Early Differentiation Research Articles

Early-phase EEG power spectrum analysis may differentiate acute encephalopathy with biphasic seizures and late reduced diffusion from prolonged febrile seizures.

Acute encephalopathy with biphasic seizures and late reduced diffusion (AESD) is clinically characterized by biphasic seizures associated with mild to severe neurological sequelae and is the most common subtype of acute encephalopathy in Japan, accounting for around 30 % of cases. The present study retrospectively analyzed the utility of electroencephalography (EEG) in determining the optimal method of diagnosing AESD at the early stage. This study explores early power value differences to differentiate acute encephalopathy from prolonged febrile seizure (FS). The subjects were patients aged six months to four years who had received intensive care for febrile status epilepticus and had a continuous EEG record up to 24 h after seizure onset. Power spectrum analysis of consecutive, 30-minute recordings at two-hour intervals were conducted, and the power values (alpha, beta, delta, theta waves) were compared between AESD group and FS group. Eight patients with AESD and 69 patients with FS were identified retrospectively. The emergence of delta waves in the frontal region was significantly higher in the AESD group at six to ten hours after onset. The emergence of beta waves across all the regions was lower in the AESD group from six hours after seizure onset. Frequency analysis of EEG in the early period after febrile status epilepticus onset demonstrated a significant difference between the AESD and FS groups. Delta wave power values in the frontal region at six to ten hours after onset might be useful for the early differentiation of AESD from FS.

Early osteogenic differentiation of human dental stem cells by gelatin/calcium phosphate- Punica granatum nanocomposite scaffold

BackgroundTissue engineering for bone regeneration aims to heal severe bone injuries. This study aimed to prepare and assess the early osteogenic differentiation effects of a gelatin/calcium phosphate- Punica granatum nanocomposite scaffold on stem cells from human exfoliated deciduous (SHED) and human dental pulp stem cells (HDPSCs).MethodsThe electrospinning method was used to prepare a gelatin/calcium phosphate nanocomposite scaffold containing pomegranate (Punica granatum) extract. The physicochemical properties of the scaffold were evaluated. The effect of the scaffold on the selected cells was done by the cell viability evaluation. A special alkaline phosphatase (ALP) kit was utilized to investigate the early osteogenic differentiation effects of the prepared scaffold on HDPSCs and SHED.ResultsThe results showed that the scaffold had uniformly accumulated in the networked form. Besides, the prepared scaffold did not have beads (structural defects). No new interactions were observed in the spectroscopic spectra of the scaffold and these peaks showed the successful formation of the fibrous nanocomposite as well. Furthermore, cell viability percentage was significantly higher for the scaffold compared with the control group (cells without any material) for both HDPSCs and SHED. Early osteogenic differentiation results specified that the ALP activity was significantly higher for the scaffold compared with the control group (cells without any material) for both HDPSCs and SHED.ConclusionThe appropriate physicochemical assay and cellular results (cell viability and early osteogenic differentiation) for the prepared fibrous nanocomposite showed that the use of this nanocomposite can be considered in the construction of various scaffolds in bone and dental tissue engineering.

Novel Alleles of the Potato Leaf Gene Identified in Italian Traditional Varieties Conferring Potato-like Leaf Shape in Tomato

The genetic diversity of tomato in Italy and the growing interest in high-quality food products highlight the importance of establishing varietal distinctiveness through molecular strategies to ensure agrifood product quality and traceability. In this study, four Italian potato-like leaf (PL) landraces were analyzed: “Spagnoletta di Formia e di Gaeta” (SPA) from southern Lazio, “Giagiù” (GIA) and “Patanara” (PTN) from Campania, and “Pomodoro di Mola” (MOL) from Apulia. These landraces were genotyped for the potato leaf gene (C), with two PL American genotypes and a non-allelic PL mutant line included as outgroups. Nagcarlang served as control. An allelism test confirmed C as determinant gene. The SCAR marker for C revealed that the Italian landraces presented determinants other than the most representative one responsible for PL. Whole-genome sequencing of SPA identified a private novel nonsense SNP variant allele, confirmed through dCAPS marker analysis. Additionally, two novel PL alleles responsible for missense variations were identified in GIA/PTN and MOL. In silico protein analysis suggested that novel C alleles could be functional determinants for the protein activity. Overall, PL mutations identified for the first time could serve as molecular tools for agrifood chain traceability, enabling early differentiation and recognition of genotypically similar varieties.

