Search Results (380)

Background: Coronary artery bypass grafting (CABG) is a well-established revascularization strategy for patients with multivessel coronary artery disease. The effectiveness of CABG is significantly influenced by antiplatelet therapy aimed at maintaining graft patency and reducing thrombotic complications. However, substantial inter-individual variability exists in platelet function responses to standard therapies such as aspirin and clopidogrel, leading to antiplatelet resistance. This variability has been linked to increased risks of myocardial infarction, stroke, and early graft failure. Platelet function testing (PFT) offers a potential strategy to identify resistance and guide more personalized antiplatelet therapy. This study aims to evaluate the association between perioperative platelet function test results and clinical outcomes following CABG. By assessing platelet responsiveness at multiple timepoints and correlating findings with postoperative events, the study seeks to determine whether PFT can stratify risk and improve patient management. Methods: This is a prospective, single-centre, observational cohort study conducted at a tertiary NHS cardiac surgery centre. Patients having elective or urgent isolated CABG will be enrolled and undergo perioperative PFT using the TEG6s system. Clinical outcomes will be monitored for 12 months postoperatively, with primary endpoints assessing the correlation between platelet function results and major adverse cardiovascular and cerebrovascular events (MACCE). Secondary endpoints will include the prevalence of antiplatelet resistance, demographic predictors, and the feasibility of integrating PFT into clinical workflows. Results: This study will report the prevalence of aspirin and clopidogrel resistance in CABG patients based on TEG6s PFT, as well as the correlation between platelet function results and MACCE, postoperative bleeding, and the need for surgical re-exploration. Additionally, it will examine the associations between demographic and clinical factors—such as diabetes status, renal function, BMI, and surgical technique—and variability in platelet responsiveness. The feasibility of incorporating PFT into perioperative workflows will also be evaluated, assessing whether results could support personalized antiplatelet management in future clinical trials. Conclusions: Findings from this study will provide real-world evidence regarding platelet function variability in CABG patients and suggest that PFT may identify those at increased risk of thrombotic complications. This exploratory analysis supports the need for larger interventional trials aimed at optimizing individualized postoperative antiplatelet therapy to improve surgical outcomes. Full article

The human body’s ability to recover from bone injuries is remarkable; however, in specific conditions, interventions are required to restore function and prevent complications. To accelerate osteogenesis, several strategies have been explored, including grafts, biomaterials, and adjuvant therapies. The aim of this narrative review was to analyze the preclinical evidence regarding the combination of particulate biomaterials and fibrin derivatives for bone regeneration. Publications using hydroxyapatite, bovine bone, β-tricalcium phosphate, and bioglass in association with fibrin glue, heterologous fibrin sealants, or platelet-rich fibrin were examined to identify recurrent experimental patterns and biological outcomes. According to the studies, hydroxyapatite and bovine bone were the most frequently investigated scaffolds, whereas fibrin glue and heterologous fibrin sealants showed consistent adhesion and favorable host response profiles in animal models. β-tricalcium phosphate demonstrated faster remodeling but lower volumetric stability, and bioglass showed high bioactivity in isolated reports. Despite heterogeneity in particle size, fibrin formulations, defect models, and follow-up periods, most studies reported enhanced bone deposition, vascularization, and integration when particulate biomaterials were combined with fibrin-based matrices. Overall, the evidence suggests that these combinations promote more organized and biologically favorable bone healing under experimental conditions. Future translational and clinical research is required to standardize protocols and determine the therapeutic applicability of these strategies in human bone repair. Full article

