Hemostatic Application Research Articles

Fabrication of dual cross-linked gelatin/oxidized regenerated cellulose/polyphosphate porous sponges and nanofibrous mats for hemostatic application

Fabrication of dual cross-linked gelatin/oxidized regenerated cellulose/polyphosphate porous sponges and nanofibrous mats for hemostatic application

Comprehensive Analysis of Sulfated Flavonoids in Eclipta prostrata for Quality Evaluation

Eclipta prostrata (Asteraceae) is employed as a hemostatic agent in many traditional medicines, owing to its sulfated flavonoid content. In this study, we obtained crude drug samples from three provinces collected in different years and analyzed their sulfated flavonoid contents using liquid chromatography–mass spectrometry (LC–MS) for quality evaluation. Because sulfated flavonoids are unstable and difficult to isolate from extracts, this study first synthesized a variety of sulfated flavonoids and accumulated spectral data in order to identify the compounds in E. prostrata. The LC–MS analysis of six crude drug samples revealed the presence of luteolin 7-sulfate, apigenin 7-sulfate, diosmetin 7-sulfate, and diosmetin 3′-sulfate. The samples without luteolin 3′-sulfate featured high apigenin 7-sulfate content. Although the samples were collected from the same locality, their compositions differed depending on the year of collection. Further, they were classified according to three patterns: (1) samples with luteolin 7-sulfate as the main component, (2) samples with apigenin 7-sulfate as the main component, and (3) samples with relatively high diosmetin sulfate content. Luteolin 7-sulfate typically exhibits relatively high erythrocyte aggregation efficiency and fibrinogen aggregation rate. These results demonstrate that the analysis of sulfated flavonoids is beneficial for the quality evaluation of E. prostrata for hemostatic applications.

Chitosan/oxidized-dextran dressings containing mesoporous bioglass nanoparticles for hemostatic applications

Chitosan/oxidized-dextran dressings containing mesoporous bioglass nanoparticles for hemostatic applications

Novel 3D Printed Biomaterials Based on Fish Scale Collagen and Plant Extracts and Its Hemostatic Efficacy

AbstractThis study focused on 3D printing‐preparation of novel biomaterials based on fish scale collagen and the following plant extracts: Eclipta alba L. Hassk leaf (EA), Piper betle L. leaf (PB), Perilla frutescens L. Britt leaf (PF), and Styphnolobium japonicum L. Schott flower bud (SJ). The characterizations of the obtained biomaterials were analyzed using infrared spectroscopy and stereo microscopy. Additionally, assessments of the biomaterials were conducted on the water contact angle, simulated body fluid contact angle, scratch resistance, color changes, and swelling degree in simulated body fluid. Furthermore, in‐vitro blood clotting time and the anti‐inflammatory ability of these biomaterials were also performed. The results indicated that the plant extracts contributed to modifying collagen, significantly improving the scratch resistance and water contact angle of the collagen/plant extract biomaterials. The plant extracts were dispersed consistently with the collagen matrix, impacting the vibrations of specific functional groups of collagen. Collagen/plant extract dressings exhibited great biocompatibility and anti‐inflammatory. The in‐vitro tests demonstrated that the ethanol EA extract notably enhanced the hemostatic effectiveness of collagen, suggesting the potential of collagen/plant extract biomaterials in hemostatic applications.

Synergistic Procoagulant Mechanism and Application of Kaolin-Zeolite Composite Hemostat for Effective Hemorrhage Control.

