CN112553883B - Biocellulosic fibres, hemostatic dressings comprising said fibres and related applications - Google Patents
Biocellulosic fibres, hemostatic dressings comprising said fibres and related applications Download PDFInfo
- Publication number
- CN112553883B CN112553883B CN201910909680.1A CN201910909680A CN112553883B CN 112553883 B CN112553883 B CN 112553883B CN 201910909680 A CN201910909680 A CN 201910909680A CN 112553883 B CN112553883 B CN 112553883B
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- China
- Prior art keywords
- biological cellulose
- hemostatic dressing
- cellulose fiber
- based compound
- chitosan
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Classifications
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- D06M11/32—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising with oxygen, ozone, ozonides, oxides, hydroxides or percompounds; Salts derived from anions with an amphoteric element-oxygen bond
- D06M11/36—Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with inorganic substances or complexes thereof; Such treatment combined with mechanical treatment, e.g. mercerising with oxygen, ozone, ozonides, oxides, hydroxides or percompounds; Salts derived from anions with an amphoteric element-oxygen bond with oxides, hydroxides or mixed oxides; with salts derived from anions with an amphoteric element-oxygen bond
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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Abstract
The invention provides a biological cellulose fiber, a hemostatic dressing containing the fiber and related applications. The biological cellulose fiber is obtained by purifying and mechanically homogenizing cellulose obtained by bacterial fermentation, and has the diameter of 20-50 nm and the length of 30-100 mu m. The biological cellulose fiber is used as a template, the zeolite molecular sieve is synthesized by a hydrothermal method, and can be compounded with chitosan at the same time to prepare the hemostatic dressing which can rapidly stop bleeding and has good mechanical property, good biocompatibility and good bacteriostatic property, and can promote wound healing.
Description
Technical Field
The invention relates to a biological cellulose fiber, a hemostatic dressing containing the fiber and related applications, in particular to the biological cellulose fiber which can be used for manufacturing the hemostatic dressing with rapid hemostasis and good mechanical property, the prepared hemostatic dressing, and preparation methods and related applications of the biological cellulose fiber and the hemostatic dressing, and relates to the field of hemostatic materials of medical instruments.
Background
Uncontrolled traumatic bleeding is a major cause of emergency medical treatment, medical surgery, or battlefield death. As shown by the American military data, more than 80% of casualties on a battlefield are caused by excessive bleeding within 1h after the trauma. Therefore, quick-acting hemostasis is the first problem to be solved for wound emergency treatment. The clinical commonly used hemostatic materials such as hemostatic gauze, hemostatic fiber, hemostatic bandage, etc. have certain limitations in use: the hemostasis time is longer; the dressing is easy to adhere to the wound, so that secondary injury is caused during dressing change; lack of antibacterial components often causes infection and suppuration of the wound.
In order to solve the above problems, novel rapid hemostatic materials such as porous zeolite materials, polysaccharide-based hemostatic materials, and polypeptide-based hemostatic materials have appeared in the field at present. Wherein:
(1) The porous zeolite material is composed of porous zeolite without any biological components, thus avoiding disease transmission and anaphylactic reaction among species. When the powder is applied directly to the bleeding site, a layer of hemostatic scab is rapidly formed on the surface of the wound to prevent the internal blood from overflowing. However, the porous zeolite material absorbs the moisture in the blood and then emits a large amount of heat, which causes inflammation of the wound, so that the porous zeolite material must be controlled to generate heat to reduce the injury to the wound during the hemostasis process. In addition, when the zeolite hemostatic bag is used, zeolite particles are remained on the wound surface, are not degraded and are easy to cause foreign body reaction.
(2) Trauma DEX, manufactured by medafer corporation, usa, is a spongy polysaccharide-based hemostatic material based on potato starch. The sponge material can absorb a large amount of water in blood at a bleeding point to promote the accelerated coagulation of blood platelets and blood proteins, thereby achieving the aim of hemostasis. However, the material is only suitable for small wounds, and the temperature rise phenomenon is accompanied in the hemostasis process.
(3) Chitosan is a polysaccharide-based hemostatic material with positive charges, which is mainly extracted from shrimp and crab shells, and can attract blood cells with negative charges and promote blood coagulation. Furthermore, after contact with blood, it is sticky and adheres tightly to the wound. Chitosan's unique hemostatic, bacteriostatic, biocompatible, wound healing promoting and gel-forming properties, giving it good performance as a hemostatic material. However, the chitosan material has poor solubility and needs to be strengthened in mechanical properties.
(4) The effective components of the fibrin dressing are fibrinogen powder, thrombin, calcium ions and the like. Such dressings are absorbable by the human body. Its hemostatic mechanism involves the dissolution of blood coagulation proteins in plasma, and the fibrin layer formed by the enzymatic reaction of fibrinogen and thrombin will adhere tightly to the damaged tissue. On the other hand, however, the material is brittle, has poor flexibility and is easily broken, which greatly limits its application.
Based on the above problems of the rapid hemostatic materials, there is still a need to develop new rapid hemostatic materials.
Disclosure of Invention
An object of the present invention is to provide a bio-cellulose fiber which can be used for preparing a hemostatic dressing having rapid hemostasis and good mechanical properties.
The invention also aims to provide a preparation method of the biological cellulose fiber.
