CN113425889B - Antibacterial hemostatic sponge and preparation method and application thereof - Google Patents
Antibacterial hemostatic sponge and preparation method and application thereof Download PDFInfo
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- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
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- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
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- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
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- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0036—Porous materials, e.g. foams or sponges
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- A—HUMAN NECESSITIES
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- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
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- A—HUMAN NECESSITIES
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/10—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing inorganic materials
- A61L2300/102—Metals or metal compounds, e.g. salts such as bicarbonates, carbonates, oxides, zeolites, silicates
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/204—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials with nitrogen-containing functional groups, e.g. aminoxides, nitriles, guanidines
- A61L2300/208—Quaternary ammonium compounds
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/21—Acids
- A61L2300/214—Amino acids
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/23—Carbohydrates
- A61L2300/232—Monosaccharides, disaccharides, polysaccharides, lipopolysaccharides
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
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- A—HUMAN NECESSITIES
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/418—Agents promoting blood coagulation, blood-clotting agents, embolising agents
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/45—Mixtures of two or more drugs, e.g. synergistic mixtures
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- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/04—Materials for stopping bleeding
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention relates to the technical field of biomedical nano materials, in particular to an antibacterial hemostatic sponge and a preparation method and application thereof, wherein the antibacterial hemostatic sponge is prepared according to the following steps: dissolving the chitosan compound, adjusting the pH value to be neutral, adding 6-aminocaproic acid and nano zinc oxide particles, uniformly stirring, and performing low-temperature freeze drying and irradiation sterilization in a mold to obtain the antibacterial hemostatic sponge. The preparation method of the antibacterial hemostatic sponge is simple and convenient, the operation is simple, the antibacterial hemostatic sponge is green and environment-friendly, and the prepared antibacterial hemostatic sponge material has high-efficiency antibacterial and hemostatic properties, can be used for preparing medical sponge and is expected to be applied to clinic.
Description
Technical Field
The invention belongs to the technical field of biomedical nano materials, and particularly relates to an antibacterial hemostatic sponge and a preparation method and application thereof.
Background
The tissue damage is easy to cause bleeding and breed bacteria, and the effective hemostasis and antibiosis means can reduce unnecessary casualties. Therefore, the development of a hemostatic material which can stop bleeding and resist bacteria has important significance in the process of trauma treatment and operation.
Chitosan is a natural polycation polysaccharide, has the advantages of no toxicity, good biocompatibility, no immunogenicity and capability of being degraded by lysozyme, and generates glucosamine after being degraded. Meanwhile, chitosan has the functions of resisting bacteria, stopping bleeding, regulating immunity, promoting wound healing, absorbing wound exudates, reducing scar formation and the like.
6-aminocaproic acid is a traditional hemostatic drug, can inhibit the activation factor of plasminogen, so that the plasminogen is not activated into plasmin, and fibrinolysis is inhibited, thereby achieving the purpose of hemostasis, 6-aminocaproic acid is mainly used for bleeding or local bleeding during various surgical operations, bleeding caused by the activity increase of the plasmin, such as gynecological bleeding, bleeding after internal organs such as prostate, liver, pancreas and lung, and the like, and 6-aminocaproic acid has no good hemostatic effect on cancer bleeding and bleeding caused by non-fibrinolysis, thereby limiting the application of hemostasis.
The nano zinc oxide material has good sterilization and bacteriostasis performance, and is widely applied to the development of sterilization and bacteriostasis functions of medical materials at present, the nano zinc oxide has good bacteriostasis effects on escherichia coli and staphylococcus aureus, but is easy to agglomerate, and special attention needs to be paid to dispersion in the preparation process, so that the nano zinc oxide needs to be subjected to complicated pretreatment steps when being used.
Based on the consideration of the technical defects, it is necessary to design a material which is convenient to use and can improve the hemostasis application of 6-aminocaproic acid, and at present, no published literature reports at home and abroad that the antibacterial hemostasis chitosan sponge is made of nano zinc oxide added with hemostatic drugs and broad-spectrum antibacterial property.
Disclosure of Invention
In order to solve the problems of the prior art, the invention provides an antibacterial hemostatic sponge and a preparation method and application thereof.