Achieving timely and effective hemorrhage control is imperative for the survival of individuals with severe bleeding. Hemostatic materials, by enhancing the natural cell-based coagulation response, are essential tools in modern and military medical practice for controlling bleeding, especially in emergency and surgical settings. Here, we report a new type of composite hemostatic material with two different aluminosilicate-based components, kaolin and zeolite, which synergistically work together in different stages of the coagulation cascade reactions. Kaolin can effectively activate the clotting factor FXII in the early stage, and zeolite can accumulate and assemble FXa and FVa on its surface and thereafter lead to the formation of highly active thrombin in the later stage. The synergistic action mechanism between kaolin and zeolite significantly boosts the levels of FXIIa and FXa, and it also greatly enhances plateau thrombin activity. For practical application, a kaolin-modified zeolite gauze is fabricated, and it demonstrates excellent hemostatic effectiveness. Compared to the combat gauze currently used in front-line treatment, it reduces blood loss by 75% and shortens hemostasis time by 33% in a rabbit femoral artery injury model. In addition, this kaolin-zeolite gauze has no heat release problem and a nearly zero particle shedding rate, which greatly decreases the safety risk compared to current commercial inorganic-based hemostatic gauzes.

Evaluation of Full Thickness Wounds Following Application of a Visco-Liquid Hemostat in a Swine Model.

Wound healing is a complex dynamic biomechanical process as the body attempts to restore the integrity of traumatized or devitalized tissues. There are four stages of wound of healing that begins with hemostasis followed by inflammation, proliferation and finally weeks later wound remodeling. Full thickness wounds usually are covered with a dressing material after hemostasis, which allows for controlled hydration. We investigated the potential of a visco-liquid hemostat, polyhedral oligomeric silsesquioxane (POSS), for providing hemostasis and to maintain a microenvironment in the wound bed that would maintain moisture content and promote early re-epithelialization. We hypothesized that the hemostatic agent POSS if left in the wound bed would maintain a protective barrier and accelerate wound healing similar to using saline to irrigate the wound to keep the wound moist. We compared the early phase of wound repair (3-7 days) in a porcine full thickness wound model to evaluate the efficacy of the material. Biopsies were taken after 3 and 7 days to determine the acute response of the POSS hemostat or saline on inflammation, cell migration, concentrations of metalloproteinase (MMPs), and tissue inhibitors of metalloproteinase (TIMPs). Accelerated healing was observed in POSS treated wounds by changes in wound contraction, keratinocyte migration, and development of granulation tissue in comparison to saline treated wounds. Increased concentrations at day 3 of MMP-2, MMP-3, and in MMP-1 at day 7 in POSS treated wounds compared to saline coincide with keratinocyte migration observed in the tissue histology and changes in wound contraction. Tissue concentrations of TIMP-1 and TIMP-2 in POSS treated wounds appear to coordinate the sequence of MMP events in the healing tissue. Matrix metalloproteinase-13, a marker for tissue remodeling, was not upregulated in the early wound healing cascade in either POSS or saline treated wounds at 3 or 7 days. Overall, the data suggests POSS treatment contributed to enhanced early cell migration and wound closure compared to saline treatment.

Chitosan microflower-embedded gelatin sponges for advanced wound management and hemostatic applications

The study explored the antimicrobial, antibiofilm, and hemostatic properties of chitosan microflowers (CMF) in sponge form. The main objective was to enhance the preparation of CMF by employing varying quantities of calcium chloride (CaCl2) and tripolyphosphate (TPP). CMF was then combined with gelatin (GE) in different proportions to produce three sponge samples: CMF0@GE, CMF1@GE, and CMF2@GE. The CMF had a morphology like that of a flower and produced surfaces with a porous sponge-like structure. The antibacterial activity, as determined by the zone of inhibition (ZOI), increased with greater doses of CMF. Among the tested samples, CMF2@GE had the greatest activity against Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, and Enterococcus faecium. CMF2@GE successfully suppressed biofilm formation, decreased clotting time to an average of 212.67 s, and exhibited excellent biocompatibility by preserving over 90 % viability of human skin fibroblast cells at dosages below 100 μg/mL. The results indicated that gelatin sponges filled with CMF have considerable promise as flexible medical instruments for wound healing and infection control.