Differential diagnosis of thrombosis and myxoma in unusual heart positions by dual-energy CT multiple parameter imaging: A case series.

Thrombus is the most common occupying lesion in the cardiac chambers, it is often distinguished from cardiac neoplastic occupations. Among them, the most common is cardiac myxoma, whose imaging manifestations are often confused with thrombus. However, the 2 types of lesions have different therapeutic strategies and are both potentially high-risk sources of embolism, so early differentiation between intracardiac thrombus and cardiac tumor is essential. In this study, we intend to investigate the value of dual-energy computed tomography (CT) in the differential diagnosis of cardiac thrombus and myxoma by retrospectively analyzing the dual-energy CT-related parameters of 2 cases of intracardiac thrombus and 1 case of cardiac myxoma. Three cases of masses located in uncommon areas of the heart with comparable imaging characteristics are presented in this study. Echocardiography revealed an isoechoic mass in the cardiac chambers, while CT scans showed a hypodense occupancy with varying morphologies. Postoperative pathology or follow-up after treatment confirmed 1 case as a right ventricular thrombus, another as a right atrial thrombus, and the third as a right ventricular myxoma. In this study, we conducted a retrospective analysis of dual-energy CT-related parameters in 2 cases of intracardiac thrombus and 1 case of cardiac myxoma. Our findings indicate notable differences in the slopes of the energy spectral curves, mean iodine density, and effective atomic number between the intracardiac thrombus and myxoma cases. Drawing upon existing literature, we propose combining different quantitative analysis methodologies to create a more objective foundation for distinguishing between cardiac thrombosis and myxoma.

Factors secreted from the stem cells of human exfoliated deciduous teeth inhibit osteoclastogenesis through the activation of the endogenous antioxidant system.

Systemic administration of conditioned medium (CM) from stem cells derived from human exfoliated deciduous teeth (SHED-CM) in mouse models of rheumatoid arthritis, osteoporosis, and osteoarthritis suppresses excessive osteoclast activity and restores bone integrity. However, the mechanism through which SHED-CM regulates osteoclastogenesis remains largely unknown. In the present study, we examined the anti-osteoclastogenic mechanism of SHED-CM in vitro. Bone marrow macrophages and RAW264.7 cells were treated with receptor activator of nuclear factor kappa-Β ligand (RANKL) in the presence of SHED-CM or CM from bone marrow mesenchymal stem cells (BMSC-CM). Osteoclast differentiation was assessed using tartrate-resistant acid phosphatase staining, actin ring formation, and expression of osteoclast-specific markers. RANKL-induced reactive oxygen species (ROS) production was analyzed as a critical mediator of osteoclastogenesis. The activation of endogenous antioxidant gene expression was examined using reverse transcription quantitative PCR. Liquid chromatography with tandem mass spectrometry (LC-MS) was used to identify proteins enriched in SHED-CM, and neutralizing antibodies were used to evaluate their functional roles. Compared to BMSC-CM, SHED-CM effectively inhibited RANKL-induced early osteoclast differentiation and late maturation. Notably, SHED-CM but not BMSC-CM suppressed RANKL-induced ROS production. SHED-CM increased the expression of genes encoding antioxidant enzymes. The LC-MS analysis identified seven proteins uniquely enriched in SHED-CM that activated the endogenous antioxidant system. Neutralizing antibodies against some of these proteins restore RANKL-induced ROS production and osteoclast differentiation. SHED-CM inhibited osteoclastogenesis, partially through the activation of multiple antioxidant enzymes in osteoclast precursors, highlighting its potential for treating bone-destructive diseases.

Association between maternal androgen levels and early fetal sex differentiation: Anogenital distance and genital tubercle length in the first trimester.

