Search Results (321)

Background: Uncontrolled hemorrhage, especially at non-compressible sites, remains a major cause of preventable trauma deaths. This study reports the development of fibrinogen-loaded calcium carbonate (CaCO₃) microparticles that combine hemostatic activity with self-propelling capability for targeted delivery against blood flow, with a focus on understanding formulation-dependent trade-offs among particle yield, protein loading, clotting performance, and transport behavior. Methods: Microparticles were synthesized via a precipitation method using different carbonate sources and characterized for yield, morphology, size, and fibrinogen encapsulation. Hemostatic function was assessed using rotational thromboelastometry (ROTEM) in fibrinogen-deficient plasma. Propulsion behavior was evaluated following exposure to protonated tranexamic acid (TXA⁺), which triggers CO₂ generation. Particle size and encapsulation were examined by microscopy and fluorescence imaging. Results: The precipitation method produced spherical micrometer-sized particles, with fibrinogen inclusion reducing yield and particle size relative to unload controls. Fluorescence microscopy confirmed successful encapsulation. Encapsulation efficiency varied with formulation, with sodium carbonate-based particles showing higher relative fibrinogen loading. ROTEM analysis demonstrated that fibrinogen-loaded particles significantly improved clot formation, increasing maximum clot firmness compared to fibrinogen-free particles, although performance remained formulation-dependent. TXA⁺-triggered propulsion achieved maximum speeds up to 4.221 cm/s. Fibrinogen-loaded particles exhibited longer activation lag times than unloaded particles, indicating a trade-off between hemostatic functionality and propulsion kinetics. Conclusions: Fibrinogen-loaded CaCO₃ microparticles exhibit both hemostatic activity and chemically triggered motion in vitro. The study identifies key formulation-dependent trade-offs between particle yield, fibrinogen loading, clotting performance, and propulsion behavior. While these findings support the feasibility of combining localization and clot stabilization mechanisms, further studies under physiologically relevant flow conditions and in vivo models are required to evaluate their potential for active delivery in non-compressible hemorrhage. Full article

Proteases, enzymes that catalyze the hydrolysis of peptide bonds in peptides and proteins, have widespread industrial applications, particularly in milk coagulation for cheese production. Microbial enzymes have been employed as alternatives to animal rennet, offering advantages such as cost-effectiveness, availability, and compliance with dietary, cultural, and religious requirements. Solid-state fermentation (SSF) is widely employed for microbial enzyme production because of its low operational costs, reduced water and energy requirements, high product concentrations, and the ability to utilize agro-industrial residues as low-cost substrates, thereby contributing to both process sustainability and waste valorization. We report the production and characterization of a novel milk-clotting protease produced by Trichoderma koningii FFT13. The protease was produced by SSF using wheat bran as the substrate, an agro-industrial residue. It was classified as a chymotrypsin-like serine protease and exhibited a specific caseinolytic activity of 9861 U/mg. The enzyme coagulated both reconstituted skim milk and pasteurized whole milk in the presence or absence of calcium. Coagulation was enhanced by increasing temperature, reaction time, enzyme concentration, and calcium levels. Scanning electron microscopy revealed destabilization of casein micelles, their progressive aggregation, and the formation of a well-defined gel network, confirming the effectiveness of the protease in milk coagulation. Therefore, these results demonstrate that the chymotrypsin-like protease from T. koningii is a promising enzyme for milk coagulation, with potential application in cheese production. The enzyme obtained constitutes an alternative to traditional coagulants, overcoming limitations related to animal rennet while potentially offering additional advantages in terms of process sustainability and industrial scalability. Full article