Cuttlefish bone derived hydroxyapatite-integrated alginate membrane for hemostasis and regeneration

Biomaterials are playing a pivotal role in facilitating hemostasis and regeneration. Fabrication of cuttlefish bone derived hydroxyapatite (HAp) infused alginate composite membrane for enhanced hemostasis and tissue regeneration. Cuttlefish bone HAp revealed Ca5(PO4)3(OH) and Ca3(PO4)2 crystalline phases with elongated spherical morphology. This HAp infused alginate membrane was fabricated using lyophilization, carboxylic as well as phosphate groups (HAp) was confirmed the formation of membrane. Graded as well as porous morphology was observed in pure alginate (ALG) and HAp alginate respectively. Enhanced hydrophilic (28.44°) and mechanical properties (23.7 MPa) were observed in HAp-ALG membrane. Optimal compatibility features was observed with the treatment of blood and fibroblast cells. Cuttlefish bone derived HAp-ALG membrane revealed rapid hemostatic behavior that was observed by clot initiation and morphological analysis. Physico-chemical, mechanical, and biological properties of cuttlefish bone derived HAp-ALG are ascertain its suitability for regeneration and hemostatic applications.

Nature-Inspired Gelatin-Based Adhesive Hydrogel: A Rapid and User-Friendly Solution for Hemostatic Applications.

Conventional hemostatic agents face challenges in achieving rapid hemostasis and effective tissue repair due to limited hemostatic scenarios, suboptimal efficacy, and inadequate adhesion to wet tissues. Drawing inspiration from nature-sourced materials, a gelatin-based adhesive hydrogel (AOT) is designed, easily prepared and quick to form, driven by Schiff base and multiple hydrogen bonds for applications in arterial and liver bleeding models. AOT exhibits exceptional adhesion to wet tissues (48.67±0.16kPa) and displays superior hemostatic properties with reduced blood loss and hemostatic time compared to other hydrogels and conventional hemostatic materials. Moreover, AOT exhibits good biocompatibility and biodegradability. In summary, this easily prepared adhesive hydrogel has the potential to supplant traditional hemostatic agents, offering a novel approach to achieve swift sealing of hemostasis and facilitate wound healing and repair in broader application scenarios, owing to its unique advantages.

Polydopamine-Coated Silk Fiber with Controllable Length for Enhanced Hemostatic Application.

The development of highly effective hemostatic materials with high biocompatibility and outstanding performance is vital to the field of biomaterials. In this study, we develop a hemostatic fiber material that exhibits high biocompatibility and excellent performance. By incorporating polydopamine (PDA) into the alkaline treatment of silk fibroin (SF), we achieve PDA-coated SF fibers with lengths that can be controlled by the alkaline concentration. The PDA coating significantly enhances the hemostatic ability of the silk fibers and exhibits superior performance in both in vitro and ex vivo experiments. By performing animal studies involving a mouse liver puncture model and a femoral vein incision model, we demonstrate the remarkable hemostatic capability of the PDA-coated SF fibers, as evidenced by the lower blood loss compared to that of a commercial hemostat powder. These findings highlight the potential of applying a PDA-assisted alkaline treatment to SF fibers to efficiently create hemostatic fibers with controllable lengths, which would be promising candidates for clinical hemostatic applications.