Cell membrane-coated nanoparticles represent a biomimetic drug delivery approach that integrates biological membrane functions with synthetic nanomaterials. Among the various membrane sources, those derived from blood cells such as red blood cells, platelets, and leukocytes offer distinctive advantages, including immune evasion, prolonged systemic circulation, and selective tissue targeting. These properties collectively enable efficient and biocompatible delivery of therapeutic agents to diseased tissues, minimizing off-target effects and systemic toxicity. This review focuses on blood cell membrane-derived nanocarriers as drug delivery and immune-regenerative platforms, in which membrane-mediated immunomodulation synergizes with therapeutic payloads to address inflammatory or degenerative pathology. We discuss recent advances in blood cell membrane coating technologies, including membrane isolation, nanoparticle core selection, fabrication techniques, and the development of hybrid and engineered membrane systems that enhance therapeutic efficacy through integrated immune regulation and localized drug action. To illustrate these advances, we also compile membrane type-specific nanocarrier systems, summarizing their core nanoparticle designs, coating strategies, therapeutic cargoes, and associated disease models. Challenges related to biological source variability, scalability, safety, and regulatory standardization remain important considerations for clinical translation. In this review we systematically address these issues and discuss emerging solutions and design strategies aimed at advancing blood cell membrane-based nanocarriers toward clinically viable immune-regenerative therapies. Full article

Bovine gammaherpesvirus 4 (BoGHV-4) is an opportunistic uterine pathogen whose reactivation is associated with postpartum inflammation and bacterial lipopolysaccharide (LPS). Platelet-rich plasma (PRP) is a regenerative biotherapeutic capable of modulating inflammatory responses, although its effects may vary depending on BoGHV4 genotype. In this study, primary bovine endometrial cells (BECs) were cultured in medium containing 10% PRP instead of fetal bovine serum, infected with two genetically divergent BoGHV-4 isolates (07-435, genotype 3; 10-154, genotype 2), and subsequently stimulated with bacterial lipopolysaccharide (LPS, 100 ng/mL). Expression of the viral immediate-early gene IE-2 and host immune genes (TLR4, TNF-α, CXCL8, and IFN-γ) were quantified by RT-qPCR from 4 to 48 h after stimulation. Isolate 07-435 induced a sustained activation of IE-2 and gradual cytokine upregulation, while isolate 10-154 elicited an early but transient inflammatory response followed by gene downregulation. PRP did not modify the strain-specific patterns of viral and inflammatory gene expression but established a common inflammatory baseline, whereas the magnitude and temporal profile of the response continued to be dictated by the viral genotype. These findings indicate that BoGHV-4 genotypic diversity remained the main determinant of response intensity and duration, supporting PRP as a context-dependent rather than a universal antiviral modulator. Full article

Dragon’s Blood, from the Dracaena cochinchinensis plant, is known for enhancing blood circulation. Its main components are Dragon’s Blood phenolic extracts (DBE). To pinpoint the active DBE constituents that are effective against thrombosis and understand their mechanism of action, the PT-stroke model was employed to assess DBE’s antithrombotic effects on cerebral blood flow and platelet aggregation. This investigation demonstrates that DBE enhances cerebral blood flow and inhibits ADP-induced platelet aggregation in photothrombotic (PT) stroke models. An FeCl₃-induced carotid artery thrombosis model was developed to test the antithrombotic activity of four DBE fractions. Through screening with this model, the ethyl acetate (EA) and methanol fractions were identified as the principal active components that effectively reduced thrombus weight and improved hemodynamics. Furthermore, the EA fraction was found to preserve the integrity of the blood–brain barrier. Phytochemical isolation allowed for the identification of compounds in the EA fractions, and UHPLC-MS was performed to characterize DBE and its active components in the bloodstream. In vitro ADP-induced platelet aggregation assays highlighted the active compounds. Through phytochemical analysis, Loureirin D (compound 17) was identified as a predominant constituent present in plasma. In vitro assays revealed that compounds 1 and 17 possess strong antiplatelet activity, with Loureirin D being confirmed as a selective P2Y12 receptor antagonist via molecular docking and cellular thermal shift assays. These findings substantiate Loureirin D as a pivotal antithrombotic component in DBE and its potential as a P2Y12-targeting therapeutic agent for thrombosis treatment. Full article