Background/Objectives: Early thrombosis of systemic-to-pulmonary artery shunts (SPS) remains a major cause of morbidity and mortality in neonates with duct-dependent pulmonary circulation. Despite advances in surgical technique, no universally accepted perioperative thrombosis-prevention protocol exists. We evaluated the early outcomes of a standardized perioperative thrombosis-prevention protocol applied in neonates undergoing SPS placement. Methods: This single-center case series included nine consecutive neonates undergoing primary modified Blalock–Taussig shunt placement between January 2024 and July 2025. A predefined and standardized perioperative thrombosis-prevention protocol was uniformly applied, incorporating preoperative aspirin when feasible, intraoperative systemic heparinization targeting activated clotting time (ACT) > 300 s, meticulous shunt flushing and de-airing, preferential distal anastomosis to the main pulmonary artery when anatomically suitable, and early postoperative continuous heparin infusion followed by enteral aspirin. The primary endpoint was early shunt thrombosis within 30 days. Results: Median age at surgery was 28 days (range 14–35), and median operative weight was 3.2 kg (range 2.8–3.6). Cardiopulmonary bypass was required in 33.3% of patients. Delayed sternal closure was performed in 22.2%. Despite recognized prothrombotic risk factors—including complex anatomy, hypoplastic pulmonary arteries, and low cardiac output syndrome (33.3%)—no early shunt thrombosis occurred (0/9). There were no reinterventions, no early mortality, and no major bleeding or intracranial hemorrhage. Conclusions: In this single-center neonatal series, implementation of a standardized perioperative thrombosis-prevention protocol was associated with preserved early shunt patency without increased bleeding risk. Although limited by a small sample size, these findings support the feasibility and short-term safety of a standardized perioperative management strategy in neonatal systemic-to-pulmonary shunt surgery. These findings should be considered hypothesis-generating and not evidence of definitive effectiveness. Full article

Uncontrolled bleeding, coagulation disorders, and infection-related complications still present substantial challenges in emergency medicine and trauma care. Developing multifunctional hemostatic materials represent an effective strategy for addressing clinical hemostasis problems. In this study, Galla chinensis polyphenols, the effective extract of Galla chinensis, were loaded onto calcium alginate-mesoporous silica granules (CMS-GC). The CMS granules were prepared by in situ liquid-phase technology and GC was loaded by impregnation methods. In vitro and in vivo studies showed that CMS-GC not only activate the endogenous coagulation pathway via GC, but also the multi-level interconnected pores of CMS granules can promote the cross-linking of GC with plasma proteins and formation of a three-dimensional network structure, which further enhances the coagulation effect and shortens the blood clotting time to less than 80 s. In rat liver and femoral artery hemorrhage models, CMS-GC significantly shortened hemostasis time and reduced blood loss, demonstrating superior hemostatic performance. Moreover, within the moist environment sustained by alginate, GC mitigates inflammatory responses via its antibacterial and free-radical clearance properties, and synergistically facilitates wound healing. This CMS-GC multifunctional granule provides an efficient new strategy for traumatic bleeding and subsequent repair. Full article

Background: Tranexamic acid (TXA) is a synthetic lysine analog widely used as an antifibrinolytic agent. Large randomized trials have demonstrated life-saving benefits when TXA is administered early in acute hemorrhage, but results regarding prophylactic administration have been conflicting, and several trials have not shown improved clinical outcomes. The mechanisms underlying this discrepancy remain incompletely understood. Objectives: To investigate the molecular and structural mechanisms that determine TXA efficacy in purified fibrin clots under conditions mimicking therapeutic versus prophylactic administration. Methods: We examined fibrinolysis induced by tissue plasminogen activator (tPA) in vitro using confocal microscopy, viscoelastic testing (ClotPro), turbidimetry, and plasmin generation assays at physiologically and therapeutically relevant concentrations of plasminogen and TXA. Scanning electron microscopy (SEM) was employed to assess fibrin structure. Results: When TXA was incorporated into fibrin clots before the addition of tPA, physiological plasminogen concentrations (2.5 µM) reversed the antifibrinolytic effect, resulting in paradoxical acceleration of lysis. By contrast, when clotting and fibrinolysis occurred simultaneously in the presence of TXA and tPA, TXA consistently prolonged lysis time irrespective of plasminogen concentration. SEM demonstrated that TXA, even at concentrations as low as 16 µM, doubled the top-quartile values of the fibrin fiber diameter, altering susceptibility to plasmin-mediated degradation without accelerating plasminogen activation. Conclusions: TXA efficacy is determined not only by dose but also by timing and the plasminogen availability in the clot microenvironment. These findings provide mechanistic insight into the failure of prophylactic TXA administration and highlight the importance of context in optimizing its clinical use. Full article

