WO2020220300A1 - 生物纤维素纤维、包含所述纤维的止血敷料与相关应用 - Google Patents

生物纤维素纤维、包含所述纤维的止血敷料与相关应用 Download PDF

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WO2020220300A1
WO2020220300A1 PCT/CN2019/085277 CN2019085277W WO2020220300A1 WO 2020220300 A1 WO2020220300 A1 WO 2020220300A1 CN 2019085277 W CN2019085277 W CN 2019085277W WO 2020220300 A1 WO2020220300 A1 WO 2020220300A1
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biocellulose
fiber
hemostatic dressing
hemostatic
chitosan
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PCT/CN2019/085277
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English (en)
French (fr)
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钟春燕
钟宇光
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钟春燕
海南光宇生物科技有限公司
钟宇光
海南椰国食品有限公司
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Application filed by 钟春燕, 海南光宇生物科技有限公司, 钟宇光, 海南椰国食品有限公司 filed Critical 钟春燕
Priority to JP2021564853A priority Critical patent/JP7320078B2/ja
Priority to PCT/CN2019/085277 priority patent/WO2020220300A1/zh
Publication of WO2020220300A1 publication Critical patent/WO2020220300A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B15/00—Preparation of other cellulose derivatives or modified cellulose, e.g. complexes
    • C08B15/02—Oxycellulose; Hydrocellulose; Cellulosehydrate, e.g. microcrystalline cellulose
    • C08B15/04—Carboxycellulose, e.g. prepared by oxidation with nitrogen dioxide

Definitions

  • the present invention relates to a kind of biocellulose fiber, a hemostatic dressing containing the fiber and related applications, and in particular to a kind of biocellulose fiber, which can be used to make a hemostatic dressing with fast hemostasis and good mechanical properties.
  • the prepared hemostatic dressing, the biocellulose fiber and the preparation method and related applications of the hemostatic dressing relate to the field of hemostatic materials for medical devices.
  • Uncontrollable traumatic bleeding is the main cause of sudden emergency treatment, medical operation or death on the battlefield. According to US military data, more than 80% of the people killed on the battlefield were caused by excessive bleeding within 1 hour after trauma. Therefore, quick-acting hemostasis is the primary problem to be solved in trauma first aid.
  • Commonly used clinical hemostatic materials such as hemostatic gauze, hemostatic fiber, hemostatic bandage, etc., have certain limitations in use: long hemostasis time; easy to adhere to the wound and cause secondary damage during dressing change; lack of antibacterial ingredients, often Causes wound infection and suppuration.
  • the porous zeolite material is composed of porous zeolite without adding any biological components, which prevents the spread of diseases and allergic reactions between species.
  • a layer of hemostatic scab will quickly form on the surface of the wound, preventing internal blood from spilling.
  • the porous zeolite will release a lot of heat after absorbing the water in the blood and cause inflammation of the wound. Therefore, the heating of the porous zeolite material must be controlled to reduce the damage to the wound during hemostasis.
  • the zeolite hemostatic package is used, the zeolite particles remain on the wound surface, do not degrade, and easily cause foreign body reactions.
  • Trauma DEX produced by Medafor, USA, is a spongy polysaccharide-based hemostatic material based on potato starch. This spongy material can absorb a large amount of water in the blood at the bleeding point, promote the accelerated coagulation of platelets and blood proteins, so as to achieve the purpose of hemostasis. However, this material is only suitable for small wounds, and the temperature rises during hemostasis.
  • Chitosan is a positively charged polysaccharide-based hemostatic material extracted from shrimp and crab shells, which can attract negatively charged blood cells and promote blood coagulation. In addition, after contact with blood, it is sticky and can adhere tightly to the wound. Chitosan's unique hemostatic, antibacterial, biocompatibility, wound healing and easy gel formation properties give it good performance as a hemostatic material. However, the solubility of chitosan material itself is poor, and the mechanical properties need to be strengthened.
  • fibrin dressings are fibrinogen powder, thrombin and calcium ions.
  • This dressing can be absorbed by the body. Its hemostasis mechanism includes the dissolution of coagulation protein in plasma, and the fibrin layer formed by the enzymatic reaction of fibrinogen and thrombin will tightly adhere to the damaged tissue. But on the other hand, the material is fragile, not flexible, and easy to break, which largely limits its application.
  • An object of the present invention is to provide a biocellulose fiber, which can be used to prepare a hemostatic dressing with rapid hemostasis and good mechanical properties.
  • Another object of the present invention is to provide a method for preparing biocellulose fibers.
  • Another object of the present invention is to provide a hemostatic dressing prepared with the biocellulose fiber.
  • Another object of the present invention is to provide a method for preparing the hemostatic dressing.
  • Another object of the present invention is to provide an application of the hemostatic dressing.
  • the present invention provides a biocellulose fiber, which can be used to prepare a hemostatic dressing with rapid hemostasis and good mechanical properties.
  • the biocellulose fiber provided by the present invention has a diameter of 20-50 nm and a length of 30-100 ⁇ m.
  • the biocellulose fiber of the present invention is obtained by cooking cellulose obtained by bacterial fermentation through a sodium hydroxide aqueous solution.
  • the cellulose obtained by bacterial fermentation can be obtained commercially, or can be prepared and obtained by itself according to the records of the prior art.
  • the cellulose obtained by fermentation of the bacteria suitable for the present invention may be, for example, cellulose obtained by fermentation of the following strains: Acetobacter xylinum, Rhizobium, Sarcina, Pseudomonas, Achromobacter, Alkali-producing One or more of the genus Bacteria, Aerobacter, or Azotobacter.
  • the process of obtaining cellulose by fermentation of these strains is a well-known technology in the art, and will not be repeated in the present invention.