It is another object of the present invention to provide a hemostatic dressing prepared with the bio-cellulosic fibers.
Another object of the present invention is to provide a method for preparing the hemostatic dressing.
It is another object of the present invention to provide the use of such a hemostatic dressing.
In one aspect, the present invention provides a bio-cellulose fiber that can be used to prepare a hemostatic dressing that rapidly stanchs and has good mechanical properties. The diameter of the biological cellulose fiber provided by the invention is 20-50 nm, and the length of the biological cellulose fiber is 30-100 mu m. The biological cellulose fiber is obtained by cooking cellulose obtained by bacterial fermentation with an aqueous solution of sodium hydroxide.
According to a specific embodiment of the present invention, the cellulose obtained by the bacterial fermentation can be obtained commercially or can be prepared according to the records of the prior art. The cellulose obtained by fermentation of the bacteria suitable for use in the present invention may be, for example, cellulose obtained by fermentation of: one or more of Acetobacter xylinum, rhizobium, sarcina, pseudomonas, achromobacter, alcaligenes, aerobacter, and Azotobacter. The process of obtaining cellulose by fermentation with these species is known in the art and is not described further herein.
According to a particular embodiment of the invention, the biocellulose fibres of the invention are obtained by cooking the cellulose obtained by fermentation of said bacteria with an aqueous sodium hydroxide solution. Wherein the process of the sodium hydroxide aqueous solution cooking treatment comprises the following steps: the biological cellulose is steamed and boiled for 10 to 30min at high temperature by 10 to 20 weight percent of sodium hydroxide aqueous solution. The biological cellulose fiber provided by the invention is mainly used for preparing a hemostatic dressing with rapid hemostasis and good mechanical property. The bio-cellulose raw material after bacterial fermentation has a large amount of bacterial residues which will affect the structural performance of the prepared hemostatic dressing. In the invention, the bacterial protein and the residual culture medium adhered to the cellulose membrane can be thoroughly removed by adopting sodium hydroxide solution with specific concentration for cooking, so that the high purity of the cellulose of the bacterial cellulose material can be ensured, and the purity can reach more than 99.9 percent, or 100 percent. Meanwhile, the sodium hydroxide can play a certain role in the activation treatment of the subsequent preparation of the hemostatic dressing.
In another aspect, the present invention also provides a method for preparing the bio-cellulose fiber, comprising:
the cellulose obtained by bacterial fermentation is steamed and boiled for 10 to 30min at high temperature by 10 to 20 weight percent of sodium hydroxide aqueous solution;
mechanically homogenizing to prepare the fiber with the diameter of 20-50 nm and the length of 30-100 mu m.
In another aspect, the invention also provides the use of the biological cellulose fiber in the preparation of a hemostatic dressing.
According to the specific embodiment of the invention, the biological cellulose fiber is used as a template, the zeolite molecular sieve is synthesized by a hydrothermal method, and can be compounded with chitosan to prepare the hemostatic dressing which can rapidly stanch and has good mechanical properties.
Thus, in another aspect, the present invention also provides a method of preparing a hemostatic dressing, the method comprising:
mixing the biological cellulose fiber, tetrapropylammonium hydroxide, a silicon-based compound and water to obtain a mixed solution; wherein the corresponding molar ratio of the biological cellulose fiber, the tetrapropylammonium hydroxide and the silicon-based compound is (0.001-0.2) to (0.01-1) to 1 in every 1L of water;
adding an aluminum-based compound into the mixed solution, and slowly stirring for 5-8 h to obtain mixed gel, wherein the molar ratio of the aluminum-based compound to the silicon-based compound is (0.05-0.1): 1;
slowly heating the mixed gel to 170-175 ℃, and keeping the temperature for 6-24 h to obtain a reaction sample; wherein the heating rate is 5-10 ℃/min;
washing the obtained sample with deionized water, drying, heating to 550-600 ℃ for carbonization treatment, keeping the temperature for 5-8 h, and cooling to obtain a zeolite molecular sieve and biocellulose-based carbon nanofiber composite material;
mixing the above composite material with biological cellulose fiber and chitosan water solution, lyophilizing, and sterilizing (conventional sterilization, such as high pressure steam sterilization or radiation sterilization) to obtain hemostatic dressing. The hemostatic dressings of the present invention may be cut as needed (typically prior to sterilization) and packaged.
According to the preparation method of the hemostatic dressing, the zeolite is obtained by taking the biological cellulose as a template and performing in-situ synthesis through a hydrothermal method, and a large number of hydroxyl groups on the surface of the biological cellulose can adsorb zeolite precursors (silicon-based and aluminum-based compounds) in the process to play a role of an in-situ template; meanwhile, the surface of the biological cellulose can also adsorb some metal ions which influence the synthesis of the zeolite in the raw materials, and the biological cellulose plays a role of a metal chelating agent; secondly, during high-temperature calcination, the nano-cellulose fibers are carbonized into the bio-cellulose-based nano-carbon fibers, the nano-carbon fibers have strong binding force with zeolite, and the nano-carbon fibers and the bio-cellulose fibers have a reinforcing effect. The method has the advantages of simple and easy process, convenient operation and low cost, and the obtained hemostatic dressing has quick hemostatic performance and good mechanical property, and the zeolite molecular sieve has strong binding force with a matrix and is not easy to fall off when in use.