Two hemostatic bionanocomposite dressings were developed using natural, semi-natural (or semi-synthetic) and synthetic polymers. The first system consisted of chitosan (CS), polyvinyl alcohol (PVA), and carboxymethyl cellulose (CMC) (CS/PVA/CMC), while the second was based on CS, PVA, and starch (SR) (CS/PVA/SR). Zinc oxide (ZnO) nanoparticles and bicyclic monoterpene camphor (CP) ketone were incorporated as bioactive agents in order to enhance antimicrobial and hemostatic performance. FTIR spectroscopy confirmed the successful solvent casting synthesis of the dressings and the interactions between the biopolymers and additives. XRD analysis indicated a predominantly amorphous structure, while SEM images and EDS analysis revealed uniform dispersion of ZnO particles within the polymer matrices without aggregation. Furthermore, the CS/PVA/CMC-1ZnO/CP sample exhibited a water sorption of 12,666 ± 126%, while CS/PVA/SR-1ZnO/CP reached 7013 ± 215%. ZnO incorporation also improved mechanical performance, with CS/PVA/SR-2ZnO/CP displaying the highest tensile strength (39.18 ± 0.2 MPa) and elongation at break (9.54 ± 1.04%). ZnO incorporation also led to a concentration-dependent increase in antibacterial activity, with SR-based dressings achieving near-complete bacterial reduction at higher ZnO loadings. All the dressings demonstrated good biocompatibility, while CS/PVA/SR-1ZnOCP showed the fastest clotting time (420 s ± 40), highlighting its potential for hemostatic applications. Full article

Objectives: Perioperative blood transfusion for acute type A aortic dissection (ATAAD) lacks clinical guidelines. This study aims to investigate the application of a thromboelastography (TEG)-based risk-stratified transfusion protocol in these patients. Methods: We conducted a two-stage study. Firstly, a retrospective analysis of ATAAD patients undergoing surgery in 2023 was performed to identify predictors of postoperative/perioperative excessive bleeding and develop a predictive model. Subsequently, a single-center prospective validation study was conducted in 2024, comparing a TEG-based risk-stratified transfusion protocol against conventional empirical transfusion. Results: In the retrospective phase (n = 57), 18 patients (31.6%) developed perioperative excessive bleeding. Preoperative activated clotting time (ACT) and TEG parameters (K-time) were independent predictors. A predictive model incorporating these variables achieved an AUC of 0.788. In the prospective phase (n = 47), 21 patients received the TEG-based risk-stratified transfusion protocol. Compared to the conventional group, the TEG risk-stratified group exhibited significantly lower postoperative drainage volume (p = 0.046), a reduced incidence of perioperative excessive bleeding (4.8% vs. 34.6%, p = 0.033), and lower transfusion costs (p = 0.029), without an increase in total transfusion volume. Conclusions: Preoperative ACT and TEG parameters effectively predict perioperative excessive bleeding in ATAAD patients. Implementing a TEG-based risk-stratified transfusion protocol optimizes blood product utilization, improves clinical outcomes, and reduces costs, offering a promising evidence-based approach for perioperative management. Full article

The Cape cobra (Naja nivea), one of Africa’s most lethal snakes, can cause rapid, life-threatening paralysis. However, the impact of this venom on platelet function and blood coagulation remains poorly understood. To address this gap, we investigated the enzymatic profiles and the impacts of N. nivea venom on multiple aspects of haemostasis using human whole blood. Our results illustrate that Cape cobra venom significantly increases clotting time in rotational thromboelastometry without affecting other coagulation parameters. This venom significantly inhibits platelet aggregation and activation yet does not exert cytotoxic effects on platelets. The venom was subsequently fractionated using reverse-phase high-performance liquid chromatography, and the most potent purified fraction was identified as a cytotoxin (three-finger toxin) through mass spectrometry. This purified fraction showed an inhibitory effect on platelet activity. These findings highlight that N. nivea venom can induce haemotoxicity in addition to neurotoxicity. Moreover, three-finger toxins may be promising candidates for bioprospecting to develop novel antithrombotic agents. Full article

