor pathogenesis. However, the effect of miR-455 on the progression of liver metastases in colorectal cancer remains controversial. As an antagonist of bone morphogenetic protein(BMP), Gremlin 1 (GREM1) may impact organogenesis, body patterning, and tissue differentiation. Nevertheless, the role of miR-455 in regulating GREM1 in colorectal cancer liver metastases and how miR-455/GREM1 axis influences tumour immune microenvironment is unclear. Methods: Bioinformatics analysis shows that miR-455/GREM1 axis plays crucial role in liver metastasis of intestinal cancer and predicts its possible mechanism. To investigate the impact of miR-455/GREM1 axis on the proliferation, invasion, and migration of colorectal cancer cells, colony formation assay, wound healing and transwell assay were examined in vitro. The Dual-Luciferase reporter gene assay and RNA pull-down assay confirmed a possible regulatory effect between miR-455 and GREM1. In vivo, colorectal cancer liver metastasis(CRLM) model mice was established to inquiry the effect of miR-455/GREM1 axis on tumor growth and macrophage polarization. The marker of macrophage polarization was tested using immunofluorescence(IF) and quantitative real-time polymerase chain reaction(qRT-PCR). By enzyme-linked immunosorbent assay (ELISA), cytokines were detected in culture medium supernatants. Results: We found that miR-455 and BMP6 expression was increased and GREM1 expression was decreased in liver metastase compared with primary tumor. miR-455/GREM1 axis promotes colorectal cancer cells proliferation, migration, invasion via affected PI3K/AKT pathway. Moreover, downregulating GREM1 augmented BMP6 expression in MC38 cell lines, inducing M2 polarization of macrophages, and promoting liver metastasis growth in CRLM model mice. Conclusion: These data suggest that miR-455/GREM1 axis promotes colorectal cancer progression and liver metastasis by affecting PI3K/AKT pathway and inducing M2 macrophage polarization. These results offer valuable insights and direction for future research and treatment of CRLM. [ABSTRACT FROM AUTHOR]

This article provides an overview of the contributions of Alexander Friedenstein, a prominent Russian scientist, to the field of stromal stem cell biology. Friedenstein's research focused on osteogenic stem cells, which play a crucial role in bone tissue generation. Inspired by the work of Alexander Maximov, Friedenstein's groundbreaking research on osteogenic cell differentiation and bone induction led to a collaboration with the MRC Bone Research Laboratory in Oxford. Despite facing challenges due to his Jewish background, Friedenstein was able to visit Oxford in the early 1990s, where he discussed his research and its potential applications with Dr. Maureen Owen. This visit fostered increased research on stromal stem cells in the Oxford laboratory, and Friedenstein's legacy in regenerative medicine is highly regarded. [Extracted from the article]

The maintenance and variation of ploidy in plants are influenced by environmental factor, and the tissue culture regeneration can often cause changes of chromosome structures and ploidy levels. In order to explore the ploidy variation of Camellia germplasm under tissue culture conditions, the callus induction system of Camellia germplasm was used to analyze the ploidy changes by flow cytometry, and the stability and variation of the ploidy under tissue culture generation were analyzed in combination with colchicine treatment. The results were as follows: (1)Six of ten of Camellia germplasms were diploid, two of them were tetraploid, one hexaploid and one decaploid, and the germplasm materials largely maintained stable ploidy levels during the tissue regeneration processes. (2)The optimum conditions for colchicine treatment were obtained, that is, the callus was cultured in treatment medium containing colchicine at 20 mg·L-1 for 10 d, followed by regeneration in tissue-differentiation culture. (3)The ploidy analyses were carried out on the treated tissues in different differentiation states, and the results showed that most of the independent tissues(including 56 callus and shoots)were found to be aneuploids, 38 tissues had ploidy between 1.5 to 2.5, and 11 tissues produced less than 1.5. This study further explores the ploidy relationship between different Camellia germplasms, and provides a reference for an in-depth study of ploidy regulation and polyploid induction of Camellia. [ABSTRACT FROM AUTHOR]