Despite treatments such as pentosan polysulfate, hyaluronic acid, botulinum toxin A, and platelet-rich plasma, many interstitial cystitis/bladder pain syndrome (IC/BPS) patients experience persistent symptoms. Urinary extracellular vesicles (uEVs) carry molecular cargo reflecting disease pathophysiology, yet their proteomic profiles in treated IC/BPS remain unexplored. This pilot study examined uEV proteomics in refractory IC/BPS cases to test the “Toxic Urine Hypothesis”—a vicious cycle, whereby urothelial dysfunction enables EV-mediated toxin penetration, triggering inflammation that further impairs the bladder barrier. Urinary EVs were isolated from six female IC/BPS patients on active treatments and four healthy female controls. Mass spectrometry-based proteomics identified differential protein expressions, followed by pathway enrichment analysis and functional validation using NF-κB reporter assays in HEK293T cells and Western blot in primary human bladder epithelial cells. IC/BPS EVs exhibited 31 upregulated proteins (including HPGD, KRT8, HSPA4, 14-3-3 family members) and 19 downregulated proteins (including neutrophil granule proteins MPO and ELANE), indicating suppressed acute neutrophil inflammation but enriched homeostatic, metabolic, and regenerative pathways. Patient EVs induced significantly higher NF-κB activation than in the controls, with upregulated 14-3-3ζ and phosphorylated NF-κB p65 in bladder epithelial cells. These findings support the “Toxic Urine Hypothesis”, revealing persistent NF-κB-mediated chronic epithelial stress despite suppressed acute inflammation in treated IC/BPS patients, suggesting that therapies targeting inflammation and regeneration may help break this vicious cycle. Full article

The recognition of trauma-induced coagulopathy (TIC) as an endogenous response to traumatic injuries rather than a consequence of therapeutic interventions has shifted the clinical approach toward an early and physiologically based hemostatic resuscitation. Prompt identification and correction of fibrinolysis and fibrinogen level derangements, dysregulated thrombin generation, and platelet dysfunction represent the cornerstones of the treatment strategies. Currently available viscoelastic hemostatic assays (VHAs) are point-of-care devices able to rapidly assess the phases of clot initiation, propagation, stabilization, and degradation, as well as isolate the contribution of specific elements—e.g., fibrinogen—to the coagulation process in fully automated analyses by multi-channel single-use cartridges. As a result, in the last decade, VHAs have been widely investigated as tools to implement individualized protocols of hemostatic resuscitation. Current guidelines support their use to optimize transfusion load in a goal-directed strategy. Nevertheless, contrasting evidence has emerged regarding the improvement in main clinical outcomes induced by the VHA-based algorithm of hemostatic resuscitation compared with those guided by conventional coagulation tests, and their place in the management of this peculiar population is still a matter of debate. We propose a narrative review ranging from TIC physiopathology as a proper substrate for viscoelastic diagnostic technique, through the strengths and weaknesses of VHAs, to their application in clinical practice. Full article

Immunothrombosis can substantially affect the course and outcomes of severe infections and immune-mediated diseases. While inflammatory thrombi are neutrophil-rich, impact of neutrophils on clot contraction, a key modulator of thrombus stability and obstructiveness, was unknown. This study investigated how neutrophils and neutrophil extracellular traps (NETs) affect the rate and extent of platelet-driven clot contraction. Isolated human neutrophils were stimulated with phorbol-12-myristate-13-acetate (PMA) to induce NETosis, confirmed by fluorescence microscopy and scanning electron microscopy. Thrombin-induced clots, formed from whole blood or platelet-rich plasma, were supplemented with non-activated or PMA-activated neutrophils. Clot contraction kinetics and viscoelasticity were analyzed. PMA-activated neutrophils significantly enhanced the rate and final extent of clot contraction compared to controls. This promoting effect was abolished by deoxyribonuclease (DNAse) I, confirming that it was mediated by NETs embedded in the fibrin network. The factor Xa inhibitor rivaroxaban also abrogated this effect, indicating a role for NET-induced endogenous thrombin generation and platelet hyperactivation. Thromboelastography revealed that NETs made clots softer and more deformable. We conclude that activated neutrophils promote clot contraction via NETs embedded in the fibrin network, which enhance platelet contractility via endogenous thrombin production and increase clot deformability, suggesting that inflammatory thrombosis may require treatments addressing this enhanced contractility. Full article