  • the biocellulose fiber of the present invention is the cellulose obtained by the bacterial fermentation through a sodium hydroxide aqueous solution cooking process.
  • the process of the said sodium hydroxide aqueous solution cooking treatment includes: high temperature cooking of the biocellulose with 10-20wt% sodium hydroxide aqueous solution for 10-30 minutes.
  • the biocellulose fiber provided by the present invention is mainly used to prepare a hemostatic dressing with rapid hemostasis and good mechanical properties. There are a large number of bacterial residues in the biocellulose raw materials fermented by bacteria, and these residues will affect the structural properties of the prepared hemostatic dressing.
  • the use of a sodium hydroxide solution of a specific concentration for cooking can completely remove the bacterial protein and the residual culture medium adhering to the cellulose membrane, ensuring the high purity of the cellulose of the bacterial cellulose material, and the purity can reach more than 99.9%. Or you can say 100%.
  • sodium hydroxide can play a certain activating effect in the subsequent activation treatment for preparing hemostatic dressings.
  • the present invention also provides a method for preparing the biocellulose fiber, the method comprising:
  • the cellulose obtained by bacterial fermentation is steamed for 10-30 minutes with a 10-20wt% sodium hydroxide aqueous solution at high temperature;
  • the present invention also provides the use of the biocellulose fiber in preparing a hemostatic dressing.
  • the biocellulose fiber is used as a template to synthesize zeolite molecular sieve by hydrothermal method, which can be compounded with chitosan at the same time to prepare a fast hemostatic and good mechanical properties. Hemostatic dressing.
  • the present invention also provides a method for preparing a hemostatic dressing, the method comprising:
  • biocellulose fiber, tetrapropylammonium hydroxide, and silicon-based compound of the present invention are mixed with water to obtain a mixed solution; wherein the corresponding moles of biocellulose fiber, tetrapropylammonium hydroxide, and silicon-based compound are added to each 1L of water
  • the ratio is (0.001 ⁇ 0.2):(0.01 ⁇ 1):1;
  • the above mixed gel is slowly heated to 170 ⁇ 175°C and kept at constant temperature for 6 ⁇ 24h to obtain a reaction sample; wherein the heating rate is 5 ⁇ 10°C/min;
  • the samples obtained above are washed with deionized water, dried, and then heated to 550°C to 600°C for carbonization treatment, kept at a constant temperature for 5 to 8 hours, and then cooled to obtain a zeolite molecular sieve and biocellulose-based carbon nanofiber composite material;
  • the above composite material is mixed with biocellulose fiber and chitosan aqueous solution, freeze-dried, and sterilized (conventional sterilization is sufficient, for example, high-pressure steam sterilization or radiation irradiation sterilization) to obtain a hemostatic dressing.
  • sterilized is sufficient, for example, high-pressure steam sterilization or radiation irradiation sterilization
  • the hemostatic dressing of the present invention can be cut (usually performed before sterilization) and packaged as required.
  • the preparation method of the hemostatic dressing of the present invention uses biocellulose as a template and hydrothermally synthesizes zeolite in situ. During this process, a large number of hydroxyl groups on the surface of biocellulose can adsorb zeolite precursors (silicon-based, aluminum-based compounds) , Plays the role of in-situ template; at the same time, the surface of biocellulose can also adsorb some metal ions in the raw materials that affect the synthesis of zeolite, and act as a metal chelating agent; secondly, while high-temperature calcination, nano-cellulose fibers are carbonized into biological Cellulose-based carbon nanofibers, carbon nanofibers and zeolite have strong binding force, and carbon nanofibers and biocellulose fibers have a reinforcing effect.
  • the method of the invention has simple process, convenient operation and low cost, the obtained hemostatic dressing has fast hemostasis performance and good mechanical properties, the zeolite molecular sieve has strong binding force with the matrix,
  • the silicon-based compound in the preparation method of the hemostatic dressing of the present invention, includes silica gel, white carbon black, sodium silicate, methyl orthosilicate, ethyl orthosilicate, and orthosilicic acid. Any one or more of propyl ester, isopropyl orthosilicate and butyl orthosilicate.
  • the aluminum-based compound in the preparation method of the hemostatic dressing of the present invention, includes sodium metaaluminate, aluminum hydroxide, aluminum sulfate, aluminum sol, aluminum isopropoxide and aluminum sec-butoxide. Any one or more of.
  • the drying is preferably freeze drying.
  • the freeze drying is preferably to freeze the sample at -20°C to -80°C for 12 to 24 hours, and then vacuum dry for 24 to 48 hours.
  • the carbonization treatment means that the material is heated from room temperature to 550°C to 600°C under the protection of vacuum, argon or nitrogen, of which 100 to 300
  • the temperature rise rate in the temperature range is 5-10°C/min
  • the temperature rise rate in the 300-500°C range is 1 to 5°C/min
  • the temperature rise rate in the 500-600°C range is preferably 25-30°C/min.
  • the cooling means that the sample after the temperature rise is slowly cooled to room temperature in an atmosphere furnace or an activation furnace.
  • the zeolite molecular sieve and the biocellulose-based carbon nanofiber composite material are mixed with the biocellulose fiber and the chitosan aqueous solution, the chitosan, the zeolite molecular sieve,
  • the mixing mass ratio of biocellulose-based carbon fiber and biocellulose fiber is 1:(0.01 ⁇ 0.1):(0.01:0.1):(0.01 ⁇ 0.1), and the better ratio is 1:(0.04 ⁇ 0.06):(0.04 ⁇ 0.06): (0.3 ⁇ 0.6).
  • the biocellulose fiber mixed with the composite material and the chitosan aqueous solution is mixed with the biocellulose fiber, tetrapropylammonium hydroxide, and silicon-based compound.
