KR102325543B1 - Hemostatic composition having improved mechanical strength and absorptiveness for effeecitve hemostasis - Google Patents

Abstract

The present invention relates to a hemostatic composition comprising a polyvinyl alcohol moiety and a composite polymer matrix in which oxidized cellulose moieties are cross-linked. The hemostatic composition has excellent swelling properties and water absorption ability, and has no side effects on the body. The hemostatic composition of the present invention can be safely applied to a living body in a crisis situation requiring rapid hemostasis treatment.

Description

Hemostatic composition with improved mechanical strength and absorption for effective hemostasis

This patent application is part of the "Civil-Military Combined Technology Development Project" by the Ministry of National Defense and Ministry of Trade, Industry and Energy of the Republic of Korea for the "Development of a biomimetic polymer rapid hemostatic agent" (organization: Inha University Hospital, project number: 16-CM-SS-07) , "Development of multidisciplinary applied medical technology and platform technology for aerospace medicine" as part of "University-focused research institute support project in science and engineering field" of the Ministry of Science and ICT As part of the Ministry of Education's "Medium Researcher Support Project", "Design and Evaluation of Stimulation Response Drug Release Controlled Stent Medical Devices for the Treatment of Malignant and Benign Human Luminal Stenosis" (Organizer: Inha University, Project No.: 2017R1A2A2A07001272) It's about results.

The present invention relates to a hemostatic composition, and more particularly, to a hemostatic composition, including a polymer matrix with improved mechanical strength and absorbency suitable for inserting into recessed hemostatic sites such as gunshot wounds, stab wounds, abdominal bleeding, intranasal bleeding, etc. to exhibit an effective hemostatic effect, It relates to a hemostatic composition that can be used as an effective hemostatic agent in a weightless space environment.

Medically, bleeding (hemorrhage) generally refers to external bleeding, which refers to a case in which blood leaks out of the body due to damage or destruction of blood vessels due to external shock. Conversely, bleeding that flows into a living tissue or body cavity rather than a wound that exits the body is called internal bleeding. Bleeding occurs very quickly and in large amounts as the blood pressure is strong and the wound is large. In addition to bleeding due to trauma, bleeding occurs frequently during surgery for treatment, and in some cases, death due to excessive bleeding during or after surgery.

Therefore, it is very important clinically, publicly, and socially to induce effective treatment by controlling bleeding. Accordingly, hemostatic agents applied with chemical or biological materials with various functions for hemostasis are being researched and developed. Hemostatic agents using chemical or biological materials developed so far have been mainly developed for the purpose of controlling bleeding occurring during surgery. Therefore, the development or product of a hemostatic agent that can be used for traumatic bleeding is relatively scarce.

For example, in Korean Patent No. 10-1201056, when a first component, which is a polysaccharide polymer in which aldehyde is introduced through hydrolysis, and a second component, which is a polymer containing an amino group, are simultaneously added to a bleeding site, living tissue It is described that it has adhesive and/or hemostatic properties by forming a schiff base. However, there is a problem with the toxicity of aldehydes and polymer amino groups, and the effect is very limited when bleeding occurs continuously or the amount of bleeding is large.

In addition, in the case of a cyanoacrylate-based glue (commercially available under trade names such as "Dermabond" and "Indermil"), it can be used as an excellent hemostatic agent because of its strong adhesion. However, cyanoacrylate-based glue has a disadvantage of being highly toxic, such as releasing formaldehyde as a decomposition product, which is known to adversely affect the human body and the environment, and is currently applied only to limited areas such as the skin.

In addition, a fibrin polymer may be used as a liquid hemostatic agent. In this case, there is a disadvantage that the thrombin activator and calcium ions, which are essential for coagulation of fibrin, cannot be used at the same time, and the room temperature conditions must be met before storage and use. In addition, since it contains protein components such as fibrin, there are significant concerns about secondary infection with biological products extracted from blood due to factors such as immune problems, lack of convenience in storage and use, and high cost.

It is an object of the present invention to provide a hemostatic composition having excellent physical properties such as inflation pressure and water absorption capacity.

Another object of the present invention is to provide a hemostatic composition with minimal side effects to the living body.

The present invention provides a hemostatic composition comprising a polyvinyl alcohol (PVA) moiety and a composite polymer matrix in which an oxidized cellulose moiety is cross-linked.

The oxidized cellulose is characterized in that it is modified (modification) to have a carboxyl group on the surface.

For example, the oxidized cellulose may be one in which a hydroxy group bonded to the 6-position carbon of glucose, which is a constituent unit of cellulose, is modified with a carboxy group.

In the composite polymer matrix, the polyvinyl alcohol moiety and the oxidized cellulose moiety may be mixed in a weight ratio of 5:1 to 50:1, preferably 5:1 to 20:1.

The composite polymer matrix may have a porous structure.

For example, the composite polymer matrix may be molded in the form of a powder, granules, sol, gel, nonwoven fabric, fabric, sponge or sheet. can

For example, it is characterized in that the composite polymer matrix is prepared in a block form, cut again, and then compressed and molded into a compressed sponge form.

The hemostatic agent according to the present invention has excellent swelling properties and water absorption properties, and does not cause unintended side reactions in vivo, for example, immune responses.

According to the present invention, in the case of a hemostatic agent with mechanical strength because it has a function of expansion through blood absorption, it may occur due to gunshot wounds, cuts, or various accidents, as well as bleeding during surgical procedures that require hemostasis compression. It can be applied to bleeding that appears in depressions in the human body, such as intra-abdominal bleeding, intra-nasal bleeding, and intra-sinus bleeding.

In addition, hemostatic agents with excellent mechanical strength can be effectively used in environments where hemostasis is difficult due to weightlessness, such as in a space environment.

More specifically, the polymer hemostatic agent according to the present invention exhibits a compression hemostatic effect by exhibiting an inflation pressure of up to 70 to 80 kPa, and at the same time has an absorption hemostatic effect showing a blood absorption capacity of 1200 to 1600% of the maximum.