Neutrophils, key effector cells of the innate immune system, combat pathogens through mechanisms including the production of reactive oxygen species (ROS) and the release of neutrophil extracellular traps (NETs). While these responses are critical for host defense, prolonged elevation of ROS and dysregulated NETosis mediated by neutrophils have been implicated in autoimmune diseases, chronic inflammation, and cancer. In pancreatic ductal adenocarcinoma (PDAC), a highly aggressive and inflammatory malignancy, an increase in neutrophils infiltrating the tumor microenvironment promotes cancer progression and metastasis through increased ROS production and NET release. Using bioluminescence imaging with the reporter L-012 and NET assays, we assessed ROS and NET release, respectively, induced by phorbol myristate acetate and platelet-activating factor in bone-marrow-isolated neutrophils from wild-type and syngeneic myeloperoxidase (MPO)-deficient mice ex vivo. MPO deficiency impaired both ROS generation and NET release, establishing a positive correlation between these processes. In vivo analyses using subcutaneous and spontaneous murine PDAC models revealed elevated ROS in tumors, which were significantly reduced upon genetic deletion of host MPO or peptidyl arginine deiminase 4, an essential enzyme for NET formation, or after treatment with hydroxychloroquine, a NET inhibitor. Furthermore, luminol and 4-[¹⁸F]fluoro-1-naphthol ([¹⁸F]4FN), functional L-012 analogs, also enabled non-invasive detection of intratumoral ROS by bioluminescence and PET imaging in vivo, respectively; [¹⁸F]4FN PET showed a three-fold increased uptake in PDAC tumors versus muscle. PDAC tissues and blood-isolated neutrophils obtained from PDAC patients exhibited elevated ROS compared to controls ex vivo. Importantly, ROS levels correlated strongly with NET formation in patient samples. These findings reveal a bidirectional relationship between ROS and NETs and highlight the potential utility of L-012- and [¹⁸F]4FN-based PET imaging for monitoring NET-associated inflammation in PDAC in vivo. Full article

Background/Objectives: As bioactive extracellular vesicles, exosomes participate in cellular communication and disease mechanisms, yet their structural complexity continues to challenge standard analytical methodologies. This review summarizes published studies reporting exosome concentrations in human plasma, serum, and platelet-rich plasma from healthy individuals and highlights methodological differences. Methods: A comprehensive PubMed search (1986–31 August 2025) was performed using terms related to exosomes and their quantification, excluding cancer- and disease-related studies. Eligible articles reported exosome concentrations in plasma, serum, or platelet-rich plasma using particle-counting techniques such as nanoparticle tracking analysis, flow cytometry, or tunable resistive pulse sensing. Results: Twenty-two articles, including 167 healthy donors, met the inclusion criteria. The following mean concentration ranges were reported: plasma (n = 18), ranged from 4.50 × 10⁸ to 6.70 × 10¹¹ particles/mL with differences by quantification method; serum (n = 10), from 5.30 × 10⁸ to 2.13 × 10¹¹ particles/mL; non-activated platelet-rich plasma (n = 1), 7.52 × 10⁹ particles/mL; activated platelet-rich plasma (n = 3), 4.87 × 10¹⁰ to 7.16 × 10¹⁰ particles/mL; and preconditioned platelet-rich plasma with photothermal biomodulation (n = 2), 2.53 × 10¹¹ to 2.99 × 10¹¹ particles/mL. Conclusions: Isolation and quantification methods exhibit high variability, which strongly influences the overall quantity and quality of the exosomes obtained. Characteristics, including cargo composition, purity, and exosome integrity, must be considered when developing validated methods. Furthermore, emerging evidence suggests that PTBM preconditioning can increase exosome release from cells. In summary, rigorous standardization of protocols is essential to advance the scientific understanding and the clinical potential of exosome-based therapies. Full article