  • the size of the biocellulose fibers in the mixed solution may be the same or different, and may be within the range of 20-50 nm in diameter and 30-100 ⁇ m in length required by the present invention.
  • the addition of this part of the biocellulose fiber further strengthens the zeolite particles and makes them difficult to fall off.
  • the nano cellulose fibers on the surface of the chitosan porous material can improve the adsorption efficiency of the material to blood, which can connect the external and internal zeolite particles.
  • the purpose is to significantly accelerate the rate of hemostasis.
  • the chitosan aqueous solution is obtained by dissolving chitosan in a 2wt% acetic acid aqueous solution.
  • the mass concentration of chitosan is 3-10 wt%, and the molecular weight of the chitosan is 100,000-350,000.
  • the zeolite molecular sieve and the biocellulose-based carbon nanofiber composite material are mixed with the biocellulose fiber and the chitosan aqueous solution, and the temperature is -20°C to -80°C. It is frozen for 12-24 hours, then vacuum dried for 24 to 48 hours, and sterilized to obtain the hemostatic dressing of the present invention.
  • the present invention also provides a hemostatic dressing prepared with the biocellulose fiber.
  • the final product of the hemostatic dressing of the present invention is a porous foam hemostatic material composed of chitosan, biocellulose fiber, nano carbon fiber (obtained by carbonization of biocellulose), and zeolite.
  • chitosan is a porous material matrix, in which biocellulose fibers, nano-carbon fibers and zeolite particles are evenly distributed.
  • the material porosity of the hemostatic dressing is 80-95%, the pore diameter is 50-500 ⁇ m, the water absorption rate is 200-1000 times of the material's own weight, and the tensile mechanical strength is 0.5-4 GPa.
  • the present invention also provides the application of the hemostatic dressing.
  • carbon fiber and biocellulose fiber improve the mechanical strength of the material; the close combination of zeolite and carbon fiber avoids the problem that zeolite particles enter the wound surface when the hemostatic material is used and affect wound healing; the addition of biocellulose fiber , It has a certain entanglement effect on the combination of zeolite and carbon fiber, and further strengthens the zeolite particles to make it not easy to fall off; at the same time, the nano cellulose fiber on the surface of the chitosan porous material can improve the adsorption efficiency of the material to the blood and play a guiding role.
  • the purpose of communicating the external and internal zeolite particles can significantly accelerate the rate of hemostasis.
  • the hemostatic dressing of the present invention has good biocompatibility, good antibacterial performance, and can promote wound healing.
  • the present invention provides the application of the hemostatic dressing in the preparation of drugs for hemostasis and wound healing.
  • bio-nanocellulose is used as a template, and the zeolite molecular sieve is synthesized in situ by a hydrothermal method.
  • the nano-cellulose fibers are carbonized into bio-cellulose-based nano-carbon fibers.
  • the carbon nanofiber and the zeolite have a strong binding force, and the carbon nanofiber and the biocellulose fiber have a reinforcing effect, and finally a hemostatic dressing that can quickly stop bleeding and has good mechanical properties is obtained.
  • the preparation process of the present invention is simple, easy to operate, convenient to operate, and low in cost.
  • the obtained hemostatic dressing has good hemostatic and mechanical properties; the zeolite molecular sieve has a stable and uniform structure, strong binding force to the matrix, and is not easy to fall off during use; and good biocompatibility , Has good antibacterial properties, and can promote wound healing.
  • biocellulose fiber tetrapropylammonium hydroxide, silicon-based compound and water to obtain a mixed solution.
  • biocellulose fiber, tetrapropylammonium hydroxide, and silicon-based compounds are added to 1L of water.
  • the corresponding molar ratio of the three substances is 0.001:0.01:1; among them, the biocellulose fiber is a combination of Acetobacter xylinum and Rhizobium
  • the biocellulose obtained by fermentation is digested with a 10wt% sodium hydroxide aqueous solution at high temperature for 10 minutes, and then mechanically homogenized to obtain cellulose fibers with a diameter of 20nm and a length of 30 ⁇ m; the silicon-based compound refers to silica gel.
  • the above composite material is mixed with biocellulose fiber and chitosan aqueous solution, freeze-dried, cut, sterilized, and packaged to finally obtain a hemostatic dressing that can quickly stop bleeding and has good mechanical properties.
  • the chitosan aqueous solution is prepared from chitosan with a molecular weight of 350,000, and its mass concentration is 3wt%.
  • Table 1 lists part of the experimental data of the effects of different contents of zeolite, carbon fiber, and biocellulose fiber on the performance of the hemostatic dressing in the research process of the present invention.
  • Example 1 obtained a hemostatic dressing that can quickly stop bleeding and has good mechanical properties.
  • the main performance test results are as follows:
  • Hemostasis A rabbit fatal femoral artery bleeding wound model was used to evaluate the effect of the hemostatic dressing, and the control sample was gauze. The results show that the hemostatic dressing of the present invention has a hemostatic rate of 100% within 1 minute, and the hemostatic rate of gauze within 8 minutes is 37%.
  • the hemostatic dressing of this embodiment contains chitosan, which is a non-dissolvable bacteriostatic material.
  • chitosan which is a non-dissolvable bacteriostatic material.
  • Appendix C5 Non-dissolvable antibacterial (antibacterial) product antibacterial performance test method, according to the standard, the antibacterial rate of the test sample group and the antibacterial rate of the control sample group The difference is greater than 26%, indicating that the product has antibacterial properties.
  • the results show that the hemostatic dressing of the present invention has a difference of 90.09% and 89.02% in the inhibition rate against Escherichia coli ATCC 8739 and Staphylicoccus aureus ATCC 6538, respectively, indicating that the hemostatic dressing of the present invention has good properties.