The active form of vitamin D3, 1α,25-dihydroxyvitamin D3 [1,25(OH)2D3], is a principal regulator of calcium homeostasis through activation of the vitamin D receptor (VDR). Previous studies have shown that 2α-(3-hydroxypropyl)-1,25D3 (O1C3) and 2α-(3-hydroxypropoxy)-1,25D3 (O2C3), vitamin D derivatives resistant to inactivation enzymes, can activate VDR, induce leukemic cell differentiation, and increase blood calcium levels in rats more effectively than 1,25(OH)2D3. In this study, to further investigate the usefulness of 2α-substituted vitamin D derivatives, we examined the effects of O2C3, O1C3, and their derivatives on VDR activity in cells and mouse tissues and on osteoblast differentiation of dedifferentiated fat (DFAT) cells, a cell type with potential therapeutic application in regenerative medicine. In cell culture experiments using kidney-derived HEK293 cells, intestinal mucosa-derived CaCO2 cells, and osteoblast-derived MG63 cells, and in mouse experiments, O2C2, O2C3, O1C3, and O1C4 had a weaker effect than or equivalent effect to 1,25(OH)2D3 in VDR transactivation and induction of the VDR target gene CYP24A1, but they enhanced osteoblast differentiation in DFAT cells equally to or more effectively than 1,25(OH)2D3. In long-term treatment with the compound without the medium change (7 days), the derivatives enhanced osteoblast differentiation more effectively than 1,25(OH)2D3. O2C3 and O1C3 were more stable than 1,25(OH)2D3 in DFAT cell culture. These results indicate that 2α-substituted vitamin D derivatives, such as inactivation-resistant O2C3 and O1C3, are more effective than 1,25(OH)2D3 in osteoblast differentiation of DFAT cells, suggesting potential roles in regenerative medicine with DFAT cells and other multipotent cells. [ABSTRACT FROM AUTHOR]

Objectives: The micro‐nano structure of 3D‐printed porous titanium (Ti) alloy with excellent performance in avoiding stress shielding and promoting bone tissue differentiation provides a new opportunity for the development of bone implants, but it necessitates higher requirements for bone tissue differentiation and the antibacterial properties of bone implants in clinical practice. Methods: This study investigated the preparation, antimicrobial properties, and osteogenesis‐promoting ability of the 3D printed porous Ti alloy anodic oxidized Ag‐carrying (Ag@3D‐TiO2) scaffolds. The 3D printed porous Ti alloy (3D‐Ti), anodized 3D printed porous Ti alloy (3D‐TiO2), and Ag@3D‐TiO2 scaffolds were synthesized using electron beam melting. The antimicrobial properties of the scaffolds were examined using antibacterial tests and their cytocompatibility was assessed using a cell proliferation assay and acridine orange/ethidium bromide (AO/EB) staining. In vitro cellular assays were used to investigate the effects of the scaffold microstructural features on cell activity, proliferation, and osteogenesis‐related genes and proteins. In vivo animal experiments were used to evaluate the anti‐inflammatory and osteogenesis‐promoting abilities of the scaffolds. Results: The Ag@3D‐TiO2 scaffolds exhibited sustained anti‐microbial activity over time, enhanced cell proliferation, facilitated osteogenic differentiation, and increased extracellular matrix mineralization. In addition, alkaline phosphatase (ALP), collagen type I (COL‐I), and osteocalcin (OCN)‐related genes and proteins were upregulated. In vivo animal implantation experiments, the anti‐inflammatory effect of the Ag@3D‐TiO2 scaffolds were observed using histology, and a large amount of fibrous connective tissue was present around it; the Ag@3D‐TiO2 scaffolds were more bio‐compatible with the surrounding tissues compared with 3D‐Ti and 3D‐TiO2; a large amount of uniformly distributed neoplastic bone tissue existed in their pores, and the chronic systemic toxicity test showed that the 3D‐Ti, 3D‐TiO2, and Ag@3D‐TiO2 scaffolds are biologically safe. Conclusion: The goal of this study was to create a scaffold that exhibits antimicrobial properties and can aid bone growth, making it highly suitable for use in bone tissue engineering. [ABSTRACT FROM AUTHOR]