Prosthechea karwinskii is an orchid endemic to Mexico used for medicinal purposes. The objective of this study was to determine the anticoagulant potential ex vivo of the extract isolated using green solvents. Coagulometric assays were performed to evaluate the anticoagulant activity: activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). For each assay, different concentrations of the extract were evaluated (0.5, 1.0, 1.5, 2.5, 3.5, 4.5, 5.5, 7.5, and 8.5 mg/mL) using platelet-poor plasma from healthy donors. The P. karwinskii leaves extract showed an anticoagulant effect by significantly prolonging (p < 0.05) the APTT and TT from a concentration of 3.5 and 2.5 mg/mL, respectively, compared to basal. The anticoagulant activity was concentration dependent. In addition, the hydroethanolic extract of P. karwinskii leaves inhibited factor XI activity by 86.10 ± 2.3%. The compounds in the extract were identified by ultra-high-performance liquid chromatography coupled with electrospray ionization and quadrupole time-of-flight mass spectrometry (UPLC-ESI-qTOF-MS/MS). The compounds identified were quinic acid, malic acid, succinic acid, L (-)-phenylalanine, guanosine, neochlorogenic acid, chlorogenic acid, rutin, kaempferol-3-O-rutinoside, azelaic acid, sebacic acid, pinellic acid, and embelin. Full article

Kaempferitrin (KAE) is a natural flavonol dirhamnopyranoside with various pharmacological activities, isolated from the antithrombotic fraction of Celastrus orbiculatus Thunb. This study aimed to investigate the antithrombotic activity and “effective forms” of KAE. The results showed that KAE significantly prolonged rabbit plasma recalcification time in vitro. In the FeCl₃-induced rat arterial thrombosis model, KAE demonstrated antithrombotic effects by inhibiting coagulation, platelet aggregation, and fibrinolysis, with a lesser risk of bleeding compared to aspirin. KAE was orally administered to rats, and a total of 192 metabolites were characterized. These included 25 phase I metabolites, 8 hydroxylated and methylated metabolites, 57 sulfated metabolites, 74 glucuronidated metabolites, 26 sulfated and glucuronidated metabolites, and 2 glycosylated metabolites. Twenty-eight compounds were considered the in vivo “effective forms” of KAE for their antithrombotic activity. Network pharmacology, molecular docking, and molecular dynamics simulations collectively predict that these “effective forms” may exert antithrombotic effects by suppressing the SRC/PI3K/AKT pathway. This study provides a foundation for a better understanding of the in vivo “effective forms” and mechanisms underlying KAE’s antithrombotic activity, which is essential for understanding of “hexue” traditional efficacy of C. orbiculatus. Full article

Sepsis is a clinical syndrome caused by abnormal host response to infection. Thrombocytopenia and platelet dysfunction are common findings in sepsis and associated with worse outcomes. The innate immune single-stranded RNA sensor, Toll-like Receptor-7 (TLR7), plays a key role in thrombocytopenia in sepsis. This study investigated whether TLR7 signaling also contributes to platelet dysfunction in sepsis, and whether the bioactivity of downstream inflammatory mediators, specifically extracellular vesicles (EVs), is impacted by the TLR7 signaling pathway. Sepsis was induced in wild-type (WT) and TLR7-deficient (TLR7^−/−) mice by cecal ligation and puncture. Blood was collected at twenty-four hours for platelet and plasma isolation, and platelet function was assessed using aggregation, adhesion, and calcium flux assays. EVs were isolated from plasma and used in vitro to evaluate their impact on platelet–leukocyte aggregate (PLA) formation. We found that septic platelets are highly activated and more adhesive, yet show markedly impaired aggregation and reduced calcium signaling, indicating functional exhaustion despite activation. Notably, mice lacking TLR7 maintained stronger platelet aggregation, enhanced adhesion, and preserved calcium release in the septic state compared to wild-type controls, suggesting a protective effect of TLR7 deficiency. Plasma EVs increased in abundance and size during sepsis and promoted clot and PLA formation in vitro. Notably, EV-mediated platelet activation was reduced with EVs derived from TLR7-deficient mice. Our results demonstrate that while sepsis drives persistent platelet activation and dysfunction, TLR7 deficiency preserves platelet function and modulates the pathogenic activity of EV-mediated platelet activation, highlighting TLR7 as a key regulator and potential therapeutic target in sepsis-induced platelet dysfunction. Full article