  • the antibacterial properties show that the hemostatic dressing of the present invention has a difference of 90.09% and 89.02% in the inhibition rate against Escherichia coli ATCC 8739 and Staphylicoccus aureus ATCC 6538, respectively, indicating that the hemostatic dressing of the present invention has good properties.
  • the antibacterial properties show that the hemostatic dressing of the present invention has a difference of 90.09% and 89.02% in the inhibition rate against Escherichia coli ATCC 8739 and Staphylicoccus aureus ATCC 6538, respectively.
  • Biocompatibility test According to the biological evaluation of GB/T 16886 medical devices, the hemostatic dressing was evaluated for cytotoxicity, delayed contact sensitization, skin irritation, etc. Biocompatibility evaluation: The intracellular toxicity test is tested in accordance with GB/T 16886-5 "Biological Evaluation of Medical Devices Part 5: In Vitro Cytotoxicity Test"; the guinea pig delayed contact sensitization test is in accordance with GB/T 16886-10 " Biological Evaluation of Medical Devices Part 10: Stimulus and Delayed Hypersensitivity Test" for testing, using the maximum test Magnusson and Kligman method.
  • the skin irritation test is tested in accordance with GB/T 16886-10 "Biological Evaluation of Medical Devices Part 10: Irritation and Delayed Hypersensitivity Test". The results show that the cytotoxicity of the hemostatic dressing is less than grade 2, no skin sensitization, no intradermal irritation, and good biological safety.
  • the potential cytotoxicity of the sample was determined by MTT method.
  • the toxicity of the test sample is Class II.
  • the cell viability gradually increased.
  • the cytotoxicity was all grade I. Therefore, the toxic reaction of the sample extract to L929 cells is level 2, without potential cytotoxicity.
  • the animals showed no abnormal symptoms or died. According to observations, the skin reaction on one side of the experimental group did not exceed the skin reaction on the side of the blank control group, the primary irritation index was 0 (no edema, erythema), and the positive control group animals were scored as 2 (obvious edema, erythema). The test sample had no irritation effect on rabbit skin.
  • the skin sensitization test is shown in Table 4.
  • the sample extract was injected intracutaneously into 10 guinea pigs, bandaged and tried to induce sensitization. During the recovery period, 10 test guinea pigs and 5 control guinea pigs were tested with sample extract and blank solution respectively. The scores of each part were recorded 24h and 48h after the patch was removed. In the experiment, the animals in the negative control group and the test group were scored as 0 (no significant change), and the animals in the positive control group were scored as 2 (moderate confluent erythema). It shows that no delayed contact sensitization was found in the sample.
  • biocellulose fiber tetrapropylammonium hydroxide, silicon-based compound and water to obtain a mixed solution.
  • biocellulose fiber, tetrapropylammonium hydroxide, and silicon-based compounds are added to 1L of water.
  • the corresponding molar ratio of these three substances is 0.001:0.05:1; among them, the biocellulose fiber is a combination of Sarcinia and pseudomonas The biocellulose obtained by fermentation of the spp.
  • the composite material is mixed with biocellulose fiber and chitosan aqueous solution, freeze-dried, cut, sterilized by high-energy rays, and packaged to finally obtain a hemostatic dressing that can quickly stop bleeding and has good mechanical properties.
  • the chitosan aqueous solution is prepared from chitosan with a molecular weight of 300,000, and its mass concentration is 4wt%.
  • the tensile mechanical strength of the hemostatic dressing of this embodiment is 1050 MPa; the porosity is 80-95%, the pore diameter is 50-500 ⁇ m, and the water absorption rate is 200-1000 times the weight of the material.
  • the rabbit fatal femoral artery hemorrhage wound model was used to evaluate the effect of the hemostatic dressing, and the result showed that the hemostatic dressing of the present embodiment had a hemostatic rate of 100% within 1 minute.
  • biocellulose fiber tetrapropylammonium hydroxide, silicon-based compound and water to obtain a mixed solution.
  • biocellulose fiber, tetrapropylammonium hydroxide, and silicon-based compound are added to 1L of water.
  • the biocellulose fiber is a combination of Acetobacter xylinum and Achromobacter
  • the biocellulose obtained by fermentation of the genus and Alcaligenes was digested at a high temperature for 30 minutes with a 14wt% sodium hydroxide aqueous solution, and then mechanically homogenized to obtain cellulose fibers with a diameter of 40nm and a length of 50 ⁇ m; the silicon-based compound is silicic acid
  • the mass ratio of sodium to butyl orthosilicate is 1:1.
  • the composite material is mixed with biocellulose fiber and chitosan aqueous solution, freeze-dried, cut, sterilized by high-energy rays, and packaged to finally obtain a hemostatic dressing that can quickly stop bleeding and has good mechanical properties.
  • the chitosan aqueous solution is prepared from chitosan with a molecular weight of 250,000 and its mass concentration is 5 wt%.
  • the tensile mechanical strength of the hemostatic dressing of this embodiment is 900 MPa; the porosity is 80-95%, the pore diameter is 50-500 ⁇ m, and the water absorption rate is 200-1000 times the weight of the material.
  • the rabbit fatal femoral artery hemorrhage wound model was used to evaluate the effect of the hemostatic dressing, and the result showed that the hemostatic dressing of the present embodiment had a hemostatic rate of 100% within 1 minute.
  • biocellulose fiber, tetrapropylammonium hydroxide, silicon-based compound and water are added to 1L of water, and the corresponding molar ratio of these three substances is 0.005:0.03:1; among them, the biocellulose fiber is a kind of fermentation of Aerobacteria and nitrogen-fixing bacteria.