1 is an image taken with a scanning electron microscope (SEM) of a polymer material synthesized according to an embodiment of the present invention. (a) and (b) show the low-resolution and high-resolution cotton fibers, respectively, and (c) and (d) show the TEMPO oxidation-treated cotton fibers, respectively.
2 is a photograph and graph showing the morphology of a polymer material synthesized according to an embodiment of the present invention. (a) is an image taken with a transmission electron microscope (TEM) of TEMPO-treated cellulose nanofibers, (b) and (c) are graphs showing the results of measuring the length dispersion and width dispersion of the TEMPO-treated cellulose nanofibers am.
3A and 3B are graphs showing Fourier-transform infrared spectroscopy (FTIR) analysis results of cellulose nanofibers subjected to TEMOP oxidation treatment according to an embodiment of the present invention, respectively. Figure 3a shows the FTIR analysis results at 4000 ~ 2500 cm ^-1 , Figure 3b shows the FTIR analysis results at 1800 ~ 800 cm ^-1 .
4 is a graph showing the results of measuring the maximum inflation pressure of a hemostatic agent prepared in the form of a sponge and including a composite polymer matrix according to an embodiment of the present invention.
5 is a graph showing the results of measuring the change in average expansion force over time of a hemostatic agent prepared in the form of a sponge and including a composite polymer matrix according to an embodiment of the present invention.
6 is a graph showing the results of measuring the water absorption rate of a hemostatic agent prepared in the form of a sponge and including a composite polymer matrix according to an embodiment of the present invention.
7 is an SEM image of a hemostatic agent prepared in the form of a sponge and including a composite polymer matrix according to an embodiment of the present invention.
8 is a graph showing the results of measuring the average pore size of a hemostatic agent prepared in the form of a sponge and including a composite polymer matrix according to an embodiment of the present invention.
9 is a graph showing the results of evaluating an in vitro inflammatory test of a hemostatic agent prepared in the form of a sponge and including a composite polymer matrix according to an embodiment of the present invention.
10a and 10b are photographs showing the results of immunohistologic analysis of the tissue state of an animal to which a hemostatic agent prepared in the form of a sponge and including a composite polymer matrix according to an embodiment of the present invention is applied, respectively, after application of the hemostatic agent; It is a photograph taken using H&E staining method and MT staining method at 3 days and 7 days elapsed time.
11a and 11b are photographs showing the results of immunohistologic analysis of the tissue state of an animal to which a hemostatic agent prepared in the form of a sponge and including a composite polymer matrix according to an embodiment of the present invention is applied, respectively, after application of the hemostatic agent; It is a photograph showing the results of analyzing CD31 at 3 days and neutrophils at 3 and 7 days.

Hereinafter, if necessary, it demonstrates, referring accompanying drawings.

Conventional polymer hemostatic agents have a disadvantage in that they can be applied only to suppress bleeding that appears in wounds on the surface of the human body, such as abrasions, in order to increase the simple blood absorption function. Accordingly, the present inventor has completed the present invention through research to develop a hemostatic agent that can be applied to the depressed area at the same time as having mechanical hemostatic performance.

Absorbable hemostatic agents in the form of a matrix or sponge absorb water from the blood and act as a hemostatic agent. When water is absorbed by these hemostatic agents, the viscosity of the blood rises and the concentration of solids such as albumin, platelets, and blood cells in the blood rises rapidly. As platelets attach to the bleeding site, they secrete various chemicals such as fibrinogen and thrombin, and induce vasoconstriction and platelet deposition around them. At the same time, the glycoprotein membrane on the platelet surface is activated, and fibrinogen is activated to insoluble fibrin and aggregates with neighboring platelets. If this process is repeated, hemostasis occurs. Therefore, the performance of the matrix type hemostatic agent is most affected by the strength of the matrix along with the water absorption performance.

Polyvinyl alcohol (PVA) is a synthetic polymer having a hydroxyl group and a small amount of acetate group, and is applied in various industrial fields depending on the average molecular weight and the degree of acetylation. PVA has excellent biocompatibility, chemical resistance and mechanical resistance, as well as hydrophilicity, good solubility in water, a semi-crystalline structure, and no toxicity.

Polyvinyl alcohol, which is the first polymer moiety constituting the polymer matrix according to the present invention, can be obtained by hydrolyzing polyvinyl acetate (vinyl acetate resin), and is a hydrophilic polymer that does not dissolve well in general organic solvents. Polyvinyl alcohol is industrially used as a coating agent, a thickener, and a raw material for cosmetics. Medically, it is used as a vascular embolizer, a thickener in ophthalmic preparations, a sponge implant for plastic reconstruction surgery such as breast, and is used for patients with rectal prolapse and colon hernia. It has been used for reconstructive surgery for over 40 years and is known to show no oral or transdermal toxicity.

The weight average molecular weight (Mw) of the polyvinyl alcohol that can be used according to the present invention is approximately 2,000 to 2,000,000. When polyvinyl alcohol having such a weight average molecular weight is used, crosslinking with chitosan can be easily formed. In relation to one exemplary embodiment, the polyvinyl alcohol that can be suitably used for the purpose of the present invention is typically produced by Kuraray and has the trade names ^{Poval TM} and Exceval ^{TM .} Can be used. Polyvinyl alcohol can be used by selecting a product group according to the degree of saponification.

For example, especially in the field of tissue engineering, a scaffold based on PVA is being studied as a material to replace artificial implants due to its porous structure and decomposition properties. In addition, PVA is known to show excellent affinity for extracellular matrix materials and skin. Accordingly, a sponge-type hemostatic agent using PVA as one of the three-dimensional matrix as a matrix has been researched and developed.

However, since PVA has poor inflation pressure characteristics, it has limitations as a matrix type hemostatic agent. The present invention uses oxidized cellulose nanofibers to realize a matrix-type hemostatic agent that has no side effects to the living body while improving dynamic expansion characteristics and water absorption ability. As an example, the oxidized cellulose nanofiber is 2,2,6,6-tetramethyl-piperidinyl-1-oxyl (2,2,6,6-tetramethyl-piperidinyl-1-oxyl; TEMPO) treated nanocellulose can be Hereinafter, in the present specification, TEMPO-treated cellulose nanofibers are abbreviated as TOCNs (TEMPO-oxidized cellulose nanofiber), and a composite matrix of PVA and TEMOP-treated cellulose nanofibers may be abbreviated as PVA-TOCN.

New properties can be obtained by manufacturing cellulose, the most abundant biopolymer on earth, in a nano size. Nano-sized cellulose, that is, nanocellulose, has excellent mechanical properties, low density (approximately 1.6 g/cm 3 ), high aspect ratio, and other surface properties by modification of surface functional groups. It has reactive hydroxyl groups that can be implemented.

Although there are some differences depending on the body part, the average human blood pressure is 130 mmHg (= 17.31 kPa) and the average minimum blood pressure is 85 mmHg (= 11.33 kPa). Therefore, in order to prevent bleeding using a matrix type hemostatic agent, the inflation pressure of the matrix must be greater than the bleeding pressure. Although the bleeding pressure may vary from case to case, the matrix material is preferred because the inflation pressure of the matrix is higher than the average maximum human blood pressure.

Water-based polymer materials have been mainly developed in the form of dressings for wound dressings used for the treatment of wounds, and are being developed for new uses such as hemostatic agents and anti-adhesion agents. The polymer developed as a wound dressing dressing takes the form of cotton wool or an absorbent pad, and the anti-adhesion film has a thin, transparent paper form or a polymer gel form. Therefore, even if a polymer of the same component is used, the manufacturing method and final external or physicochemical properties vary depending on the purpose of use.

