Blood Hemolysis Research Articles

Measurement of resistivity of biofluids in the presence of an electrical double layer

Abstract Detecting alterations in the electrical response of human biofluids can provide valuable information in the early stages on pathological conditions in a patient. Developing techniques for obtaining rapid information on the electrical resistivity of common ionic biofluids by non-invasive techniques can improve medicare through a rapid and more precise diagnosis. In this work, we analyze the use of an electrical device consisting of a cell formed of flat parallel steel plates to measure the resistivity of these substances. When the electrical cell is filled with a biofluid an electrical double layer (EDL) is formed on the electrodes/biofluid interface. Then, we measure the electrical response and examine the general conditions under which the EDL does not interfere with the assesment of the biofluid’s resistivity. The electrical response is analyzed in terms of an equivalent circuit model. Also, a simplified theoretical model of a suspension of biological cells considering spherical particles with a membrane is discussed to validate measurements and theoretically exhibit the contribution of the cell’s membrane to the effective resistivity. We present measurements of the resistance of the electrical cell filled with electrolyte solutions, blood plasma, and diluted suspensions of erythrocytes in a hypotonic solution. Results show that the resistance of the electrical cell is sensitive to the volume density of biological cells suspended between the parallel plate electrodes, producing a signal with a high signal to noise ratio. From the measured resistance of a suspension of erythrocytes in a isotoinc solution and the simplified theoretical model, we estimate the value of the conductivity of the interior of the erythrocytes. The results show that measured resistance varies with blood samples and hemolysis progression. The device's sensitivity to the number of erythrocytes passing between the electrodes makes it useful for measuring sedimentation kinetics.

Biological testing unification for hemodialysis membranes evaluation: A step towards standardization.

Current hemodialysis treatments can cause adverse effects, many of which are linked to the membranes used in the process. These issues are being addressed through new materials and technologies, making it urgent to establish minimum guidelines for evaluating such membranes. This review proposes standardizing the biological tests and variables to evaluate the performance of new membranes, aiming to replicate hemodialysis conditions closely. The tests were categorized into protein adsorption, protein transmission, platelet adhesion, platelet activation, blood coagulation times, hemolysis, complement activation, and cytotoxicity. For protein adsorption, static tests are recommended as an initial step to rule out membrane adhesion, followed by dynamic tests that must be conducted using a crossflow system (>250mL/min flow) and a solution mimicking real conditions (BSA, lysozyme, trypsin, pepsin, creatinine, urea, albumin, fibrinogen, and γ-globulin). Protein transmission tests must employ dynamic conditions, using human blood or platelet-rich plasma for a minimum time of 3.5h. Complement activation should be tested using human blood and ELISA assays to detect C3, C5 TCC, and SC5b-9. Blood coagulation times (APTT, TT, FT, TCT, and TAT) should be measured with platelet-poor and platelet-rich plasma. Hemolysis tests should transition from water bath to continuous mode for at least 3.5h. Cytotoxicity tests should compare the MTT assay with other methods (Alamar Blue, Lactate Dehydrogenase Assay, Flow Cytometry, or Trypan Blue Exclusion Test) and use different cell types for comprehensive validation. By implementing these minimum biological tests, membrane evaluations would more accurately reflect the real-world applications, ensuring biocompatibility, effectiveness, and efficiency.

ISO 10993-4 Compliant Hemocompatibility Evaluation of Gellan Gum Hybrid Hydrogels for Biomedical Applications.