Gene therapy is reshaping the therapeutic paradigm in hemophilia by enabling sustained endogenous clotting factor production after a single administration. This approach moves disease management beyond lifelong replacement therapy. While clinical trials have demonstrated marked reductions in bleeding rates and treatment burden, real-world implementation has revealed emerging complexities. These include interindividual variability in transgene expression reflected by a progressive reduction in circulating FVIII or FIX activity over time, uncertainty regarding the long-term durability of expression, immune-mediated constraints, and episodes of transaminase elevation. This review addresses a critical transition point in the field: the shift from proof-of-concept efficacy toward integration of gene therapy into long-term hemophilia care. We examine determinants of therapeutic stability, host–vector immune interactions, and mechanisms underlying loss or fluctuation of expression, with emphasis on monitoring strategies and post-therapy management pathways. Immunogenic processes affecting vector transduction, hepatocellular responses, and transgene persistence are discussed alongside current approaches to immune modulation. This review uniquely focuses on post-gene therapy clinical integration rather than vector design or trial outcomes. Beyond direct factor correction, evolving therapeutic concepts targeting coagulation rebalancing and immune regulation are considered within a systems-based framework. Psychosocial adaptation and patient-reported outcomes are also explored, underscoring that therapeutic success extends beyond hemostatic control. In aggregate, these perspectives position gene therapy not as a singular curative event but as a component of an evolving, biologically integrated management strategy. Long-term follow-up translational research (LTFU) and coordinated global efforts will be essential to optimize durability, safety, and equitable access. Full article

Plant-derived coagulants are increasingly explored as alternatives to animal rennet. This study provides the first evaluation of the coagulation kinetics and technological properties of Onopordum platylepis Murb. in ewe’s milk. Response surface methodology was applied to optimize temperature (30, 33, and 36 °C), pH (5.5, 6.0, and 6.5), and calcium chloride concentration (2, 4, and 6 mM). The optimal conditions for minimizing milk-clotting time were 36 °C, pH 5.5, and 6 mM CaCl₂. Under standardized activity (50 IMCU/L), the Rheological properties of gels produced by Onopordum platylepis were compared with Cynara cardunculus, Cynara humilis, animal rennet, a commercial plant coagulant, and fermentation-produced chymosin. Onopordum platylepis showed slower curd-firming rates than animal rennet and Cynara cardunculus, but similar behavior to Cynara humilis. Gels produced with Onopordum platylepis exhibited firmness comparable to commercial plant coagulants. The water-holding capacity was similar to other coagulants, though protein losses were higher for Onopordum platylepis, Cynara humilis, and Cynara cardunculus than animal rennet. Overall, Onopordum platylepis demonstrates potential for ewe’s milk cheese production, in which highly proteolytic coagulants are used. Full article

