Search Results (147)

Heparin, an essential plasma-derived therapy, acts as a naturally occurring anticoagulant and is essential in various physiological processes. Due to its complex structure, repeating units of sulfated glycosaminoglycan, it attracts attention in the field of commercial pharmaceuticals. In recent decades, significant advancements have been made in the development of economical adsorbents designed especially for the extraction of heparin from the intestinal mucosa of pigs, as evidenced by investments from various pharmaceutical industries. This requirement arises from the demand for efficient, scalable extraction methods for natural sources. In this study, we investigated the application of beta zeolites to increase the recovery of heparin from real porcine mucosa samples, emphasizing materials with greater adsorption surfaces, higher thermal stability, and increased porosity. According to our research, the zeolite CP814E’s macropores and huge surface area allow it to adsorb up to 20.6 mg·g⁻¹ (39%) of heparin from actual mucosa samples. We also investigated the adsorbent’s surface conditions, which are essential for efficient heparin recovery, and adjusted temperature and pH to enhance heparin uptake. These findings demonstrate that zeolite-based adsorbents can enhance the extraction of heparin effectively for use in medicinal applications. Full article

Background: Acute lung injury (ALI) and its severe form, acute respiratory distress syndrome (ARDS), are critical conditions lacking effective pharmacologic therapies. Lipopolysaccharide (LPS), a bacterial endotoxin, is a well-established trigger of ALI. Emerging evidence suggests that heparin derivatives may attenuate lung injury, but their mechanisms remain unclear. Methods: This study evaluated the protective effects of 2-O, 3-O desulfated heparin (ODSH) in a murine model of LPS-induced ALI. Mice received LPS intratracheally with or without ODSH pre-treatment. Lung injury was assessed by bronchoalveolar lavage fluid (BALF) analysis, Evans blue dye albumin EBDA) extravasation, and histopathology. Results: ODSH treatment significantly reduced BALF protein concentration, inflammatory cell infiltration, and EBDA leakage. ODSH preserved endothelial barrier function in vitro, as evidenced by transendothelial electrical resistance (TER) measurements in human lung microvascular endothelial cell (HLMVEC) monolayers. Histological assessment (H&E staining) and myeloperoxidase (MPO) staining demonstrated reduced lung injury and neutrophil infiltration in the ODSH group. ODSH also downregulated pro-inflammatory mediators (NF-κB, IL-6, p38 MAPK) and upregulated the anti-inflammatory cytokine IL-10. Conclusions: ODSH mitigates LPS-induced ALI by reducing vascular permeability, neutrophilic inflammation, and pro-inflammatory signaling while enhancing IL-10 expression. These findings suggest ODSH may offer a novel therapeutic approach for treating ALI. Full article

Age-related macular degeneration (AMD) is a leading cause of irreversible blindness worldwide, primarily due to pathological choroidal neovascularization (CNV). Our study investigates a chemically modified heparin derivative as a novel strategy to selectively modulate angiogenic signaling, offering a reduced anticoagulant risk and preclinical support for AMD treatment. We explored the therapeutic potential of 6-O-desulfated heparin (Hep-6Od) as an antiangiogenic agent with diminished anticoagulant activity. Synthesized via selective 6-O-desulfation and characterized using nuclear magnetic resonance (NMR), Hep-6Od demonstrated safety in retinal pigment epithelial cells with no cytotoxic effects at various concentrations. In vitro, the compound significantly inhibited endothelial cell proliferation, migration, and capillary tube formation. Differential scanning fluorimetry (DSF) assays confirmed molecular interaction between Hep-6Od and fibroblast growth factor 2 (FGF-2), suggesting interference with pro-angiogenic signaling pathways. In vivo, a laser-induced CNV model in lean Zucker rats showed a dose-dependent reduction in neovascular lesion areas after an intravitreal Hep-6Od injection. Compared to unfractionated heparin, Hep-6Od exhibited reduced anticoagulant effects in PT and aPTT assays while maintaining robust antiangiogenic properties. These findings support Hep-6Od as a promising alternative to anti-vascular endothelial growth factor (VEGF) therapies for AMD treatment, potentially expanding current retinal vascular disease interventions. The results underscore its potential to transform AMD management, pending further clinical validation and awaiting confirmation in further studies. Full article

