A novel hydrogel based on Bletilla striata polysaccharide for rapid hemostasis: Synthesis, characterization and evaluation.

The purpose of this study is to develop a new polysaccharide-based hydrogel. The Box-Behnken design was used to optimize the optimal synthesis conditions of the hydrogel, with the swelling parameters as indicators. The findings of rheologic tests confirm that free radical polymerization and the introduction of linear polymers improved the mechanical strength of the hydrogel. Combined with the characterization results, the gel mechanism of BSP-g-PAA/PVA DN hydrogel was proposed. The intermolecular association and entanglement increase, which effectively dissipates energy, thereby enhancing the mechanical properties of the hydrogel. In vitro blood compatibility experiments show that DN hydrogel has a low hemolysis rate and a good coagulation effect. The material is non-cytotoxic to L929 cells. The hepatic haemorrhage and mouse-tail amputation models of rats and mice were used to further evaluate the in vivo wound sealing and hemostatic properties of the hydrogel. The blood loss and hemostatic time were significantly lower than those of the control group, indicating that the hydrogel has excellent hemostatic effects. Therefore, the obtained BSP-g-PAA/PVA DN network hydrogel has good comprehensive properties and is expected to be used as a hemostatic material or a precursor of a drug carrier and a tissue engineering scaffold.

An in situ catechol functionalized ε-polylysine/polyacrylamide hydrogel formed by hydrogen bonding recombination with high mechanical property for hemostasis.

In situ hydrogel has attracted widely attention in hemostasis due to its ability to match irregular defects, but its application is limited by insufficient mechanical strength and long gelation time. Although some specifical in situ chemically cross-linked hydrogels could be fast formed and exhibit high mechanical strength, they unable to absorb blood. Hence their applications were further limited in emergency hemostasis usage. In this study, a robust hydrogel formed by hydration of powders was developed using multiple hydrogen bonds crosslinking. Here, catechol groups modified ε-polylysine (PL-CAT) and polyacrylamide (PAAM) were used to construct the PL-CAT/PAAM hydrogel. This hydrogel could be formed within 7 s to adhere and seal bleeding sites. The catechol groups endowed the hydrogel outstanding adhesive strength, which was 3.5 times of fibrin glue. Besides, the mechanical performance of in-situ PL-CAT/PAAM hydrogel was explored and the results showed that the hydrogel exhibited high compressive strength (0.47 MPa at 85% strain). Most importantly, the blood loss of wound treated with PL-CAT/PAAM hydrogel powders was 1/7 of untreated group, indicating the hydrogel's excellent hemostatic effect. And the cytotoxicity studies indicated that the PL-CAT/PAAM hydrogel had low toxicity. To summarize, this hydrogel could be a potential hemostatic material in emergency situations.

A chitosan hydrogel sealant with self-contractile characteristic: From rapid and long-term hemorrhage control to wound closure and repair.

Emergent and long-term hemorrhage control is requisite and beneficial for reducing global mortality and postoperative complications (e.g., second bleeding and adverse tissue adhesion). Despite recent advance in injectable hydrogels for hemostasis, achieving rapid gelation, strong tissue-adhesive property and stable mechanical strength under fluid physiological environment is still challenging. Herein, we developed a novel chitosan hydrogel (CCS@gel) via dynamic Schiff base reaction and mussel-inspired catechol chemistry. The hydrogel possessed high gelation rate (<10 s), strong wet adhesiveness, excellent self-healing performance and biocompatibility. More importantly, the CCS@gel exhibited saline-induced contractile performance and mechanical enhancement, promoting its mechanical property in moist internal conditions. In vivo studies demonstrated its superior hemostatic efficacy for diverse anticoagulated visceral and carotid bleeding scenarios, compared to commercialized fibrin glue. The hydrogel-treated rats survived for 8 weeks with minimal inflammation and postoperative adhesion. These results revealed that the promising CCS@gel would be a facile, efficient and safe sealant for clinical hemorrhage control.

Ultrafast in-situ forming halloysite nanotube-doped chitosan/oxidized dextran hydrogels for hemostasis and wound repair.