Background/Objectives: Syringa vulgaris L. (common lilac) is one of the most popular ornamental plant species. Through the ages, many parts of S. vulgaris, including fruits, flowers, leaves, and branches, have been used in folk medicine due to their beneficial biological activity. Lilac flowers are the basis of many supplements available on the market. Moreover, its petals and flowers are edible and are an aromatic ingredient in preserves and desserts. However, the data about the antioxidant properties of various parts of S. vulgaris is limited only to the in vitro antioxidant capacity of the extracts—so far, the effect of S. vulgaris flower extract on the parameters of oxidative stress in biological materials, including plasma, has not been demonstrated. Therefore, the aim of our study was to investigate the protective effects of the extract from S. vulgaris L. flowers against oxidative stress in human plasma, and its influence on the coagulation process in vitro. Methods: We measured the levels of three parameters of oxidative stress in human plasma treated with H₂O₂/Fe²⁺ (the donor of hydroxyl radicals): lipid peroxidation (based on the level of thiobarbituric acid reactive substances (TBARS)), protein carbonylation, and thiol oxidation. Ascorbic acid (vitamin C) was used as a reference antioxidant. In addition, we studied the effect of the extract on three coagulation parameters of human plasma-activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). We also compared the biological properties of the extract from S. vulgaris flowers with the properties of a phenolic extract from Taraxacum officinalis (dandelion) flowers, as they have proven antioxidant activity in both in vitro and in vivo models and can modulate hemostasis in vitro. Results: Our UHPLC-HRMS analyses of S. vulgaris extract led to a tentative identification of 50 compounds, mainly phenolics and secoiridoids. For the first time, the present study demonstrated that the extract from S. vulgaris flowers (at the concentrations of 1–50 µg/mL) significantly reduced plasma lipid peroxidation and protein carbonylation induced by H₂O₂/Fe²⁺. Moreover, the concentrations of 1–25 µg/mL significantly reduced the oxidation of thiol groups in plasma treated with H₂O₂/Fe²⁺. The anticoagulant tests also demonstrated that S. vulgaris flowers extract, at physiologically relevant concentrations (1–50 µg/mL), did not affect blood clotting times in vitro, suggesting that it is hemostatically safe. Conclusions: Despite the differences in composition, the extracts from lilac flowers and dandelion flowers exhibited similar protective effects against oxidative damage to human plasma components. However, the extract from S. vulgaris flowers had a stronger inhibitory effect on lipid peroxidation than the extract from dandelion flowers. Full article

Heparin remains a foundational parenteral anticoagulant across both acute and chronic care settings. This narrative review summarizes clinical indications and dosing of unfractionated (UFH) and low-molecular-weight heparin (LMWH). It also details laboratory monitoring using activated partial thromboplastin (APTT), anti-factor Xa (anti-Xa), activated clotting time (ACT) and viscoelastic testing (VET), including common pitfalls and interferences. We provide considerations for specific populations as well as complications including heparin resistance, heparin-induced thrombocytopenia (HIT) and heparin reversal strategies. Future research directions include harmonization of therapeutic ranges, mitigation of assay interference and prospective evaluation on monitoring, particular in extracorporeal membrane oxygenation (ECMO), pregnancy and cardiac surgical settings. Full article

Background: There is growing interest in plant-derived compounds for managing vascular diseases. Veratramine (VRT), a steroidal alkaloid isolated from plants of the Veratrum genus, exhibits diverse biological effects such as antihypertensive, analgesic, and antitumor activities, yet its influence on hemostasis and thrombus formation has not been characterized. This investigation sought to determine whether VRT exerts anticoagulant effects using integrated in vitro and murine models. Methods: VRT’s anticoagulant profile was comprehensively evaluated using integrated biochemical, cellular, and murine models, including clotting time assays (aPTT/PT), chromogenic enzymatic assays, fibrin polymerization analysis, platelet aggregometry, and endothelial modulation of PAI-1/t-PA under inflammatory conditions. Results: VRT treatment significantly prolonged both intrinsic and extrinsic coagulation times, directly inhibited enzymatic activities of thrombin and FXa, and attenuated their generation by endothelial cells. Additionally, VRT interfered with fibrin clot formation and diminished agonist-induced platelet aggregation. Ex vivo coagulation analyses confirmed its anticoagulant action, while endothelial studies revealed a reduced PAI-1/t-PA ratio following VRT exposure. Conclusions: These data establish VRT as possessing novel direct dual inhibition of thrombin and FXa alongside suppression of fibrin polymerization, platelet reactivity, and PAI-1 expression—positioning it as a promising multifunctional anticoagulant agent. While preclinical murine models preclude direct clinical translation absent pharmacokinetic data, these findings warrant further mechanistic and translational investigation. Full article