  • the obtained biocellulose was steamed for 15 minutes with a 16wt% sodium hydroxide aqueous solution at high temperature, and then mechanically homogenized.
  • the obtained nanofibers were cellulose fibers with a diameter of 50nm and a length of 60 ⁇ m; silicon-based compounds refer to methyl orthosilicate and isosilicate Propyl ester, the mass ratio is 1:1.
  • the composite material is mixed with biocellulose fiber and chitosan aqueous solution, freeze-dried, cut, sterilized by high-energy rays, and packaged to finally obtain a hemostatic dressing that can quickly stop bleeding and has good mechanical properties.
  • the chitosan aqueous solution is prepared from chitosan with a molecular weight of 200,000, and its mass concentration is 6 wt%.
  • the tensile mechanical strength of the hemostatic dressing of this embodiment is 800 MPa; the porosity is 80-95%, the pore diameter is 50-500 ⁇ m, and the water absorption is 200-1000 times the weight of the material.
  • the rabbit fatal femoral artery hemorrhage wound model was used to evaluate the effect of the hemostatic dressing, and the result showed that the hemostatic dressing of the present embodiment had a hemostatic rate of 100% within 1 minute.
  • biocellulose fiber, tetrapropylammonium hydroxide, silicon-based compound and water are added to 1L of water.
  • the corresponding molar ratio of the three substances is 0.01:0.02:1; among them, the biocellulose fiber is obtained by fermentation of Alcaligenes Biocellulose was digested at a high temperature with 18wt% sodium hydroxide aqueous solution for 28 minutes, and then mechanically homogenized to obtain cellulose fibers with a diameter of 50nm and a length of 70 ⁇ m.
  • the silicon-based compound refers to ethyl orthosilicate and propyl orthosilicate. The mass ratio is 1:1.
  • the composite material is mixed with biocellulose fiber and chitosan aqueous solution, freeze-dried, cut, sterilized by high-energy rays, and packaged to finally obtain a hemostatic dressing that can quickly stop bleeding and has good mechanical properties.
  • the chitosan aqueous solution is prepared from chitosan with a molecular weight of 150,000, and its mass concentration is 7wt%.
  • the tensile mechanical strength of the hemostatic dressing of this embodiment is 850 MPa; the porosity is 80-95%, the pore diameter is 50-500 ⁇ m, and the water absorption is 200-1000 times the weight of the material.
  • the rabbit fatal femoral artery hemorrhage wound model was used to evaluate the effect of the hemostatic dressing, and the result showed that the hemostatic dressing of the present embodiment had a hemostatic rate of 100% within 1 minute.
  • biocellulose fiber tetrapropylammonium hydroxide, silicon-based compound and water to obtain a mixed solution.
  • biocellulose fiber, tetrapropylammonium hydroxide, and silicon-based compounds are added to 1L of water, and the corresponding molar ratio of the three substances is 0.16:0.5:1; among them, the biocellulose fiber is a mixture of Acetobacter xylinum and Aerobacter.
  • the biocellulose obtained by fermentation with nitrogen-fixing bacteria was digested with a 20wt% sodium hydroxide aqueous solution at high temperature for 10 minutes, and then mechanically homogenized to obtain a cellulose fiber with a diameter of 50nm and a length of 100 ⁇ m; silicon-based compounds refer to silica gel and white carbon
  • the ratio of black, methyl orthosilicate and butyl orthosilicate is 1:1:1:1.
  • the composite material is mixed with biocellulose fiber and chitosan aqueous solution, freeze-dried, cut, sterilized by high-energy rays, and packaged to finally obtain a hemostatic dressing that can quickly stop bleeding and has good mechanical properties.
  • the chitosan aqueous solution is prepared from chitosan with a molecular weight of 100,000, and its mass concentration is 10wt%.
  • the tensile mechanical strength of the hemostatic dressing of this embodiment is 750 MPa; the porosity is 80-95%, the pore diameter is 50-500 ⁇ m, and the water absorption is 200-1000 times the weight of the material.
  • the rabbit fatal femoral artery hemorrhage wound model was used to evaluate the effect of the hemostatic dressing, and the result showed that the hemostatic dressing of the present embodiment had a hemostatic rate of 100% within 1 minute.