A series of halloysite nanotube (HNT)-doped chitosan (CS)/oxidized dextran (ODEX) adhesive hydrogels were developed through a Schiff base reaction. The resultant CS/ODEX/HNT hydrogels could not only form in situ on wounds within only 1 s when injected, but could also adapt to wounds of different shapes and depths after injection. We established four rat and rabbit hemorrhage models and demonstrated that the hydrogels are better than the clinically used gelatin sponge for reducing hemostatic time and blood loss, particularly in arterial and deep noncompressible bleeding wounds. Moreover, the natural antibacterial features of CS and ODEX provided the hydrogels with strong bacteria-killing effects. Consequently, they significantly promoted methicillin-resistant Staphylococcus aureus -infected-wound repair compared to commercial gelatin sponge and silver-alginate antibacterial wound dressing. Hence, our multifunctional hydrogels with facile preparation process and utilization procedure could potentially be used as first-aid biomaterials for rapid hemostasis and infected-wound repair in emergency injury events.

Design and Preclinical Evaluation of Chitosan/Kaolin Nanocomposites with Enhanced Hemostatic Efficiency.

In the current study, hemostatic compositions including a combination of chitosan and kaolin have been developed. Chitosan is a marine polysaccharide derived from chitins, a structural component in the shells of crustaceans. Both chitosan and kaolin have the ability to mediate a quick and efficient hemostatic effect following immediate application to injury sites, and thus they have been widely exploited in manufacturing of hemostatic composites. By combining more than one hemostatic agent (i.e., chitosan and kaolin) that act via more than one mechanism, and by utilizing different nanotechnology-based approaches to enhance the surface areas, the capability of the dressing to control bleeding was improved, in terms of amount of blood loss and time to hemostasis. The nanotechnology-based approaches utilized to enhance the effective surface area of the hemostatic agents included the use of Pluronic nanoparticles, and deposition of chitosan micro- and nano-fibers onto the carrier. The developed composites effectively controlled bleeding and significantly improved hemostasis and survival rates in two animal models, rats and rabbits, compared to conventional dressings and QuikClot® Combat Gauze. The composites were well-tolerated as demonstrated by their in vivo biocompatibility and absence of clinical and biochemical changes in the laboratory animals after application of the dressings.

A novel injectable starch-based tissue adhesive for hemostasis.

Hemorrhage remains one of the direct causes of high mortality. The development of ideal hemostatic materials with sound ability to deal with severe wound is urgent needed. Although starch-based hemostatic powder has been widely used, hydrous physiological environments severely hamper its binding to the target tissue, thereby limiting the effectiveness in hemostasis. Herein, inspired by mussel adhesive protein, a novel injectable tissue-adhesive hydrogel (St-Dopa hydrogel) composed of starch, succinic anhydride and dopamine was developed in situ by enzymatic crosslinking. The results show that St-Dopa hydrogels were intimately integrated with biological tissue and formed robust barriers to reduce blood loss. St-Dopa hydrogels exhibited superior capacity for in vitro and in vivo hemostasis as compared with chitin hydrogels. In addition to the ease of operation, St-Dopa hydrogels exhibited rapid sol-gel transition, porous microscopic morphology, good swelling ratio and biodegradability, tissue-like elastomeric mechanical properties and excellent cyto/hemo-compatibility. These results suggest that this newly developed St-Dopa hydrogel is a promising biological adhesive and hemostatic material.

Mechanically and functionally strengthened tissue adhesive of chitin whisker complexed chitosan/dextran derivatives based hydrogel.

Schiff base reaction crosslinking hydrogels are advantageous by rapid formation and absence of external crosslinkers. However, poor mechanical hindered their broader applications. Here, a mechanically strengthened tissue adhesive was constructed through incorporation of chitin nano-whiskers (CtNWs) with a Schiff base crosslinking hydrogel of carboxymethyl chitosan (CMCS) and dextran dialdehyde (DDA). The optimal formulation of complexed hydrogel exhibited 1.87 folds higher compressive stress than non-complexed and 1.51 time higher adhesive strength on porcine skin. The complexed hydrogel exhibited negligible cytotoxicity, anti-swelling performance in PBS, optimum antibacterial and hemostatic capacities. In vivo implantation studies confirmed the complexed hydrogel was degradable without long-term inflammatory responses. Desirable efficacy of injectable complexed hydrogel as hemostat was demonstrated in rat liver injury model, which could avoid severe postoperative adhesion and necrosis as observed in the treatment with commercial 3 M™ vetbond™ tissue adhesive. The results highlighted that the complexed hydrogel potentiated rapid hemostasis and wound repair applications.

In-vitro and in-vivo evaluation of modified sodium starch glycolate for exploring its haemostatic potential.