According to a specific embodiment of the present invention, in the preparation method of the hemostatic dressing of the present invention, the silicon-based compound includes any one or more of silica gel, white carbon black, sodium silicate, methyl orthosilicate, ethyl orthosilicate, propyl orthosilicate, isopropyl orthosilicate, and butyl orthosilicate.
According to a specific embodiment of the present invention, in the preparation method of the hemostatic dressing of the present invention, the aluminum-based compound comprises any one or more of sodium metaaluminate, aluminum hydroxide, aluminum sulfate, aluminum sol, aluminum isopropoxide and aluminum sec-butoxide.
According to a specific embodiment of the present invention, in the method for preparing the hemostatic dressing of the present invention, the drying is preferably freeze-drying. The freeze drying is preferably to freeze the sample at-20 to-80 ℃ for 12 to 24 hours and then to vacuum dry for 24 to 48 hours.
According to a specific embodiment of the invention, in the preparation method of the hemostatic dressing, the carbonization treatment refers to heating the material from room temperature to 550-600 ℃ under vacuum, argon or nitrogen protection, wherein the heating rate is 5-10 ℃/min in the range of 100-300 ℃, the heating rate is 1-5 ℃/min in the range of 300-500 ℃, and the heating rate is preferably 25-30 ℃/min in the range of 500-600 ℃.
According to a specific embodiment of the present invention, in the preparation method of the hemostatic dressing of the present invention, the cooling refers to slowly cooling the sample after the temperature rise to room temperature in an atmosphere furnace or an activation furnace.
According to a specific embodiment of the invention, in the preparation method of the hemostatic dressing, when the zeolite molecular sieve and the biocellulose-based nano carbon fiber composite material are mixed with the biocellulose fibers and the chitosan aqueous solution, the mixing mass ratio of the chitosan, the zeolite molecular sieve and the biocellulose-based carbon fibers to the biocellulose fibers is 1: (0.01-0.1): (0.01:0.1): (0.01-0.1), and the more preferable ratio is 1: (0.04-0.06): (0.04-0.06): (0.3-0.6).
According to a specific embodiment of the present invention, in the method for preparing the hemostatic dressing of the present invention, the size of the bio-cellulose fiber mixed with the composite material and the chitosan aqueous solution may be the same as or different from the size of the bio-cellulose fiber in the mixed solution of the bio-cellulose fiber, the tetrapropylammonium hydroxide and the silicon-based compound, and may be within the range of 20 to 50nm in diameter and 30 to 100 μm in length as required by the present invention. The addition of the part of biological cellulose fibers further strengthens zeolite particles, so that the zeolite particles are not easy to fall off, and meanwhile, the nano cellulose fibers on the surface of the chitosan porous material can improve the adsorption efficiency of the material to blood, play a role in communicating the external zeolite particles with the internal zeolite particles and obviously accelerate the hemostasis speed.
According to a specific embodiment of the present invention, in the preparation method of the hemostatic dressing of the present invention, the chitosan aqueous solution is obtained by dissolving chitosan in 2wt% acetic acid aqueous solution. Preferably, in the chitosan aqueous solution, the mass concentration of chitosan is 3-10 wt%, and the molecular weight of chitosan is 10-35 ten thousand.
According to the specific embodiment of the invention, in the preparation method of the hemostatic dressing, the zeolite molecular sieve and the biocellulose-based nano carbon fiber composite material are mixed with the biocellulose fiber and the chitosan aqueous solution, then the mixture is frozen for 12 to 24 hours at the temperature of between 20 ℃ below zero and 80 ℃ below zero, and then the mixture is dried for 24 to 48 hours in vacuum, and sterilized to obtain the hemostatic dressing.
In another aspect, the invention also provides a hemostatic dressing prepared from the bio-cellulose fiber. The final product of the hemostatic dressing is a porous foam hemostatic material consisting of chitosan, biological cellulose fiber, nano carbon fiber (obtained by carbonizing biological cellulose) and zeolite. Structurally, chitosan is a matrix of porous material, in which biological cellulose fiber, nano carbon fiber and zeolite particles are uniformly distributed. The material porosity of the hemostatic dressing is 80-95%, the pore diameter is 50-500 μm, the water absorption rate is 200-1000 times of the material weight, and the tensile mechanical strength is 0.5-4 GPa.
In another aspect, the invention also provides the application of the hemostatic dressing. According to the hemostatic dressing, the mechanical strength of the material is improved by the carbon fibers and the biological cellulose fibers; the tight combination of the zeolite and the carbon fiber avoids the problem that the healing of the wound is affected because zeolite particles enter the wound surface when the hemostatic material is used; the addition of the biological cellulose fiber has a certain entanglement effect on the combination of the zeolite and the carbon fiber, so that zeolite particles are further reinforced and are not easy to fall off; meanwhile, the nano cellulose fibers on the surface of the chitosan porous material can improve the adsorption efficiency of the material to blood, achieve the aim of communicating the external zeolite particles with the internal zeolite particles and obviously accelerate the hemostasis speed. The hemostatic dressing has good biocompatibility and good bacteriostatic performance, and can promote wound healing. Therefore, the invention provides the application of the hemostatic dressing in preparing medicines for stopping bleeding and promoting wound healing.