In order to improve the hemostatic action and biocompatibility, cellulose without an oxidized structure in its molecular structure, i.e., a regenerated cellulose block, can be considered. In general, in order to produce regenerated cellulose, natural cellulose pulp is treated with a strong alkali such as 20% (w/v) sodium hydroxide, chemically transformed into alkali cellulose, and then pulverized and stored at room temperature, followed by sulfur dioxide treatment to obtain high-viscosity. Eggplant prepares a cellulose solution. The high-viscosity cellulose solution is again treated with sodium hydroxide, and fibrous regenerated cellulose is obtained through spinning and sulfuric acid treatment. At this time, the cellulose in which the carbonyl group structure in the sugar structure is reduced back to the hydroxyl group structure through sulfuric acid treatment is called regenerated cellulose.

However, there are many problems when applying regenerated cellulose as a medical material. As a medical polymer material, it is necessary to easily control the molecular weight of the polymer material in order to obtain a desired shape and physicochemical properties, and in particular, it is necessary to obtain a material having a uniform molecular weight. However, as described above, regenerated cellulose is obtained by treating natural pulp with a strong alkali or strong acid. In particular, due to the characteristics of the cellulose material, it is effective for blood absorption and hemostasis through wound sealing or pasting.

Oxidized cellulose is sometimes used to overcome the disadvantages of cellulose material, but in the case of a polymer matrix or sponge using oxidized cellulose alone, it absorbs blood and transforms it into a hydrogel form, losing the mechanical strength required for hemostasis. many. According to the Republic of Korea Patent 10-1318421, in order to overcome this, carboxymethylcellulose foam for hemostasis and wound healing and a manufacturing method thereof are presented. The above patent is an invention for a polymer matrix that does not gel using poorly water-soluble carboxymethyl cellulose. Although the inventor describes that poorly water-soluble carboxymethyl cellulose can be obtained by controlling the degree of oxidation of cellulose, it can be said that it is technically inferior to the present invention because the expansion pressure for effective hemostasis is not considered. Referring to FIG. 4 presented by the inventor, it can be confirmed that most of the polymer matrix forms a hydrogel after absorption, so it can be expected that the physical strength is insufficient to exhibit a sufficient hemostatic effect.

In addition, in order to coat the regenerated cellulose block with chitosan, the cellulose block is washed, immersed in an acetic acid solution in which chitosan is dissolved, and dried to prepare an expandable cellulose block. Basically, there is no complete clinical confirmation for the biocompatibility of chitosan with acetic acid, and it is difficult to solve various problems of free chitosan flowing from the bleeding site. In particular, recently, reports of patients showing an acute allergic reaction to chitosan are rapidly increasing, and appropriate supplementary measures are required for this.

According to the present invention, in order to improve the mechanical strength of a hemostatic agent having a matrix type, oxidized cellulose nanofibers are used together with PVA as a material of a composite matrix to increase the swelling pressure and water absorption capacity, and to have no cytotoxicity to the human body. It is possible to implement a safe hemostatic agent that does not exist.

Cellulose, a raw material for oxidized cellulose nanofibers, is the most abundant biopolymer on earth, and is a straight-chain polysaccharide in which D-glucose is linked through β-1,4 glycoside bonds. Cellulose accounts for most of the cell wall of higher plants, especially the xylem, and nanocrystalline cellulose, ie, cellulose nanocrystals (CNCs), makes up a significant portion of the xylem of higher plants.

Since intra- and inter-molecular hydrogen bonds are formed between the hydroxyl group and oxygen of D-glucose composed of 6 carbons, cellulose maintains a stable structure. Within cellulose fibrils, highly regular regions (crystalline parts) and irregular regions (non-crystalline parts) are distinguished. In particular, crystalline cellulose has excellent mechanical properties.

In one exemplary embodiment, a purification and homogenization process may be required prior to isolating the cellulosic microfibers and/or crystalline components. For example, if cellulose is of wood or plant origin, matrix materials such as hemicelluloses and lignin are impurities and must be removed. Also, if the cellulose is from tunicates such as sea squirts, the mantle and protein matrix need to be removed. Also, when bacterial cellulose is used, bacteria and other cultures must be removed. Cellulose can also be obtained from the cell wall of algae. In this case, the matrix material and the culture solution present in the cell wall of the algae must be removed. Methods such as physical methods (mechanical treatment) and chemical methods (acid hydrolysis and enzymatic hydrolysis) may be used to separate microfibers, nanofibers and purified cellulose of crystal structure. For example, physical methods include high pressure homogenizer, high intensity ultra-sonication, micro fluidization, cryo-crushing and/or grinder/refiner. /refiner) and the like may be used. If necessary, mechanical treatment may be performed several times to obtain nano-sized cellulose, and a separation process such as filtration or centrifugation may be performed after mechanical treatment to remove impurities (metal powder, etc.) resulting from the separation process. may be

On the other hand, acid hydrolysis removes the amorphous region from the cellulose microfibers using an acid. Since the crystalline region of cellulose is relatively resistant to hydrolysis by acid, the amorphous region with weak acid resistance is relatively removed, leaving only the crystalline region. Cellulose purified by hydrolysis with an acid can be mixed with deionized water and treated to have an appropriate acid concentration. Dilute sulfuric acid, hydrochloric acid and/or maleic acid are the acids commonly used in connection with the hydrolysis of cellulose, and the mixture is subjected to a separation process (filtration and / or centrifugation), washed, dialyzed, and sonication may be performed to disperse the crystalline cellulose.

In enzymatic hydrolysis, cellulases are used, A- and B-type cellulases called cellobiohydrolases attack the crystalline domains in cellulose, and C- and D-types called endoglucanase Cellulolytic enzymes mainly attack and hydrolyze irregular regions of cellulose. When these two cellobiohydrolases and endoglucanase are used together, there is an advantage in that purified cellulose having a good structure can be obtained compared to other methods.

A part of the matrix of the present invention uses cellulose modified by oxidizing hydroxyl groups present on the surface of the nanocellulose obtained by the above method. As described above, hydrogen bonds are formed due to the abundance of hydroxyl groups on the surface of D-glucose constituting the nanocellulose. Due to these hydrogen defects, the structure of the cellulose is stable but insoluble in a solvent. On the other hand, in the present invention, the surface-modified nanocellulose is used as a matrix together with PVA to increase the mechanical strength of the matrix and at the same time improve physical properties such as expansion pressure.

