PH12016000337A1 - Hemostatic granules - Google Patents

Abstract

The present invention relates to a hemostatic granule (or hemostat) which is used to reduce or stop bleeding in wounds or openings made during medical procedures, and the methods of making such hemostat. The hemostat comprises of one or more polymer base crosslinked by radiation.

Description

Irradiated hydrogels formed from the combinations of these materials were : used in creating granular hemostat prototypes with improved properties from its components. CMKC, CMC and PEO blends were found to have the most potential in terms of BCI values in whole blood clotting assay. Blood variation studies were made to see how the samples will hold up at higher blood volume and showed that the potential hydrogel hemostats were more effective than the commercial product Gelfoam. The lowest BCI values were recorded for PEO-

KC, CMC 0.80 and CMKC-2s. Quikclot was found to produce competitive results and inspired the creation of a gauze-type hemostat coated with the hydrogel solutions. ;

Concentration and irradiation dose were optimized to the lowest possible i values for economical considerations. Addition of other components was also no considered to enhance mechanical properties. Consolidation of all analysis - identified 20% w/w CMC 0.80 irradiated at a dose of 40 kGy to be the best formulation for the hemostat granules. It was found to have comparable effectiveness as the Celox granules.

The gauze-type was prepared either by soaking or pressing hydrogel solution to a gauze backing material. Pressed PEO blend (5% PEO, 2.5% KC and 2.5%

PEG irradiated at 10 kGy) and soaked CMC blend (10% CMC dried to final concentration of 20%) showed promising efficiency compared to Celox gauze and Quikclot gauze in platelet adhesion and coagulation time tests.

CMC is believed to work by absorbing fluids from the blood sample to concentrate protein factors while providing a structure with a negative charge : density that can charge-activate the coagulation cascade. Like Celox, it can cause clot formation of blood even in the presence of anti-coagulants such as heparin and acid citrate dextrose. PEQ-KC-PEG, on the other hand, combines the absorptive ability of KC, the negative activating charge of PEO and the binding/sealing property of PEG to create a network that also encourages platelet adhesion.

The hemostatic agent is preferably PEO, KC, carboxymethyl cellulose (CMC) and combinations thereof. One combination is PEO and KC which show good results. The hemostatic agent may constitute at least 2.5% by weight of the hemostatic material or at least 5% when in combination. Typically, the hemostatic agents constitutes from 5-30% by weight of the hemostat. 40 The hemostat may further comprise a medical surfactant, which refers to any surfactant that is pharmaceutically acceptable for contact with or administration to a human or animal body and does not cause any significant detrimental effects to the human or animal body. Examples of this are ethylene oxide and propylene oxide block copolymers, polyethylene glycol, propylene glycol, fatty 45 acids and their salts, and silicon based surfactants. Typically, the medical surfactant used is polyethylene glycol. It may constitute 2-5% by weight of a hemostat.

The hemostat is prepared by dissolving the polymer or combinations thereof in water. A medical surfactant may be added. The mixture is then spread on both Lo sides of a dressing material, typically cotton gauze. The mixture is pressed onto the dressing at 1 bar using a 1 mm mold.

The mixture is then subjected to irradiation to induce crosslinking and produce = what is essentially a hydrogel. Typically, gamma irradiation is employed at a dose range between from 10-40 kGy. At 20-30 kGy, sterilization of the hemostat also happens simultaneously. The hydrogel is dried and ground into 5 particulate, granular, powder or flake form. When in granular form, the granules are kept at a size between 0.5-1.0 mm.

The hemostat is provided in a sterile form. Sterilization may be carried out using any of the conventionally known methods such as gamma irradiation, electron beam treatment, ethylene oxide sterilization etc. Typically, gamma irradiation is used.