In the above-described embodiment of the present invention, the process conditions not specified in detail can be carried out according to the conventional procedures in the art.
The invention has the beneficial effects that:
the zeolite molecular sieve is synthesized in situ by a hydrothermal method by using biological nano-cellulose as a template. While calcining at high temperature, the nano-cellulose fiber is carbonized into the bio-cellulose-based nano-carbon fiber. The binding force of the nano carbon fiber and the zeolite is strong, and meanwhile, the nano carbon fiber and the biological cellulose fiber have a reinforcing effect, so that the hemostatic dressing which can rapidly stop bleeding and has good mechanical properties is finally obtained. The preparation process is simple and easy to implement, convenient to operate and low in cost, and the obtained hemostatic dressing has good hemostatic performance and mechanical property; the zeolite molecular sieve has stable and uniform structure, strong binding force with a matrix and difficult shedding when in use; has good biocompatibility and good bacteriostatic property, and can promote wound healing.
Detailed Description
The following detailed description is provided for the purpose of illustrating the embodiments and the advantageous effects thereof, and is not intended to limit the scope of the present disclosure.
Example 1
The preparation method of the hemostatic dressing of this example:
(1) Mixing the biological cellulose fiber, the tetrapropylammonium hydroxide, the silicon-based compound and water to obtain a mixed solution. Wherein 1L of water is added with biological cellulose fiber, tetrapropylammonium hydroxide and a silicon-based compound, and the corresponding molar ratio of the three substances is 0.001: 0.01: 1; wherein the biological cellulose fiber is prepared by fermenting Acetobacter xylinum and Rhizobium to obtain biological cellulose, steaming with 10wt% sodium hydroxide water solution at high temperature for 10 min, and mechanically homogenizing to obtain cellulose fiber with nanofiber diameter of 20 nm and length of 30 μm; the silicon-based compound refers to silica gel.
(2) Adding sodium metaaluminate into the mixed solution, and slowly stirring for 5 hours to obtain mixed gel, wherein the molar ratio of the aluminum-based compound to the silicon-based compound is 0.05;
(3) Slowly heating the mixed gel in a high-pressure kettle to 170 ℃, and keeping the temperature for 6 hours, wherein the heating rate is 5 ℃/min;
(4) Washing the obtained sample with deionized water, freezing for 24h at-20 ℃, then drying for 24h in vacuum, then putting the dried sample into an atmosphere furnace for carbonization treatment and heating to 550 ℃, keeping the temperature for 5h, and slowly cooling the sample to room temperature in the atmosphere furnace to obtain the zeolite molecular sieve and biocellulose-based carbon nanofiber composite material; wherein, the carbonization treatment refers to that the temperature of the material is raised from room temperature to 550 ℃ under the protection of nitrogen, wherein the temperature raising rate in the range of 100-300 ℃ is 5 ℃/min, the temperature raising rate in the range of 300-500 ℃ is 1 ℃/min, and the temperature raising rate in the range of 500-550 ℃ is 30 ℃/min.
(5) The composite material is mixed with biological cellulose fiber and chitosan aqueous solution, and the hemostatic dressing which can rapidly stop bleeding and has good mechanical property is finally obtained after freeze drying, cutting, sterilization and packaging. Wherein the chitosan water solution is prepared from chitosan with the molecular weight of 35 ten thousand, and the mass concentration of the chitosan water solution is 3wt%.
The table shows part of experimental data of the influence of different contents of zeolite, carbon fiber and biological cellulose fiber on the performance of the hemostatic dressing in the research process.
Watch 1
The above example 1 obtains a hemostatic dressing with rapid hemostasis and good mechanical properties, and the main performance test results are as follows:
hemostasis: the effect of the hemostatic dressing is evaluated by adopting a rabbit fatal femoral artery bleeding wound model, and a control sample is gauze. The result shows that the hemostatic dressing of the invention has the hemostatic rate of 100 percent within 1min and the hemostatic rate of 37 percent within 8 min.
Bacteriostasis: the hemostatic dressing of this example contains chitosan, which is a non-leaching bacteriostatic material. According to the test method of the bacteriostatic performance of the non-dissolution antibacterial (bacteriostatic) product in appendix C5 of GB15979-2002 hygienic Standard for Disposable sanitary articles, the difference between the bacteriostatic rate of the tested sample group and the bacteriostatic rate of the control sample group is more than 26 percent according to the standard specification, which indicates that the product has antibacterial performance. The result shows that the difference of the bacteriostatic rates of the hemostatic dressing on Escherichia coli (ATCC 8739) and staphylococcus aureus (ATCC 6538) respectively reaches 90.09% and 89.02%, which indicates that the hemostatic dressing has good bacteriostatic performance.
Biocompatibility experiment: with reference to the biological evaluation of GB/T16886 medical instruments, the hemostatic dressings were evaluated for cytotoxicity, delayed contact sensitization of guinea pigs, skin irritation, and the like. Evaluation of biocompatibility: intracellular toxicity test according to GB/T16886-5 "evaluation of medical devices biology part 5: in vitro cytotoxicity test "; guinea pig delayed contact sensitization test part 10 of the "biological evaluation of medical devices" in GB/T16886-10: stimulation and delayed type hypersensitivity tests were performed using the Maxinusson and Kligman methods for the maximum tests. Skin irritation test according to GB/T16886-10, part 10 of the biological evaluation of medical devices: stimulation and delayed type hypersensitivity tests. The results show that: the hemostatic dressing has cytotoxicity less than grade 2, no skin sensitization reaction, no skin irritation reaction and good biological safety.
