Search Results (617)

Extracellular vesicles (EVs), nanoscale membrane-enclosed particles, are natural carriers of proteins and nucleic acids. Microalgae are widely used as a source of bioactive substances in the food and cosmetic industries and definitely have a potential to be used as the producers of EVs for biomedical applications. In this study, the extracellular vesicles isolated from the culture medium of two unicellular microalgae, Chlamydomonas reinhardtii (Chlamy-EVs) and Parachlorella kessleri (Chlore-EVs), were characterized by atomic force microscopy (AFM), cryo-electronic microscopy (cryo-EM), and nanoparticle tracking analysis (NTA). The biocompatibility with human cells in vitro (HEK-293T, DF-2 and A172) and biodistribution in mouse organs and tissues in vivo were tested for both microalgal EVs. An exogenous therapeutic protein, human heat shock protein 70 (HSP70), was successfully loaded to Chlamy- and Chlore-EVs, and its efficient delivery to human glioma and colon carcinoma cell lines has been confirmed. Additionally, in order to search for potential therapeutic biomolecules within the EVs, their proteomes have been characterized. A total of 105 proteins were identified for Chlamy-EVs and 33 for Chlore-EVs. The presence of superoxide dismutase and catalase in the Chlamy-EV constituents allows for considering them as antioxidant agents. The effective delivery of exogenous cargo to human cells and the possibility of the particle yield optimization by varying the microalgae growth conditions make them favorable producers of EVs for biotechnology and biomedical application. Full article

Primary aldosteronism (PA), the most common cause of secondary hypertension, is increasingly recognized as an independent driver of adverse cardiac remodeling, mediated through mechanisms beyond elevated blood pressure alone. Chronic aldosterone excess leads to myocardial fibrosis, left ventricular hypertrophy, and diastolic dysfunction via mineralocorticoid receptor activation, oxidative stress, inflammation, and extracellular matrix dysregulation. These changes culminate in a distinct cardiomyopathy phenotype, often underrecognized in early stages. Multimodality cardiac imaging, led primarily by conventional and speckle-tracking echocardiography, and complemented by exploratory cardiac magnetic resonance (CMR) techniques such as T1 mapping and late gadolinium enhancement, enables non-invasive assessment of structural, functional, and tissue-level changes in aldosterone-mediated myocardial damage. While numerous studies have established the diagnostic and prognostic relevance of imaging in PA, several gaps remain. Specifically, the relative sensitivity of different modalities in detecting subclinical myocardial changes, the long-term prognostic significance of imaging biomarkers, and the differential impact of adrenalectomy versus medical therapy on cardiac reverse remodeling require further clarification. Moreover, the lack of standardized imaging-based criteria for defining and monitoring PA-related cardiomyopathy hinders widespread clinical implementation. This narrative review aims to synthesize current knowledge on the pathophysiological mechanisms of aldosterone-induced cardiac remodeling, delineate the strengths and limitations of existing imaging modalities, and critically evaluate the comparative effects of surgical and pharmacologic interventions. Emphasis is placed on early detection strategies, identification of imaging biomarkers with prognostic utility, and integration of multimodal imaging into clinical decision-making pathways. By outlining current evidence and highlighting key unmet needs, this review provides a framework for future research aimed at advancing personalized care and improving cardiovascular outcomes in patients with PA. Full article

Background/Objectives: Myelofibrosis (MF) is a myeloproliferative neoplasm characterized by the replacement of healthy bone marrow (BM) with malignant and fibrotic tissue. In a healthy state, bone marrow is composed of approximately 60–70% fat cells, which are replaced as disease progresses. Proton density fat fraction (PDFF), a non-invasive and quantitative MRI metric, enables analysis of BM architecture by measuring the percentage of fat versus cells in the environment. Our objective is to investigate variance in quantitative PDFF-MRI values over time as a marker of disease progression and response to treatment. Methods: We analyzed existing data from three cohorts of mice: two groups with MF that failed to respond to therapy with approved drugs for MF (ruxolitinib, fedratinib), investigational compounds (navitoclax, balixafortide), or vehicle and monitored over time by MRI; the third group consisted of healthy controls imaged at a single time point. Using in-house MATLAB programs, we performed a voxel-wise analysis of PDFF values in lower extremity bone marrow, specifically comparing the variance of each voxel within and among mice. Results: Our findings revealed a significant difference in PDFF values between healthy and diseased BM. With progressive disease non-responsive to therapy, the expansion of hematopoietic cells in BM nearly completely replaced normal fat, as determined by a markedly reduced PDFF and notable reduction in the variance in PDFF values in bone marrow over time. Conclusions: This study validated our hypothesis that the variance in PDFF in BM decreases with disease progression, indicating pathologic expansion of hematopoietic cells. We can conclude that disease progression can be tracked by a decrease in PDFF values. Analyzing variance in PDFF may improve the assessment of disease progression in pre-clinical models and ultimately patients with MF. Full article