This study examines the hemocompatibility of gellan-gum-based hybrid hydrogels, with varying gellan-gum concentrations and constant sodium alginate and silk fibroin concentrations, respectively, in accordance with ISO 10993-4 standards. While previous studies have focused on cytocompatibility, the hemocompatibility of these hydrogels remains underexplored. Hydrogels were formulated with 0.3%, 0.5%, 0.75%, and 1% gellan gum combined with 3% silk fibroin and 4.2% sodium alginate separately, using physical and ionic cross-linking. Swelling behavior was analyzed in phosphate (pH 7.4) and acetic (pH 1.2) buffers and surface morphology was examined by scanning electron microscopy (SEM). Hemocompatibility tests included complete blood count (CBC), coagulation assays, hemolysis index, erythrocyte morphology, and platelet adhesion analysis. Results showed that gellan gum-sodium alginate hydrogels exhibited faster swelling than gellan gum-silk fibroin formulations. SEM indicated smoother surfaces with sodium alginate, while silk fibroin increased roughness, further amplified by higher gellan-gum concentrations. Hemocompatibility assays confirmed normal profiles in formulations with 0.3%, 0.5%, and 0.75% gellan gum, while 1% gellan gum caused significant hemolytic and thrombogenic activity. These findings highlight the excellent hemocompatibility of gellan-gum-based hydrogels, especially the sodium alginate variants, supporting their potential in bioengineering, tissue engineering, and blood-contacting biomedical applications.

Post-mortem diagnosis of malaria in decomposed and embalmed body.

The diagnosis of malaria during the autopsy of a decomposed corpse may prove challenging. Macroscopic changes are non-specific and may include, among others, cerebral oedema, pulmonary oedema, hepatosplenomegaly and, on occasion, the presence of petechiae. The most effective diagnostic tools for malaria are the examination of blood smears and the use of rapid immunochromatographic tests. As a result of the progressive putrefaction of the corpse and blood hemolysis, classical tests are no longer viable. Consequently, the sole remaining option is the utilisation of real-time reaction (RT-PCR) to ascertain the presence of plasmodium DNA in specific organs. This study concerns the diagnosis of a fatal form of cerebral malaria in a 23-year-old Caucasian male who had travelled to Africa. The autopsy was conducted at a local hospital, after which the body was embalmed and stored in cold storage for a period of 8.5 months. Subsequently, the corpse was transported to Poland, where a further forensic autopsy was conducted. A significant challenge was to confirm the presence of malaria in a corpse that had been embalmed several months prior to the investigation. Samples were obtained from internal organs for genetic analysis to determine the presence of parasite DNA. An RT-PCR test was conducted on genetic material obtained from the brain, heart, lungs, kidney, liver, and spleen. The presence of Plasmodium falciparum genetic material was identified in samples obtained from the brain, lungs, kidney, liver, and spleen. These findings substantiated the post-mortem diagnosis of a severe form of cerebral malaria, which was the underlying cause of death.

Detection of Increased Activity of Human Parvovirus B19 Using Commercial Laboratory Testing of Clinical Samples and Source Plasma Donor Pools - United States, 2024.

In most persons, human parvovirus B19 (B19) causes a mild respiratory illness, but infection can result in adverse health outcomes in persons who are pregnant, immunocompromised, or who have chronic hemolytic blood disorders. During the first quarter of 2024, several European countries reported increases in B19 activity. In the United States, there is no routine surveillance for B19. To assess increases in B19 activity in the United States, trends in testing and results from two independent populations were examined: 1) the presence of immunoglobulin (Ig) M antibodies, a marker of recent infection, in clinical specimens ordered by physicians and 2) B19 nucleic acid amplification testing (NAAT) in pooled donor source plasma from a large commercial laboratory during 2018-2024. The proportion of IgM-positive clinical specimens reached 9.9% in the second quarter (Q2) of 2024 after remaining <1.5% during 2020-2023 and was higher than Q2 peaks in 2018 (3.8%, p<0.001) and 2019 (5.1%, p<0.001). The prevalence of B19-NAAT-positive donor pools (512 donations per pool) reached 20% in June 2024 after remaining <2% during 2020-2023 and was higher than peaks in 2018 (6.7%, p<0.001) and 2019 (7.3%, p<0.001). Considering the B19 activity increase in the United States in 2024, promotion of measures to prevent respiratory viruses and monitor for adverse B19-related outcomes by health care providers and public health authorities might reduce adverse health outcomes in pregnant persons and others at increased risk.

Investigating the efficacy of uncrosslinked porcine collagen coated vascular grafts for neointima formation and endothelialization.