The cytotoxicity test in vitro is shown in the second table.
TABLE II, cell viability%
After the sample-soaked solution was cultured (37 ℃ C., 5% by weight of CO2) with the vigorously growing L-929 cells for 24 hours, the potential cytotoxicity of the samples was determined by MTT method. When the soaking solution is 100%, the toxicity of the test sample is class II. Along with the dilution of the soak solution, the cell activity is gradually improved, and after the soak solution is diluted to 50%, the cytotoxicity is I grade. Therefore, the toxic reaction of the sample leaching solution to the L929 cells is grade 2, and the sample leaching solution has no potential cytotoxicity.
Skin irritation: the samples are respectively pasted on the backs of 3 New Zealand white rabbits, and after 1h, 24h, 48h and 72 h, the reaction conditions of skin erythema, edema and the like are observed and graded and scored according to the skin irritation reaction degree. The results of skin reactions on the samples are shown in Table III.
TABLE III, observation of skin reaction results of samples
The animals did not show abnormal symptoms or death during the test. According to the observation, the skin reaction on one side of the experimental group did not exceed that on one side of the blank control group, the primary stimulation index was 0 (edema-free, erythema), and the positive control group animals were scored as 2 (significant edema, erythema). The test samples were non-irritating in rabbit skin reaction type.
The skin sensitization test is shown in Table four.
TABLE IV clinical observations of sensitization-stimulated skin reactions in sample guinea pigs
The sample leachate was injected intradermally into 10 guinea pigs, bandaged and attempted to induce sensitization. During the recovery period, 10 tested guinea pigs and 5 control guinea pigs were subjected to the challenge patch test using the sample leach solution and the blank solution, respectively. The site scores were recorded 24h and 48h after patch removal. In the test, the scores of the animals in the negative control group and the animals in the test group are both 0 (no significant change), and the score of the animals in the positive control group is 2 (moderate fusion erythema). Indicating that no delayed contact sensitization was found in the samples.
Example 2
The preparation method of the hemostatic dressing of this example:
(1) Mixing the biological cellulose fiber, the tetrapropylammonium hydroxide, the silicon-based compound and water to obtain a mixed solution. Wherein 1L of water is added with biological cellulose fiber, tetrapropylammonium hydroxide and a silicon-based compound, and the corresponding molar ratio of the three substances is 0.001: 0.05:1; wherein the biological cellulose fiber is prepared by fermenting biological cellulose obtained from Sarcina and Pseudomonas, steaming at high temperature for 20 min with 12 wt% sodium hydroxide water solution, and mechanically homogenizing to obtain cellulose fiber with nanofiber diameter of 30 nm and length of 40 μm; the silicon-based compound refers to white carbon black and silica gel, and the mass ratio is 1.
(2) Adding aluminum hydroxide into the mixed solution, and slowly stirring for 6 hours to obtain mixed gel, wherein the molar ratio of the aluminum-based compound to the silicon-based compound is 0.06;
(3) Slowly heating the mixed gel in an autoclave to 175 ℃, and keeping the temperature for 8 hours, wherein the heating rate is 10 ℃/min;
(4) Washing the obtained sample with deionized water, freezing at-30 ℃ for 12 hours, then vacuum drying for 48 hours, then putting the dried sample into an atmosphere furnace for carbonization treatment, heating to 600 ℃, keeping the temperature for 6 hours, and then slowly cooling the sample to room temperature in the atmosphere furnace to obtain the zeolite molecular sieve and biocellulose based carbon nanofiber composite material; wherein, the carbonization treatment refers to that the temperature of the material is raised from room temperature to 600 ℃ under the vacuum protection, wherein the temperature raising rate in the range of 100-300 ℃ is 10 ℃/min, the temperature raising rate in the range of 300-500 ℃ is 5 ℃/min, and the temperature raising rate in the range of 500-600 ℃ is 25 ℃/min.
(5) The composite material is mixed with biological cellulose fiber and chitosan aqueous solution, and the hemostatic dressing which can rapidly stop bleeding and has good mechanical property is finally obtained after freeze drying, cutting, high-energy ray sterilization and packaging. Wherein the chitosan aqueous solution is prepared from chitosan with the molecular weight of 30 ten thousand, and the mass concentration of the chitosan aqueous solution is 4 wt%.
The hemostatic dressing of the present example has a tensile mechanical strength of 1050MPa; the porosity is 80-95%, the pore diameter is 50-500 μm, and the water absorption is 200-1000 times of the material weight. The effect of the hemostatic dressing is evaluated by adopting a rabbit fatal femoral artery bleeding wound model, and the result shows that the hemostatic dressing has the hemostatic rate of 100% within 1 min.