Sepsis is a clinical syndrome characterized by a dysregulated host response to infection, frequently resulting in septic shock and multi-organ failure. Emerging evidence highlights the critical role of neutrophil extracellular traps (NETs) in the pathophysiology of sepsis. NETs are extracellular structures composed of chromatin DNA, histones, and granular proteins released by neutrophils through a specialized form of cell death known as NETosis. While NETs contribute to the containment of pathogens, their excessive or dysregulated production in sepsis is associated with endothelial damage, immunothrombosis, and organ dysfunction. Several NET-associated biomarkers have been identified, including circulating cell-free DNA (cfDNA), histones, MPO-DNA complexes, and neutrophil elastase–DNA complexes, which correlate with the disease severity and prognosis. Therapeutic strategies targeting NETs are currently under investigation. Inhibition of NET formation using PAD4 inhibitors or ROS scavengers has shown protective effects in preclinical models. Conversely, DNase I therapy facilitates the degradation of extracellular DNA, reducing the NET-related cytotoxicity and thrombotic potential. Additionally, heparin and its derivatives have demonstrated the ability to neutralize NET-associated histones and mitigate coagulopathy. Novel approaches include targeting upstream signaling pathways, such as TLR9 and IL-8/CXCR2, offering further therapeutic promise. Full article

Tuberculosis (TB) is a major global health threat, with the current Bacillus Calmette–Guérin (BCG) vaccine having limited efficacy against adult pulmonary disease. Trained immunity (TI) is a form of innate immune memory that enhances antimicrobial defense. It is characterized by the epigenetic and metabolic reprogramming of innate immune cells and holds promise as a promising approach to prevent TB. In this study, we investigated the capacity of heparin-binding hemagglutinin (HBHA), a methylated antigen of Mycobacterium tuberculosis, to induce TI in murine RAW264.7 macrophages, human-derived THP-1 macrophages, and human peripheral blood mononuclear cells (hPBMCs). HBHA-trained macrophages exhibited the enhanced expression of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) following secondary lipopolysaccharide stimulation. The epigenetic profiling indicated elevated levels of H3K4me1 and H3K4me3 histone marks at cytokine gene loci. Further, metabolic analysis revealed heightened lactate production and the increased expression of glycolytic enzymes. Functionally, HBHA-trained macrophages exhibited improved control of intracellular mycobacteria, as evidenced by a significant reduction in colony-forming units following BCG infection. These findings elucidate that HBHA induces a functional TI phenotype via coordinated epigenetic and metabolic changes, and suggest HBHA may serve as a valuable tool for studying TI and its relevance to host defense against mycobacterial infections, pending further in vivo and clinical validation. Full article

Heparin, a widely used polysaccharidic anticoagulant of animal origin, is associated with risks of contamination and adverse effects, notably bleeding and thrombocytopenia. These limitations have prompted interest in alternative sulfated polysaccharides with anticoagulant properties and improved safety profiles. This study explored the anticoagulant potential of two marine bacterial exopolysaccharides (EPS), infernan and diabolican. It assessed whether chemical modifications (depolymerization, oversulfation) could enhance their anticoagulant properties compared to unfractionated and low molecular weight heparins. Native EPS were depolymerized to generate different molecular weights and then chemically oversulfated to increase negative charge density. Anticoagulant activities were evaluated using clotting and thrombin generation assays (TGA). Molecular docking was performed to model interactions with antithrombin and heparin cofactor II. Only highly sulfated derivatives significantly prolonged activated partial thromboplastin time while showing negligible effect on thrombin time and anti-factor Xa activity. They present different structures, and their binding to antithrombin is not achieved via the classic pentasaccharide motif. In TGA, these derivatives inhibited thrombin formation at higher doses than heparin but induced a marked delay in clot generation. Docking analyses supported their ability to bind serpins, albeit with lower specificity than heparin. Their limited anti-Xa activity and non-animal origin position them as promising anticoagulant candidates. Full article