The invention is made from biocompatible material which does not damage the wound site or inflict burning sensation. In granular form, it can come into contact with bleeding points easily and adjust to the size of bleeding site accordingly. it will act as both a stopper by forming a gel-blood clot and concentrate natural coagulation factors to promote physiological clotting processes. The particles do not easily disperse in the wind and can be easily removed in clumps or by irrigation from wounds.

Hemostat may be used both in wound trauma found in emergency cases or in incisions made during surgical procedures. As a first aid, it can be used to stop reduce or stop bleeding until hospital care can be provided. One target beneficiary is the military as such injuries are often sustained in their service and immediate medical care is either unavailable or found with substantial distance from their post.

The preparation of the hemostatic granule is summarized as follows: * weighing the raw materials comprising polyethylene oxide (PEQ) at 4- 12%; kappa carrageenan (KC) at 1.5-6%; and polyethylene glycol (PEG) at 1.5-6%; 40 ¢ preparing PEO first by mixing in ample amount of water and letting to dissolve overnight; e mixing together the separate components and adding remaining water; ¢ mixing until the mixture becomes a smooth paste; * vacuum sealing in a foil pouch; and s irradiating at 10-30 Gy for crosslinking and sterilization to produce a hydrogel; and . ¢ drying and grounding the hydrogel! into granular form.

Preferably, the hemostatic granule is comprised of PEO at 5%; KC at 2.5%; = and PEG at 2.5%; and irradiation is done at 25 Gy. :

An acute systemic toxicity test of the hemostatic granules showed there were . no observed signs of systemic toxicity such as changes in body weight, ; reduction of feed and water intake and behavioral, respiratory and neurologic changes on healthy adult 8-week old male and female Sprague-Dawley rats.

Blood chemistry assay showed that the sample was neither hepatotoxic nor - nephrotoxic. Thus, the hemostat granules extract can be considered non-toxic and are safe to use. The LD50 for the hemostatic granules is zero (LD50 = 0). :

In the MTT Cytotoxicity Assay, hemostat granules were tested against L-929

Mouse Fibroblasts, ATCC CCLI, NCTC 929 of Strain L. Cell viability was observed to be 87% which is considered non-cytotoxic.

» [v1

HEMOSTATIC GRANULES

TECHNICAL FIELD

The present invention relates to a hemostatic material (or hemostat) which is used to reduce or stop bleeding in wounds or openings made during medical procedures, and the methods of making such hemostat. The hemostat comprises of one or more polymer base crosslinked by radiation and can be in oo ! particulate, granular, powder or flake form.

BACKGROUND

Profuse bleeding induced by civilian or military trauma has been the cause of so many fatalities. Uncontrolled hemorrhage has been reported as the leading cause of preventable pre-hospital death. In any surgical procedure, hemostasis is also a primary concern because rapid clotting helps prevent bicod loss, infection and initiates the process of tissue repair and healing. The ability to control the flow of blood following vascular injury is critical to the survival of these types of emergency casualty situations. An anti-hemorrhagic agent, otherwise known as hemostatic agent, is widely used to arrest this bleeding.

Continuous pressure to the wound is the traditional method to stop bleeding.

Constriction of wound site allows clotting factors to collect and form congealed blood to act as plug. This, however, becomes ineffective when there is severe . haemorrhaging or multiple points of bleeding. Hemostatic agents aid in addressing this problem, and for which substantial prior art exists.

JP2009502749A used the particulate hemostatic agent formed from chitosan in the form of granules or particles mainly held in the poly-4-hydroxybutyrate that can be used to treat intraluminal bleeding. US20080097271A1 disclosed devices and methods for the delivery of hemostatic agents to bleeding wounds, comprising a first component (zeolite) and a second component (kaolin clay), both components being in particle form and commingled with each other, and both components having hemostatic properties. A device incorporating such an agent comprises a receptacle wherein at least a portion of the receptacle have mesh openings therein through which the blood may flow to come into contact with the particles of the hemostatic agent. A bandage applicable to a bleeding wound is defined by a substrate, a mesh mounted on the substrate, and the hemostatic agent retained in the mesh.