To evaluate the use of anogenital distance (AGD) and genital tubercle length (GTL) between 11 and 13+6weeks of gestation for fetal sex determination and to assess the impact of maternal androgen levels on these measurements. A cross-sectional study was conducted from February to June 2017 with patients undergoing first trimester Down syndrome screening. Inclusion criteria were: (1) female age 18-49, (2) gestational age between 11 and 13+6weeks, (3) optimal visualization of AGD and GTL, and (4) nonsmoking status. Maternal androgen levels (total testosterone, free testosterone, androstenedione, and dehydroepiandrostenedione) were measured simultaneously with ultrasound. AGD was significantly higher in male fetuses compared to females. With a cutoff of 5.0mm, fetal sex was identified in 82% of female fetuses and 70% of male fetuses after the 12th week. Linear regression analysis showed that only AGD was a significant predictor of fetal sex (β: 0.54, p<0.001). In the 12-12+6week group, a positive correlation between AGD and maternal androstenedione was found in female fetuses (r: 0.23, p=0.038). In contrast, negative correlations were observed between AGD and both androstenedione (r: -0.475, p<0.001) and total testosterone (r: -0.282, p=0.026) in male fetuses. AGD and GTL measurements show sexual dimorphism in the first trimester, with AGD correlating positively with maternal androstenedione in females and negatively in males. These findings suggest that AGD and GTL are reliable, non-invasive methods for early fetal sex determination.

Inhibiting Autophagy by Chemicals During SCAPs Osteodifferentiation Elicits Disorganized Mineralization, While the Knock-Out of Atg5/7 Genes Leads to Cell Adaptation.

SCAPs (Stem Cells from Apical Papilla), derived from the apex of forming wisdom teeth, extracted from teenagers for orthodontic reasons, belong to the MSCs (Mesenchymal Stromal Cells) family. They have multipotent differentiation capabilities and are a potentially powerful model for investigating strategies of clinical cell therapies. Since autophagy-a regulated self-eating process-was proposed to be essential in osteogenesis, we investigated its involvement in the SCAP model. By using a combination of chemical and genetic approaches to inhibit autophagy, we studied early and late events of osteoblastic differentiation. We showed that blocking the formation of autophagosomes with verteporfin did not induce a dramatic alteration in early osteoblastic differentiation monitored by ALP (alkaline phosphatase) activity. However, blocking the autophagy flux with bafilomycin A1 led to ALP repression. Strikingly, the mineralization process was observed with both compounds, with calcium phosphate (CaP) nodules that remained inside cells under bafilomycin A1 treatment and numerous but smaller CaP nodules after verteporfin treatment. In contrast, deletion of Atg5 or Atg7, two genes involved in the formation of autophagosomes and essential to trigger canonical autophagy, indicated that both genes could be involved differently in the mineralization process with a modification of the ALP activity while final mineralization was not altered.

NUBP2 deficiency disrupts the centrosome-check point in the brain and causes primary microcephaly.

Microcephaly affects 1 in 2,500 babies per year. Primary microcephaly results from aberrant neurogenesis leading to a small brain at birth. This is due to altered patterns of proliferation and/or early differentiation of neurons. Premature differentiation of neurons is associated with defects in the centrosome and/or primary cilia. In this study, we report on the first patients identified with NUBP2 -deficiency and utilize a conditional mouse model to ascertain the molecular mechanisms associated with NUBP2 -deficient primary microcephaly. We identified homozygous NUBP2 variants in these patients who displayed profound primary microcephaly in addition to intrauterine growth restriction, cervical kyphosis, severe contractures of joints, and facial dysmorphia. We then generated a mouse model using Emx1-Cre to ablate Nubp2 from the forebrain. The mice presented with severe microcephaly starting at E18.5. Neurospheres generated from the forebrain of Emx1-Cre; Nubp2 flox/flox conditional deletion mice were used to support the pathogenicity of the patient variants. We show that loss of Nubp2 increases both canonical and non-canonical cell death, but that loss of p53 fails to rescue microcephaly in the mouse model. Examination of neurogenesis in Emx1-Cre; Nubp2 flox/flox mice revealed distinct alterations in proliferation and cellular migration accompanied by supernumerary centrosomes and cilia. We therefore propose that NUBP2 is a novel primary microcephaly-related gene and that the role of Nubp2 in centrosome and cilia regulation is crucial for proper neurogenesis.

Potential risk factors for delayed neurological sequelae and myocardial injury following acute carbon monoxide poisoning: A retrospective study.