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Abstract

一种生物纤维素纤维、包含该纤维的止血敷料与相关应用。生物纤维素纤维是细菌发酵得到的纤维素经纯化处理、机械均质而得到的，其直径为20～50nm，长度为30～100μm。以该生物纤维素纤维作为模板，水热法合成沸石分子筛，可同时与壳聚糖复合，制备得到一种可以快速止血并且力学性能良好的止血敷料，其生物相容性好，具有良好的抑菌性能，且能够促进伤口愈合。

Description

生物纤维素纤维、包含所述纤维的止血敷料与相关应用 技术领域
本发明是关于一种生物纤维素纤维、包含所述纤维的止血敷料与相关应用，具体地说，特别是指一种可应用于制作快速止血并且力学性能良好的止血敷料的生物纤维素纤维、所制备的止血敷料、所述生物纤维素纤维及止血敷料的制备方法以及相关应用，涉及医疗器械止血材料领域。
背景技术
难以控制的创伤出血是突发性急救治疗、医疗手术或战场死亡的主要原因。如美军资料显示，战场上有80％以上的阵亡人员，是创伤后1h内出血过多导致的。因此速效止血是创伤急救需解决的首要问题。临床常用的止血材料如止血纱布、止血纤维、止血绷带等，在使用中都有一定的局限性：止血时间较长；易与伤口粘连从而导致换药时的二次伤害；缺少抗菌成分，常造成伤口的感染与化脓。
针对以上问题，目前领域中已出现了多孔沸石材料、多糖基止血材料、多肽基止血材料等新型快速止血材料。其中：
(1)多孔沸石材料是由多孔沸石组成，未添加任何生物成分，这就避免了物种间疾病传播和过敏反应。当把这种粉末直接施加到出血点后，伤口表面会迅速形成一层止血痂，阻止内部血液的外溢。但多孔沸石吸收血液中的水分后会放出大量的热，导致伤口炎症，因此必须控制多孔沸石材料发热，以减轻止血过程中的对伤口的损伤。此外，沸石止血包在使用时，沸石颗粒残留在创面上，不降解，易引起异物反应。
(2)由美国Medafor公司生产的Trauma DEX是一种以马铃薯淀粉为基质的海绵状多糖基止血材料。这种海绵状材料可以在出血点吸收血液中的大量水分，促使血小板和血蛋白的加速凝结，从而达到止血目的。但该材料只适用于小创伤，且止血过程中伴有温度升高现象。
(3)壳聚糖是一种主要从虾蟹壳中提取的带正电的多糖基止血材料，能吸引带负电的血细胞并促进血液凝固。此外在与血液接触后，它具有粘性，能紧紧地粘附于创口。壳聚糖特有的止血、抑菌、生物相容性、促进伤口愈合以及易于形成凝胶的性质，赋予它用作止血材料的良好性能。但壳聚糖材料本身溶解性能差，力学性能有待加强。
(4)纤维蛋白敷料的有效成分是纤维蛋白原粉末、凝血酶和钙离子等。这种敷料可被人体吸收。它的止血机理包括血浆中的凝血蛋白的溶解，纤维蛋白原与凝血酶的酶促反应 形成的纤维蛋白层将紧紧附着在损伤的组织上。但另一方面，该材料易碎，柔韧性不好，容易折断，很大程度上限制了它的应用。
基于以上快速止血材料所存在的各种问题，仍有必要研发新型的快速止血材料。
发明内容
本发明的一个目的在于提供一种生物纤维素纤维，其可用于制备快速止血并且力学性能良好的止血敷料。
本发明的另一目的在于提供一种生物纤维素纤维的制备方法。
本发明的另一目的在于提供以所述生物纤维素纤维制备的止血敷料。
本发明的另一目的在于提供所述止血敷料的制备方法。
本发明的另一目的在于提供所述止血敷料的应用。
一方面，本发明提供了一种生物纤维素纤维，其可用于制备快速止血并且力学性能良好的止血敷料。根据本发明所提供的生物纤维素纤维，其直径为20～50nm，长度为30～100μm。本发明的生物纤维素纤维是细菌发酵得到的纤维素经氢氧化钠水溶液蒸煮处理而得到的。
根据本发明的具体实施方案，本发明中，所述的细菌发酵得到的纤维素可以商购获得，也可按照现有技术的记载自行制备获得。适用于本发明的细菌发酵得到的纤维素，例如可以是以下菌种发酵得到的纤维素：木醋杆菌、根瘤菌属、八叠球菌属、假单胞菌属、无色杆菌属、产碱菌属、气杆菌属或固氮菌属中的一种或几种。以这些菌种发酵得到纤维素的过程是所属领域的已知技术，本发明不再赘述。
根据本发明的具体实施方案，本发明的生物纤维素纤维是所述细菌发酵得到的纤维素经过氢氧化钠水溶液蒸煮处理的。其中所述的氢氧化钠水溶液蒸煮处理的过程包括：生物纤维素经10～20wt％的氢氧化钠水溶液高温蒸煮10～30min。本发明提供的生物纤维素纤维主要是用于制备快速止血并且力学性能良好的止血敷料。经细菌发酵的生物纤维素原材料中有大量的细菌残留体，这些残留物将影响所制备的止血敷料的结构性能。本发明中，采用特定浓度的氢氧化钠溶液蒸煮能够彻底去除菌体蛋白和粘附在纤维素膜上的残余培养基，保证细菌纤维素材料的纤维素高纯度，纯度可达99.9％以上，或者可以说100％。同时，氢氧化钠在后续制备止血敷料的活化处理中可以起到一定的活化作用。
另一方面，本发明还提供了所述生物纤维素纤维的制备方法，该方法包括：
将细菌发酵得到的纤维素经10～20wt％的氢氧化钠水溶液高温蒸煮10～30min；
机械均质，以制备得到直径20～50nm、长度30～100μm的纤维。
另一方面，本发明还提供了所述生物纤维素纤维在制备止血敷料中的用途。
根据本发明的具体实施方案，本发明中，是将所述生物纤维素纤维作为模板，水热法合成沸石分子筛，可同时与壳聚糖复合，制备得到一种可以快速止血并且力学性能良好的止血敷料。