Reparative tertiary dentinogenesis requires the recruitment and odontogenic differentiation of dental pulp stem cells (DPSCs). Extracellular vesicles (EVs) as bioactive molecules have gained attention in regenerative medicine for their ability to mediate tissue repair through intercellular communication, influencing cell recruitment, proliferation, and differentiation. This study aimed to evaluate the effects of EVs on DPSC homing and odontogenic differentiation for dentin regeneration. DPSC-derived EVs were cultured in either growth (EV-G) or odontogenic differentiation (EV-O) conditions and isolated using a modified precipitation method. EVs were characterized by nanoparticle tracking analysis, scanning electron microscopy, antibody array, and cellular uptake assay. Treatment with 5 × 10⁸ EVs/mL significantly enhanced DPSC chemotaxis and proliferation compared with a no-treatment control and a lower dosage of EV (5 × 10⁷ EVs/mL). Gene expression and biochemical analyses revealed that EV-O up-regulated odontogenic markers including collagen type 1A1 (COL1A1), runt-related transcription factor 2 (RUNX2), and alkaline phosphatase (ALP). EV-O enhanced dentin regeneration by approximately 55% over vehicle controls in a rabbit partial dentinotomy/pulpotomy model. We identified key microRNAs (miR-21-5p, miR-221-3p, and miR-708-3p) in EV-O involved in cell homing and odontogenesis. In conclusion, our EV-based cell homing and odontogenic differentiation strategy has significant therapeutic potential for dentin regeneration. Full article

Suaeda salsa is relatively tolerant to cadmium (Cd) contamination. In order to investigate the bioaccumulation and stress responses of S. salsa under chronic exposure, we explored the growth, accumulation, and changes in antioxidant enzymes and glutathione (GSH) under different Cd concentrations over a 30-day soil culture experiment. Seedling height and weight in the 13.16 mg/kg Cd group were 13.26 cm and 0.21 g, significantly higher than the control group. Growth was significantly inhibited under high Cd concentration exposure, with a seedling and root length of 9.65 cm and 3.77 cm. The Cd concentration in all tissues was positively related to Cd treatment concentration, with the Cd contents in the roots being higher than in the other tissues. At a subcellular level, Cd was mainly concentrated in the cell walls, organelles, and soluble components within the range of 0.05–8.29, 0.02–2.40 and 0.08–1.35 μg/g, respectively. The accumulation of Cd in the roots tracked its proportion in the cell walls. The malondialdehyde (MDA) content of the plant tissues increased with increasing Cd concentration, indicating that Cd stress caused oxidative damage. The GSH content increased with increasing Cd concentration, with maximum values of 0.515 μmol/g in the stem in the 66.07 mg/kg Cd group. The catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD) activity showed different change trends under Cd exposure. The results in this study could provide useful information on the tolerance mechanism of Cd in S. salsa, which provides information for exploiting S. salsa as a candidate for phytoremediation of Cd contamination. Full article

Background: Dynamic computer-aided navigation systems are a real-time motion tracking technology widely applied in oral implantology and endodontics to enhance precision and reduce complications. However, their reliability, accuracy, and usability in dentoalveolar surgery and maxillary bone augmentation remain underinvestigated. Methods: A systematic review following PRISMA guidelines was conducted and registered on PROSPERO (CRD42024610153). PubMed, Scopus, Web of Science, and Cochrane Library databases were searched until October 2024 to retrieve English eligible studies, without restrictions on the publication year, on dynamic computer-assisted navigation systems in dentoalveolar and bone augmentation surgeries. Exclusion criteria were surgery performed without dynamic computer-assisted navigation systems; dental implant placement; endodontic surgery; and maxillo-facial surgery. The outcomes were reliability, accuracy, post-operative course, surgical duration, complications, patient- and clinician-reported usability, acceptability, and satisfaction. Included studies were qualitatively synthetized and judged using dedicated tools for the different study designs. Results: Twenty-nine studies with 214 patients were included, showing high reliability in dentoalveolar and bone augmentation surgeries comparable to or superior to freehand surgeries, higher accuracy in dentoalveolar surgery compared to maxillary bone augmentation, and reduced complication rates across all surgeries. While overall surgical duration slightly increased due to technology installation, operative time was reduced in third molar extractions. Patient-reported outcomes were poorly investigated. Clinician-reported outcomes were mixed, but difficulties in the differentiation of soft tissue from hard tissue were recorded, especially in sinus floor elevation. Conclusions: Dynamic computer-assisted navigation systems enhance accuracy and safety in dentoalveolar and bone augmentation surgery. Further studies are needed to assess the underinvestigated patient-reported outcomes and standardize protocols. Full article