In one exemplary embodiment, a part of the hydroxyl group on the surface of the nanocellulose fiber, for example, the hydroxyl group connected to carbon (C6) at position 6 of D-glucose, which is a repeating unit of cellulose, may be modified with a carboxy group. have. For example, in order to obtain nanocellulose modified with a carboxyl group, 2,2,6,6-tetramethyl-piperidinyl-1-oxyl (2,2,6,6-tetramethyl-piperidinyl-1-oxyl; TEMPO) is It is used as a catalyst, and as a primary oxidizing agent, hypochlorite, for example, NaClO may be used, but the present invention is not limited thereto. For example, cellulose nanofibers modified by treatment with TEMPO and an oxidizing agent may be obtained by treating cellulose present in plants such as cotton with TEMPO. In particular, in the case of performing TEMPO treatment, the hydroxy group bonded to the 6-position carbon of D-glucose, which is a cellulose constituent unit, is regioselectively converted to carboxy and modified even under mild conditions. If necessary, hydrobromic acid or phosphoric acid may be used instead of hypochlorite. In another alternative embodiment, the unit unit of cellulose is NaBr together with TEMPO and NaClO, a type of hypochlorite, at about pH 10-11 in order to convert the primary hydroxyl group present at a specific position of glucose into a carboxy group. can be used In this case, the primary hydroxyl group of glucose may be converted into an aldehyde and then converted into a carboxyl group or a carboxyl salt form. In this case, NaBr is used as a second catalyst used to increase the yield.

In an exemplary embodiment, the carboxyl group may be adsorbed on the surface of the nanocellulose and connected via electrostatic bonding. Using electrostatic bonding, the high surface charge of the nanocellulose surface can be advantageous. In this regard, when separating and purifying cellulose, chemical separation may be preferable.

In addition, suitable surfactants may also be used to stabilize the nanocellulose. The surfactant is not particularly limited, and fatty acids such as stearic acid, cetyltetramethylammoniumbromide (CTAB), polyethyleneamine (PEI), or components that can be used as pore formers to be described later are also surfactants. It can be used as an activator. In another alternative embodiment, a chemical modification reaction may be performed via covalent bonding to the nanocellulose.

According to the present invention, the oxidized form of nanocellulose, for example, nanocellulose modified to convert certain hydroxyl groups into carboxyl groups, is used together with PVA as a common matrix in the hemostatic composition. When a small amount of modified nanocellulose is added to PVA, the physical properties can be improved, such as increasing the expansion pressure and the expansion rate. In one exemplary embodiment, the PVA and oxidized nanocellulose constituting the composite matrix of the hemostatic composition are in a weight ratio of 5:1 to 50:1, preferably in a weight ratio of 5:1 to 20:1, more preferably in a weight ratio of 5:1 to 20:1. It may be mixed in a weight ratio of 5:1 to 15:1, but the present invention is not limited thereto. For example, if the blending ratio of PVA based on the oxidized cellulose exceeds 50 times, it is difficult to expect an increase in the improvement of the physical properties of the PVA according to the blending of the oxidized cellulose. On the other hand, if the blending ratio of PVA based on the oxidized cellulose is less than 5 times, there is a possibility that the water absorption power is excessively lowered. That is, when the PVA and the oxidized nanocellulose meet the above-mentioned mixing ratio, it can have sufficient swelling pressure and water absorption ability to prevent bleeding, so it can be utilized as an astringent hemostatic agent (styptic).

In one exemplary embodiment, the composite matrix of PVA-TEMOP-treated cellulose nanofibers may be molded into various shapes. In one example, the composite matrix may be formed into a sponge, woven or non-woven, particulate, granular material or sheet shape. For example, when molded in a sponge shape, it is further molded to have a coin shape, thereby improving water absorption and inflation pressure. For example, the hemostatic agent having PVA and oxidized nanocellulose as a common matrix according to the present invention may have a maximum inflation pressure of 40 to 70 kPa, and a water absorption rate of 600 to 1600%, for example 800 to 1600%, The present invention is not limited thereto.

As described above, the present invention is a hemostatic composition using polyvinyl alcohol and oxidized nanocellulose as a composite matrix. The present inventors have found that mixing polyvinyl alcohol with oxidized nanocellulose with excellent mechanical properties can improve the expansion pressure and expansion rate, while maintaining the high water absorption capacity of PVA, and does not cause side effects such as cytotoxicity or inflammatory reaction. It was confirmed that it can be used as a safe hemostatic composition because it is not.

According to an exemplary embodiment of the present invention, a cross-linking agent may be used for crosslinking between the polyvinyl alcohol molecules and the oxidized nanocellulose fiber molecules. For example, a crosslinking agent having an aldehyde group may be used, but any crosslinking agent capable of forming a crosslink by reacting with an amine group constituting the chitosan molecule may be used.

For example, glutar aldehyde, dextran polyaldehyde, dimethyl adipimidate dihydrochloride (dimethyl adipimidate2HCl), which are 'homo-bifunctional' crosslinking agents having the same functional group at both ends of the molecule , DMA), dimethyl suberimidate dihydrochloride (DMS), and disuccinimidyl suberate (DSS) may be used. In addition, succinimidyl-4 (N-maleimidomethyl)-cyclohexane-1-carboxylate (succinimidyl-4 (N-maleimidomethyl)-cyclohexane-1-carboxylate, SMCC), N-succinimidyl-3- (2-pyridyldithio-propionate) (N-succinimidyl-3-(2-pyridyldithio)-propionate), N-succinimidyl (4-iodoacetyl)-aminobenzoate (N-succinimidyl (4 'Hetero-bifunctional' crosslinking agents such as -iodoacetyl)-aminobenzoate (SIAB) and m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) may be used. have.

The amount of crosslinking agent used can be adjusted according to the molecular weight and input amount of polyvinyl alcohol and oxidized nanocellulose fibers used. It can be removed through a masking reaction, washing, or purification process. For example, the crosslinking agent may be added at a concentration of approximately 0.05 to 0.5 mL/g, preferably 0.1 to 0.2 mL/g, based on the total weight of the polymer comprising the polyvinyl alcohol-oxidized nanocellulose matrix constituting the polymer matrix. , the present invention is not limited thereto.

If necessary, the polymer matrix comprising polyvinyl alcohol moieties and oxidized nanocellulose moieties may have a porous structure. In order to form a porous polymer matrix, an appropriate porogen may be added in the process of blending the polyvinyl alcohol moiety and the oxidized nanocellulose moiety.

Pore formers that can be used include ether-based surfactant/emulsifier, ester-based surfactant/emulsifier, cationic surfactant/emulsifier, alkaline carbonate and/or starch (starch) can be. As the ether-based surfactant/emulsifier, polyethylene glycol tert-octylphenyl ether (trade name: Triton™ X-100), which is a non-ionic surfactant having a hydrophilic polyethylene oxide chain, may be used. Further, examples of the ester surfactant/emulsifier include fatty acid esters, for example, polyethoxylated ester of sorbitans. These fatty acid esters are commercially sold under trade names such as Span or Tween.