从而，另一方面，本发明还提供了一种止血敷料的制备方法，该方法包括：
将本发明的生物纤维素纤维、四丙基氢氧化铵、硅基化合物与水混合，得到混合液；其中每1L水中加入生物纤维素纤维、四丙基氢氧化铵、硅基化合物对应的摩尔比为(0.001～0.2):(0.01～1):1；
向上述的混合液中加入铝基化合物，缓慢搅拌5～8h，得到混合凝胶，其中铝基化合物与硅基化合物的摩尔比为(0.05～0.1):1；
上述混合凝胶缓慢加热至170～175℃，恒温6～24h，得到反应样品；其中升温速率为5～10℃/min；
上述所得样品用去离子水清洗，干燥，然后加热至550℃～600℃碳化处理，恒温5～8h后，冷却，得到沸石分子筛与生物纤维素基纳米碳纤维复合材料；
将上述复合材料与生物纤维素纤维、壳聚糖水溶液混合，经冷冻干燥，灭菌(常规灭菌即可，例如可以是高压蒸汽灭菌，或者射线辐照灭菌)，得到止血敷料。本发明的止血敷料可根据需要进行切割(通常在灭菌前进行)，包装。
本发明的止血敷料的制备方法，以生物纤维素为模板、水热法原位合成得到沸石，在这一过程中生物纤维素表面的大量羟基能够吸附沸石前驱体(硅基、铝基化合物)，起到原位模板的作用；同时，生物纤维素表面也可以吸附原料中影响沸石合成的一些金属离子，起到金属螯合剂的作用；其次，高温煅烧的同时，纳米纤维素纤维碳化为生物纤维素基纳米碳纤维，纳米碳纤维与沸石结合力强，同时纳米碳纤维与生物纤维素纤维具有增强作用。本发明的方法工艺简单易行、操作方便、成本低，得到的止血敷料具有快速止血性能并且力学性能良好，沸石分子筛与基体结合力强，使用时不易脱落。
根据本发明的具体实施方案，本发明的止血敷料的制备方法中，所述的硅基化合物包括硅胶、白炭黑、硅酸钠、正硅酸甲酯、正硅酸乙酯、正硅酸丙酯、正硅酸异丙酯和正硅酸丁酯中的任意一种或几种。
根据本发明的具体实施方案，本发明的止血敷料的制备方法中，所述的铝基化合物包括偏铝酸钠、氢氧化铝、硫酸铝、铝溶胶、异丙醇铝和仲丁醇铝中的任意一种或几种。
根据本发明的具体实施方案，本发明的止血敷料的制备方法中，所述的干燥优选为冷 冻干燥。所述的冷冻干燥优选是将样品在-20℃～-80℃下冷冻12～24h，然后真空干燥24～48h。
根据本发明的具体实施方案，本发明的止血敷料的制备方法中，所述的碳化处理是指材料在真空、氩气或氮气保护下，由室温升温至550℃～600℃，其中100～300℃区间升温速率为5～10℃/min，300～500℃区间升温速率为1～5℃/min，500～600℃区间升温速率优选为25～30℃/min。
根据本发明的具体实施方案，本发明的止血敷料的制备方法中，所述的冷却是指升温结束后的样品在气氛炉或活化炉中缓慢冷却至室温。
根据本发明的具体实施方案，本发明的止血敷料的制备方法中，沸石分子筛与生物纤维素基纳米碳纤维复合材料与生物纤维素纤维、壳聚糖水溶液混合时，其中壳聚糖、沸石分子筛、生物纤维素基碳纤维与生物纤维素纤维的混合质量比为1：(0.01～0.1)：(0.01:0.1)：(0.01～0.1)，更佳的比例为1：(0.04～0.06)：(0.04～0.06)：(0.3～0.6)。
根据本发明的具体实施方案，本发明的止血敷料的制备方法中，与复合材料、壳聚糖水溶液混合的生物纤维素纤维，与生物纤维素纤维、四丙基氢氧化铵、硅基化合物的混合液中的生物纤维素纤维的尺寸可以相同或不同，在本发明所要求的直径20～50nm、长度30～100μm范围内即可。此部分生物纤维素纤维的加入，进一步加固了沸石颗粒，使其不易脱落，同时壳聚糖多孔材料表面的纳米纤维素纤维能够提高材料对血液的吸附效率，起到导通外部与内部沸石颗粒的目的，显著加快止血速率。
根据本发明的具体实施方案，本发明的止血敷料的制备方法中，所述的壳聚糖水溶液是将壳聚糖溶解于2wt％的乙酸水溶液中得到的。优选地，所述的壳聚糖水溶液中，壳聚糖质量浓度为3～10wt％，所述的壳聚糖分子量为10～35万。
根据本发明的具体实施方案，本发明的止血敷料的制备方法中，沸石分子筛与生物纤维素基纳米碳纤维复合材料与生物纤维素纤维、壳聚糖水溶液混合后，在-20℃～-80℃下冷冻12～24h，然后真空干燥24～48h，灭菌，得到本发明的止血敷料。
另一方面，本发明还提供了以所述生物纤维素纤维制备的止血敷料。本发明的止血敷料，最终产品是由壳聚糖、生物纤维素纤维、纳米碳纤维(由生物纤维素碳化得到)、沸石构成的多孔泡沫状止血材料。结构上，壳聚糖为多孔材料的基体，其中均匀分布着生物纤维素纤维、纳米碳纤维以及沸石颗粒。该止血敷料的材料孔隙率为80～95％，孔径为50～500μm，吸水率为材料自身重量的200～1000倍，拉伸力学强度为0.5～4GPa。
另一方面，本发明还提供了所述止血敷料的应用。本发明的止血敷料，其中碳纤维与 生物纤维素纤维提高了材料的力学强度；沸石与碳纤维的紧密结合，避免了止血材料使用时沸石颗粒进入创面，影响伤口愈合的问题；生物纤维素纤维的加入，对沸石与碳纤维的结合体有一定的缠结作用，进一步加固了沸石颗粒，使其不易脱落；同时壳聚糖多孔材料表面的纳米纤维素纤维能够提高材料对血液的吸附效率，起到导通外部与内部沸石颗粒的目的，显著加快止血速率。本发明的止血敷料生物相容性好，具有良好的抑菌性能，且能够促进伤口愈合。从而，本发明提供了所述止血敷料在制备止血、促进伤口愈合的药物中的应用。
本发明的上述方案中，未详细注明的方法条件可以按照所属领域中的常规操作进行。