This study evaluates the efficacy of uncrosslinked porcine collagen coated vascular grafts (UPCCVG) in facilitating neointima formation and endothelialization. Prior to coating, the uncrosslinked porcine collagen underwent comprehensive characterization employing SDS-PAGE, image analysis, circular dichroism and immunogenicity. The PET substrate of the vascular graft was coated with collagen solution utilizing the dip-coating method. Water permeability, blood leakage resistance, radial compliance, hemolysis, cytotoxicity and cell proliferation of UPCCVG in vitro were studied. Subsequent in vivo evaluation involved the implantation of UPCCVG as a substitute for the porcine abdominal aorta. Digital subtraction angiography (DSA) was employed to evaluate UPCCVG patency post-implantation, while histology, immunohistochemistry, and scanning electron microscopy were utilized to assess neointima formation and endothelialization. The in vivo thrombosis of UPCCVG was analyzed simultaneously to further characterize its blood compatibility. The uncrosslinked collagen demonstrated high purity, maintaining its triple helix structure and molecular weight akin to the type I bovine collagen standard substrate, indicative of preserved biological activity and low immunogenicity. UPCCVG exhibited water permeability, blood leakage resistance, radial compliance and blood compatibility comparable to commercial grafts. DSA revealed satisfactory patency of UPCCVG without evidence of stenosis or swelling at the 3-week post-implantation mark. Histological analysis illustrated well-developed neointima with appropriate thickness and controlled proliferation. Immunohistochemistry confirmed the presence of endothelial cells (VWF positive) and smooth muscle cells (α-SMA positive) within the neointima, indicating successful endothelialization. Moreover, the morphology of the neointima surface closely resembled that of the natural artery tunica intima, oriented along the direction of blood flow. UPCCVG, composed of uncrosslinked porcine collagen, demonstrates promising potential in fostering neointima formation and endothelialization while mitigating intimal hyperplasia. This biocompatible uncrosslinked porcine collagen merits further investigation for its clinical applications in vascular reconstruction.

Comparative Analysis of Blood Transfusion Accuracy and Hemolysis Rate of Transfusion Cartridge Set Between Conventional Infusion Pumps and Cylinder-Type Infusion Pumps.

Infusion pumps are critical in delivering fluids, including medications and blood products, in controlled amounts. However, conventional pumps can cause hemolysis and other issues such as flow variations and infection risks, especially during blood transfusions. To address these limitations, a novel cylinder-type infusion pump, the Anyfusion H-100, was developed, which includes a specialized blood transfusion cartridge set that combines syringe and peristaltic infusion methods. This study evaluates the accuracy and hemolysis rates of the Anyfusion H-100 compared to conventional pumps, aiming to confirm its viability as a safe and effective medical device for blood transfusions. This study evaluated six different infusion rates (10-180 cc/hr) and conducted 57 transfusion trials, 20 of which used a 1:1 blood-saline dilution. Blood transfusion accuracy was measured using research-grade packed red blood cells, and hemolysis rates were assessed before and after transfusion by chi-square tests and independent sample t-tests. Anyfusion demonstrated an average transfusion error rate of 3.77%, compared to 4.00% for the Terufusion, with no statistically significant difference in hemolysis rates (p = 0.697). Bland-Altman plots confirmed their equivalent performance, with hemolysis rates of 0.566 ± 0.095% for Anyfusion and 0.518 ± 0.126% for Terufusion. Anyfusion provides an accurate and reliable blood transfusion performance comparable to that of Terufusion, with no significant difference in hemolysis rates; its integration of syringe and infusion methods shows a potential for safer and more efficient transfusion practices, especially in pediatric and emergency settings.

Extract from Psidium guineense Sw leaves: An alternative against resistant strains of Staphylococcus aureus