Background/Objectives: Heparosan is a component of the capsular polysaccharide in Escherichia coli K5 and Pasteurella multocida Type D. It shares a similar glycan structure with heparin and can be enzymatically modified to produce bioactive heparin. Methods: In this study, the probiotic strain E. coli Nissle 1917 (EcN), which naturally produces heparosan, was genetically engineered to utilize sucrose as a carbon source for growth while achieving high-yield heparosan biosynthesis. Results: By expressing the sucrose hydrolase genes sacA (from Bacillus subtilis) or spI (from Bifidobacterium adolescentis), EcN was enabled to utilize sucrose, achieving heparosan titers of 131 mg/L and 179 mg/L, respectively. Further metabolic engineering was performed to block the glycolytic and pentose phosphate pathways, thereby redirecting sucrose-derived glucose-6-phosphate and fructose-6-phosphate toward heparosan biosynthesis, while glycerol was supplemented as an auxiliary carbon source to support cell growth. Finally, the key biosynthesis genes galU, kfiD, and glmM were overexpressed, resulting in an engineered strain with a heparosan titer of 622 mg/L. Conclusions: This study represents the first successful engineering of EcN to utilize sucrose as the carbon source for growth, while achieving enhanced heparosan production through synergistic carbon source utilization. These findings establish a foundational strategy for employing this strain in the sucrose-based biosynthesis of other glycosaminoglycans. Full article

hiPSC-derived blood–brain barrier (BBB) models are valuable for pharmacological and physiological studies, yet their translational potential is limited due to insufficient cell phenotypes and the neglection of the complex environment of the BBB. This study evaluates the plasma compatibility with hiPSC-derived microvascular endothelial-like cells to enhance the translational potential of in vitro BBB models. Therefore, plasma samples (sodium/lithium heparin, citrate, EDTA) and serum from healthy donors were tested on hiPSC-derived microvascular endothelial-like cells at concentrations of 100%, 75%, and 50%. After 24 h, cell viability parameters were assessed. The impact of heparin-anticoagulated plasmas was further evaluated regarding barrier function and endothelial phenotype of differentiated endothelial-like cells. Finally, sodium-heparin plasma was tested in an isogenic triple-culture BBB model with continuous TEER measurements for 72 h. Only the application of heparin-anticoagulated plasmas did not significantly alter viability parameters compared to medium. Furthermore, heparin plasmas improved barrier function without increasing cell density and induced a von Willebrand factor signal. Finally, continuous TEER measurements of the triple-culture model confirmed the positive impact of sodium-heparin plasma on barrier function. Consequently, heparin-anticoagulated plasmas were proven to be compatible with hiPSC-derived microvascular endothelial-like cells. Thereby, the translational potential of BBB models can be substantially improved in the future. Full article

Current research demonstrates the expanding therapeutic potential of heparin derivatives in oncology, extending beyond traditional anticoagulation mechanisms. This systematic analysis examines the structural characteristics, molecular mechanisms, and therapeutic applications of heparin-based compounds in malignancy treatment. The essential antithrombin binding pentasaccharide sequence has enabled development of specialized molecular variants, particularly fractionated heparins and their non-anticoagulant counterparts. These agents exert antineoplastic effects via multiple pathways, particularly through modulation of heparanase enzymatic activity and specific protein–glycosaminoglycan interactions. Evidence from pivotal clinical trials (FRAGMATIC, MAGNOLIA, GASTRANOX) confirms efficacy in managing cancer-associated thrombosis while indicating potential enhancement of chemotherapeutic outcomes. The preparation methods utilize enzymatic cleavage reactions and selective chemical derivatization to generate structurally modified heparins exhibiting unique molecular characteristics and biological activities. Analysis of the glycosaminoglycan analog dociparstat sodium reveals significant activity in myeloid malignancies, mediated by specific interference with CXCL12/CXCR4 signaling cascades. Significant challenges remain in manufacturing scale-up, analytical validation, and long-term safety assessment. Future studies must address dose optimization, combination strategies, and controlled clinical trials to determine the full therapeutic potential of these compounds in clinical oncology. Full article