Natural polyphenols, particularly tannins, are of interest due to their complex composition and multi-target biological activities. A highly purified tannin fraction was isolated from Aronia melanocarpa fruits, and its composition was characterized by HPLC-MS and IR spectroscopy. The Aronia tannin fraction exhibited comprehensive antioxidant properties, demonstrating superior DPPH radical scavenging activity compared to quercetin and a membrane-protective effect exceeding reference antioxidants. In vivo, Aronia tannins showed a delayed but potent antioxidant effect against cyclophosphamide (CP)-induced oxidative stress, significantly reducing malondialdehyde (MDA) levels, with the maximum effect observed at days 14–21. The immunomodulatory effect involved a complex regulation of the phagocytic system: selective activation of the monocytic arm with simultaneous modulation of neutrophilic activity. Crucially, a high phagocytic completion rate was maintained, indicating support for both bacterial uptake and intracellular killing. Tannins accelerated recovery post-CP, restoring leukocyte and platelet counts. Modulation of neutrophil oxidative metabolism, measured by chemiluminescence, indicates an ability to balance defense activation with prevention of excessive oxidative stress. These findings confirm the potential of the Aronia melanocarpa tannin fraction for correcting oxidative stress and immune dysfunction. Full article

Background: Diabetic wound healing has always been a clinical challenge with minimal response or efficacy to standard treatment. This study aims to assess the therapeutic potential of hypoxia-induced extracellular vesicles (hy-EVs) produced by human umbilical cord mesenchymal stem cells (HUCMSCs) to treat diabetic wounds. Methods: HUCMSCs were isolated from umbilical cord tissue, cultured under hypoxic conditions to induce the release of extracellular vesicles (EVs) and compared with normoxia-induced extracellular vesicles (n-EVs). We assessed the functions of hy-EVs on human skin fibroblasts (HSFs) and human umbilical vein endothelial cells (HUVECs) in vitro. Simultaneously, we analyzed the pro-angiogenic effects of hy-EVs, their effects on macrophage polarization, and their ability to scavenge endogenous reactive oxygen species (ROS). In addition, a diabetic wound model was established to assess the curative effect of hy-EVs in diabetic wound healing. Results: We found by in vitro study that hy-EVs markedly improved the functional activities of HSFs, thus significantly promoting wound repair. Remarkably, it was determined that hy-EVs greatly enhanced the proliferation and migration ability as well as the angiogenic ability of HUVECs, while promoting the expression of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial-generation-associated factor A (VEGFA), and platelet endothelial adhesion molecule (CD31), which suggested that hy-EVs can effectively activate the HIF-1α pathway to promote angiogenesis. Above all, we found that hy-EVs promoted the expression of CD206 while decreasing the expression of CD86, suggesting that hy-EVs could induce macrophages to shift from M1-type (pro-inflammatory) to M2-type (anti-inflammatory), thereby modulating the inflammatory response. Additionally, hy-EVs inhibited ROS production in both HSFs and HUVECs to reduce oxidative stress. In vivo results showed that hy-EVs enhanced collagen deposition and angiogenesis, modulated macrophage polarization, and inhibited immune response at the wound spot, which significantly enhanced diabetic wound healing. Conclusions: Our study shows that hy-EVs significantly promote angiogenesis through activation of the HIF-1α pathway, modulate macrophage polarization and attenuate cellular oxidative stress, possibly through delivery of specific miRNAs and proteins. Our discoveries offer a key theoretical basis and potential application to develop novel therapeutic strategies against diabetes-related tissue injury. Full article