Design and application of distal radial artery hemostat

Transradial approach is the classical access for coronary angiography and percutaneous coronary intervention (PCI). With the increase in the number of interventional procedures, some disadvantages of the transradial approach have also been found, it is easy to lead to various complications, such as radial artery occlusion, radial nerve injury, and puncture difficulties after radial artery spasm. Therefore, some experts put forward the approach of distal radial artery approach for interventional therapy, which has the advantages of convenient positioning, easy postoperative hemostasis, less damage to the proximal radial artery and improving patients' comfort. However, there is no special distal radial artery hemostat in clinic, which limits the development of this approach to a certain extent. Therefore, based on the principles of anatomy and physics, cardiovascular physician at Jiading District District Central Hospital in Shanghai designed and invented a distal radial artery hemostatic device, which is convenient for clinical hemostasis of distal radial artery puncture, and obtained the National Utility Model Patent (patent number: ZL 2021 2 2097829.6). The hemostatic device consists of a glove body with a silicone gasket protruding towards the skin on the inner surface and a binding component. The patient's hand is inserted into the glove body, and after being fixed by the restraint component, the silicone gasket can effectively compress the location of the radial artery puncture point, and play a good hemostatic effect with less pressure, avoid the common complications of proximal radial artery hemostatic, and reduce the discomfort of the patient. Has good application value.

Microwave-assisted synthesis of crosslinked ureido chitosan for hemostatic applications

In this study, we present a facile method for introducing hydrophilic ureido groups (NH2-CO-NH-) into chitosan using a microwave-assisted reaction with molten urea, with the aim of enhancing chitosan's interaction with blood components for improved hemostasis. The formation of the ureido groups through nucleophilic addition reaction between the amine groups in chitosan and in situ generated isocyanic acid was confirmed by FTIR, CP/TOSS 13C NMR, and CP/MAS 15N NMR spectroscopic techniques. However, in stark contrast to the glucans, the said modification introduced extensive crosslinking in chitosan. Spectroscopic studies identified these crosslinks as carbamate bridges (-NH-COO-), which were likely formed by the reaction between the ureido groups and hydroxyl groups of adjacent chains through an isocyanate intermediate. These carbamate bridges improved ureido chitosan's environmental stability, making it particularly resistant to changes in pH and temperature. In comparison to chitosan, the crosslinked ureido chitosan synthesized here exhibited good biocompatibility and cell adhesion, rapidly arrested the bleeding in a punctured artery with minimal hemolysis, and induced early activation and aggregation of platelets. These properties render it an invaluable material for applications in hemostasis, particularly in scenarios that necessitate stability against pH variations and degradation.

Preparation and assessment of agar/TEMPO-oxidized bacterial cellulose cryogels for hemostatic applications.

In this work, we have demonstrated agar and oxidized bacterial cellulose cryogels as a potential hemostatic dressing material. TEMPO-oxidized bacterial cellulose (OBC) was incorporated into the agar matrix, improving its mechanical and hemostatic properties. The oxidation of bacterial cellulose (BC) was evidenced by chemical characterization studies, confirming the presence of carboxyl groups. The in vitro blood clotting test conducted on agar/OBC composite cryogels demonstrated complete blood clotting within 90 seconds, indicating their excellent hemostatic efficacy. The cryogels exhibited superabsorbent properties with a swelling degree of 4200%, enabling them to absorb large amounts of blood. Moreover, the compressive strength of the composite cryogels was appreciably improved compared to pure agar, resulting in a more stable physical structure. The platelet adhesion test proved the significant ability of the composite cryogels to adhere to and aggregate platelets. Hemocompatibility and cytocompatibility tests have verified the safety of these cryogels for hemostatic applications. Finally, the material exhibited remarkable in vivo hemostatic performance, achieving clotting times of 64 seconds and 35 seconds when tested in the rat tail amputation model and the liver puncture model, respectively. The experiment results were compared with those of commercial hemostat, Axiostat, and Surgispon, affirming the potential of agar/OBC composite cryogel as a hemostatic dressing material.

The In Vivo Fate of Polycatecholamine Coated Nanoparticles Is Determined by a Fibrinogen Enriched Protein Corona.