The invention is realized by the following technical scheme:
a preparation method of an antibacterial hemostatic sponge comprises the following steps:
(1) Preparing chitosan gel: dissolving a chitosan compound and preparing into chitosan gel with the mass fraction of 2-4%;
(2) Preparing an antibacterial hemostatic sponge: adjusting the pH value of the chitosan gel obtained in the step (1) to be neutral, adding 6-aminocaproic acid and nano zinc oxide particles into the chitosan gel, uniformly stirring, freezing the mixture at the temperature of between 10 ℃ below zero and 20 ℃ below zero for 8 to 12 hours, and then performing freeze drying and sterilization to obtain the antibacterial hemostatic sponge;
wherein the mass ratio of the chitosan compound to the 6-aminocaproic acid is 20-5; the mass ratio of the chitosan compound to the nano zinc oxide is 20.
Preferably, the chitosan compound in the step (1) is chitosan or water-soluble chitosan quaternary ammonium salt or water-soluble carboxymethyl chitosan, and the deacetylation degree of the chitosan in the chitosan compound is more than or equal to 95%.
Preferably, when the chitosan compound is chitosan, the chitosan compound is dissolved in an acetic acid solution with the volume fraction of 1% to prepare chitosan gel;
and when the chitosan compound is water-soluble chitosan quaternary ammonium salt or water-soluble carboxymethyl chitosan, dissolving the chitosan compound in deionized water to prepare chitosan gel.
Preferably, the pH value of the step (2) is adjusted to be neutral by using 0.1mol/L NaOH and 1% volume fraction acetic acid solution.
Preferably, the low-temperature freeze-drying conditions in the step (2) are as follows: freeze drying at-40 deg.C for 6-10 hr.
Preferably, the sterilization method in step (2) is: by using Co 60 Sterilizing by irradiation for 3-5h with the radiation intensity of 15-20kGy.
The invention also protects the antibacterial hemostatic sponge prepared by the preparation method.
The invention also protects the application of the antibacterial hemostatic sponge in preparing medical sponge.
Compared with the prior art, the invention has the following beneficial effects:
1. the pure chitosan has unsatisfactory sterilization and bacteriostasis effects, so the hemostatic and antibacterial effects are achieved by adding hemostatic 6-aminocaproic acid and antibacterial nano zinc oxide; furthermore, the chitosan quaternary ammonium salt has better sterilization and bacteriostasis performance, and the added nano zinc oxide also has better sterilization and bacteriostasis performance, so that the antibacterial and hemostatic performance of the chitosan quaternary ammonium salt is improved by adding the hemostatic medicament 6-aminocaproic acid and the antibacterial nano zinc oxide. The carboxymethyl chitosan has certain hemostatic performance, and better hemostatic and antibacterial effects can be achieved by adding hemostatic 6-aminocaproic acid and antibacterial nano zinc oxide.
2. According to the preparation method, the reaction raw materials and the reaction steps are controlled, so that the antibacterial hemostatic biological sponge with excellent hemostatic and antibacterial properties is prepared, the antibacterial hemostatic biological sponge is directly used, pretreatment of nano zinc oxide is not needed, and the antibacterial hemostatic sponge is of a porous structure and can be applied to hemostasis of cancer hemorrhage and hemorrhage caused by non-fibrinolysis.
3. The preparation method is easy to implement, and compared with the existing reported chitosan sponge, the prepared high-efficiency antibacterial hemostatic biological sponge has more outstanding hemostatic and antibacterial effects; the following results are obtained through experimental layers: the material of the invention has good treatment effect on wound hemostasis and wound infection, and is expected to be applied to clinic.
Drawings
FIG. 1 is an optical photograph of an antibacterial hemostatic sponge according to example 3 of the present invention, wherein (a) is a sterilized package, (b) is a plan view, and (c) is a cross-sectional view;
FIG. 2 is a scanning electron microscope image of the antibacterial hemostatic sponge prepared in example 3 of the present invention at different magnifications, wherein (a) is a 30-fold magnified image and (b) is a 100-fold magnified image;
FIG. 3 is a chart showing the hemostatic effect of the antibacterial hemostatic sponge prepared in example 3 of the present invention in a rabbit liver hemorrhage model;
wherein (a) is a liver model, (b) is a liver bleeding model, and (c) is a schematic diagram of the hemostasis of the antibacterial hemostatic sponge on the liver bleeding model;
fig. 4 is a comparison chart of the bacteriostatic performance test of the antibacterial hemostatic sponges prepared in example 4, example 5, comparative example 2 and comparative example 3.