Background/Objectives: The skin is an important barrier that protects against invasion by foreign substances and retains water in the body. Several skin diseases involve dry skin due to a disrupted skin barrier, and most skin diseases that appear on dry skin are accompanied by itch. Dry skin-induced itch and mechanical alloknesis reduce quality of life. Sulfated hemicellulose (i.e., pentosan polysulfate sodium), similar to heparin, is a compound belonging to the sulfated polysaccharide family; however, in contrast to heparin, it is derived from plant materials. We herein investigate the effects of the topical application of OJI-204, a sulfated hemicellulose made by purifying and chemically synthesizing hemicellulose, on dry skin in a mouse model. Methods: The mouse model of dry skin was generated using a mixture of acetone and ether with water. Either OJI-204 (3% or 10%) or 0.3% heparinoid, PBS (control), was applied twice a day to the acetone and diethyl ether/water (AEW)-treated area. The degree of skin dryness was evaluated by measuring transepidermal water loss and stratum corneum hydration. Scratching behavior was recorded the day before AEW treatment and the day after the final day, and an alloknesis assay was performed on the day after the final day. Results: We found that 3% or 10% OJI-204 attenuated dry skin conditions (erythema/hemorrhage, scarring/dryness, edema, and excoriation/erosion) and itchiness more effectively than 0.3% heparinoid. Furthermore, the degree of dryness improved to the same degree as that with heparinoid. OJI-204 also significantly reduced dry skin-induced spontaneous itch and mechanical alloknesis. Conclusions: These results suggest the potential of OJI-204 as a therapeutic or preventive agent for dry skin. Full article

Background: Laboratory monitoring of the effect of low-molecular-weight heparins (LMWHs) is generally not necessary. However, prompt evaluation of heparin inhibitory effects (i.e., anti-Xa activity) is important in cases of life-threatening bleeding, need for urgent surgery or acute thromboembolism under LMWH treatment. We aimed to establish a simple and reliable point-of-care method for the detection of enoxaparin. Methods: Eighty patients under enoxaparin therapy and ten healthy volunteers without any anticoagulant treatment were enrolled. Simultaneous measurements of anti-Xa activity using the chromogenic method and clotting times in the absence and presence of polybrene using viscoelastometric assays containing Russell’s viper venom (RVV-test) were performed on the ClotPro device. Results: Among the measured and derived RVV-test parameters, the ratio of the RVV clotting times (RVV CT) detected in the absence and presence of polybrene showed the best statistically significant correlation with anti-Xa activity (r = 0.774, p < 0.001). Based on ROC analysis, we designated RVV CT ratios of 1.02, 1.23 and 1.6 as the best cut-off values for separating anti-Xa ranges below and above 0.3 and 0.6 IU/mL, respectively. If the RVV CT ratio is below or above 1.23, the anti-Xa activity is suggested to be below 0.6 IU/mL or above 0.3 IU/mL with high certainty, respectively. Further differentiation is possible if the RVV CT ratio is measured below 1.02 or above 1.6. In these cases, the measured anti-Xa values are below 0.3 IU/mL or above 0.6 IU/mL, respectively, with high probability and good predictive values. Conclusions: Our method can provide semiquantitative information on the effect of enoxaparin and the expected anti-Xa activity within 10 min in real clinical situations. Full article

Background and aims: Enhanced cell proliferation is crucial for reducing production time and cost in cell therapy, and human platelet lysate (HPL) is often used to boost cell proliferation due to its favorable safety profile. Understanding the roles of different HPL components and their effects on cell culture can lead to more informed choices in medium formulation, which in turn can influence cell behavior and outcomes. Therefore, this study aimed to investigate the effects of two types of HPL, i.e., heparin-supplemented HPL (He-HPL) and fibrinogen-depleted HPL without heparin (Fd-HPL), on human osteoblasts. Materials and Methods: He-HPL and Fd-HPL were prepared from expired platelet concentrates. The presence of growth factors, i.e., brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF), and cytokines, i.e., interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α), in HPL was evaluated. Human fetal osteoblast (hFOB) cells were cultured in Dulbecco’s Modified Eagle Medium supplemented with either He-HPL or Fd-HPL. The cell morphology, viability, calcium deposition, and expression of osteogenic genes were assessed. Results: Comparable levels of BDNF (p > 0.05), VEGF (p > 0.05), and IL-6 (p > 0.05) were detected in both types of HPL, whereas He-HPL exhibited significantly higher levels of TNF-α (p < 0.05). However, there were no notable differences in cell morphology, viability, population doubling time, or total cell yield between the two HPL types. Similarly, no differences were observed in the mineralization of cells treated with He-HPL compared to Fd-HPL. Nonetheless, hFOB cells cultured with He-HPL demonstrated significantly higher expression of osteogenic markers Runx2 and ALP (p < 0.05) compared to those cultured with Fd-HPL. Conclusions: He-HPL and Fd-HPL demonstrate comparable performance in promoting osteoblast proliferation and mineralization, making both usable for bone tissue engineering. However, He-HPL might have a slight edge as it enhances osteogenic gene expression. Full article