The control of blood flow from breached blood vessels during surgery or trauma is challenging. With the existing treatment options being either expensive or ineffective, the development of a haemostat that overcome such drawbacks would be beneficial. With an aim to develop an ideal haemostat, the potential of sodium starch glycolate (SSG), a commonly used pharmaceutical disintegrant was modified to obtain porous microparticles (pSSG). The biodegradability, cyto-compatibility and haemo-compatibility of the modified particles were confirmed using appropriate studies. In comparison to starch and SSG, the irregular shaped pSSG demonstrated spontaneous and significant fluid absorption (3500+500 %) and formed a physical barrier to blood flow. In addition, significant blood cells aggregation and platelet activation was observed in the modified micoparticles leading to rapid clot formation. In-vivo studies on liver and abdominal artery injury models in rats indicated the superior haemostatic potential of pSSG over SSG and starch. The results indicated that pSSG can be explored further in clinical evaluation as a hemostat.

Polysaccharide Based Hemostatic Strategy for Ultrarapid Hemostasis.

Bleeding complications usually cause significant morbidity and mortality in civilian and military populations. In clinical application, hemostatic sponges, gauzes, hydrogel, and bandages are widely used as the traditional effective hemostatic products for hemorrhage. However, the traditional hemostatic devices or agents cannot meet the requirement for treatment of massive bleeding. Therefore, the excellent hemostatic performance of hemostatic products are of great significance for saving lives. Natural polysaccharides, as the main chemical component, have been widely used in the preparation of hemostasis due to their perfect biocompatibility and biodegradability. Polysaccharide based hemostatic products are available in variety of forms, such as, hydrogel, sponges, gauze and microspheres. The purpose of the present review is to report the research progress on polysaccharide hemostatic products and technology.

Chitosan-Based Bioactive Hemostatic Agents with Antibacterial Properties-Synthesis and Characterization.

Massive blood loss is responsible for numerous causes of death. Hemorrhage may occur on the battlefield, at home or during surgery. Commercially available biomaterials may be insufficient to deal with excessive bleeding. Therefore novel, highly efficient hemostatic agents must be developed. The aim of the following research was to obtain a new type of biocompatible chitosan-based hemostatic agents with increased hemostatic properties. The biomaterials were obtained in a quick and efficient manner under microwave radiation using l-aspartic and l-glutamic acid as crosslinking agents with no use of acetic acid. Ready products were investigated over their chemical structure by FT-IR method which confirmed a crosslinking process through the formation of amide bonds. Their high porosity above 90% and low density (below 0.08 g/cm3) were confirmed. The aerogels were also studied over their water vapor permeability and antioxidant activity. Prepared biomaterials were biodegradable in the presence of human lysozyme. All of the samples had excellent hemostatic properties in contact with human blood due to the platelet activation confirmed by blood clotting tests. The SEM microphotographs showed the adherence of blood cells to the biomaterials' surface. Moreover, they were biocompatible with human dermal fibroblasts (HDFs). The biomaterials also had superior antibacterial properties against both Staphylococcus aureus and Escherichia coli. The obtained results showed that proposed chitosan-based hemostatic agents have great potential as a hemostatic product and may be applied under sterile, as well as contaminated conditions, by both medicals and individuals.

In situ synthesis of poly (γ- glutamic acid)/alginate/AgNP composite microspheres with antibacterial and hemostatic properties.

In the present work, a poly(γ-glutamic acid)/alginate/silver nanoparticle (PGA/Alg/AgNP) composite microsphere with excellent antibacterial and hemostatic properties was prepared by the in situ UV reduction and emulsion internal gelation method, and its potential application for antibacterial hemostatic dressing was explored. Well dispersed AgNPs were in situ synthesized by a UV reduction method with alginate as stabilizer and reductant. The AgNPs showed excellent antibacterial activities against both gram-negative and gram-positive bacteria. Additionally, the AgNPs prepared by the in-situ UV reduction exhibited better biocompatibility and antibacterial effects than those prepared by the conventional chemical reduction method. PGA/Alg/AgNP composite microspheres were then prepared with the AgNPs by an emulsion internal gelation method. Such microspheres were found to be a porous and hollow network with pH-sensitive swelling properties and excellent hemostatic performance, indicating its application potentials as an advanced antibacterial hemostatic material.

Biomimetic chitosan-graft-polypeptides for improved adhesion in tissue and metal.