Specifically, fatty acid esters that can be used as pore formers according to the present invention include (polyethylene glycol) sorbitan monolaurate ((polyethylene glycol) sorbitan monolaurate, Tween 20 or Span 20), sorbitan monopalmitate, Span 40), (polyethylene) sorbitan monostearate ((polyethylene) sorbitan monostearate, Tween 40), (polyoxyethylene) sorbitan monostearate (Tween 60), sorbitan monooleate (Span 80) , sorbitan seqquioleate (Tween 61), sorbitan trioleate (Tween 65), sorbitan isostearate (sorbitan isostearate, Tween 80), and the like.

The cationic surfactant/emulsifier may be, for example, an alkyl ammonium salt, an alkylpyridinium salt, an alkylimidazolinium salt, a quaternary ammonium salt and/or a primary to tertiary aliphatic amine salt. Specifically, the cationic surfactant is tyltrimethylammonium bromide (CTAB), tetradecylmethylammonium bromide (TTAB), cetylmethylammonium chloride (CTAC), hexadecyltrimethylammonium bromide (hexadecyltrimethylammonium bromide) HTAB), N-dodecyl pyridinium chloride, and/or benzalkonium chloride (alkyldimethyl benzylammonium chloride) may be used.

In addition, alkaline carbonates that can be used as pore formers include, for example, sodium bicarbonate (NaHCO ₃ ), sodium carbonate (Na ₂ CO ₃ ), potassium bicarbonate (KHCO ₃ ), potassium carbonate (K ₂ CO ₃ ), calcium bicarbonate (Ca( HCO ₃ ) ₂ ), calcium carbonate (CaCO ₃ ), magnesium carbonate (MgCO ₃ ), magnesium bicarbonate (Mg(HCO ₃ ) ₂ ), ammonium bicarbonate (NH ₄ HCO ₃ ), ammonium carbonate ((NH ₄ ) ₂ CO ₃ ), bicarbonate, gadolinium (Gd (HCO _{3) 3,} carbonate, gadolinium _{(Gd 2 (CO 3) 3} ), bicarbonate, lithium (LiHCO _3), lithium carbonate (LiCO _3), sodium bicarbonate, rubidium (RbHCO _3), carbonate, rubidium (Rb ₂ CO ₃ ), zinc carbonate (ZnCO ₃ ), zinc bicarbonate (Zn(HCO ₃ ) ₂ ), iron (II) carbonate (FeCO ₃ ), iron (II) bicarbonate (Fe(HCO ₃ ) ₂ ), silver carbonate (Ag) ₂ CO ₃ ), silver bicarbonate (AgHCO ₃ ), gold (III) carbonate (Au ₂ (CO ₃ ) ₃ ), gold (I) carbonate (Au ₂ CO ₃ ), and combinations thereof. have.

In one exemplary embodiment, each pore in the porous structure formed in the composite polymer matrix may have a microporous porous structure, a mesoporous porous structure, and/or a macroporous porous structure. have. As used herein, the microporous pore means a porous structure having an average pore diameter of less than 2 nm, and the mesoporous pore means a porous structure having an average pore diameter of 2 nm or more and less than 50 nm. means a porous structure with an average diameter of 50 nm or more.

Although the amount of the pore former used to form the porous polymer matrix is not particularly limited, the pore former is approximately 0.001 to 1.0 g/ g, preferably at a concentration of 0.001 to 0.5 g/g, but the present invention is not limited thereto.

If necessary, when preparing a porous polymer matrix by treatment with a pore former, an inorganic strong acid such as hydrochloric acid, sulfuric acid, or nitric acid is added to the reactant to adjust the pH of the reactant to perform a crosslinking reaction with chitosan, a cationic polymer, and a crosslinking agent can promote In one exemplary embodiment, the inorganic strong acid may be added at a concentration of 0.05 to 0.5 mL/g based on the total weight of the polymer of polyvinyl alcohol and chitosan to adjust the pH of the reactant to about 4 to 5.5, but the present invention provides this It is not limited.

According to another aspect of the present invention, the present invention relates to a hemostatic agent comprising a polyvinyl alcohol moiety and a polymer matrix in which a chitosan moiety is bonded through cross-linking as an active ingredient. For example, a hemostatic agent including a polymer matrix is variously molded in the form of powder, granule, sol, gel, nonwoven fabric, fabric, sponge or sheet depending on the processing method. can be At this time, the polymer matrix according to the present invention serves as a base material for supporting the hemostatic agent, and can act as a hemostatic agent by covering the bleeding site and contracting the tissue around the blood vessel.

If necessary, the hemostatic agent according to the present invention, in addition to the above-mentioned polymer matrix, one or more hemostasis promoting components, for example, thrombin, thromboplastin, fibrinogen, casein kinase II, It may include a hemostasis promoting component selected from the group consisting of collagen, gelatin, and combinations thereof. The hemostatic agent can be prepared by dissolving these hemostasis promoting components in physiological saline or the like, immersing the above-mentioned nonwoven fabric or fabric of the polymer matrix in the solution, and drying the solution.

Hereinafter, the present invention will be described through exemplary embodiments, but the present invention is not limited to the technical ideas described in the following examples.

Synthesis Example 1: Preparation of TEMPO-oxidized cellulose nanofibers

100% cosmetic cotton (cotton beauty) natural cellulose was added to TEMPO (2,2,6,6-tetramethyl-piperidinyl-1-oxyl)/NaBr solution, and mixed using a magnetic stirrer at room temperature. When the noodles were released, NaCl was added to the mixture to regioselectively convert the cellulose C6 primary hydroxyl groups to carboxyl groups. Until the pH reached 10-11, 1M NaOH was intermittently added dropwise to the mixture to induce efficient oxidation of C6 primary hydroxyl groups. Using centrifugation and a vacuum filtration system, a salt free tempo-oxidized cellulose filter cake was obtained, which was then dried and dispersed in distilled water. To prepare a TEMPO-oxidized cellulose nanofiber solution (TOCN solution), the solution was treated three times with probe sonication. After centrifuging the solution to remove iron powder as a probe, a supernatant, a TOCN solution without iron powder, was obtained.