CN101687056A is about a clay-based hemostatic agents and devices for the 40 delivery thereof, comprising a gauze substrate, and also a polyol such as glycerol or the like disposed on the gauze substrate to bind the clay material.

US 9,352,066 discloses hemostatic devices for promoting blood clotting that fF eeee—_——,—,—m—m,,eeneneoeY/sy mm mmm meme mm > , can include a substrate (e.g., gauze, textile, sponge, sponge matrix, one or more fibers, etc.), a hemostatic material disposed thereon such as kaolin clay, and a binder material such as crosslinked calcium alginate with a high guluronate monomer molar percentage disposed on the substrate to substantially retain the hemostatic material. oo

JP2014523312A provides a peptide analogs sequence, hemostatic or tissue sealing material, the absorbent polymer of which may be natural or genetically ~ engineered. It covers polysaccharide, cellulose, alkyl celluloses, methyl cellulose, alkyl hydroxyalkyl cellulose, a hydroxyalkyl cellulose, cellulose sulfate, and salts of carboxymethyl cellulose, chitin, carboxymethyl chitin hyaluronic acid, and salts of hyaluronic acid, alginate, and propylene glycol alginate, glycogen, dextran, dextran sulfate, and curdlan, pectin, pullulan, xanthan, chondroitin, chondroitin suifate If, and carboxymethyl dextran, and o: carboxymethyl chitosan, and chitosan, and heparin, and heparin sulfate, and heparan, and heparan sulfate, and dermatan sulfate, and keratan sulfate, and carrageenan, and chitosan, and starch, and amylose, amylopectin If, poly-N- and glucosamine, and polymannuronic acid, and polyguluronic acid, said a derivative of polysaccharide, and combinations thereof, selected from a group consisting of , hemostasis or tissue according to claim 3 seal material.”

Fan et al (2011) synthesized and characterized the properties of carboxymethyl kappa carrageenan to develop a wound healing material possessing anticoagulant activity, antimicrobial activity and moisture absorbability and moisture-retention capacity. Carboxymethyl k-carrageenan (CMKC) was synthesized by the reaction of k-carrageenan with monochloroacetic acid.

A wide range of different forms of hemostatic agents are commercially available, each adequately proven effective in their various applications.

However, the cost of these hemostatic agents is quite prohibitive as they are mostly imported and not locally available. These imported products are sold on the Philippine market at prices ranging from PhP 1,000 to PhP 6,000 for single use packs. Development of a hemostat that can be produced and made available locally will significantly help in lowering healthcare cost in the country.

Hemostat under the brand name Quikclot comprises a zeolite compound that absorbs water from wound site to concentrate coagulation factors to hasten physiological hemostasis. Although effective, water absorption of zeolite generates heat thereby causing thermal injury and tissue damage in the wound 40 site. It also causes discomfort to the person applying pressure to the wound or anyone standing nearby in wet and windy conditions.

The invention seeks to provide hemostatic materials made from radiation modified-polymers that can be comparable or superior to existing imported 45 products present in the market. This can be widely used especially in 2 ] a emergency response situations like those in military battle fields, disasters, ~ household accidents and even in surgical medical operations. j

DETAILED DESCRIPTION .

A hemostat agent is a substance that promotes hemostasis. The hemostatic agent may be one or more polymer with hemostatic property. Examples of : polymers include polyvinyl pyrrolidone (PVP), polyethylene oxide (PEO), chitosan, cellulose and its derivatives and kappa-carrageenan (KC) and its . derivatives.