Psidium guineense SW., commonly known as sour guava, is used in traditional medicine to address diverse health issues, such as inflammation, infections, and gastrointestinal disorders. Although previous research has highlighted the antimicrobial properties of the root extract and the antioxidant potential of its fruits, information regarding the activities associated with the leaf extract is lacking. The aim of the study was to use the traditional knowledge of the biological properties of P. guineense and investigate its antimicrobial activity, mechanism of action, synergistic effects, and antivirulence activity against Staphylococcus aureus. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) were determined for S. aureus, including antibiotic-resistant isolates. The synergy between the extract and antibiotics was evaluated. A growth curve was constructed for S. aureus treated with the extract. The anti-hemolytic activity was assessed by inhibiting hemolysis in human blood, and the inhibition of staphyloxanthin (STX) was examined. The extract effectively inhibited all strains with MIC and MBC values ranging from 256 to 512 µg/mL and 512 to 1024 µg/mL, respectively. Antimicrobial synergy experiments demonstrated the enhanced effectiveness of antibiotics, especially ciprofloxacin, when combined with the P. guineense extract, especially with ciprofloxacin, resulting in complete synergy against all S. aureus strains. In growth curve tests, the extract inhibited growth at MIC of 256 µg/mL within 8 h and significantly delayed it at MIC/2 (128 µg/mL) within 12 h. In antivirulence tests, the extract effectively reduced hemolysis and STX production by S. aureus at MIC and MIC/2, indicating its potential as an antimicrobial agent with antivirulence properties. The extract effectively inhibited S. aureus, including drug-resistant strains with low MIC and MBC values, exhibiting strong antimicrobial and synergistic properties with antibiotics. also In addition, the extract accelerated recovery by limiting bacterial growth and exhibiting antivirulence capabilities, reducing the severe symptoms of S. aureus infections.

A-343 Novel In-Line Hemolysis Detection on a Blood Gas Analyzer for Capillary Samples

Abstract Background Point-of-care (POC) blood gas and electrolyte analyzers utilizing whole blood as the predominant sample type are blinded by the presence of hemolysis until today. Moreover, capillary samples are particularly vulnerable to in-vitro hemolysis due to excessive force applied during sample collection. Here, we introduce the GEM Premier 7000 with iQM3 (GEM 7000) which incorporates automatic, in-line hemolysis detection in capillary blood gas samples without a need for additional sample volume or reagents. Methods The hemolysis detection module which is integrated into the disposable GEM cartridge uses acoustophoretic technology and photometric measurements to detect hemolysis in whole blood. In GEM 7000, hemolysis of ≥ 116 mg/dL was found to have significant interference with potassium results. A hemolysis flag is presented as the degree of hemolysis: none (0 - 50 and 51 - 115), mild (116 - 200 and 201 - 300), moderate (301 - 400), and gross (≥401). Due to volume limitations with native capillary samples and challenges in assessing hemolysis levels with a comparator assay, venous blood spiked volumetrically with hemolysate and transferred to capillary devices was used to verify the hemolysis flag performance. An agreement analysis was performed using the Roche cobas hemolysis value. The device's (GEM 7000) ability to flag hemolyzed capillary samples is demonstrated in a simulated clinical setting using native and contrived fingerstick capillary samples. Results Hemolysis verification with transferred capillary samples demonstrated that 98.2% (274/279) of samples were correctly flagged based on the cobas hemolysis value of spiked hemoglobin. Hemolysis in the native capillary samples (N = 235) was primarily distributed in the 0-50 mg/dL range. Additionally, native capillary samples altered using vigorous flea mixing (&amp;gt; 1 minute) showed hemolysis distribution over a wide hemolysis degree (none to gross), and their potassium results were flagged accordingly. Conclusions By providing integrated hemolysis detection in capillary blood sample measurements, the GEM Premier 7000 with iQM3 system can provide quality and confidence in the clinical interpretation of critical results such as potassium.

Evaluation of IgG and Complement Component C4 Levels in Low-Income Countries, Yemen Republic in Light of Their Proposed Role in the Hemolysis of Stored CPDA-1 Whole Blood.