Detailed Description
The following detailed description of specific embodiments of the invention is provided, but it should be understood that the scope of the invention is not limited to the specific embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention. The experimental methods described in the examples of the present invention are all conventional methods unless otherwise specified.
Example 1
A preparation method of an antibacterial hemostatic sponge comprises the following steps:
(1) Preparing chitosan gel: 2g of chitosan quaternary ammonium salt is dissolved in 50mL of deionized water to prepare chitosan gel;
(2) Preparing an antibacterial hemostatic sponge: adjusting pH of chitosan gel to neutral with 0.1mol/L NaOH and 1% acetic acid solution, adding 10% 6-aminocaproic acid hemostatic, stirring for 1h, adding 2% nano zinc oxide, stirring for 2h to obtain mixed gel, freezing at-10 deg.C for 12h, freeze drying at-40 deg.C for 10h, sterilizing by irradiation, and sterilizing with Co 60 Sterilizing for 3h by irradiation, wherein the radiation intensity is 20kGy, and obtaining the antibacterial hemostatic sponge containing 6-aminocaproic acid and nano zinc oxide.
Example 2
A preparation method of an antibacterial hemostatic sponge comprises the following steps:
(1) Preparing chitosan gel: 2g of carboxymethyl chitosan is dissolved in 70mL of deionized water to prepare chitosan gel;
(2) Preparing an antibacterial hemostatic sponge: adjusting pH of chitosan gel to neutral with 0.1mol/L NaOH and 1% acetic acid solution, adding 10% 6-aminocaproic acid hemostatic, stirring for 1h, adding 2% nano zinc oxide, stirring for 2h to obtain mixed gel, freezing at-15 deg.C for 10h, freeze drying at-40 deg.C for 8h, sterilizing by irradiation, and sterilizing with Co 60 Sterilizing by irradiation for 4h with the radiation intensity of 20kGy to obtain the antibacterial hemostatic sponge containing 6-aminocaproic acid and nano zinc oxide.
Example 3
A preparation method of an antibacterial hemostatic sponge comprises the following steps:
(1) Preparing chitosan gel: dissolving 2g of chitosan in 50mL of acetic acid solution with volume fraction of 1% to prepare chitosan gel;
(2) Preparing an antibacterial hemostatic sponge: adjusting pH of chitosan gel to neutral with 0.1mol/L NaOH and 1% acetic acid solution, adding 10% 6-aminocaproic acid hemostatic, stirring for 1 hr, adding 2% nanometer zinc oxide, stirring for 2 hr to obtain mixed gel, and mixingFreezing at-20 deg.C for 8 hr, freeze drying at-40 deg.C for 12 hr in a freeze dryer, and sterilizing the freeze-dried sponge with Co 60 Sterilizing by irradiation for 5h with the radiation intensity of 15kGy to obtain the antibacterial hemostatic sponge containing 6-aminocaproic acid and nano zinc oxide.
Example 4
The same preparation procedure as in example 3 was followed, except that the mass fraction of nano zinc oxide was changed from 2% to 3%.
Example 5
The same preparation procedure as in example 3 was followed, except that the mass fraction of nano zinc oxide was changed from 2% to 5%.
Example 6
The preparation method is the same as that of example 1, except that 0.1g of 6-aminocaproic acid hemostatic is added and stirred for 1 hour, 0.1g of nano zinc oxide is added and stirred for 2 hours to obtain mixed gel.
Example 7
The preparation method is the same as the preparation method of the example 2, except that 0.5g of 6-aminocaproic acid hemostatic is added and stirred for 1 hour, 0.5g of nano zinc oxide is added and stirred for 2 hours to obtain mixed gel.