Electrical stimulation (ES) is a widely discussed topic in the field of cartilage tissue engineering due to its ability to induce chondrogenic differentiation (CD) and proliferation. It shows promise as a potential therapy for osteoarthritis (OA). In this study, we stimulated mesenchymal stem cells (MSCs) incorporated into collagen hydrogel (CH) scaffolds, consisting of approximately 500,000 cells each, for 1 h per day using a 2.5 V p p (119 mV/mm) 8 Hz sinusoidal signal. We compared the cell count, morphology, and CD on days 4, 7, and 10. The results indicate proliferation, with an increase ranging from 1.86 to 9.5-fold, particularly on day 7. Additionally, signs of CD were observed. The stimulated cells had a higher volume, while the stimulated scaffolds showed shrinkage. In the ES groups, up-regulation of collagen type 2 and aggrecan was found. In contrast, SOX9 was up-regulated in the control group, and MMP13 showed a strong up-regulation, indicating cell stress. In addition to lower stress levels, the control groups also showed a more spheroidic shape. Overall, scaffold-based ES has the potential to achieve multiple outcomes. However, finding the appropriate stimulation pattern is crucial for achieving successful chondrogenesis. [ABSTRACT FROM AUTHOR]

Regenerative medicine aims to address substantial defects by amplifying the body's natural regenerative abilities and preserving the health of tissues and organs. To achieve these goals, materials that can provide the spatial and biological support for cell proliferation and differentiation, as well as the micro-environment essential for the intended tissue, are needed. Scaffolds such as polymers and metallic materials provide three-dimensional structures for cells to attach to and grow in defects. These materials have limitations in terms of mechanical properties or biocompatibility. In contrast, biominerals are formed by living organisms through biomineralization, which also includes minerals created by replicating this process. Incorporating biominerals into conventional materials allows for enhanced strength, durability, and biocompatibility. Specifically, biominerals can improve the bond between the implant and tissue by mimicking the micro-environment. This enhances cell differentiation and tissue regeneration. Furthermore, biomineral composites have wound healing and antimicrobial properties, which can aid in wound repair. Additionally, biominerals can be engineered as drug carriers, which can efficiently deliver drugs to their intended targets, minimizing side effects and increasing therapeutic efficacy. This article examines the role of biominerals and their composite materials in regenerative medicine applications and discusses their properties, synthesis methods, and potential uses. [ABSTRACT FROM AUTHOR]

The enzymes performing protein post-translational modifications (PTMs) form a critical post-translational regulatory circuitry that orchestrates literally all cellular processes in the organism. In particular, the balance between cellular stemness and differentiation is crucial for the development of multicellular organisms. Importantly, the fine-tuning of this balance on the genetic level is largely mediated by specific PTMs of histones including lysine methylation. Lysine methylation is carried out by special enzymes (lysine methyltransferases) that transfer the methyl group from S-adenosyl-L-methionine to the lysine residues of protein substrates. Set7/9 is one of the exemplary protein methyltransferases that however, has not been fully studied yet. It was originally discovered as histone H3 lysine 4-specific methyltransferase, which later was shown to methylate a number of non-histone proteins that are crucial regulators of stemness and differentiation, including p53, pRb, YAP, DNMT1, SOX2, FOXO3, and others. In this review we summarize the information available to date on the role of Set7/9 in cellular differentiation and tissue development during embryogenesis and in adult organisms. Finally, we highlight and discuss the role of Set7/9 in pathological processes associated with aberrant cellular differentiation and self-renewal, including the formation of cancer stem cells. [ABSTRACT FROM AUTHOR]

Multicellular organisms are composed of many tissue types that have distinct morphologies and functions, which are largely driven by specialized proteomes and interactomes. To define the proteome and interactome of a specific type of tissue in an intact animal, we developed a localized proteomics approach called Methionine Analog-based Cell-Specific Proteomics and Interactomics (MACSPI). This method uses the tissue-specific expression of an engineered methionyl-tRNA synthetase to label proteins with a bifunctional amino acid 2-amino-5-diazirinylnonynoic acid in selected cells. We applied MACSPI in Caenorhabditis elegans, a model multicellular organism, to selectively label, capture, and profile the proteomes of the body wall muscle and the nervous system, which led to the identification of tissue-specific proteins. Using the photo-cross-linker, we successfully profiled HSP90 interactors in muscles and neurons and identified tissue-specific interactors and stress-related interactors. Our study demonstrates that MACSPI can be used to profile tissue-specific proteomes and interactomes in intact multicellular organisms. [ABSTRACT FROM AUTHOR]