Materials screened for potential hemostatic agents included refined kappa- carrageenan (KC MW 916 kDa), carboxymethylated «-carrageenan - synthesized in the laboratory (CMKC-1S MW 504 kDa and CMKC-2S 538 kDa), poly(vinyl pyrrolidone) (PVP, MW 360 kDa), poly(vinyl alcohol) (PVA),

I5 poly(ethylene oxide) (PEO 400 kDa), carboxymethyl cellulose (CMC) with 1.29 and 2.34 degree of substitution, and « and B chitosan obtained from VAEC,

Vietnam (a-CTS and B-CTS MV 72 kg/mol). Known hemostatic agents and wound dressings like gauze and QuikClot (powder) were also used for comparison. The materials were powdered to maintain uniformity among the solids. Equivalent weight was used for the liquid materials.

The lowest blood clot index (BCI) recorded was observed with KC and its carboxymethylated form, CMKC. This was followed by PEO, CMC and u-CTS.

QuikClot powder, a commercially available hemostat, was surprisingly relatively ineffective compared to the polymers. In fact, the powder and the portion that was soaked in water to counteract its reported exothermic effect on wounds were only as good as simple cotton gauze.

Granule-type formulations (from hydrogels)

Initial formulations for the hydrogels were prepared by mixing a combination of polymers which performed most efficiently in the raw screening. Various permutations of a base polymer (PVP, PEO and CMC 2.34) with a second polymer (KC and chitosan) in a constant proportion were constituted appropriately in deionized water. CMKC was prepared by itself with the same total weight. All the samples were irradiated at the Cobalt-60 facility of the

Philippine Nuclear Research Institute at a uniform dose of 20 kGy. The samples were frozen and subsequently freeze-dried for 12 to 24 hrs. One portion of the dried gel was grinded to granule form for tests. Concentration and/or irradiation dose were optimized as needed. Later, other components, 40 like kaolin, the active ingredient in QuikClot, polyethylene glycol (PEG) and starch, were added to contribute additional properties. Oven drying was also employed later for a more economical process.

EE

The blending of synthetic and natural polymers in hydrogel production has fd produced materials with better mechanical properties and biocompatibility. The - raw materials that passed the initial screening were combined to improve their properties. -

Result of the whole blood clotting experiment on a constant area (1.0x0.5cm) of the dried hydrogels show significantly higher BCI values. The samples were therefore grinded for a greater surface area. As expected, the BCI values in . general were lower. The lowest BCl values were observed in the PEO-KC on blend. The negative charges present in the PEO-KC polymers may contribute : to the contact activation of the coagulation pathway.

Between KC and CTS, KC was found to produce better results, evident especially in conjunction with PEO. CTS is a popular ingredient in commercially on

I5 available hemostat. Analysis of CTS films showed that its positive charge promoted erythrocyte, fibrinogen and platelet adhesion and activation but inhibited the contact system (intrinsic pathway) and failed in promoting non- adherent platelet activation (He, et al., 2013). These contradictory mechanisms may somewhat affect its performance, allowing KC to outshine it in the blood clotting assay. As such, further formulations were prepared using KC only.

Given that the BCI of the samples were within range with one another, blood was increased while the sample weight was maintained to see how they will hold up. The same samples with 12.5% w/w concentration and irradiated at 20 kGy were used except CMKC and CMC. CMKC concentration and dose increased to 30% w/w and 30 kGy respectively while CMC with different degrees of substitution were prepared at 20% and 40 kGy.

All of the samples showed higher coagulation efficiency against gauze and Gelfoam, the commercially available hemostat frorh Pfizer, The mode of action of Gelfoam is not fully understood although the effects believed to be more physical, as it provides a mechanical matrix to facilitate clotting. This is also a possible mode of action with the powdered hydrogels. In addition, they are able to absorb water thereby concentrating the necessary proteins responsible for the coagulation cascade. Such proteins are plasma protease factor XII (FXil), prekallikrein and FXI, which can be activated when blood comes into contact with negatively-charged surfaces like those in PEO, KC and CMKC, initiating a series of proteolytic reactions collectively known as contact activation reaction.