Inspired by the mussel foot protein and chitosan-based macromolecular adhesives, a series of chitosan-graft-polypeptides were synthesized by ring-opening polymerization of three N-carboxyanhydrides (NCAs) - 3,4-dihydroxyphenylalanine-N-carboxyanhydride (DOPA-NCA), cysteine-NCA (Cys-NCA) and arginine-NCA (Arg-NCA) - using partial-NH2-protected chitosan as an initiator. These copolymers demonstrated good biodegradability and low cytotoxicity. The results of lap-shear adhesion test showed that the maximum lap-shear adhesion strength on the porcine skin and aluminum sheet were 195.97 ± 21.1 kPa and 3080 ± 320 kPa, respectively, and the maximum tensile adhesion strength on bone was 642.70 ± 61.1 kPa. The rat experiment in vivo showed that these copolymers exhibited good hemostatic performance and can promote the healing of skin wound and bone fracture. It is expected that thesecopolymeric adhesives will have broad applications in hemostasis and soft tissue adhesions.

Biodegradable N, O-carboxymethyl chitosan/oxidized regenerated cellulose composite gauze as a barrier for preventing postoperative adhesion.

Tissue adhesion is one of the most common complications after surgery (especially after abdominal surgery), causing notable influences after the damaged tissue has healed. A physical barrier placed between the wound site and the adjacent tissues is a convenient and highly effective technique to minimize or prevent abdominal adhesions. In this work, the N, O-carboxymethyl chitosan/oxidized regenerated cellulose (N, O-CS/ORC) composite gauze was prepared. The N, O-CS/ORC composite gauze is degradable; in addition, the gauze exhibits excellent antimicrobial functionality against S. aureus and E. coli bacteria. Moreover, the notable hemostatic efficacy of the N, O-CS/ORC composite gauze was confirmed in rabbit livers/ears as models. The results showed that the N, O-CS/ORC composite gauze is nontoxic toward normal cells and can restrain the adhesion of fibroblast cells, thereby indicating its potential use in preventing tissue adhesion. In addition, the rat models for abdominal defect-cecum abrasion were used to evaluate the efficacy of N, O-CS/ORC composite gauze in preventing tissue adhesions after surgery. The results indicated that the N, O-CS/ORC composite gauze can significantly prevent postsurgical peritoneal adhesions. Finally, the potential anti-adhesion mechanism of the N, O-CS/ORC composite gauze, which may attribute to the combination of barrier function and instinct activity of N, O-CS and ORC, was investigated.

Investigation of the Effects of Molecular Parameters on the Hemostatic Properties of Chitosan.

Hemorrhea is one of the major problems in war, trauma care, and surgical operation that threaten the life of the injured and patients. As a novel polymeric hemostatic agent, biodegradable chitosan can stop bleeding through a variety of approaches. In this paper, chitosan with various molecular parameters was prepared from chitin as raw material through deacetylation, oxidative degradation, hydrophilic modification, and salt formation reactions. The influence of different polymer parameters on the hemostatic effects of chitosan was investigated by in vitro coagulation time and dynamic coagulation assay. The results showed that when the molecular weights were high (105⁻106) and approximate, the coagulation effect of chitosan improved with a decrease of the deacetylation degree and achieved a prominent level in a moderate degree of deacetylation (68.36%). With the same degree of deacetylation, the higher the molecular weight of chitosan, the better the procoagulant effect. The substituent derivatives and acid salts of chitosan showed significant procoagulant effects, especially the acid salts of chitosan. In addition, the hemostasis mechanism of chitosan with various parameters was preliminarily explored by analyzing the plasma recalcification time (PRT). The efforts in this paper laid a basis for further study of the structure⁻activity relationship and the mechanism of chitosan hemostasis.

Hemostatic properties of in situ gels composed of hydrophobically modified biopolymers.

Hemorrhaging often occurs during cardiac surgery, and postoperative bleeding is associated with medical complications or even death. Medical complications resulting from hemorrhaging can lead to longer hospital stays, thus increasing costs. Hemostatic agents are the main treatment for bleeding. In the present study, hemostatic agents composed of aldehyde groups and hydrophobically modified with hyaluronic acid (ald-hm-HyA) and hydrophobically modified gelatin (hm-ApGltn) were developed and their hemostatic effects were evaluated. These modified hemostatic agents formed more stable blood clots compared with the nonhydrophobically modified HyA-based hemostatic agent. The bulk strength of the whole blood clot using the aldehyde and stearoyl group-modified hyaluronic acid (ald-C18-HyA)/hm-ApGltn-based hemostatic agent was higher than that of the aldehyde group only modified HyA (ald-HyA)/hm-ApGltn-based hemostatic agent. Rheological experiments using α-cyclodextrin showed that hydrophobic modification of HyA with C18 groups effectively enhanced anchoring to the red blood cell surface. Therefore, the ald-hm-HyA/hm-ApGltn-based hemostatic agent has potential applications in cardiac surgery.