Example 1: TEMPO-oxidized cellulose nanofiber characterization (characterization)

(1) SEM image

The morphology of the TEMPO-oxidized cellulose nanofibers (TOCN) synthesized in Synthesis Example 1 was examined with a high-resolution scanning electron microscope (HR-SEM; SU8010, Hitachi) having a 3 kV acceleration voltage. A sample was placed on a glass substrate, the glass substrate on which the sample was placed was fixed to a carbon tape, and Pt was sputtered thereon for 180 seconds. 1 is a SEM image of the synthesized TEMPO-oxidized cellulose nanofiber (TOCN). For comparison, SEM images of non-TEMPO-treated cotton cellulose are shown in (a) and (b), and SEM images of TOCN are shown in (c) and (d). Both the cotton cellulose and TOCN had a rod shape with a long length compared to the width. Cotton cellulose consists of micro fibrils larger than 50 μm, in which disordered amorphous domains are removed by chemical treatment, leaving regular nano-sized crystalline portions to form crystalline nanofibers. changed

(2) TME and stomatal analysis

A transmission microscope (TEM; CM200) was used to observe the more detailed morphology of TOCN. The TEM image is shown in (a) on the left side of FIG. 2, and length dispersion and width dispersion are shown on the right side of FIG. 2 (b) and (c), respectively. The length dispersion graph was prepared with an interval of 20 nm, and the TOCN prepared in Synthesis Example 1 was analyzed to have an average length of 231.75 (±97.57) nm. The width dispersion graph was prepared with an interval of 3 nm, and the average width of the TOCN prepared in Synthesis Example 1 was analyzed to be 16.48 (±8.30) nm. Based on the length and width analysis results of TOCN, the aspect ratio (length/width) of TOCN was 14.06. The microfiber width is dependent on the cellulose source, and C6 primary hydroxyl groups on the surface of untreated cellulose microfibers are selectively converted to C6 carboxylate ester groups by TEMPO/NaBr/NaCIO oxidation, so that cotton fibers was successfully converted into nano-sized fibers by TEMPO-oxidation.

(3) FTIR analysis

In addition, in order to confirm whether the hydroxyl group of cellulose C6 was converted to a functional group or modified, Fourier transform infrared spectroscopy (FTIR) was performed. A sample was prepared in the form of a thin film, and the sample was analyzed in a wavelength range of ^{400 cm -1} to 4000 cm ^{-1 .} The FTIR analysis results are shown in FIGS. 3A and 3B . ^{Referring to Fig. 3a, the peaks of 3345 cm -1} and 2916 cm ^-1 observed in cotton fiber and TOCN are related to OH stretching and CH stretching. Referring to Figure 3b, the peaks at 1164, 1107, 1056 and 1027 cm ^-1 are, respectively, the asymmetric stretching of the COC bridge, the asymmetric stretching of the anhydroglucose ring, the stretching of CO in the β-glycosidic bond, and the CO stretching of the ethanol group and the CO assigned to the deformation. Unlike cotton fibers, TOCN showed a peak at ^{1605 cm -1} , which was assigned to the ^{asymmetric carboxylate COO-vibration.} This characteristic peak means that the C6 carboxyl group was converted to a C6 carboxyl group due to oxidation of the C6 primary hydroxyl group on the surface of the cotton fiber under TEMPO/NaBr/NaClO conditions.

Synthesis Example 2: PVA sponge production

Polyvinyl alcohol (weight average molecular weight: 125,000 Da; Sigma-Aldrich) was dissolved in distilled water and heat-treated in a water bath at 80° C. while stirring at 330 rpm for 5 hours to prepare a 10 wt% PVA aqueous solution. To form pores, 20 g of PVA aqueous solution, 1.3 g of triton X-100, and 3.3 mL of 37% formaldehyde were mixed, and centrifuged at 3000 rpm for 2 minutes. HCl was added directly to the foaming solution and mixed again for 2 minutes at 3000 rpm. Finally, the foamed solution was poured into a pre-heated conical tube and transferred to a 65° C. oven to form crosslinks. After 6 hours, the solid foam was washed with tap water while repeatedly removing the unreacted residue. After putting the solid foam in an excess of distilled water, the distilled water was heated for 1 hour, and residues (particularly, triton-X 100) trapped in the pores were completely removed using the evaporated distilled water bubbles. To remove unreacted formaldehyde, the sample was placed in 200 mL of 1% glycine and stirred overnight. The solid foam was washed with excess water and dried at room temperature for 3 days to prepare a PVA matrix solid foam.

Synthesis Example 3: PVA-TOCN sponge preparation

Polyvinyl alcohol (weight average molecular weight: 125,000 Da; Sigma-Aldrich) was dissolved in the TOCN solution obtained in Synthesis Example 1, and heat-treated in a water bath at 80° C. while stirring at 330 rpm for 5 hours, PVA and TOCN were 10:1 A PVA-TOCN solution mixed in a weight ratio was prepared. Then, the same procedure as described in Synthesis Example 2 was repeated to prepare a sponge-type hemostatic agent having a PVA-TOCN composite matrix.

Synthesis Examples 4 to 5: Preparation of PVA-TOCN sponge

PVA- A hemostatic agent in the form of a sponge having a TOCN composite matrix was prepared. The prepared sponge was cut into a rectangular cylindrical shape so that it could be applied to the depressed bleeding site, and it was compressed again to have a coin shape with a final diameter of 0.5 to 1.0 cm and a height of 1 to 1.5 cm. The size (diameter and height) of the final compressed coin form is not limited by this practice.

Example 2: Evaluation of the inflation pressure of a composite matrix hemostatic agent

The inflation pressure of the hemostatic agent molded into a sponge shape including the composite matrix prepared in Synthesis Examples 4 to 5 was evaluated. For comparison, the inflation pressure of the sponge-shaped hemostatic agent having a matrix composed of the PVA matrix prepared in Synthesis Example 2 was also evaluated. In order to evaluate the expansion force, each sample was dried at room temperature for 2-3 days, and the sample was cut with a unicore punch to have a cylindrical shape (diameter 12 mm, height 12 mm). After putting the sponge in a cylindrical press mold (diameter 12 mm), a pressure of 5 MPa was applied using a compressor to form a coin shape. The expansion pressure was calculated from the blocked force of the tensile tester compression mode (BMS tech). The zero point of force was set by lowering the tensile test jig until it reached the sample in the Petri dish. Distilled water at 37° C. was added into the Petri dish to allow the sample to absorb moisture, and the expansion force over time was measured. To obtain the expansion pressure, the measured expansion force was divided by the sample area. Since bleeding is difficult to occur when the inflation pressure is high, the inflation pressure is the most important factor in the present invention. 4 shows the maximum expansion pressure for each sponge, and FIG. 5 shows the average expansion force change over time from when the sponge starts to absorb distilled water. The PVA sponge showed an average maximum inflation pressure of 28.12 kPa, but the PVA-TOCN (10:1) sponge showed an average maximum inflation pressure of 67.37 kPa. As the content of TOCN increased, the maximum inflation pressure increased. In addition, as shown in FIG. 5 , when TOCN was added, the expansion rate to reach a constant state of expansion was increased. These results suggest that the added TOCN affects the mechanical properties of the sponge. It seems that some hydroxyl groups present on the TOCN surface form a bridge with the hydroxyl groups of PVA, increasing the density and thickening of the PVA sponge skeleton.