Introduction: A drug-specific chromogenic assay is not immediately available, so it hampers the treatment of patients who present in a clinical emergency. In this pilot study, we aimed to create a formula to predict a plasma edoxaban level based on the international normalized ratio (INR) and heparin-calibrated anti-Xa activity and derive a novel workflow for routine laboratory diagnosis. Method: Forty-two patients prescribed edoxaban were recruited and randomized to a testing or validation cohort. Plasma levels from the testing cohort were used to create a prediction formula that was then validated in a validation cohort and real-world clinical requests. Results: The INR-derived formula had high sensitivity (95.8–100%) to predict the plasma edoxaban level > 50 ng/mL and >100 ng/mL but with low specificity. However, the specificity of predicting the plasma edoxaban level of ≥100 ng/mL was 100% by using an INR ≥ 1.5 as cut-off. Heparin-calibrated anti-Xa-derived formula had a high sensitivity (90.9–100%) and specificity (93.8–100%) in real clinical situations. A two-tier approach of combining INR-derived and heparin-calibrated anti-Xa-derived formulae can overcome the low specificity and utilize the advantages of wide availability and a short turnaround time of the INR-derived formula. Conclusions: Both INR-derived and heparin-calibrated anti-Xa-derived formulae can be applied to calculate the plasma edoxaban level. A two-tier workflow of combining these two formulae greatly helps streamline the treatment of patients prescribed edoxaban who present in a clinical emergency. Adoption of this framework is feasible for routine diagnostic laboratories. Full article

The depolymerized products and oligosaccharide fractions from sea cucumber fucosylated glycosaminoglycans (FGs) are promising anticoagulant candidates, and more novel FG-derived oligosaccharides from low-priced sea cucumbers are expected to be obtained. This study isolated 5−12 oligomers (OF1−OF3) with unusual branches from β-eliminative depolymerized products of Colochirus quadrangularis FG (CqFG). Detailed NMR analyses showed that OF1−OF3 consisted of a chondroitin 4,6-sulfates backbone and some sulfated fucosyl branches (FucS), including monosaccharides (α-l-Fuc_2S4S, α-l-Fuc_3S, α-l-Fuc_4S, α-l-Fuc_2S3S4S, and α-l-Fuc_2S) and a disaccharide D-Gal_3S4S-α1,3-l-Fuc_2S4S with the ratio of ~36:35:10:7:3:9, attached to the C-3 position of β-d-GlcA or its derivatives, such as α-l-Δ4,5GlcA and β-d-GlcA-ol. Unusually, α-l-Fuc_3S was the main FucS branch; no α-l-Fuc_3S4S branch was found, and α-l-Fuc_2S3S4S and α-l-Fuc_2S branches were also found in OF1–OF3. The OF2 and OF3 could strongly inhibit the intrinsic and common coagulation pathways. Intrinsic FXase is a target of OF2 and OF3 inhibiting the intrinsic coagulation pathways, and the unusual side chains may increase the intrinsic FXase inhibitory activity. OF2 and OF3 showed negligible bleeding risk, and less bleeding than heparin (HP), low-molecular-weight heparins (LMWHs), and CqFG. These findings support novel FG oligosaccharides with some unusual branches from low-priced sea cucumbers to be prepared as safer anticoagulants. Full article

Lactoferrin is a highly safe antibacterial protein found in the human body and in foods. Calcium phosphate (CaP) nanoparticles with immobilized lactoferrin could therefore be useful as intraoral disinfectants for the prevention and treatment of dental infections because CaP is a mineral component of human teeth. In this study, we fabricated CaP nanoparticles with co-immobilized lactoferrin and heparin using a simple one-step coprecipitation process. Heparin, a negatively charged polysaccharide, was used as both an immobilizing agent for lactoferrin and a particle-dispersing agent. The immobilization efficiency for lactoferrin in the CaP nanoparticles depended on the concentrations of both the lactoferrin and heparin in the reaction solution and was over 90% under optimal conditions. The nanoparticles had a hydrodynamic diameter of about 150–200 nm and could be well dispersed in water, owing to their relatively large negative zeta potential derived from heparin. They were found to exhibit antibacterial activity against Actinomyces naeslundii, which is involved in the initial formation of dental plaque that consequently leads to dental caries and periodontal disease. These results indicate the potential of the proposed nanoparticles as intraoral disinfectants. Full article