Fabrication of photo-crosslinkable glycol chitosan hydrogel as a tissue adhesive.

In this work, an in situ gelling system composed of glycol chitosan (GC) was fabricated and evaluated regarding its tissue-adhesive, anti-bacterial and hemostatic properties. GC conjugated with 3-(4-hydroxyphenyl) propionic acid gelled immediately after illumination with blue light in the presence of ruthenium complex. The phenolic GC hydrogel was investigated regarding its mechanical property, hydration, degradation rate, cytotoxicity, tissue adhesiveness, and hemostatic ability. The hydrogel was shown to glue two pieces of tissues tightly in an egg-membrane model. The antibiotic-incorporated hydrogel killed bacteria effectively. When the hydrogel was applied to a wound in a mouse liver model, bleeding was reduced quickly and greatly. All the promising results show that the photo-chemically crosslinkable GC hydrogel could be used as a tissue adhesive, controlled drug release, and a hemostat.

Synthesis and biological evaluation of a new series of cinnamic acid amide derivatives as potent haemostatic agents containing a 2-aminothiazole substructure.

Ten new cinnamic acid derivatives containing a 2-aminothiazole substructure were designed and synthesized. This series of compounds exhibited good thermostabilities as demonstrated by thermogravimetric analysis. In coagulation assays (prothrombin time, activated partial thromboplastin time and thrombin time) in vitro, most compounds demonstrated excellent activities to promote blood coagulation. Among the studied series, compounds N1, N4, N5 and W5 exhibited a significant coagulation activity. Further studies indicated that compound N5 (IC50=1.87µmol/L) displayed the most suitable efficacy of promoting platelet aggregation than the clinically used haemostatic drug etamsylate (IC50=46.22µmol/L). Furthermore, the relationship between the functional groups of the compounds and the corresponding blood coagulant activity was explored in this study.

In situ thiolated alginate hydrogel: Instant formation and its application in hemostasis.

An in situ formed hydrogel was synthesized by sodium alginate and cysteine methyl ester, which turned the sodium alginate into thiolated alginate (SA-SH). SA-SH can in situ formed into hydrogel (SA-SS-SA) with a large amount of water through covalent bond in less than 20 s. The structure characterization showed that the mechanism of SA-SH gelation was thiol-disulfide transformation. The rheology and cytotoxicity experiments of SA-SS-SA hydrogel were also investigated, which indicated that SA-SS-SA hydrogel had an appropriate mechanical strength as well as an excellent biocompatibility. The SA-SS-SA hydrogel would degrade under certain conditions after a few days and its mechanism was disulfide alkaline reduction. Finally, the hemostatic property of SA-SH was tested by rat tail amputation experiment. The time to hemostasis of rat reduced from 8.26 min to 3.24 min, which proved that SA-SH had an excellent hemostatic property.

Hydrogelation of the Short Self-Assembling Peptide I3QGK Regulated by Transglutaminase and Use for Rapid Hemostasis.

The self-assembly of short peptides is a promising route to the creation of smart biomaterials. To combine peptide self-assembly with enzymatic catalysis, we design an amphiphilic short peptide I3QGK that can self-assemble into long nanoribbons in aqueous solution. Upon addition of transglutaminase (TGase), the peptide solution undergoes a distinct sol-gel transition to form a rigid hydrogel, which shows strong shear-thinning and immediate recovery properties. Transmission electron microscopy (TEM) and atomic force microscopy (AFM) measurements indicate the occurrence of considerable nanofibers in addition to the original nanoribbons. Liquid chromatography and mass spectrometry analyses reveal the enzymatic formation of peptide dimers from monomers through intermolecular ε-(γ-glutamyl)lysine isopeptide bonding. The dimers rapidly self-assemble into flexible and entangled nanofibers, and the coexistence of the original nanoribbons and the newly created nanofibers is responsible for hydrogelation. Factor XIII in blood is converted by thrombin to an active TGase (Factor XIIIa) during bleeding, so the peptide solution shows a more rapid and effective hemostasis via a combination of gelling blood and promoting platelet adhesion, relative to other hemostasis methods or materials. These features of I3QGK, together with its low cytotoxicity against normal mammalian cells and noninduction of nonspecific immunogenic responses, endow it with great potential for future clinical hemostasis applications.