These processes trigger the intrinsic pathway of coagulation. These modes of 40 action may work in tandem resulting in the evident superior performance of the synthesized hydrogels. CMKC-2s was found to exhibit the most hemostatic effect even at higher blood volume, again possibly owing to its ability to charge- activate protein factors and high absorbing capacity. Quikclot Combat Gauze, the second generation of the product line, was found to have similar hemostatic 45 capability as CMKC-2s at higher blood volume (NKMCT p>0.05). Quikclot is made of non-woven gauze steeped with kaolin, an inert negatively charged material known to activate factor Xil and platelet factor 3 which initiates intrinsic coagulation. Similarity in BCI values and suggested mode of action coincided i. with those of CMKC-2s.

Utilizing the active component of Quikclot, 1% kaolin was added to the PVP-KC and CMC-KC hydrogel mixtures. Kaolin granules were shown to have w comparatively poor hemostatic effect suggesting the need for a matrix to house the material as was done in Quikclot. Kaolin was added to the PVP and CMC hydrogel formulation. Analysis using 0.65 mL of blood showed a marked increase in coagulation efficiency of PVP impregnated with kaolin but a . decrease in CMC with kaolin. The difference seen with PVP could be accounted by the addition of the negatively charged kaolin to an otherwise - neutral compound, thereby allowing the contact activation mechanism to : proceed. CMC has a negative net charge density, and adding kaolin increases : the charge density that should have made it as efficient if not more so. Results however showed increase in charge density actually lowers pro-coagulant activity in the case of CMC.

Upon acquisition of CMC with DS 0.8, hydrogels of CMC with varying degree of substitution were prepared and irradiated at 40 kGy. Results showed an increase in hemostatic capacity with the decrease in the degree of substitution.

A higher degree of substitution generally translates to a higher charge density.

Again, this followed the trend of decreasing coagulation efficiency with increasing charge density of CMC at higher blood volume.

Blood clotting index is predicted to increase at a higher blood volume when the material is at a fixed weight. Although this is observed in most of the hydrogels, there are some samples that did not conform to this trend. Gauze, PEO-KC,

Gelfoam showed a distinct drop in BC! values at 0.50 mL of blood. Quikclot, on the other hand, showed the same decrease at 0.35 mL. It is possible that the breaks in the trend are caused by mechanisms that work predominately or in tandem at certain blood volumes. There is also the possible difference between the interaction of blood with the sample surface and with its matrix when absorbed. If this is true, then the other samples must also exhibit this occurrence only at a much higher blood level since their BCI remained relatively low at the blood volumes used. Three samples were subjected to the blood variation experiment until 1.0 mL of whole blood was added. Another drop in BCl was seen in PEO-KC at 1.0 mL. CMKC-2s also showed a decrease in BCI value at this volume. Meanwhile, there was a drop at 0.85 mL for CMC 40 0.80. The differences in the trend breaks may be caused by their different physical properties. The exact mode of action of this apparent phenomenon is still unknown.

Based on experiments so far, three of the most effective formulations were 45 PEO-KC, CMKC-2s and CMC-0.8. However, despite the very promising results for CMKC-2s, it was opted to be set aside at the due to economical considerations. CMKC-2s can be synthesized by subjecting KC through a b lengthy process of carboxymethylation twice (2s). As such, subsequent experiments were focused only on PEO-KC and CMC-0.8. ;

Modification of concentration and dose, extraction of soluble fraction and ~ addition of PEG and starch

Concentration and/or irradiation dose were optimized and certain additions (PEG and starch) were made to the base formulation to control structural properties. It was previously observed that PEO-KC hydrogel became tough a after the drying process. Poly(ethylene glycol) and starch were added to the - mixture in order to increase flexibility. Addition of PEG showed significant improvement compared to the base formulation. On the other hand, the on addition of starch resulted in weaker hemostatic performance. Although starch - is known to aid in gel strength and flexibility, it can also decrease swelling oy capacity because of its poor hydrophilicity, which may explain this behaviour.