Background: Many patients harbor minimal residual disease (MRD)—small clusters of residual tumor cells that survive therapy and evade conventional detection but drive recurrence. Although advances in molecular and computational methods have improved circulating tumor DNA (ctDNA)-based MRD detection, these approaches face challenges: ctDNA shedding fluctuates widely across tumor types, disease stages, and histological features. Additionally, low levels of driver mutations originating from healthy tissues can create background noise, complicating the accurate identification of bona fide tumor-specific signals. These limitations underscore the need for refined technologies to further enhance MRD detection beyond DNA sequences in solid malignancies. Methods: Profiling circulating cell-free mRNA (cfmRNA), which is hyperactive in tumor and non-tumor microenvironments, could address these limitations to inform postoperative surveillance and treatment strategies. This study reported the development of OncoMRD BREAST, a customized, gene signature-informed cfmRNA assay for residual disease monitoring in breast cancer. OncoMRD BREAST introduces several advanced technologies that distinguish it from the existing ctDNA-MRD tests. It builds on the patient-derived gene signature for capturing tumor activities while introducing significant upgrades to its liquid biopsy transcriptomic profiling, digital scoring systems, and tracking capabilities. Results: The OncoMRD BREAST test processes inputs from multiple cutting-edge biomarkers—tumor and non-tumor microenvironment—to provide enhanced awareness of tumor activities in real time. By fusing data from these diverse intra- and inter-cellular networks, OncoMRD BREAST significantly improves the sensitivity and reliability of MRD detection and prognosis analysis, even under challenging and complex conditions. In a proof-of-concept real-world pilot trial, OncoMRD BREAST’s rapid quantification of potential tumor activity helped reduce the risk of incorrect treatment strategies, while advanced predictive analytics contributed to the overall benefits and improved outcomes of patients. Conclusions: By tailoring the assay to individual tumor profiles, we aimed to enhance early identification of residual disease and optimize therapeutic decision-making. OncoMRD BREAST is the world’s first and only gene signature-powered test for monitoring residual disease in solid tumors. Full article

Tracking T cells in vivo using MRI is a major challenge due to the difficulty of labeling these non-phagocytic cells with a sufficient contrast agent to generate a detectable signal change. In this study, we explored CD4-Superparamagnetic iron oxide nanoparticles (SPION), which is commonly used in magnetic cell sorting, as a potential receptor-mediated, specific CD4⁺ T cell MRI labeling agent. We optimized the labeling protocol for maximal CD4⁺ cell labeling and viability. Cell health was confirmed with trypan blue assay, and labeling efficacy was confirmed with Prussian blue staining, transmission electron microscopy, and MRI of labeled cell pellets. Key cell functionality was assessed by flow cytometry. Next, CD4-SPION-labeled T cells or unlabeled T cells were delivered via intravenous injection in naïve mice. Liver MRIs pre-, 24 h, and 72 h post-T cell injection were performed to determine in vivo tracking ability. Our results show that CD4-SPION induces significant attenuation of T2 signals in a concentration-dependent manner, confirming their potential as an effective MRI contrast agent. In vitro, analyses showed that CD4⁺ T cells were able to uptake CD4-SPION without affecting cellular activity and key functions, as evidenced by Prussian blue staining and flow cytometric analysis of IL-2 receptor and the IL-7 receptor α-chains, CD69 upregulation, and IFN-γ secretion. In vivo, systemically distributed CD4-SPION-labeled T cells could be tracked in the liver at 24 and 72 h after injection, contrary to controls. Histological staining of tissue sections validated the findings. Our results showed that SPION CD4⁺ T cell sorting coupled with longitudinal MR imaging is a valid method to track CD4⁺ T cells in vivo. This safe, specific, and sensitive approach will facilitate the use of SPION as an MRI contrast agent in clinical practice, allowing for non-invasive tracking of adoptive cell therapies in multiple disease conditions. Full article