Non-typeable Haemophilus influenzae (NTHi) and Moraxella catarrhalis (Mcat) are two common respiratory tract pathogens often associated with acute exacerbations in Chronic Obstructive Pulmonary Disease (COPD) as well as with otitis media (OM) in children. Although there is evidence that these pathogens can adopt persistence mechanisms such as biofilm formation, the precise means through which they contribute to disease severity and chronicity remains incompletely understood, posing challenges for their effective eradication. The identification of potential vaccine candidates frequently entails the characterization of the host-pathogen interplay in vitro even though this approach is limited by the fact that conventional models do not permit long term bacterial infections. In the present work, by using air-liquid-interface (ALI) human airway in vitro models, we aimed to recreate COPD-related persistent bacterial infections. In particular, we explored an alternative use of the ALI system consisting in the assembly of an inverted epithelium grown on the basal part of a transwell membrane with the aim to enable the functionality of natural defense mechanisms such as mucociliary clearance and cellular extrusion that are usually hampered during conventional ALI infection experiments. The inversion of the epithelium did not affect tissue differentiation and considerably delayed NTHi or Mcat infection progression, allowing one to monitor host-pathogen interactions for up to three weeks.Notably, the use of thesemodels, coupledwith confocal and transmission electron microscopy, revealed unique features associated with NTHi and Mcat infection, highlighting persistence strategies including the formation of intracellular bacterial communities (IBCs) and surface-associated biofilm-like structures. Overall, this study demonstrates the possibility to perform long term host-pathogen investigations in vitro with the aim to define persistence mechanisms adopted by respiratory pathogens and individuate potential new vaccine targets. [ABSTRACT FROM AUTHOR]

Application of optical coherence tomography (OCT) in neurosurgery mostly includes the discrimination between intact and malignant tissues aimed at the detection of brain tumor margins. For particular tissue types, the existing approaches demonstrate low performance, which stimulates the further research for their improvement. The analysis of speckle patterns of brain OCT images is proposed to be taken into account for the discrimination between human brain glioma tissue and intact cortex and white matter. The speckle properties provide additional information of tissue structure, which could help to increase the efficiency of tissue differentiation. The wavelet analysis of OCT speckle patterns was applied to extract the power of local brightness fluctuations in speckle and its standard deviation. The speckle properties are analysed together with attenuation ones using a set of ex vivo brain tissue samples, including glioma of different grades. Various combinations of these features are considered to perform linear discriminant analysis for tissue differentiation. The results reveal that it is reasonable to include the local brightness fluctuations at first two wavelet decomposition levels in the analysis of OCT brain images aimed at neurosurgical diagnosis. [ABSTRACT FROM AUTHOR]

Microtubules are an indispensable component of all eukaryotic cells due to their role in mitotic spindle formation, yet their organization and number can vary greatly in the interphase. The last common ancestor of all eukaryotes already had microtubules and microtubule motor proteins moving along them. Sponges are traditionally regarded as the oldest animal phylum. Their body does not have a clear differentiation into tissues, but it contains several distinguishable cell types. The choanocytes stand out among them and are responsible for creating a flow of water with their flagella and increasing the filtering and feeding efficiency of the sponge. Choanocyte flagella contain microtubules, but thus far, observing a developed system of cytoplasmic microtubules in non-flagellated interphase sponge cells has been mostly unsuccessful. In this work, we combine transcriptomic analysis, immunofluorescence, and electron microscopy with time-lapse recording to demonstrate that microtubules appear in the cytoplasm of sponge cells only when transdifferentiation processes are activated. We conclude that dynamic cytoplasmic microtubules in the cells of sponges are not a persistent but rather a transient structure, associated with cellular plasticity. [ABSTRACT FROM AUTHOR]

Glioblastoma is a fast‐growing and aggressive form of brain cancer. Even with maximal treatment, patients show a low median survival and are often subjected to a high recurrence incidence. The currently available treatments require multimodal management, including maximal safe surgical resection, followed by radiation and chemotherapy. Because of the infiltrative glioblastoma nature, intraoperative differentiation of cancer tissue from normal brain parenchyma is very challenging, and this accounts for the low rate of complete tumor resection. For these reasons, clinicians have increasingly used various intraoperative adjuncts to improve surgical results, such as fluorescent agents. However, most of the existing fluorophores show several limitations such as poor selectivity, photostability, photosensitization and high costs. This could limit their application to successfully improve glioblastoma resection. In the present perspective, we highlight the possibility to develop next‐generation fluorescent tools able to more selectively label cancer cells during surgical resection. [ABSTRACT FROM AUTHOR]