Comparative example 1
A preparation method of an antibacterial hemostatic sponge comprises the following steps:
(1) Preparing gel: dissolving 2g of chitosan in 50mL of acetic acid solution with volume fraction of 1% to prepare chitosan gel;
adding 0.5g of sodium carboxymethylcellulose into 25mL of deionized water to prepare sodium carboxymethylcellulose gel, and uniformly mixing the chitosan gel and the sodium carboxymethylcellulose gel to prepare gel;
(2) Preparing an antibacterial hemostatic sponge: adjusting pH of gel to neutral with 0.1mol/L NaOH and 1% acetic acid solution, adding 10% 6-aminocaproic acid hemostatic, stirring for 1h, adding 2% nanometer zinc oxide, stirring for 2h to obtain mixed gel, freezing at-10 deg.C for 12h, freeze drying at-40 deg.C for 10h, and freeze dryingIrradiating freeze-dried sponge with Co for sterilization 60 And (5) performing irradiation sterilization for 3 hours, wherein the radiation intensity is 20kGy, and obtaining the antibacterial hemostatic sponge containing 6-aminocaproic acid and nano zinc oxide.
Comparative example 2
The same procedure as in example 3 was followed except that 6-aminocaproic acid and nano zinc oxide were not added.
Comparative example 3
The same procedure as in example 3 was followed except that no nano-zinc oxide was added.
Comparative example 4
The prior art uses a common hemostatic sponge available on the market.
Comparative example 1 also produced an antibacterial hemostatic sponge, but it was less flexible and less elastic than the hemostatic sponge of example 3, indicating that the addition of sodium carboxymethyl cellulose affected the performance of the hemostatic sponge.
The antibacterial hemostatic biological sponges with excellent hemostatic and antibacterial performances are prepared in the examples 1 to 7 of the invention, and the following comparative studies are carried out on the antibacterial hemostatic biological sponges in the examples 3, 4 and 5 and the comparative examples 2 to 4, wherein the specific study methods and results are as follows:
FIG. 2 is a scanning electron microscope observation of the antibacterial hemostatic sponge prepared in example 3, wherein the sponge has a three-dimensional porous structure with continuous sheets, and the porous structure is favorable for absorbing liquid, namely blood;
fig. 3 is a diagram of the hemostatic effect of the hemostatic sponge prepared in example 3 of the present invention in a rabbit liver hemorrhage model, and it can be seen that the hemostatic sponge has hemostatic effect.
The hemostatic effect research result is as follows:
the research method comprises the following steps: 15 healthy rabbits were randomly divided into three groups, one group was used to test the antibacterial hemostatic biological sponge prepared in example 3, one group was used to test the sponge of comparative example 2, and the other group was used to test the ordinary hemostatic sponge of comparative example 4.
After anesthesia skin preparation, a 1cm wound is made on the surface of the rabbit liver, the wound is pressed by a sample after 10s of free bleeding, whether hemostasis exists or not is observed every 10s, and if bleeding continues, the pressing is continued until hemostasis exists. The results show that: the sponge hemostasis time is closely related to the content of 6-aminocaproic acid, the higher the content of 6-aminocaproic acid is, the faster the hemostasis time is, when the 10 percent addition amount is reached (example 3), hemostasis can be realized within 35 seconds, and the content of ZnO has no obvious influence on the chitosan hemostasis time; chitosan without any drug required 98 seconds for hemostasis (comparative example 2) and a common hemostatic sponge (comparative example 4) required 120 seconds.
The research of the antibacterial effect:
respectively adopting the sample leaching liquor of the embodiment 4, the embodiment 5, the comparative example 2 and the comparative example 3 to carry out bacteriostasis experiments, taking an agarose culture dish, adjusting escherichia coli (E.coli) and staphylococcus aureus (S.aureus) to be 105/m, placing the sample leaching liquor in the culture dish, placing in a constant-temperature incubator at 37 ℃ for 24 hours, and calculating bacteriostasis rate;
fig. 4 shows the bacteriostatic rates of the hemostatic sponges of example 4, example 5, comparative example 2 and comparative example 3 on staphylococcus aureus and escherichia coli, respectively, and as can be seen from fig. 4, the chitosan and chitosan/6-aminocaproic acid sponges with single purity have no obvious bacteriostatic action, and the sponges with 3% of nano zinc oxide added have the bacteriostatic rate of 35% on staphylococcus aureus and 46% on escherichia coli; the sponge added with 5% of nano zinc oxide has 58% of bacteriostasis rate to staphylococcus aureus and 64% of bacteriostasis rate to escherichia coli.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, it is intended that such changes and modifications be included within the scope of the appended claims and their equivalents.