Example 3
(1) Mixing the biological cellulose fiber, the tetrapropylammonium hydroxide, the silicon-based compound and water to obtain a mixed solution. Wherein 1L of water is added with biological cellulose fiber, tetrapropylammonium hydroxide and a silicon-based compound, and the corresponding molar ratio of the three substances is 0.2: 1; wherein the biological cellulose fiber is prepared by fermenting Acetobacter xylinum, achromobacter and Alcaligenes, steaming biological cellulose with 14 wt% sodium hydroxide water solution at high temperature for 30min, and mechanically homogenizing to obtain cellulose fiber with nanofiber diameter of 40 nm and length of 50 μm; the silicon-based compound is sodium silicate and n-butyl silicate, and the mass ratio is 1.
(2) Adding aluminum sulfate into the mixed solution, and slowly stirring for 7 hours to obtain mixed gel, wherein the molar ratio of the aluminum-based compound to the silicon-based compound is 0.07: 1;
(3) Slowly heating the mixed gel in a high-pressure kettle to 170 ℃, and keeping the temperature for 10 hours, wherein the heating rate is 6 ℃/min;
(4) Washing the obtained sample with deionized water, freezing for 18 h at-40 ℃, then vacuum-drying for 48h, then putting the dried sample into an atmosphere furnace for carbonization treatment and heating to 570 ℃, keeping the temperature for 7h, and slowly cooling the sample to room temperature in the atmosphere furnace to obtain the zeolite molecular sieve and biocellulose-based carbon nanofiber composite material; wherein, the carbonization treatment refers to that the temperature of the material is raised from room temperature to 570 ℃ under the protection of argon, wherein the temperature raising rate in the range of 100-300 ℃ is 6 ℃/min, the temperature raising rate in the range of 300-500 ℃ is 4 ℃/min, and the temperature raising rate in the range of 500-570 ℃ is 28 ℃/min.
(5) The composite material is mixed with biological cellulose fiber and chitosan aqueous solution, and the hemostatic dressing which can rapidly stanch and has good mechanical property is finally obtained after freeze drying, cutting, high-energy ray sterilization and packaging. Wherein the chitosan aqueous solution is prepared from chitosan with the molecular weight of 25 ten thousand, and the mass concentration of the chitosan aqueous solution is 5 wt%.
The hemostatic dressing of the present example has a tensile mechanical strength of 900MPa; the porosity is 80-95%, the pore diameter is 50-500 μm, and the water absorption is 200-1000 times of the material weight. The effect of the hemostatic dressing is evaluated by adopting a rabbit fatal femoral artery bleeding wound model, and the result shows that the hemostatic dressing has the hemostatic rate of 100% within 1 min.
Example 4
(1) Mixing the biological cellulose fiber, the tetrapropylammonium hydroxide, the silicon-based compound and water to obtain a mixed solution. Wherein 1L of water is added with biological cellulose fiber, tetrapropylammonium hydroxide and a silicon-based compound, and the corresponding molar ratio of the three substances is 0.005: 0.03: 1; wherein the biological cellulose fiber is prepared by fermenting biological cellulose obtained by Aerobacter and azotobacter, steaming at high temperature with 16 wt% sodium hydroxide water solution for 15 min, and mechanically homogenizing to obtain cellulose fiber with nanofiber diameter of 50nm and length of 60 μm; the silicon-based compound is methyl orthosilicate and isopropyl orthosilicate in a mass ratio of 1.
(2) Adding aluminum sol and aluminum sec-butoxide into the mixed solution according to the mass ratio of 1:1, slowly stirring for 8 hours to obtain mixed gel, wherein the molar ratio of the aluminum-based compound to the silicon-based compound is 0.08;
(3) Slowly heating the mixed gel in a high-pressure kettle to 170 ℃, and keeping the temperature for 12 h, wherein the heating rate is 8 ℃/min;
(4) Washing the obtained sample with deionized water, freezing for 24h at-50 ℃, then vacuum-drying for 48h, then putting the dried sample into an atmosphere furnace for carbonization treatment and heating to 550 ℃, keeping the temperature for 8h, and slowly cooling the sample to room temperature in the atmosphere furnace to obtain the zeolite molecular sieve and biocellulose-based carbon nanofiber composite material; wherein, the carbonization treatment refers to that the temperature of the material is raised from room temperature to 550 ℃ under the protection of nitrogen, wherein the temperature raising rate in the range of 100-300 ℃ is 10 ℃/min, the temperature raising rate in the range of 300-500 ℃ is 3 ℃/min, and the temperature raising rate in the range of 500-550 ℃ is 26 ℃/min.
(5) The composite material is mixed with biological cellulose fiber and chitosan aqueous solution, and the hemostatic dressing which can rapidly stanch and has good mechanical property is finally obtained after freeze drying, cutting, high-energy ray sterilization and packaging. Wherein the chitosan aqueous solution is prepared from chitosan with the molecular weight of 20 ten thousand, and the mass concentration of the chitosan aqueous solution is 6 wt%.
The hemostatic dressing of the present example has a tensile mechanical strength of 800 MPa; the porosity is 80-95%, the pore diameter is 50-500 μm, and the water absorption is 200-1000 times of the material weight. The effect of the hemostatic dressing is evaluated by adopting a rabbit fatal femoral artery bleeding wound model, and the result shows that the hemostatic dressing has the hemostatic rate of 100% within 1 min.