Polycatecholamine coatings have attracted significant attention in the past 10 years owing to their ability to functionalize a wide range of materials. Here we apply the use of such coatings to drug nanocrystals, made from a poorly soluble drug compound, to postfunctionalize the nanocrystal surface with the aim of providing steric stabilization and extending their circulation time after intravenous injection. We show that both polydopamine and polynorepinephrine can be used to successfully modify drug nanocrystals and subsequently incorporate end-functionalized PEG to the surface. Even though high grafting densities of PEG were achieved, we observed rapid clearance and increased liver uptake for polycatecholamine functionalized drug nanocrystals. Using both surface sensitive model systems and protein corona profiling, we determine that the rapid clearance was correlated with an increase in adsorption of proteins involved in coagulation to the polycatecholamine surface, with fibrinogen being the most abundant. Further analysis of the most abundant proteins revealed a significant increase in thiol-rich proteins on polycatecholamine coated surfaces. The observed interaction with coagulation proteins highlights one of the current challenges using polycatecholamines for drug delivery but might also provide insights to the growing use of these materials in hemostatic applications.

Tranexamic acid (class I drug) reduced and capped gold nanoparticles as a potential hemostatic agent with enhanced performance

External hemostatic agents play a crucial role in stabilizing an impaired process during pathological conditions. The idea is to stabilize the in vivo system as soon as possible. This study uses a class I hemostatic drug tranexamic acid as a reducing and capping agent for synthesizing the gold nanoparticles (Tr-AuNPs). Being the synthetic analogue of lysine and a biologically inspired alkylamine molecule, the chemistry can be fine-tuned for stable material that can simultaneously target the intrinsic and extrinsic hemostatic pathway, making it promising for hemostatic applications. The Tr-AuNPs of hydrodynamic diameter ∼46 nm were synthesized and evaluated physio-chemically using various analytical techniques wherein they showed hemocompatibility and increased thrombus weight compared to the native drug. The decrease in prothrombin time (PT) and international normalized ratio supported by the dynamic thromboelastography (TEG) study indicates the prepared nano-conjugate’s potential in reducing time for attaining hemostasis as compared to the native tranexamic acid drug. At a 9 μg ml−1 concentration, Tr-AuNPs had a procoagulant effect, shown by decreased reaction time (R) and coagulation time (K) with improved α angle and MA. There was a significant increase in the rate of coagulation in vivo by Tr-AuNPs, i.e. (52 s) compared to the native tranexamic acid (360 s). Radiolabelling studies ascertained the in vivo biocompatibility (non-invasive distribution, residence, clearance, and stability) of the Tr-AuNPs. The short-term toxicity studies were conducted to establish a proof of concept for the biomedical application of the material. The results highlighted the use of biologically alkyl amine molecules as capping and reducing agents for the synthesis of nanoparticles, which have shown a synergistic effect on the coagulation cascade while holding the potential for also acting as potential theranostic agents.

A 3D-Printed Cuttlefish Bone Elastomeric Sponge Rapidly Controlling Noncompressible Hemorrhage.

Developing a self-expanding hemostatic sponge with high blood absorption and rapid shape recovery for noncompressible hemorrhage remains a challenge. In this study, a 3D-printed cuttlefish bone elastomeric sponge (CBES) is fabricated, which combined ordered channels and porous structures, presented tunable mechanical strength, and shape memory potentials. The incorporation of cuttlefish bone powder (CBp) plays key roles in concentrating blood components, promoting aggregation of red blood cells and platelets, and activating platelets, which makes CBES show enhanced hemostatic performance compared with commercial gelatin sponges in vivo. Moreover, CBES promotes more histiocytic infiltration and neovascularization in the early stage of degradation than gelatin sponges, which is conducive to the regeneration and repair of injured tissue. To conclude, CBp loaded 3D-printed elastomeric sponges can promote coagulation, present the potential to guide tissue healing, and broaden the hemostatic application of traditional Chinese medicine.

Influence of Lavender Essential Oil on the Physical and Antibacterial Properties of Chitosan Sponge for Hemostatic Applications.