This study aimed to quantify horizontal and vertical bone gain using superimposition of preoperative and postoperative cone beam computed tomography (CBCT) in severe alveolar ridge defects treated with a modified guided bone regeneration (GBR) technique based on customized titanium occlusive barriers with a window design, combined with autologous blood clot and β-tricalcium phosphate (β-TCP). In this prospective case series, 13 patients (28 defects) were treated. Customized titanium barriers were digitally designed based on CBCT data and manufactured by laser sintering. The barriers were fixed over the defects and filled with a mixture of an autologous blood clot and β-TCP, providing an osteoconductive scaffold within a stable regenerative compartment. A standardized window-based follow-up protocol was applied during healing, including irrigation and controlled deepithelialization. Primary outcomes were horizontal and vertical bone gain, assessed by pre- and postoperative CBCT superimposition. Histological evaluation was performed at the time of implant placement. After 8 months, significant bone gain was observed, with a mean horizontal gain of 4.50 ± 2.02 mm and a mean vertical gain of 4.40 ± 2.82 mm (p < 0.0001), confirmed by linear mixed-effects models and patient-level sensitivity analyses (p < 0.001). Histological analysis revealed well-vascularized newly formed bone with active osteoblasts and no inflammatory response. Keratinized gingiva formation was observed at all sites. One minor complication (mild screw loosening) was recorded and successfully resolved. This study is presented as a prospective case series; therefore, the results should be interpreted as exploratory evidence and do not allow direct comparisons or conclusions regarding equivalence or superiority over other GBR techniques. The present report specifically evaluates the regenerative phase prior to functional loading; therefore, although implants were placed according to protocol, implant survival and long-term functional outcomes were not assessed and cannot be inferred from these data. Within the limitations of this prospective case series, customized titanium occlusive barriers with a window design demonstrated promising results for horizontal and vertical bone augmentation and keratinized gingiva formation, without the need for autologous bone grafts or primary wound closure. Full article

Background: Unfractionated heparin (UFH) is routinely administered during transcatheter aortic valve replacement (TAVR) to prevent thromboembolic complications. However, there are no clear evidence-based guidelines defining optimal heparin dosing or target activated clotting time (ACT) values. This study aimed to evaluate the association between intraprocedural UFH dosing, ACT values, and peri-procedural stroke risk in the overall population of patients undergoing TAVR, with a prespecified stratified analysis according to body mass index (BMI ≥ 30 vs. <30 kg/m²). Methods: This analysis enrolled consecutive individuals with severe aortic stenosis (AS) who were treated with TAVR using either balloon-expandable or self-expanding valves. The primary outcome was the occurrence of stroke during the periprocedural period in the overall population and according to BMI (<30 vs. ≥30 kg/m²). Secondary endpoints included periprocedural parameters, clinical outcomes (in-hospital and 1-year mortality), and safety outcomes. Subgroup analysis was performed to assess stroke risk according to ACT values. Patients with atrial fibrillation or receiving chronic oral anticoagulation were excluded. Results: A total of 1045 patients underwent TAVR between 2022 and 2024, including 827 with BMI < 30 and 218 with BMI ≥ 30. The study population had a mean age of 82 ± 6 years, and 56% of patients were male. In the overall study population, the mean heparin dose was 47 U/kg and the mean ACT value was 218 s. Patients with lower BMI received higher heparin doses (50 vs. 40 U/kg, p < 0.01) and had higher ACT values (221 vs. 208 s, p < 0.01). Protamine use was low and similar between groups. Periprocedural stroke rates were low overall (1.1%) and comparable between study groups (1.2% vs. 0.9%, p = 0.71). One-year mortality was also similar (3% vs. 4%, p = 0.53), with no significant differences in other safety outcomes. Subgroup analysis by ACT (≤250 vs. >250 s) showed no difference in stroke rates (1% vs. 1.5%, p = 0.60). Conclusions: In this single-center cohort, differences in heparin dosing and ACT values were not associated with differences in peri-procedural stroke or overall procedural outcomes. However, given the low number of stroke events, these findings should be interpreted cautiously. Prospective randomized studies are needed to define optimal anticoagulation strategies during TAVR. Full article