Cultivated meat (CM) offers a sustainable and ethical alternative to conventional animal agriculture, involving cell maturation in a controlled environment. To emulate the structural complexity of traditional meat, the development of animal-free and edible scaffolds is crucial, providing vital physical and biological support during tissue development. The aligned vascular bundles of the decellularised asparagus scaffold were selected to facilitate the attachment and alignment of murine myoblasts (C2C12) and porcine adipose-derived mesenchymal stem cells (pADMSCs). Muscle differentiation was assessed through immunofluorescence staining with muscle markers, including Myosin heavy chain (MHC), Myogenin (MYOG), and Desmin. The metabolic activity of Creatine Kinase in C2C12 differentiated cells significantly increased compared to proliferated cells. Quantitative PCR analysis revealed a significant increase in Myosin Heavy Polypeptide 1 (MYH1) and MYOG expression compared to Day 0. These results highlight the application of decellularised plant scaffold (DPS) as a promising, edible material conducive to cell attachment, proliferation, and differentiation into muscle tissue. To create a CM prototype with biological mimicry, pADMSC-derived muscle and fat cells were also co-cultured on the same scaffold. The co-culture was confirmed through immunofluorescence staining of muscle markers and LipidTOX staining, revealing distinct muscle fibres and adipocytes containing lipid droplets respectively. Texture profile analysis conducted on uncooked CM prototypes and pork loin showed no significant differences in textural values. However, the pan-fried CM prototype differed significantly in hardness and chewiness compared to pork loin. Understanding the scaffolds' textural profile enhances our insight into the potential sensory attributes of CM products. DPS shows potential for advancing CM biomanufacturing. [ABSTRACT FROM AUTHOR]

Association of circulating glycoprotein acetyls (GlycA), a systemic inflammation biomarker, with lung function and respiratory diseases remain to be investigated. We examined the genetic correlation, shared genetics, and potential causality of GlycA (N = 115,078) with lung function and respiratory diseases (N = 497,000). GlycA showed significant genetic correlation with FEV1 (rg = −0.14), FVC (rg = −0.18), asthma (rg = 0.21) and COPD (rg = 0.31). We consistently identified ten shared loci (including chr3p21.31 and chr8p23.1) at both SNP and gene level revealing potential shared biological mechanisms involving ubiquitination, immune response, Wnt/β-catenin signaling, cell growth and differentiation in tissues or cells including blood, epithelium, fibroblast, fetal thymus, and fetal intestine. Genetically elevated GlycA was significantly correlated with lung function and asthma susceptibility (354.13 ml decrement of FEV1, 442.28 ml decrement of FVC, and 144% increased risk of asthma per SD increment of GlycA) from MR analyses. Our findings provide insights into biological mechanisms of GlycA in relating to lung function, asthma, and COPD. Here, the authors observed significant genetic correlation of GlycA with lung function, asthma and COPD and identified ten shared loci revealing potential shared biological mechanisms involving ubiquitination and Wnt/β-catenin signaling. [ABSTRACT FROM AUTHOR]

Millions of women worldwide are infertile due to gynecological disorders, including premature ovarian insufficiency, polycystic ovary syndrome, Asherman syndrome, endometrial atrophy, and fallopian tube obstruction. These conditions frequently lead to infertility and have a substantial impact on the quality of life of the affected couples, primarily because of their psychological implications and high financial costs. Recently, using platelets to stimulate cell proliferation and tissue differentiation has emerged as a promising approach in regenerative medicine. Platelet-rich plasma (PRP) shows considerable potential for promoting endometrial hypertrophy and follicle development, making it a promising therapeutic option for tissue repair or replacement. This review provides an overview of the recent advancements and underlying mechanisms of PRP therapy for various female reproductive diseases and presents new therapeutic options for addressing female infertility. [ABSTRACT FROM AUTHOR]