When compacted to form a coin tissue shape in a composite matrix sponge, the dense skeletal density prevents the matrix structure from collapsing and improves resiliency when water absorbs. The strong interaction between PVA and TOCN is induced due to the other free hydroxy and carboxyl groups present on the TOCN surface, enhancing the expansion pressure. In addition, due to the presence of carboxyl groups on the surface, TOCN is well dispersed in the PVA solution, which affects the improvement of the expansion force. On the other hand, the improvement of the expansion rate can be explained by the presence of functional groups present on the TOCN surface. When water is added to the pressed sponge, there is an interaction between the polymer and the solvent, ie, hydration and hydrogen bond formation. The carboxyl group present on the TOCN surface helps to attract the polar solvent, and due to the negative charge of the carboxyl group, hydrogen electrons are pulled toward the matrix polymer, reducing the energy of the system and increasing the entropy.

Example 3: Evaluation of water absorption rate of the composite mattress hemostatic agent

The water absorption rate of each of the composite matrix hemostatic agents prepared in Synthesis Examples 2 to 4 was evaluated as follows. Each sample molded into a coin shape was dried at room temperature for 2-3 days in a chamber rich in silica gel. The dried sample was weighed on a scale, and the coin-shaped sample was immersed in distilled water at 37° C. for 10 minutes. The sample was removed from the water and the sample was placed on a filter paper for 1 minute to remove excess water. For the moisture absorption weight of the sample, the weight of the sample to which moisture was adsorbed was measured on a scale. The water uptake ratio was calculated by weight analysis using the following formula.

W _r is the water absorption percentage, W _w is the weight of the sample after absorption of water, and W _d is the weight of the dry sample.

The measurement result is shown in FIG. The rate of water absorption depends on the vacancy of the sponge to receive the water and the capillary pressure. The capillary pressure can be roughly calculated using the following equation.

where γ is the surface tension in the liquid, θ is the contact angle between the liquid and the solid surface, and r is the pore radius. As the content of TOCN increases, the radius of the sponge pore size decreases, and the capillary pressure increases. Capillary pressure is proportional to the ability to receive water, with increasing TOCN content increasing water absorption.

Example 4: Morphology Evaluation of Composite Mattress Hemostatic Agent

(1) SEM image

The morphologies of the PVA sponge prepared in Synthesis Example 2 and the PVA-TOCN (10:1) sponge prepared in Synthesis Example 2 were evaluated with a high-resolution scanning electron microscope (HR-SEM; SU8010; Hitachi) at an acceleration voltage of 1.5 kV. . Each sample was cut and Pt was sputtered onto the sample for 180 seconds. An evaluation result is shown in FIG. The left side of FIG. 7 is an SEM image of the PVA sponge, and the right side of FIG. 7 is an SEM image of the PVA-TOCN sponge. At a low resolution (see (a) and (b) of Fig. 7), the number of pores in the PVA sponge appears to be larger than the number of pores in the PVA-TOCN sponge. In addition, the pore size of the PVA sponge also appears to be larger than the pore size of the PVA-TOCN sponge. However, the thickness of the skeleton of the PVA-TOCN sponge is thicker than that of the PVA sponge. At 20k magnification and 22k magnification (see Fig. 7(c), (d), (e), (f)), the PVA sponge shows a smooth surface, but the PVA-TOCN shows a rough surface. When magnified to 50k (see (g) and (h) of Figure 7), the PVA sponge skeleton does not show the bundle, but it can be seen that the TOCN bundles arranged in the PVA-TOCN sponge skeleton cover the inside of the PVA skeleton. Due to the bundle-like structure of TOCN, the PVA-TOCN skeleton sponge is thickened. The mechanical properties were improved due to the reinforced PVA-TOCN framework structure. In addition, it seems that the adsorption rate is improved as the capillary pressure is increased due to the thickened PVA-TOCN framework structure.

(2) pore size dispersion

Pore size variance was analyzed using low-resolution SEM images. ImageJ program was used to analyze the sponge pore size. The scale was set using the scale bar of the SEM image, and the image was set as a threshold value. Then, pore size dispersion was performed using ImageJ's particle analysis program. The pore size analysis was set at _{500 μm 2 to infinity based on references.} The analysis results are shown in FIG. 8 . PVA sponge and PVA-TOCN sponge have similar pore size dispersion morphology. However, the average pore size of the PVA-TOCN sponge is smaller than that of the PVA sponge, and the difference in water absorption rate may be explained by this average pore size dispersion. That is, as the average pore size of the PVA-TOCN sponge decreased, the capillary pressure increased, and the moisture adsorption rate was improved.

Example 5: In vitro toxicity test

Since cytotoxicity is very important in a medical device, an in vitro toxicity test was performed to indirectly confirm the safety of the sample prepared in Synthesis Example. The in vitro toxicity test was performed using the effluent of L-929 mouse fibroblast-bearing samples, and was performed according to the ISO 10993-5:2009(E) method. PVA-TOCN sponge, 0.1% ZDEC (zinc diethyldithiocarbamate) polyurethane film (positive control), and high-density polyethylene film (negative control) were used for the test. Each sample was placed at 24±2° C. in a Minimum Essential Medium (MEM) solution containing Fetal Bovine Serum (FBS) to prepare an effluent. L-929 fibroblast cells (ATCC CCL1, NCTN Clone 929) ^{were inoculated at a density of 1 X 10 5} cells/mL in 6-well plast containing 2 mL MEM containing 10% FBS, 37±1℃, 5 Incubated for 24 hours at ±1 CO _{2 concentration.} After selecting a well suitable for the test, removing the culture medium from the well, adding each effluent to each well and incubating for 48 hours. Qualitative analysis was performed using a microscope. Cell morphology, vacuole formation, cell adhesion, cell lysis (cytolysis), and membrane lysis (membranolysis) were observed, and grades were evaluated according to the standards in Table 1 below.

Status of cells (ISO 10993-5:2009(E) Qualitative morphological grade of cytolysis of extracts) ranking Reactivity status of all cultures 0 does not exist Individual intracytoplasmic granules: no lysis, no decrease in cell growth One Slight Less than 20% of cells are round in shape, loosely attached, no granules in the cytoplasm; or change in morphology; Some lysed cells visible; Slight sexual inhibition observed 2 Mild Up to 50% of cells have a spherical shape, no granules in the cytoplasm (devoid), no extensive cell lysis; <50% growth inhibition observed 3 Moderate up to 70% of cells are round in shape, including round or lysed cells; Although the cell layer was not completely destroyed, more than 50% growth inhibition was observed. 4 Severe Almost complete destruction in the cell layer

Quantitative analysis was performed by calculating Relative cell count (RCC %). Each well was washed twice with PBS, and 300-500 μl trypsin-EDTA was added to each well to detach the cells (detached). After collecting the supernatant cells in a 15 mL conical tube, using a hemocytometer, the cell density (cells/mL) was measured based on the following equation.

The results of the qualitative analysis according to this example are shown in Table 2 below. As shown in Table 2, there was no confluent monolayer, intracellular granulation, cell rounding, or cell lysis in the PVA-TOCN sponge effluent.