Example 5
(1) Mixing the biological cellulose fiber, the tetrapropylammonium hydroxide, the silicon-based compound and water to obtain a mixed solution. Wherein 1L of water is added with biological cellulose fiber, tetrapropylammonium hydroxide and a silicon-based compound, and the corresponding molar ratio of the three substances is 0.01: 0.02: 1; wherein the biological cellulose fiber is prepared by subjecting biological cellulose obtained by fermentation of Alcaligenes to high temperature cooking with 18 wt% sodium hydroxide water solution for 28 min, and mechanically homogenizing to obtain cellulose fiber with nanofiber diameter of 50nm and length of 70 μm; the silicon-based compound refers to ethyl orthosilicate and propyl orthosilicate in a mass ratio of 1:1.
(2) Adding aluminum isopropoxide into the mixed solution, and slowly stirring for 5 hours to obtain mixed gel, wherein the molar ratio of the aluminum-based compound to the silicon-based compound is 0.09;
(3) Slowly heating the mixed gel in an autoclave to 175 ℃, and keeping the temperature for 24 hours, wherein the heating rate is 7 ℃/min;
(4) Washing the obtained sample with deionized water, freezing for 24h at-60 ℃, then drying for 24h in vacuum, then putting the dried sample into an atmosphere furnace for carbonization treatment and heating to 600 ℃, keeping the temperature for 8h, and then slowly cooling the sample to room temperature in the atmosphere furnace to obtain the zeolite molecular sieve and biocellulose-based carbon nanofiber composite material; wherein, the carbonization treatment refers to that the temperature of the material is raised from room temperature to 600 ℃ under the protection of argon, wherein the temperature raising rate in the range of 100-300 ℃ is 9 ℃/min, the temperature raising rate in the range of 300-500 ℃ is 4 ℃/min, and the temperature raising rate in the range of 500-600 ℃ is 27 ℃/min.
(5) The composite material is mixed with biological cellulose fiber and chitosan aqueous solution, and the hemostatic dressing which can rapidly stop bleeding and has good mechanical property is finally obtained after freeze drying, cutting, high-energy ray sterilization and packaging. Wherein the chitosan aqueous solution is prepared from chitosan with the molecular weight of 15 ten thousand, and the mass concentration of the chitosan aqueous solution is 7wt%.
The hemostatic dressing of the present example has a tensile mechanical strength of 850MPa; the porosity is 80-95%, the pore diameter is 50-500 μm, and the water absorption is 200-1000 times of the material weight. The effect of the hemostatic dressing is evaluated by adopting a rabbit lethal femoral artery bleeding wound model, and the result shows that the hemostatic rate of the hemostatic dressing of the embodiment within 1min is 100%.
Example 6
(1) Mixing the biological cellulose fiber, the tetrapropylammonium hydroxide, the silicon-based compound and water to obtain a mixed solution. Wherein 1L of water is added with biological cellulose fiber, tetrapropylammonium hydroxide and a silicon-based compound, and the corresponding molar ratio of the three substances is 0.16: 0.5: 1; wherein the biological cellulose fiber is prepared by fermenting Acetobacter xylinum, aerobacter sp and azotobacter to obtain biological cellulose, steaming with 20wt% sodium hydroxide water solution at high temperature for 10 min, and mechanically homogenizing to obtain cellulose fiber with nanofiber diameter of 50nm and length of 100 μm; the silicon-based compound is silica gel, white carbon black, methyl orthosilicate and butyl orthosilicate, and the ratio is 1.
(2) Adding sodium metaaluminate, aluminum hydroxide and aluminum sulfate into the mixed solution, wherein the mass ratio of the aluminum metaaluminate to the aluminum hydroxide to the aluminum sulfate is 1;
(3) Slowly heating the mixed gel in an autoclave to 170 ℃, and keeping the temperature for 24 hours, wherein the heating rate is 5 ℃/min;
(4) Washing the obtained sample with deionized water, freezing for 24h at-80 ℃, then vacuum-drying for 48h, then putting the dried sample into an atmosphere furnace for carbonization treatment and heating to 600 ℃, keeping the temperature for 8h, and then slowly cooling the sample to room temperature in the atmosphere furnace to obtain the zeolite molecular sieve and biocellulose-based carbon nanofiber composite material; wherein, the carbonization treatment refers to that the temperature of the material is raised from room temperature to 600 ℃ under the protection of nitrogen, wherein the temperature raising rate in the range of 100-300 ℃ is 7 ℃/min, the temperature raising rate in the range of 300-500 ℃ is 3 ℃/min, and the temperature raising rate in the range of 600 ℃ is 27 ℃/min.
(5) The composite material is mixed with biological cellulose fiber and chitosan aqueous solution, and the hemostatic dressing which can rapidly stop bleeding and has good mechanical property is finally obtained after freeze drying, cutting, high-energy ray sterilization and packaging. Wherein the chitosan water solution is prepared from chitosan with the molecular weight of 10 ten thousand, and the mass concentration of the chitosan water solution is 10 wt%.
The hemostatic dressing of this example has a tensile mechanical strength of 750MPa; the porosity is 80-95%, the pore diameter is 50-500 μm, and the water absorption is 200-1000 times of the material weight. The effect of the hemostatic dressing is evaluated by adopting a rabbit fatal femoral artery bleeding wound model, and the result shows that the hemostatic dressing has the hemostatic rate of 100% within 1 min.