Background and Clinical Significance: Prosthetic rehabilitation in the aesthetic zone of periodontally compromised patients presents a complex clinical challenge, requiring a careful coordination of aesthetic, functional, and biological demands. This case highlights the benefits of digital dentistry, interdisciplinary collaboration, and regular maintenance in achieving long-term success in complex rehabilitations of periodontally compromised patients. Case Presentation: This case report describes the digital minimally invasive rehabilitation of a 39-year-old male patient with Stage III periodontitis, occlusal discrepancies, tooth mobility, and an interincisal diastema. A fully digital workflow—including intraoral scanning, aesthetic previewing, and mandibular motion analysis—was employed to guide diagnosis, treatment planning, and prosthetic execution. Conservative tooth preparations using a biologically oriented approach (BOPT) were combined with customised provisional restorations to support soft tissue conditioning and functional control throughout the provisional phases. Mandibular motion tracking facilitated the design of a personalised anterior guidance to improve occlusion and correct the deep bite. The interincisal diastema was initially maintained then closed during the advanced phase of treatment based on aesthetic simulations and patient preference. One unplanned endodontic treatment was required during the provisional phase, but no other complications occurred. Conclusions: At the four-year follow-up, the patient demonstrated stable periodontal and occlusal conditions, improved clinical indices, and high satisfaction with the aesthetic outcome. Full article

Characterized by insufficient blood supply leading to tissue hypoxia and damage, ischemia is the underlying cause of major conditions such as ischemic stroke, myocardial infarction, and peripheral artery disease. Stem cell therapy, as a regenerative strategy, demonstrates significant potential in restoring tissue blood flow and organ function in ischemic environments. This review systematically explores the latest advances in stem cell therapy for ischemic diseases, focusing on different cell types and their mechanisms of action, including direct differentiation, paracrine signaling, immunomodulation, and microenvironment regulation. Furthermore, it highlights innovations in gene editing and bioengineering technologies that enhance cell delivery, targeting, and therapeutic efficacy. Simultaneously, this article discusses the challenges faced, advances in cell tracking and delivery, and future research directions, aiming to provide insights for the development of more effective and personalized treatment strategies Full article

The global prevalence of hepatocellular carcinoma (HCC) is getting worse, leading to an urgent need for improved diagnostic and prognostic strategies. Liquid biopsy, which analyzes circulating tumor cells (CTCs), cell-free DNA (cfDNA), cell-free RNA (cfRNA), and extracellular vesicles (EVs), has emerged as a minimally invasive and promising alternative to traditional tissue biopsy. These biomarkers can be detected using sensitive molecular techniques such as digital PCR, quantitative PCR, methylation-specific assays, immunoaffinity-based CTC isolation, nanoparticle tracking analysis, ELISA, next-generation sequencing, whole-genome sequencing, and whole-exome sequencing. Despite several advantages, liquid biopsy still has challenges like sensitivity, cost-effectiveness, and clinical accessibility. Reports highlight the significance of multi-analyte liquid biopsy panels in enhancing diagnostic sensitivity and specificity. This approach offers a more comprehensive molecular profile of HCC, early detection, and tracking therapeutic treatment, particularly in those cases where single-analyte assays and imaging fail. The technological advancement in the isolation and analysis of CTC, cell-free nucleic acids, and EVs is increasing our understanding of extracting genetic information from HCC tumors and discovering mechanisms of therapeutic resistance. Furthermore, crucial information on tumor-specific transcriptomic and genomic changes can be obtained using cfRNA and cfDNA released into the peripheral blood by tumor cells. This review provides an overview of current liquid biopsy strategies in HCC and their use for early detection, prognosis, and monitoring the effectiveness of HCC therapy. Full article

Background: Pancreatic adenocarcinoma (PAAD) is an aggressive subtype of pancreatic cancer that is estimated to have a 5-year overall survival rate of only 13%. Most patients present with advanced disease with unpredictable outcomes. The identification of prognostic biomarkers is important to accurately stratify these patients. Methods: We investigated the molecular and survival-related role of ENHO in PAAD by analyzing TGCA mRNA and miRNA data. Survival analysis was conducted using TIMER2.0, “survival”, and “survminer”. Gene set enrichment analysis was conducted using enrichr, while miRNA-mRNA interactions were identified using “multiMiR”. Immune infiltration was assessed using CIBERSORT ABS and ImmuCellAI. Results: We observed that ENHO was strikingly downregulated in PAAD tissues (p = 3.68 × 10⁻⁶⁸), and patients with higher ENHO levels enjoyed significantly better overall survival (HR = 0.597; 95% CI: 0.419–0.852; p < 0.01). Pathway analysis showed that genes co-upregulated with ENHO were enriched for insulin secretion and ion channel activity, whereas those co-downregulated were related to epithelial–mesenchymal transition and extracellular matrix remodeling. Higher ENHO also tracked with increased CD8⁺ T-cell infiltration and correlated positively with PDCD1 and LAG3 but negatively with B7-H3, CD70, and NT5E. Conclusions: Our results point to a protective role for ENHO in pancreatic adenocarcinoma. Full article