Acute carbon monoxide poisoning (COP) has been a common cause of emergency hospital visits over the past decade. Besides the immediate symptoms of poisoning, carbon monoxide exposure can cause various long-term complications, especially delayed neurological sequelae (DNS) and myocardial injury (MI). This study retrospectively enrolled 502 patients with COP, including complete collection data, from the Taiwan National Poison Control Center between January 1, 2000, and December 31, 2015. After collecting the relevant clinical and laboratory data, multivariate logistic regression analysis was performed to investigate the associations between potential predictors and risk factors, hazard ratio (HR), and confidence intervals (CI). The cumulative incidence rates were 12.0% and 19.7% for DNS and MI, respectively. A Glasgow Coma Scale (GCS) score of <9 (HR 2.55; 95% CI: 1.52-4.27) and rhabdomyolysis (HR 2.68; 95% CI: 1.59-4.53) were identified as individual indicators of DNS in patients with COP. However, a greater risk for MI was associated with a GCS score of <9 (HR 2.50; 95% CI: 1.67-3.74), rhabdomyolysis (HR 4.91; 95% CI: 3.28-7.35), acute renal impairment (HR 2.43; 95% CI: 1.59-3.71), and leukocytosis (HR 9.55; 95% CI: 3.88-23.50). Hyperbaric oxygen therapy for patients with COP was more beneficial for DNS (HR 0.64; 95% CI: 0.34-1.20) than for MI (HR 1.94; 95% CI: 0.94-4.01). Early differentiation of risk factors between DNS and MI contributes to an effective evaluation of patients with acute COP and the provision of appropriate therapy.

MicroRNA in neuroexosome as a potential biomarker for HIV-associated neurocognitive disorders.

HIV-associated neurocognitive disorder (HAND) is a complication of chronic inflammation caused by HIV infection that impairs cognitive and motor functions. HAND can occur at any age, regardless of the duration of infection, even in people living with HIV (PLWH) whose blood viral load is controlled by antiretroviral therapy. The diagnosis of HAND requires a battery of neuropsychological tests, which is time-consuming and burdensome, limiting its effectiveness for screening PLWH. Here, we aimed to identify biomarkers for quantitatively diagnosing and screening for HAND using minimally invasive blood tests. Neuronal-derived exosomes (neuroexosomes) were isolated from the peripheral blood of PLWH, and the transcriptomes of their microRNAs (miRNAs) were analyzed. We identified five upregulated miRNAs (hsa-miR-16-5p, hsa-miR-26a-3p, hsa-92a-3p, hsa-miR-103a-3p, and hsa-miR-185-5p), and two downregulated miRNA (hsa-miR-3613-3p and hsa-miR-4668-5p) in PLWH diagnosed with HAND (HAND PLWH). Functional analysis of five miRNAs whose expression levels increased in HAND PLWH using the database showed that these miRNAs are involved in motor proteins and endocytosis, which are associated with nerve function. The expression levels of hsa-miR-16-5p, hsa-miR-103a-3p, and hsa-miR-185-5p were significantly higher than those in the non-HIV controls and non-HAND PLWH, suggesting that these miRNAs are potential biomarkers for HAND. Since there were no changes in known dementia miRNA biomarkers in HAND PLWH, the miRNAs identified in this study will allow for early differentiation of HAND.

Fine-tuning of gene expression through the Mettl3-Mettl14-Dnmt1 axis controls ESC differentiation.

The marking of DNA, histones, and RNA is central to gene expression regulation in development and disease. Recent evidence links N6-methyladenosine (m6A), installed on RNA by the METTL3-METTL14 methyltransferase complex, to histone modifications, but the link between m6A and DNA methylation remains scarcely explored. This study shows that METTL3-METTL14 recruits the DNA methyltransferase DNMT1 to chromatin for gene-body methylation. We identify a set of genes whose expression is fine-tuned by both gene-body 5mC, which promotes transcription, and m6A, which destabilizes transcripts. We demonstrate that METTL3-METTL14-dependent 5mC and m6A are both essential for the differentiation of embryonic stem cells into embryoid bodies and that the upregulation of key differentiation genes during early differentiation depends on the dynamic balance between increased 5mC and decreased m6A. Our findings add a surprising dimension to our understanding of how epigenetics and epitranscriptomics combine to regulate gene expression and impact development and likely other biological processes.