Hemolysis is the most severe change that occurs in stored blood and can cause severe consequences in patients after transfusion. This study examines the potential role of IgG and complement, exampled by C4, in the hemolysis of stored CPDA-1 blood under poor storage conditions in low-income countries. The study was performed on 30 whole blood units (250 mL) drawn from convenience healthy volunteer donors with CPDA-1 anticoagulant and stored at 2-6 °C for 35 days. Each well-mixed blood bag was sampled at 0, 7, 21 and 35 days and examined for CBC, plasma hemoglobin, hemolysis percent and determination of IgG and C4. The plasma hemoglobin level and hemolysis percent increased continuously to reach 1.56 g/dl and 7.05% at the end of storage time. Hemolysis increased alongside the mean IgG concentration that was increased significantly from day 0 of storage (7.68±1.75 g/L) and peaked on day 7 (11.55±1.57 g/L), then declined to reach 8.33±2.09 g/L on day 35. Also, the mean concentration of C4 increased from day 0 of storage (0.15±0.06 g/L) to a peaked on day 21 (0.18±0.04) then declined on day 35 (0.17±0.06 g/L). The coordinated action of IgG and C4 is reflected by the positive correlation of their delta changes (r=0.616, p<0.0001). Elevated hemolysis percent in whole CPDA-1 stored blood in Yemen was accompanied by initial increase of IgG and C4 followed by final decline, which indicate their activation and consumption during hemolysis. Further studies for other hemolysis markers and analyses will give a full idea about that.

Development of small-diameter vascular grafts using PCL/PLA/gelatin/PVA/hyaluronic acid nanofibers containing VEGF/enoxaparin

The extending demands for narrow-diameter vascular grafts to restore damaged arteries have captivated researcher's attention. The absence of appropriate vascular substitutes is due to the poor patency rate of small-caliber synthetic grafts caused by thrombosis and vessel hemodynamic mismatch that induce intimal hyperplasia. We developed small-diameter vascular grafts in two layers through electrospinning. The inner layer was fabricated by gelatin/polyvinyl alcohol (PVA)/hyaluronic acid/enoxaparin and vascular endothelial growth factor (VEGFs) and the outer layer was manufactured via polycaprolactone (PCL)/polylactic acid (PLA)/hyaluronic acid. The products were characterized by atomic force microscope (AFM), scanning electron microscope (SEM), water contact angle (WCA), attenuated total reflectance-fourier transform infrared spectroscopy (ATR-FTIR), swelling test, mechanical test, and release rate. Blood compatibility tests including blood clotting time, hemolysis, and human platelet sedimentation were measured. The interactions of human umbilical vein endothelial cells (HUVECs) and human endometrial stem cells (hEnSCs) with nanofibrous vascular grafts were detected by SEM, viability assay, and DAPI staining. The results exhibited the produced small-diameter vascular grafts had proper antithrombotic properties due to two factors of VEGF and enoxaparin. After two months of the subcutaneous implantation of the scaffolds on the back of rats, the tubular vascular grafts lacked any macroscopic and histological signs of acute inflammation and had appropriate degradation rates. Furthermore, lumen patency was observed after 2 months. Consequently, VEGF and enoxaparin improves cell and blood compatibility in the small-diameter vascular grafts.

Evaluation of some nonroutine cardiac biomarkers among adults and children with beta-thalassemia major.

Cardiac injury caused by iron overload is the leading cause of mortality and morbidity in patients with beta-thalassemia, owing to frequent blood transfusion, increased iron overload, and blood hemolysis. This research aimed to assess several novel cardiac biomarkers in the blood samples of children and adult patients with beta-thalassemia major (βTM), along with their respective control groups. These biomarkers included endothelin 1 (ET-1), N-terminal pro-brain natriuretic peptide (NT-proBNP), atrial natriuretic peptide (ANP), growth differentiation factor-15 (GDF-15), and renalase (RNLS). This case-control study was done on 46 patients with βTM (23 children <18 years, and 23 adults ≥18 years) from the Genetic Hematology Center in Thi-Qar province, Iraq, and 42 comparable controls in 2 groups (21 for each group) in the period from February to April 2023. Levels of ET-1, NT-proBNP, ANP, GDF-15, RNLS, and ferritin were higher in the children and adults with βTM than in the control subjects. Elevations of the novel cardiac biomarkers ET-1, NT-proBNP, ANP, GDF-15, and RNLS in the sera of children and adult patients with βTM when compared with comparable control subjects confirm that the majority of patients with βTM are at risk of cardiac and cardiovascular complications even when there are no obvious symptoms, especially in children, which gives suitable predictive biomarkers.