Extracellular vesicles (EVs) are lipid membrane-enclosed particles released by all cells and can be isolated from various sources, even from solid tissues. This study focuses on isolating and characterizing EVs from mouse lymph nodes (LNs). Male C57BL/6 mice were injected with complete Freund’s adjuvant, with or without ovalbumin. Inguinal and popliteal LNs were incised 9 days after immunization, and EV isolation was carried out using a combination of differential centrifugation and size-exclusion chromatography. The characteristic morphology of small and large EVs was confirmed by transmission electron microscopy. Particle size distribution and concentration were determined by nanoparticle tracking analysis, while protein and lipid contents were measured by bicinchoninic acid assay, and sulfo-phospho-vanillin assays, respectively, to calculate the protein-to-lipid ratio. Immune and EV markers were analyzed by using flow cytometry and Western blot assay, revealing significant changes between immunized mice compared to controls. This study establishes a novel protocol for isolating and characterizing EVs from LNs and highlights the impact of immunization on EV properties, offering insights into their roles in immune processes. Full article

Background/Objectives: Although cardiac resynchronization therapy (CRT) plays an established role in the management of heart failure, a significant proportion of patients do not respond despite appropriate candidate selection. The optimization of CRT pacing is one strategy to enhance response. Fusion pacing algorithms aim to synchronize intrinsic right ventricular (RV) conduction with paced left ventricular (LV) activation, resulting in a more physiological ventricular depolarization pattern. This approach may improve electrical synchrony and enhance left ventricular contraction compared to conventional simultaneous biventricular pacing. The aim of this study was to compare the acute, beat-to-beat effects of standard biventricular pacing versus fusion pacing on myocardial function, using both conventional and speckle-tracking echocardiography in heart failure patients with left bundle branch block (LBBB). Methods: In total, 27 heart failure patients (21 men and 6 women) with reduced ejection fraction (EF < 35%), left bundle branch block (QRS > 150 ms), and newly implanted CRT-D systems (Abbott) underwent echocardiographic assessment immediately after device implantation. Echocardiographic parameters—including left atrial strain, left ventricular strain, TAPSE, mitral and tricuspid valve function, and cardiac output—were measured at 5 min intervals under three different pacing conditions: pacing off, simultaneous biventricular pacing, and fusion pacing using Abbott’s SyncAV^® algorithm. Results: In our study, CRT led to a significant shortening of the QRS duration from 169 ± 19 ms at baseline to 131 ± 17 ms with standard biventricular pacing, and further to 118 ± 16 ms with fusion pacing (p < 0.05). Despite the electrical improvement, no significant changes were observed in global longitudinal strain (GLS: −9.15 vs. −9.39 vs. −9.13; p = NS), left ventricular stroke volume (67.5 mL vs. 68.4 mL vs. 68.5 mL; p = NS), or left atrial parameters including strain, area, and ejection fraction. However, fusion pacing was associated with more homogeneous segmental strain patterns, improved aortic valve closure time, and enhanced right ventricular function as reflected by tissue Doppler-derived S’. Conclusions: Immediate QRS narrowing observed in CRT patients—particularly with fusion pacing optimization—is associated with a more homogeneous pattern of left ventricular contractility and improvements in selected measures of mechanical synchrony. However, these acute electrical changes do not translate into immediate improvements in stroke volume, global LV strain, or left atrial function. Longer-term follow-up is needed to determine whether the electrical benefits of CRT, especially with fusion pacing, lead to meaningful hemodynamic improvements. Full article

Electric fields (EFs) are widely employed to promote tissue regeneration and accelerate wound healing. Despite extensive study, the cellular responses elicited by EFs are complex and not well understood. The present work focuses on cell migration—a process essential to organismal development, immune surveillance, and repair—and seeks to achieve its precise, closed-loop regulation. Effective control is impeded by (i) the nonlinear and stochastic nature of migratory dynamics and (ii) safety constraints that restrict the admissible EF magnitude. To address these challenges, we reformulate a neural network (NN) feedback controller previously developed for single-cell membrane-potential regulation and adapt it to guide population-level cell migration. A projection operator is embedded into the NN weight-update law to prevent maladaptive learning that arises when the control signal saturates at its EF limit. Numerical simulations confirm that the modified controller maintains accurate trajectory tracking under saturation and outperforms the original NN design. Finally, we demonstrate a proof-of-concept by implementing the controller in vitro to direct the electrotactic migration of naïve macrophages in 2D culture under a unidirectional EF. For the in vitro experiments, we compare performance to the standard proportional–integral–derivative (PID) controller. Full article