The FIRE biosensor illuminates iron regulatory protein activity and cellular iron homeostasis.

On Earth, iron is abundant, bioavailable, and crucial for initiating the first catalytic reactions of life from prokaryotes to plants to mammals. Iron-complexed proteins are critical to biological pathways and essential cellular functions. While it is well known that the regulation of iron is necessary for mammalian development, little is known about the timeline of how specific transcripts network and interact in response to cellular iron regulation to shape cell fate, function, and plasticity in the developing embryo and beyond. Here, we present a ratiometric genetically encoded dual biosensor called FIRE (Fe-IRE [iron-responsive element]) to evaluate iron regulatory protein (IRP)-binding activity and cellular iron status in live cells, allowing for the study and dissection of dynamic changes in cellular iron and IRP activity over developmental time. FIRE reveals a previously unrecognized foundational timeline of IRP activity and cellular iron homeostasis during stem cell pluripotency transition and early differentiation.

Interplay of SHH, WNT and BMP4 signaling regulates the development of the lamina propria in the murine ureter.

In the mammalian ureters, the lamina propria presents as a prominent layer of connective tissue underneath the urothelium. Despite its important structural and signaling functions, little is known how the lamina propria develops. Here, we show that in the murine ureter, the lamina propria arises at late fetal stages and massively increases by fibrocyte proliferation and collagen deposition after birth. WNT, SHH, BMP4 and retinoic acid signaling are all active in the common mesenchymal progenitor of smooth muscle cells and lamina propria fibrocytes. However, around birth, the lamina propria becomes a target for epithelial WNT and SHH signals and a source of BMP4 and retinoic acid. SHH and WNT signaling promote lamina propria and smooth muscle cell differentiation and proliferation at fetal and early postnatal stages while BMP4 signaling is required for early smooth muscle cell differentiation but not for its later maintenance. Our findings suggest that in presence of SHH and WNT signaling it is the modulation of BMP4 signaling which is the major determinant for the segregation of lamina propria and smooth muscle cells.

Sonic Hedgehog signaling regulates the optimal differentiation pace from early-stage mesoderm to cardiogenic mesoderm in mice.

Sonic Hedgehog (Shh), encoding an extracellular signaling molecule, is vital for heart development. Shh null mutants show congenital heart disease due to left-right asymmetry defects stemming from functional anomaly in the midline structure in mice. Shh signaling is also known to affect cardiomyocyte differentiation, endocardium development, and heart morphogenesis, particularly in second heart field (SHF) cardiac progenitor cells that contribute to the right ventricle, outflow tract, and parts of the atrium. Despite extensive studies, our understanding remains incomplete. Notably, Shh signaling is suggested to promote cardiac differentiation, while paradoxically preventing premature differentiation of SHF progenitors. In this study, we elucidate the role of Shh signaling in the earliest phase of cardiac differentiation. Our meta-analysis of single-cell RNA sequencing suggests that cardiogenic nascent mesoderm cells expressing the bHLH transcription factor Mesp1 interact with axial mesoderm via Hh signaling. Activation of Hh signaling using a Smoothened agonist delayed or suppressed the differentiation of primitive streak cells expressing T-box transcription factor T to Mesp1+ nascent mesoderm cells both in vitro and ex vivo. Conversely, inhibition of Hh signaling by cyclopamine facilitated cardiac differentiation. The reduction of Eomes, an inducer of Mesp1, by Hh signaling appears to be the underlying mechanism of this phenomenon. Our data suggest that SHH secreted from axial mesoderm inhibits premature differentiation of T+ cells to Mesp1+ nascent mesoderm cells, thereby regulating the pace of cardiac differentiation. These findings enhance our comprehension of Shh signaling in cardiac development, underscoring its crucial role in early cardiac differentiation.

Early differentiation of committed erythroid cells defined by miR-144/451 expression.