This time, the commercial product used for comparison is Celox granules to allow for a fair assessment given the granular structure of the samples. PEO-

KC-PEG and the CMC granules were shown to be as efficient as Celox granules, this suggests possibly similar mechanisms at play. Celox works through the electrostatic interaction of its positively charged active material, chitosan, and negatively charged red blood cells to form a gel-like clot. This mechanism is independent of the coagulation pathway and thus can work just as well on heparinized blood or when administered to people with clotting disorders or hypothermic blood, and or people treated with anti-platelet drugs like aspirin or blood thinners. Both CMC and KC delivers good water retention as well as gelling and binding capability which may work like Celox to form the clot.

The formulations chosen from this batch for further study were those which used the least amount of materials while maintaining structural integrity, namely the 5PEO-2.5KC-2.5PEG and 20CMC-40kGy. The PEO blend was subjected to three irradiation doses, one lower and one higher than the usual dose used. The resulting hydrogels were divided so that half was dried and grinded as usual and the other was swelled for 24 h in water before drying and grinding. This allowed the extraction of the soluble fraction of hydrogel.

There was no significant difference between all the samples. From this, it can be said that: (1) the least amount of dose, 10 kGy, can produce the same 40 efficiency which can later cut back on costs; and (2) removing the soluble fraction did not bear significant effect to the ability of the samples to coagulate blood.

Use of unreconstituted ACD-blood and heparinised blood 45 With the ability of Celox to work independently of the coagulation pathway, the samples were tested against blood in the presence anti-coagulant ACD and heparin. Celox indeed showed the same efficiency in the presence of anti- oe coagulants. The same trend can be seen in CMC. Once again, this further proves the possibility of similar mechanisms at play and the ability to act PL independently of the clotting cascade. The PEO blend, however, did not show - adistinctive trend in the presence of the anti-coagulants. 5

The samples were dried by freeze-drying or oven drying at 50° or 80°C. Since } they bore no significant difference, the drying method most convenient and : cost-effective for producers can be employed. -

Platelet adhesion on

Platelet adhesion test was performed using the Cytotox 96 Non-Radioactive =

Cytotoxicity Assay. The dried hydrogels were cut into 1x1 cm squares instead Co of grinding. The CMC blend was found to be more efficient than the PEO = formulations, possibly due to the coarser surface texture and porosity of the former compared to the smooth and brittle surface of the latter. CMC was found to perform much better than Gelffoam, which is understandable, given that its mechanism relies more on mechanical plugging than platelet adhesion and/or activation. Celox remains competitive but less so than CMC.

Gauze-type formulations ! A small amount of hydrogel solutions (using materials with favourable results from previous analysis) were placed on both sides of about 3x3 inches of medical gauze and pressed until a thin layer of the paste was formed on its surface. After irradiation, the samples were divided in two. Half was used as-is and the other was dried. A polyethylene nonwoven fabric was also used as a potential backing material. The resulting product was dried and cut into 1x1 cm squares.

Popular hemostat dressings include HemCon, Rapid Deployment Hemostat,

Quikclot Combat Gauze and Celox Gauze. The basic design consists of an active material embedded on a matrix such as gauze or non-woven fiber, and are usually used on minor wounds (low pressure bleeding) and abrasions.

Unfortunately, these products are not locally available and are quite expensive, ranging from USD120-1000 per 4x4 inches bandage. In view of the target application of the project, this avenue was considered and dressings made from standard gauze and the polymer solutions were created.