Qualitative analysis result of cytotoxicity test (48 hours) PVA-TOCN Solvent (control) voice control positive control A B C A B C A B C A B C fusion fault 100 100 100 100 100 100 100 100 100 5 5 5 intracellular granulation 100 100 100 100 100 100 100 100 100 0 0 0 spheroidization 0 0 0 0 0 0 0 0 0 0 0 0 Dissolution 0 0 0 0 0 0 0 0 0 95 95 95 ranking 0 0 0 4

In addition, the quantitative analysis results according to this example are shown in Table 3 below. The PVA-TOCN sponge effluent shows an RCC value of 100.6% in quantitative analysis. The PVA-TOCN sponge was grade 0 in the cytotoxic response, showing no cytotoxicity.

Cytotoxicity test quantitative analysis result (48 hours) Cell Concentration (number/mL) PVA-TOCN menstruum voice control positive control One 8.2 x 10 ⁵ 8.4 x 10 ⁵ 8.2 x 10 ⁵ 0 2 8.1 x 10 ⁵ 7.9 x 10 ⁵ 8.2 x 10 ⁵ 0 3 8.1 x 10 ⁵ 8.0 x 10 ⁵ 7.9 x 10 ⁵ 0 average 8.1 x 10 ⁵ 8.1 x 10 ⁵ 8.1 x 10 ⁵ 0 RCC (%) 100.0 100.0 100.0 0

Example 6: Inflammation test

Raw 264.7 cells, one of macrophages, were used for indirect inflammation tests. Considering the phosphorylcholine group, which is a major component of the cell membrane, alpha-GPC was used as a negative control, and LPS, which is known to induce an inflammatory response, was used as a positive control. A cell culture medium was prepared by mixing Dulbecco's Modified Eagle's Medium (DMEM) with high glucose concentration, 10% fetal bovine serum (FBS; Gibco), and 1% penicillin streptomycin (pen stem, Gibco). To prepare an effluent sample, alpha-glycerylphosphorylcholine (alpha-GPC; nootrabiolabs) E. coli-derived lipopolysaccharide (LPS; Sigma Aldrich), PVA-TOCN sponge was added to each culture medium at 1000 ng / mL, and incubated for 24 hours at 37±1° C., ±5 CO _{2 concentration.} Raw 264.7 cells (passage number 24) ^{were inoculated into each culture medium at a concentration of 5 X 10 4} cells/mL in a 24 well plate (non-additive culture medium, alpha-GPA-added culture medium; LPS-added culture medium and PVA-TOCN effluent culture medium) ), 37±1° C., and _{incubated for 24 hours at a concentration of ±5 CO 2 .} Raw 264.7 cells were activated by the medium to release cytokines. After 24 hours, each cell culture medium was centrifuged at 1,500 rpm for 3 minutes to separate the cells, and the suspension containing TNF-alpha was quantitatively analyzed using a TNF-alpha ELISA kit (Invitrogen, INV-KMC 3011). analyzed. The amount of TNF-alpha is proportional to the fluorescence intensity. The fluorescence intensity was detected using a microplate reader (Bio-Rad), and a standard TNF-alpha curve graph was obtained. Fluorescence intensity was detected three times for each sample, and the amount of TNF-alpha was measured using a standard curve graph. The analysis results are shown in FIG. 9 . The PVA-TOCN sponge effluent showed the lowest TNF-alpha concentration, confirming that the PVA-TOCN sponge did not stimulate an inflammatory response. In other words, it means that the PVA-TOCN sponge is biologically safe against the inflammatory response.

Example 7: Immunohistological verification of the hemostatic effect of PVA-TOCN sponge

(1) H&E and MT immunity

After anesthetizing an adult New Zealand rabbit (2.0 kg, male) using zoleti and rumpun, the rabbit's nose tissue was wounded for bleeding using a surgical punch with a 3 mm pore size, and the wound was removed after 48 hours. The nasal cavity was packed with a PVA sponge and a PVA-TOCN sponge. Then, 3 and 7 days after the start of bleeding, nasal tissues were analyzed using H&E (Hematoxylin & eosin) staining and MT (Masson Trichrome) staining. Negative controls were wounds and no action was taken, and positive controls were packed with Vaseline (baseline). The analysis results are shown in FIGS. 10A and 10B . In the nasal cavity of rabbits packed with PVA-TOCN, the hemostatic effect was comparable to that of the positive control using Vaseline.

(2) Assessment of angiogenesis ability

CD31 (vascular endothelial cells) and neutrophils (immunity) were analyzed in rabbit nasal tissues according to the same procedure as described above using histoimmunological methods. The analysis results are shown in FIGS. 11A and 11B , respectively. In the nasal passages of rabbits treated with PVA-TOCN, CD31 and neutrophils were produced to a similar extent to the positive controls. These results can be interpreted as the contribution of PVA-TOCN to regenerating blood vessels and increasing immunity in bleeding tissues.

In the above, the present invention has been described based on exemplary embodiments and examples of the present invention, but the present invention is not limited to the technical ideas described in the above embodiments and examples. Rather, those skilled in the art to which the present invention pertains can easily propose various modifications and changes based on the above-described embodiments and examples. However, it is clear from the appended claims that all such modifications and variations are within the scope of the present invention.

Claims (8)

A composite polymer matrix in which a polyvinyl alcohol (PVA) moiety and an oxidized cellulose moiety are cross-linked, wherein the hydroxyl group in the molecule is cross-linked by a cross-linking agent having an aldehyde group. Hemostatic composition comprising.
The hemostatic composition according to claim 1, wherein the oxidized cellulose is modified to have a carboxyl group.
The haemostatic composition according to claim 1, wherein the oxidized cellulose is a carboxyl group in which a hydroxyl group binding to the 6-position carbon of glucose, which is a constituent unit, is modified.
The hemostatic composition according to any one of claims 1 to 3, wherein the polyvinyl alcohol moiety and the oxidized cellulose moiety are mixed in a weight ratio of 5:1 to 50:1 in the composite polymer matrix. .
The haemostatic composition according to any one of claims 1 to 3, wherein the polyvinyl alcohol moiety and the oxidized cellulose moiety are mixed in a weight ratio of 5:1 to 20:1 in the composite polymer matrix. .
The haemostatic composition according to any one of claims 1 to 3, wherein the composite polymer matrix has a porous structure.
The method according to any one of claims 1 to 3, wherein the composite polymer matrix is a powder, a granule, a sol, a gel, a non-woven fabric, a fabric, a sponge, a compression Hemostatic composition, characterized in that molded in the form of a sponge or sheet.
[Claim 4] The hemostatic composition according to any one of claims 1 to 3, wherein the composite polymer matrix is prepared in a block form, cut again, and then compressed to form a compressed sponge.

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