Before committing to an erythroid cell lineage, hematopoietic stem cells differentiate along a myeloid cell pathway to generate megakaryocyte-erythroid biopotential progenitor cells in bone marrow. Recent studies suggest that erythroid progenitors (EryPs) could be generated at the level of common myeloid progenitors (CMPs). However, due to a lack of suitable markers, little is known about the early differentiation of these committed EryP cells during CMP development. Herein, using miR-144/451-eGFP knock-in mice, we found that early differentiation of committed erythroid cells could be defined by miR-144/451 expression within CMPs. Single-cell RNA sequencing showed that miR-144/451+ progenitors had obvious differentiation characteristics of erythroid lineage cells and diverged from megakaryocyte and other myeloid cell lineages. These progenitors exclusively gave rise to erythroid cells, both in vitro and in vivo, and the commitment to an erythroid cell lineage was accompanied by loss of CD53 expression. Our findings will facilitate further understanding of the molecular mechanisms governing erythroid development and support the identification of therapeutic targets for diseases related to erythrocyte development.

Transcriptional Integration of Meiotic Prophase I Progression and Early Oocyte Differentiation.

Female reproductive senescence results from the regulated depletion of a finite pool of oocytes called the ovarian reserve. This pool of oocytes is initially established during fetal development, but the oocytes that comprise it must remain quiescent for decades until they are activated during maturation in adulthood. In order for developmentally competent oocytes to populate the ovarian reserve they must successfully initiate both meiosis and oogenesis. As the factors that regulate the timing and fidelity of these early events remain elusive, we assessed the precise function and timing of the transcriptional regulator TAF4b during meiotic prophase I progression in mouse fetal oocytes. Compared to matched controls, E14.5 Taf4b-deficient oocytes enter meiosis I in a timely manner however, their subsequent progression through the pachytene-to-diplotene transition of meiotic prophase I is compromised. Moreover, this disruption of meiotic progression is associated with the reduced ability of Taf4b-deficient oocytes to repair double-strand DNA breaks. Transcriptional profiling of Taf4b-deficient oocytes reveals that between E16.5 and E18.5 these oocytes fail to coordinate the reduction of meiotic gene expression and the induction of oocyte differentiation genes. These studies reveal that TAF4b promotes the formation of the ovarian reserve in part by orchestrating the timely transition to meiosis I arrest and oocyte differentiation, which are often perceived as separate events.

Fate mapping in mouse demonstrates early secretory differentiation directly from Lgr5+ intestinal stem cells.

The intestinal epithelium has a remarkably high turnover in homeostasis. It remains unresolved how this is orchestrated at the cellular level and how the behavior of stem and progenitor cells ensures tissue maintenance. To address this, we combined quantitative fate mapping in three complementary mouse models with mathematical modeling and single-cell RNA sequencing. Our integrated approach generated a spatially and temporally defined model of crypt maintenance based on two cycling populations: stem cells at the crypt-bottom and transit-amplifying (TA) cells above them. Subsequently, we validated the predictions from the mathematical model, demonstrating that fate decisions between the secretory and absorptive lineages are made within the stem cell compartment, whereas TA cell divisions contribute specifically to the absorptive lineage. These quantitative insights provide further direct evidence for crypt-bottom stem cells as the dominant driver of the intestinal epithelium replenishment.

Deep Learning for Discrimination of Early Spinal Tuberculosis from Acute Osteoporotic Vertebral Fracture on CT.

Early differentiation between spinal tuberculosis (STB) and acute osteoporotic vertebral compression fracture (OVCF) is crucial for determining the appropriate clinical management and treatment pathway, thereby significantly impacting patient outcomes. To evaluate the efficacy of deep learning (DL) models using reconstructed sagittal CT images in the differentiation of early STB from acute OVCF, with the aim of enhancing diagnostic precision, reducing reliance on MRI and biopsies, and minimizing the risks of misdiagnosis. Data were collected from 373 patients, with 302 patients recruited from a university-affiliated hospital serving as the training and internal validation sets, and an additional 71 patients from another university-affiliated hospital serving as the external validation set. MVITV2, Efficient-Net-B5, ResNet101, and ResNet50 were used as the backbone networks for DL model development, training, and validation. Model evaluation was based on accuracy, precision, sensitivity, F1 score, and area under the curve (AUC). The performance of the DL models was compared with the diagnostic accuracy of two spine surgeons who performed a blinded review. The MVITV2 model outperformed other architectures in the internal validation set, achieving accuracy of 98.98%, precision of 100%, sensitivity of 97.97%, F1 score of 98.98%, and AUC of 0.997. The performance of the DL models notably exceeded that of the spine surgeons, who achieved accuracy rates of 77.38% and 93.56%. The external validation confirmed the models' robustness and generalizability. The DL models significantly improved the differentiation between STB and OVCF, surpassing experienced spine surgeons in diagnostic accuracy. These models offer a promising alternative to traditional imaging and invasive procedures, potentially promoting early and accurate diagnosis, reducing healthcare costs, and improving patient outcomes. The findings underscore the potential of artificial intelligence for revolutionizing spinal disease diagnostics, and have substantial clinical implications.