Platelet adhesion 40 Initially, gauze-type hemostats were prepared using dilute solutions of polymers. Medical gauze was soaked or brushed with the solution and dried in the oven until concentrated to a final %w/w that would give substantial crosslinking (10% for PEO and 20% for CMC). It was then irradiated and dried before use. Celox gauze showed an efficient performance in platelet adhesion 45 compared to the typical gauze. The same efficiency could be observed from the samples and a marked increase in 10% CMC. The evident difference between the value produced by gauze and those soaked in the hydrogel solution indicate a significant effect of the active material, making it a viable design. Quikclot gauze was noted to be more effective than regular gauze but less than Celox gauze and the samples. :

The initial gauze-type design (soak-concentrate-irradiate) resulted in an uneven distribution of the active material across the backing material. This time the dressings were created using a presser to create a uniform thin film on the gauze surface. They were either used as is or oven-dried and steamed before use. :

The CMC dressings were unremarkable showing only little difference against standard gauze. Compared to the previous design, the CMC layer on the gauze was thicker and the dressing was stiffer and less flexible than the earlier N design which maintained the flexibility of the gauze backing. Based on previous experiments, pure CMC hydrogel had enough manoeuvrability to expand creating a larger surface for platelets to adhere. On the other hand, a very thin layer of CMC on gauze retains its effectiveness in adhesion properties. The porous gauze and CMC may work in tandem to create a loose network where platelets can be trapped. However, the thicker layer pressed on the gauze diminished its efficiency possibly because the interaction occurs mainly on the

CMC surface instead of the CMC-gauze network and the backing constricts it from expanding further unlike a pure hydrogel.

PEO samples, on the other hand, were able to keep up with Celox gauze. The

PEG may play into binding the PEO to the gauze backing and the KC present loosens crosslinked networks of PEO to allow platelets to penetrate and adhere.

Coagulation time

Coagulation time (CT) was evaluated in polypropylene tubes, a non-activating surface. In all cases, addition of samples facilitated formation of coagulum compared to blank tubes. However, it can be noted that the PEO-KC-PEG coated gauze had significantly faster clotting time (30% faster than control) than both gauze and Celox. The ability to form clots in a timely manner is an important property in hemostat development and is evidently exhibited by the sample developed.

Relatively low cost and abundant materials available locally were screened for 40 their potential as hemostatic agents. These include natural polysaccharides, synthetic polymers and some plant extracts. Initial analysis of raw materials showed blood coagulation efficiency of «-carrageenan [KC] and its carboxymethylated form [CMKC], poly(ethylene oxide) [PEO], carboxymethyl cellulose [CMC] and powdered a-chitosan [o-CTS]. Suggested mechanism 45 involved primary hemostasis through water absorption (concentration of proteins), platelet adhesion and activation, and contact activation.

Claims (7)

CLAIMS: .

1. A hemostatic granule characterized in that the granule is comprised of polyethylene oxide (PEO) at 4-12%; kappa carrageenan (KC) at 1.5-6%; and polyethylene glycol (PEG) at 1.5-6%. "

2. A hemostatic granule according to Claim 1 wherein the PEO is 5%; KC is }

2.5%; and PEG is 2.5%. )

3. A hemostatic granule according to Claim 1 wherein the granule is 0.5-1 mm : in size.

4. A process of preparing a hemostatic granule comprising the following steps: e weighing the raw materials comprising polyethylene oxide (PEO) at 4- - 12%; kappa carrageenan (KC) at 1.5-6%; and polyethylene glycol (PEG) % at 1.5-6%; e preparing PEO first by mixing in ample amount of water and letting to dissolve overnight; e mixing together the separate components and adding remaining water; e mixing until the mixture becomes a smooth paste; e vacuum sealing in a foil pouch; and » irradiating at 10-30 Gy for crosslinking and sterilization to produce a hydrogel; and + drying and grounding the hydrogel into granular form.

5. A process of preparing a hemostatic granule according to Claim 4 wherein the PEQ is 5%; KC is 2.5%; and PEG is 2.5%.

6. A process of preparing a hemostatic granule according to Claim 4 wherein irradiating is done at 25 Gy.

7. The use of the hemostatic granule in treating wound trauma and stopping bleeding.