Fermentation impact: A comparative study on the functional and biological properties of Banana peel waste

Banana fruit is a highly consumed and widely cultivated world food crop that generates plenty of waste globally. In this work, the phytochemical, nutritional, scavenging and therapeutic potentials of banana peel (BP) extracts were compared before and after fermentation. Halophilic fungi (Alternaria alternata, Pleosporaceae spp., Fusarium culmorum) were used in fermentation media designated as fermented banana peel FBP1, FBP2, and FBP3, respectively. Phytochemical coumarins, terpenoids, tannins, saponins, quinones, flavonoids, alkaloids, carbohydrates, proteins and steroids were found in all extracts while anthraquinone was identified in BP extracts only. Fermented extracts showed less quantity of Carbohydrate, compared to BP (477.1 ± 28.93 mg/g). Fermentation influenced the protein concentration as FBP1 showed a maximum protein of 56.9 ± 8.91 mg/g. Decreased quantities of Total Phenolic Contents (TPC), Total Flavonoid contents (TFC), and Vitamin C were noted in fermented products. The BP contained TPC (18 ± 2.59 mg GAE/g), TFC (20.5 ± 2.11 mg QE/g), carotenoid (1.03 ± 0.19 mg/g) and vitamin C (33.46 ± 2.63 mg/L). For BP, high antioxidant activity was observed, IC50 values of DPPH scavenging and FRAP assay were 2.01 ± 0.06 mg/mL and 12.81 ± 0.03 mg/mL, respectively. All the extracts were potentially active against the Salmonella typhi, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli BP extract showed high antibacterial activity than the fermented products. Among all the above, S. aureus showed high sensitivity to BP and FBP2 with 26.33 ± 2.49 and 26.33 ± 0.97 mm zone of inhibition and S. typhi was highly inhibited by BP and FBP1 with 26.26 ± 1.77 and 26.66 ± 2.63 mm. BP was highly active against K. pneumoniae and P. aeruginosa with 31.33 ± 1.74 and 32.33 ± 1.59 mm zone of inhibition and E. coli was sensitive to FBP2 with 25.7 ± 2.33 mm zone, respectively. The BP extract possessed potent antifungal activity against Mucor mucedo (84 %), Aspergillus niger (72 %) and Aspergillus flavus (83 %), which was higher than the fermented products. The antileishmanial assay was undertaken for all extracts against promastigotes of Leishmania major, BP showed good activity IC50 = 0.763 ± 0.01 mg/g. In the anti-inflammatory assays the BP showed lowest IC50 values by protein denaturing (0.612 ± 0.01), proteinase inhibitory (0.502 ± 0.01) and blood hemolysis assay (0.515 ± 0.01 mg/g). The minimum concentration indicated that BP was highly potent in response to antileishmanial and inflammation activity.

Comparison of the Impact of NaIO4-Accelerated, Cu2+/H2O2-Accelerated, and Novel Ion-Accelerated Methods of Poly(l-DOPA) Coating on Collagen-Sealed Vascular Prostheses: Strengths and Weaknesses.

Sensitive biomaterials subjected to surface modification require delicate methods to preserve their structures and key properties. These include collagen-sealed polyester vascular prostheses. For their functionalization, coating with polycatecholamines (PCAs) can be used. PCAs change some important biological properties of biomaterials, e.g., hydrophilicity, bioactivity, antibacterial activity, and drug binding. The coating process can be stimulated by oxidants, organic solvents, or process conditions. However, these factors may change the properties of the PCA layer and the matrix itself. In this work, collagen-sealed vascular grafts were functionalized with a poly(l-DOPA) (PLD) layer using novel seawater-inspired ion combination as an accelerator, compared to the sodium periodate, Cu2+/H2O2 mixture, and accelerator-free reference methods. Then, poly(l-DOPA) was used as the interface for antibiotic binding. The coated prostheses were characterized (SEM, FIB-SEM, FTIR, UV/vis), and their important functional parameters (mechanical, antioxidant, hemolytic, and prothrombotic properties, bioactivity, stability in human blood and simulated body fluid (SBF), antibiotic binding, release, and antibacterial activity) were compared. It was found that although sodium periodate increased the strength and drug-binding capacity of the prosthesis, it also increased the blood hemolysis risk. Cu2+/H2O2 destabilized the mechanical properties of the coating and the graft. The seawater-inspired ion-accelerated method was efficient, stable, and matrix- and human blood-friendly, and it stimulated hydroxyapatite formation on the prosthesis surface. The results lead to the conclusion that selection of the PCA formation accelerator should be based on a careful analysis of the biological properties of medical devices. They also suggest that the ion-accelerated method of PLD coating on medical devices may be highly effective and safer than the oxidant-accelerated coating method.