Bisphenol S exposure interrupted human embryonic stem cell derived cardiomyocytes differentiation through ER-NF-κB/ERK signaling pathway.

Bisphenol S (BPS) has been put into production as a wide range of Bisphenol A (BPA) alternatives, while little is known regarding its cardiac developmental toxicity. To explore the effect of BPS on cardiomyocyte differentiation and its mechanism, our study established the human embryonic stem cell-cardiomyocyte differentiation model (hESC-CM), which was divided into early period of differentiation (DP1:1-8d), anaphase period of differentiation (DP2:9-16d) and whole stage of differentiation (DP3:1-16d) exposed to human-related levels of BPS. We found that the survival rate of cardiomyocytes was more sensitive to BPS at the early stage of differentiation than at the anaphase stage of differentiation, and exposure to higher than 30 µg/mL BPS throughout the differentiation period decreased the expression of cTnT. BPS may affect cardiomyocyte differentiation by activating ERβ-NF-κB/ERK signaling pathway, and the signaling pathway of each stage might be different. During DP1, 3 µg/mL of BPS may increase the inflammatory effect of cardiomyocytes mainly through the ERβ-NF-κB signaling pathway, thereby inhibiting cell proliferation, and leading to impaired cardiac function in early differentiation. During DP2, BPS may activate the ERβ-ERK signaling pathway, increase cardiomyocyte apoptosis, alter the establishment of the outer matrix, and thus affect myocardial differentiation. However, exposure to BPS throughout the differentiation stage may disrupt the immune response and cell differentiation, which in turn interrupts heart function. The benchmark dose lower confidence limit (BMDL) of the relative expression of cTnT mRNA exposed by BPS during DP3 was the lowest among all the BMDLs of a good fit, with BMDL5 of 1.96 × 10-2 µg/mL, which is lower than the current reported exposure levels of BPS in maternal serum (0.03-0.07 ng/mL) and maternal umbilical cord serum (0.03-0.12 ng/mL).

Transglutaminase-triggered dual gradients of mechanical and biochemical cues self-assembling peptide hydrogel for guiding MC3T3-E1 cell behaviors

The mechanical properties and bioactive motif densities of extracellular matrix materials play crucial roles in regulating cell behaviors, such as cell adhesion, migration, proliferation, and differentiation. However, current studies on cellular responses to ECM predominantly concentrated on polymer hydrogels featuring a single factor, such as the mechanical strength, the types of bioactive motifs, and the morphology of the polymers. This limited focus may overlook the complex interplay of multiple factors. Here, we developed dual gradient peptide Q3GT-I3K hydrogels with tunable mechanical strength (0.3–4.0 kPa) and different density of bioactive motif (0.45–3.67 mM) by enzymatic crosslinking. These hydrogels can mimic the viscoelasticity of natural soft tissues. The properties of mechanical strength and cell responsive motif density could be controlled by modulating the proportion of the substrates in the enzymatic reaction. MC3T3 cells significantly differentiated into osteoblasts after seeded on the Q3GT-I3K hydrogel (2.8 kPa, 1.83 mM Q3GT) for 21 days, identifying from the elevated expression of alkaline phosphatase and substantial calcium nodule formation. Importantly, the engineered hydrogels exert a synergistic effect on the cell behaviors such as early adhesion, late proliferation, and differentiation of MC3T3-E1 cells. This paper introduces a new strategy for designing tissue engineering scaffold materials with specific functions.