Uncontrollable bleeding continues to stand as the primary cause of fatalities globally following surgical procedures, traumatic incidents, disasters, and combat scenarios. The swift and efficient management of bleeding through the application of hemostatic agents has the potential to significantly reduce associated mortality rates. One significant drawback of currently available hemostatic products is their susceptibility to bacterial infections at the bleeding site. As this is a prevalent issue that can potentially delay or compromise the healing process, there is an urgent demand for hemostatic agents with antibacterial properties to enhance survival rates. To mitigate the risk of infection at the site of a lesion, we propose an alternative solution in the form of a chitosan-based sponge and antimicrobial agents such as silver nanoparticles (AgNPs) and lavender essential oil (LEO). The aim of this work is to provide a new type of hemostatic sponge with an antibacterial barrier against a wide range of Gram-positive and Gram-negative microorganisms: Staphylococcus epidermidis 2018 and Enterococcus faecalis VRE 2566 (Gram-positive strains) and Klebsiella pneumoniae ATCC 10031 and Escherichia coli ATCC 35218 (Gram-negative strains).

Enhancement of hemostatic properties of Cyclotella cryptica frustule through genetic manipulation

BackgroundThe silicified cell wall of diatoms, also known as frustule, shows huge potential as an outstanding bio-nanomaterial for hemostatic applications due to its high hemostatic efficiency, good biocompatibility, and ready availability. As the architectural features of the frustule determine its hemostatic performance, it is of great interest to develop an effective method to modify the frustule morphology into desired patterns to further improve hemostatic efficiency.ResultsIn this study, the gene encoding Silicalemma Associated Protein 2 (a silicalemma-spanning protein) of Cyclotella cryptica (CcSAP2) was identified as a key gene in frustule morphogenesis. Thus, it was overexpressed and knocked down, respectively. The frustule of the overexpress lines showed no obvious alteration in morphology compared to the wild type (WT), while the size, specific surface area (BET), pore volume, and pore diameter of the knockdown strains changed greatly. Particularly, the knockdown frustules achieved a more pronounced coagulation effect and in vivo hemostatic performance than the WT strains. Such observations suggested that silicalemma proteins are ideal genetic encoding targets for manipulating frustule morphology associated hemostatic properties. Furthermore, the Mantel test was adopted to identify the key morphologies associated with C. cryptica bleeding control. Finally, based on our results and recent advances, the mechanism of frustule morphogenesis was discussed.ConclusionThis study explores a new strategy for enhancing the hemostatic efficiency of the frustule based on genetic morphology modification and may provide insights into a better understanding of the frustule morphogenesis mechanism.

Development of Composite Sponge Scaffolds Based on Carrageenan (CRG) and Cerium Oxide Nanoparticles (CeO2 NPs) for Hemostatic Applications.

In this study, a novel absorbable hemostatic agent was developed using carrageenan (CRG) as a natural polymer and cerium oxide nanoparticles (CeO2 NPs). CRG-CeO2-0.5 and CRG-CeO2-1 composites were prepared by compositing CeO2 to CRG + CeO2 at a weight ratio of 0.5:100 and 1:100, respectively. The physicochemical and structural properties of these compounds were studied and compared with pristine CRG. Upon incorporation of CeO2 nanoparticles into the CRG matrix, significant reductions in hydrogel degradation were observed. In addition, it was noted that CRG-CeO2 exhibited better antibacterial and hemostatic properties than CRG hydrogel without CeO2 NPs. The biocompatibility of the materials was tested using the NIH 3T3 cell line, and all samples were found to be nontoxic. Particularly, CRG-CeO2-1 demonstrated superior hemostatic effects, biocompatibility, and a lower degradation rate since more CeO2 NPs were present in the CRG matrix. Therefore, CRG-CeO2-1 has the potential to be used as a hemostatic agent and wound dressing.

Optimization of Keratin Sponge Preparation Conditions for Hemostatic Application Using Response Surface Methodology (RSM)

Optimization of Keratin Sponge Preparation Conditions for Hemostatic Application Using Response Surface Methodology (RSM)