Fabrication and hemostasis evaluation of a carboxymethyl chitosan/sodium alginate/Resina Draconis composite sponge

Hemostasis is the first step of emergency medical treatment. It is particularly important to develop rapid-acting and efficacious hemostatic materials. Carboxymethyl chitosan (CMCS), sodium alginate (SA) and Resina Draconis (RD) were composited uniformly by polyelectrolyte blending. Their composite sponges (CMCS/SA/RD) were prepared by freeze-induced phase separation. CMCS/SA/RD sponges were characterized by Fourier transform infrared spectroscopy and scanning electron microscopy, and their blood absorption and hemolysis ratio were analyzed. The hemostatic effect of the composite sponges was evaluated by coagulation in vitro and in vivo. The composite sponges had a porous network structure. The water absorption ratio was >8000 %, and hemolysis ratio was <5 %. CMCS/SA/RD-II and CMCS/SA/RD-III composite sponges shortened the coagulation time in vitro by 11.33 s and 9.66 s, the hepatic hemostasis time by 13.8 % and 23.3 %, and the hemostasis time after mouse-tail amputation by 28.9 % and 23.9 %, respectively. A preliminary study on its coagulation mechanism showed that CMCS/SA/RD had significant effects on erythrocyte adsorption, platelet adhesion, and shortening of the activated partial thromboplastin time.

The influence of the disk pump of assisted circulation of the left ventricular bypass on hemolytic blood parameters

The influence of the disk pump of assisted circulation of the left ventricular bypass on hemolytic blood parameters

Cetyltrimethylammonium-chloride assisted in situ metabolic incorporation of nano-sized ROS-generating cascade-reaction containers in Gram-positive and Gram-negative peptidoglycan layers for the control of bacterially-induced sepsis

Cascade-reaction containers generating reactive oxygen species (ROS) as an alternative for antibiotic-based strategies for bacterial infection control, require endogenous oxygen-sources and ROS-generation close to or preferably inside target bacteria. Here, this is achieved by cetyltrimethylammonium-chloride (CTAC) assisted in situ metabolic labeling and incorporation of mesoporous SiO2-nanoparticles, dual-loaded with glucose-oxidase and Fe3O4-nanoparticles as cascade-reaction containers, inside bacterial cell walls. First, azide-functionalized d-alanine (D-Ala-N3) was inserted in cell wall peptidoglycan layers of growing Gram-positive pathogens. In Gram-negatives, this could only be achieved after outer lipid-membrane permeabilization, using a low concentration of CTAC. Low concentrations of CTAC had no adverse effect on in vitro blood clotting or hemolysis nor on the health of mice when blood-injected. Next, dibenzocyclooctyne-polyethylene-glycol modified, SiO2-nanoparticles were in situ click-reacted with d-Ala-N3 in bacterial cell wall peptidoglycan layers. Herewith, a two-step cascade-reaction is facilitated inside bacteria, in which glucose-oxidase generates H2O2 at endogenously-available glucose concentrations, while subsequently Fe3O4-nanoparticles catalyze generation of •OH from the H2O2 generated. Generation of •OH inside bacterial cell walls by dual-loaded mesoporous SiO2-nanoparticles yielded more effective in vitro killing of both planktonic Gram-positive and Gram-negative bacteria suspended in 10 % plasma than SiO2-nanoparticles solely loaded with glucose-oxidase. Gram-positive or Gram-negative bacterially induced sepsis in mice could be effectively treated by in situ pre-treatment with tail-vein injected CTAC and d-Ala-N3, followed by injection of dual-loaded cascade-reaction containers without using antibiotics. This makes in situ metabolic incorporation of cascade-reaction containers as described attractive for further investigation with respect to the control of other types of infections comprising planktonic bacteria. Statement of SignificanceIn situ metabolic-incorporation of cascade-reaction-containers loaded with glucose-oxidase and Fe3O4 nanoparticles into bacterial cell-wall peptidoglycan is described, yielding ROS-generation from endogenous glucose, non-antibiotically killing bacteria before ROS inactivates. Hitherto, only Gram-positives could be metabolically-labeled, because Gram-negatives possess two lipid-membranes. The outer membrane impedes direct access to the peptidoglycan. This problem was solved by outer-membrane permeabilization using a quaternary-ammonium compound. Several studies on metabolic-labeling perform crucial labeling steps during bacterial-culturing that in real-life should be part of a treatment. In situ metabolic-incorporation as described, can be applied in well-plates during in vitro experiments or in the body as during in vivo animal experiments. Surprisingly, metabolic-incorporation proceeded unhampered in blood and a murine, bacterially-induced sepsis could be well treated.