Claims (7)
1. The preparation method of the antibacterial hemostatic sponge is characterized by comprising the following steps:
(1) Preparing chitosan gel: dissolving a chitosan compound and preparing into chitosan gel with the mass fraction of 2-4%;
(2) Preparing an antibacterial hemostatic sponge: adjusting the pH value of the chitosan gel obtained in the step (1) to be neutral, adding 6-aminocaproic acid into the chitosan gel, stirring the mixture for 1h, then adding nano zinc oxide particles into the mixture, uniformly stirring the mixture, freezing the mixture for 8 to 12h at the temperature of between 10 ℃ below zero and 20 ℃ below zero, and then carrying out freeze drying and sterilization to obtain the antibacterial hemostatic sponge;
wherein the mass ratio of the chitosan compound to the 6-aminocaproic acid is 20-5; the mass ratio of the chitosan compound to the nano zinc oxide is 20;
the chitosan compound in the step (1) is chitosan or water-soluble chitosan quaternary ammonium salt or water-soluble carboxymethyl chitosan, and the deacetylation degree of the chitosan in the chitosan compound is more than or equal to 95%.
2. The method for preparing an antibacterial hemostatic sponge according to claim 1, wherein when the chitosan compound is chitosan, the chitosan compound is dissolved in an acetic acid solution with a volume fraction of 1% to prepare chitosan gel;
and when the chitosan compound is water-soluble chitosan quaternary ammonium salt or water-soluble carboxymethyl chitosan, dissolving the chitosan compound in deionized water to prepare chitosan gel.
3. The method for preparing an antibacterial hemostatic sponge according to claim 1, wherein the pH in step (2) is adjusted to neutral by using 0.1mol/L NaOH and 1% volume acetic acid solution.
4. The method for preparing an antibacterial hemostatic sponge according to claim 1, wherein the conditions of low-temperature freeze drying in step (2) are as follows: freeze drying at-40 deg.C for 6-10 hr.
5. The method for preparing an antibacterial hemostatic sponge according to claim 1, wherein the sterilization in step (2) is: by using Co 60 Sterilizing by irradiation for 3-5h with the radiation intensity of 15-20kGy.
6. An antibacterial hemostatic sponge prepared by the preparation method of any one of claims 1-5.
7. Use of an antibacterial haemostatic sponge according to claim 6 in the manufacture of a medical sponge.
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| CN101366969B (en) * | 2008-09-12 | 2012-07-18 | 武汉华纳联合药业有限公司 | Uses of zinc oxide as reinforcing agent of chitosan biomembrane |
| CN101954117A (en) * | 2010-09-27 | 2011-01-26 | 中国人民解放军第三军医大学野战外科研究所 | Hemostatic bacteriostatic biological dressing and preparation method thereof |
| RU2468129C2 (en) * | 2010-12-30 | 2012-11-27 | Государственное образовательное учреждение высшего профессионального образования "Саратовский государственный университет им. Н.Г. Чернышевского" | Biopolymeric fibre, composition of forming solution for its obtaining, method of forming solution preparation, linen of biomedical purpose, biological bandage and method of wound treatment |
| CN102604138B (en) * | 2012-03-12 | 2013-09-11 | 浙江大学 | Method for preparing chitosan substrate film by self-deposition of micron gel |
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| AU2016220560A1 (en) * | 2015-02-16 | 2017-09-21 | Ihor Volodymyrovych GAIOVYCH | Hemostatic composition and hemostatic device (variants) |
| EA026104B1 (en) * | 2015-04-02 | 2017-03-31 | Общество С Ограниченной Ответственностью "Тектум" | Haemostatic and wound healing medicine |
| WO2016176186A1 (en) * | 2015-04-27 | 2016-11-03 | KOSTRUBA, Pavel | Hemostatic composition and device |
| WO2020044237A1 (en) * | 2018-08-27 | 2020-03-05 | Advamedica Inc. | Composite dressings, manufacturing methods and applications thereof |
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