Claims (13)
1. A method of making a hemostatic dressing, the method comprising:
mixing biological cellulose fibers, tetrapropylammonium hydroxide, a silicon-based compound and water to obtain a mixed solution;
adding an aluminum-based compound into the mixed solution, and slowly stirring for 5-8 h to obtain mixed gel;
slowly heating the mixed gel to 170-175 ℃, and keeping the temperature for 6-24 h to obtain a reaction sample; wherein the heating rate is 5-10 ℃/min;
washing the reaction sample with deionized water, drying, heating to 550-600 ℃ for carbonization treatment, keeping the temperature for 5-8 h, and cooling to obtain the zeolite molecular sieve and biocellulose-based carbon nanofiber composite material;
mixing the composite material with biological cellulose fibers and a chitosan aqueous solution, and performing freeze drying and sterilization to obtain the hemostatic dressing;
the biological cellulose fiber is obtained by boiling cellulose obtained by bacterial fermentation with 10-20 wt% of sodium hydroxide aqueous solution, the diameter of the biological cellulose fiber is 20-50 nm, and the length of the biological cellulose fiber is 30-100 mu m.
2. A method of making the hemostatic dressing of claim 1, further comprising:
the cellulose obtained by bacterial fermentation is boiled for 10 to 30min by 10 to 20 weight percent of sodium hydroxide aqueous solution;
mechanically homogenizing to prepare the biological cellulose fiber with the diameter of 20-50 nm and the length of 30-100 mu m.
3. The method for preparing a hemostatic dressing according to claim 1, wherein the zeolite molecular sieve is synthesized by hydrothermal method with the biological cellulose fiber as a template.
4. A method of making a hemostatic dressing according to claim 1, comprising:
mixing the biological cellulose fiber, tetrapropylammonium hydroxide, a silicon-based compound and water to obtain a mixed solution; wherein the corresponding molar ratio of the biological cellulose fiber, the tetrapropylammonium hydroxide and the silicon-based compound is (0.001-0.2) to (0.01-1) to 1 in every 1L of water;
adding an aluminum-based compound into the mixed solution, and slowly stirring for 5-8 h to obtain mixed gel, wherein the molar ratio of the aluminum-based compound to the silicon-based compound is (0.05-0.1): 1;
slowly heating the mixed gel to 170-175 ℃, and keeping the temperature for 6-24 h to obtain a reaction sample; wherein the heating rate is 5-10 ℃/min;
washing the reaction sample by using deionized water, drying, heating to 550-600 ℃ for carbonization treatment, keeping the temperature for 5-8 h, and cooling to obtain a zeolite molecular sieve and biocellulose-based carbon nanofiber composite material;
and mixing the composite material with biological cellulose fiber and chitosan aqueous solution, and performing freeze drying and sterilization to obtain the hemostatic dressing.
5. The method according to claim 4, wherein the silicon-based compound comprises one or more of silica gel, white carbon black, sodium silicate, methyl orthosilicate, ethyl orthosilicate, propyl orthosilicate, isopropyl orthosilicate and butyl orthosilicate.
6. The method of claim 4, wherein the aluminum-based compound comprises any one or more of sodium metaaluminate, aluminum hydroxide, aluminum sulfate, aluminum sol, aluminum isopropoxide and aluminum sec-butoxide.
7. The method of claim 4, wherein the drying is freeze-drying; the freeze drying is to freeze the sample at-20 to-80 ℃ for 12 to 24 hours and then to carry out vacuum drying for 24 to 48 hours.
8. The method according to claim 4, wherein the carbonization treatment is to heat the material from room temperature to 550-600 ℃ under vacuum, argon or nitrogen protection, wherein the heating rate is 5-10 ℃/min in the range of 100-300 ℃, 1-5 ℃/min in the range of 300-500 ℃ and 30 ℃/min in the range of 500-600 ℃.
9. The method according to claim 4, wherein the cooling is performed by slowly cooling the sample after the temperature rise to room temperature in an atmosphere furnace or an activation furnace.
10. The method according to claim 4, wherein when the composite material is mixed with the biological cellulose fibers and the chitosan aqueous solution, the mixing mass ratio of the chitosan, the zeolite molecular sieve, the biological cellulose-based carbon fibers and the biological cellulose fibers is 1: (0.01-0.1): (0.01-0.1): (0.01-0.1) or the mixing mass ratio is 1: (0.04-0.06): (0.04-0.06): (0.3-0.6).
11. The method according to claim 4, wherein the chitosan solution has a chitosan mass concentration of 3-10 wt% and a chitosan molecular weight of 10-35 ten thousand.
12. A hemostatic dressing prepared from biological cellulose fibers, which takes chitosan as a matrix of a porous material, wherein the biological cellulose fibers, carbon nanofibers and zeolite particles are uniformly distributed, the material porosity of the hemostatic dressing is 80-95%, the pore diameter is 50-500 mu m, the water absorption is 200-1000 times of the weight of the material, and the tensile mechanical strength is 0.5-4 GPa, and the hemostatic dressing is prepared by the method according to any one of claims 1-11.
13. Use of the hemostatic dressing of claim 12 in the manufacture of a medicament for hemostasis and/or wound healing promotion.
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