Construction of GPC3-modified Lipopolymer SiRNA Delivery System.

Gene therapy has been widely concerned because of its unique therapeutic mechanism. However, due to the lack of safe and effective carries, it has not been widely used in clinical practice. Glypican 3 (GPC3) is a highly specific proteoglycan for hepatocellular carcinoma and is a potential diagnostic and therapeutic target for hepatocellular carcinoma. Herein, to monitor the effect of gene therapy and enhance the transfection efficiency of gene carriers, GPC3-modified lipid polyethyleneimine-modified superparamagnetic nanoparticle (GLPS), a type of visualized carrier for siRNA (small-interfering RNA) targeting the liver, was prepared. We performed in vitro gene silencing, cytotoxicity, and agarose gel electrophoresis to identify the optimal GLPS formulation. In vitro MRI and Prussian blue staining verified the liver-targeting function of GLPS. We also analyzed the biocompatibility of GLPS by co-culturing with rabbit red blood cells. Morphological changes were evaluated using HE staining. The GLPS optimal formulation consisted of LPS and siRNA at a mass ratio of 25:1 and LPS and DSPE-PEG-GPC3 at a molar ratio of 2:3. GLPS exhibited evident liver-targeting function. In vitro, we did not observe morphological changes in red blood cells or hemolysis after co-culture. In vivo, routine blood analysis revealed no abnormalities after GLPS injection. Moreover, the tissue morphology of the kidney, spleen, and liver was normal without injury or inflammation. GLPS could potentially serve as an effective carrier for liver-targeted MRI monitoring and siRNA delivery.

A simple algorithm based on complete blood count and haemolysis index to enable potassium determination in haemolysed samples

A simple algorithm based on complete blood count and haemolysis index to enable potassium determination in haemolysed samples

Antibacterial, Anti-Biofilm, and Anti-Inflammatory Properties of Gelatin-Chitosan-Moringa-Biopolymer-Based Wound Dressings towards Staphylococcus aureus and Escherichia coli.

In contemporary times, the sustained aspiration of bioengineering and biomedical applications is the progressive advancement of materials characterized by biocompatibility and biodegradability. The investigation of the potential applications of polymers as natural and non-hazardous materials has placed significant emphasis on their physicochemical properties. Thus, this study was designed to investigate the potential of gelatin-chitosan-moringa leaf extract (G-CH-M) as a novel biomaterial for biomedical applications. The wound-dressing G-CH-M biopolymer was synthesized and characterized. The blood haemolysis, anti-inflammatory, antioxidant, and antibacterial activities of the biopolymer were investigated against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacterial isolates. Our results showed that S. aureus swarming motility was drastically affected. However, the biopolymer had no significant effect on the swarming motility of E. coli. In addition, the biopolymer showed high antibacterial capacities, especially against S. aureus. Plasmid DNA was observed to be effectively protected from oxidative stresses by the biopolymer. Furthermore, the biopolymer exhibited greatly suppressed haemolysis (lower than 2%), notwithstanding the elevated concentration of 50 mg/mL. These results indicated that this novel biopolymer formulation could be further developed for wound care and contamination prevention.