本发明的有益效果：
本发明采用生物纳米纤维素作为模板，水热法原位合成沸石分子筛。高温煅烧的同时，纳米纤维素纤维碳化为生物纤维素基纳米碳纤维。纳米碳纤维与沸石结合力强，同时纳米碳纤维与生物纤维素纤维具有增强作用，最终得到一种可以快速止血并且力学性能良好的止血敷料。本发明制备工艺简单易行、操作方便、成本低，得到的止血敷料具有良好的止血性能和力学性能；沸石分子筛结构稳定均一，其与基体结合力强，使用时不易脱落；生物相容性好，具有良好的抑菌性能，且能够促进伤口愈合。
具体实施方式
以下通过具体实施例详细说明本发明的实施过程和产生的有益效果，旨在帮助阅读者更好地理解本发明的实质和特点，不作为对本案可实施范围的限定。
实施例1
本实施例的止血敷料的制备方法：
(1)将生物纤维素纤维、四丙基氢氧化铵、硅基化合物与水混合，得到混合液。其中1L水中加入生物纤维素纤维、四丙基氢氧化铵、硅基化合物，该三种物质对应的摩尔比为0.001:0.01:1；其中，生物纤维素纤维是将木醋杆菌与根瘤菌属发酵得到的生物纤维素经10wt％的氢氧化钠水溶液高温蒸煮10min，然后机械均质得到的纳米纤维直径为20nm、长度为30μm的纤维素纤维；硅基化合物是指硅胶。
(2)上述的混合液中加入偏铝酸钠，缓慢搅拌5h，得到混合凝胶，其中铝基化合物与硅基化合物的摩尔比为0.05:1；
(3)上述混合凝胶在高压釜中缓慢加热至170℃，恒温6h，其中升温速率为5℃/min；
(4)上述所得样品用去离子水清洗，在-20℃下冷冻24h，然后真空干燥24h，然 后将干燥后的样品放入气氛炉中碳化处理加热至550℃，恒温5h后，样品在气氛炉炉中缓慢冷却至室温，得到沸石分子筛与生物纤维素基纳米碳纤维复合材料；其中，碳化处理是指材料在氮气保护下，由室温升温至550℃，其中100～300℃区间升温速率为5℃/min，300～500℃区间升温速率为1℃/min，500～550℃区间升温速率为30℃/min。
(5)将上述复合材料与生物纤维素纤维、壳聚糖水溶液混合，经冷冻干燥、切割、灭菌、包装，最终得到可以快速止血并且力学性能良好的止血敷料。其中壳聚糖水溶液是由分子量为35万的壳聚糖配制成的，其质量浓度为3wt％。
表一列出了本发明在研究过程中沸石、碳纤维、生物纤维素纤维不同含量时对止血敷料性能影响的部分实验数据。
表一
Figure PCTCN2019085277-appb-000001
上述实施例1得到了一种可以快速止血并且力学性能良好的止血敷料，其主要性能测试结果如下：
止血：采用兔致命性股动脉出血伤口模型对止血敷料效果进行评估，对照样品为纱布。结果表明，本发明的止血敷料在1min内的止血率为100％，纱布在8min内的止血率为37％。
抑菌性：本实施例的止血敷料中含有壳聚糖，为非溶出性抑菌材料。参照GB15979-2002《一次性使用卫生用品卫生标准》中附录C5非溶出性抗(抑)菌产品抑菌性能试验方法，按标准规定，被试样品组抑菌率与对照样品组抑菌率差值大于26％，表示产品具有抗菌性能。结果表明，本发明的止血敷料对大肠杆菌(Escherichia coli ATCC 8739)和金黄色葡萄球菌(Staphylicoccus aureus ATCC 6538)的抑菌率差值分别达90.09％和89.02％，说明本发明的止血敷料具有良好的抑菌性能。
生物相容性实验：参照GB/T 16886医疗器械的生物学评价，分别对止血敷料进行细胞毒性、豚鼠迟发接触性致敏、皮肤刺激等评价。生物相容性评价：细胞内毒性试验按照GB/T 16886-5《医疗器械生物学评价第5部分：体外细胞毒性试验》进行测试；豚鼠迟发型接触致敏试验按照GB/T 16886-10《医疗器械生物学评价第10部分： 刺激与迟发型超敏反应试验》进行测试，采用最大限度试验Magnusson和Kligman法。皮肤刺激试验按照GB/T 16886-10《医疗器械生物学评价第10部分：刺激与迟发型超敏反应试验》进行测试。结果表明：止血敷料细胞毒性小于2级，无皮肤致敏反应，无皮内刺激反应，具有良好的生物安全性。
体外细胞毒性试验见表二。
表二、细胞活力％
Figure PCTCN2019085277-appb-000002
样品浸泡液与生长旺盛的L-929细胞继续培养(37℃,5％CO2)24h后，采用MTT法测定样品的潜在细胞毒性。当浸泡液为100％时，测试样品毒性为II级。随着浸泡液的稀释，细胞活力逐步提高，浸泡液稀释到50％后，细胞毒性均为I级。因此，样品浸提液对L929细胞的毒性反应为2级，无潜在的细胞毒性。
皮肤刺激：将样品分别贴敷在3只新西兰白兔背部，经1h、24h、48h和72h后，观察皮肤红斑和水肿等反应情况，按皮肤刺激反应程度分级记分。样品皮肤反应结果观察见表三。
表三、样品皮肤反应结果观察
Figure PCTCN2019085277-appb-000003
试验过程中动物未出现异常症状或死亡。根据观察，实验组一侧皮肤反应未超过空白对照组一侧的皮肤反应，原发性刺激指数为0(无水肿、红斑)，阳性对照组动物记分为2(明显水肿、红斑)。试验样品在兔皮肤反应类型为无刺激作用。
皮肤致敏试验见表四。
表四、样品豚鼠致敏激发皮肤反应的临床观察
Figure PCTCN2019085277-appb-000004
将样品浸提液皮内注射10只豚鼠，包扎并试图诱发致敏。恢复期内，10只受试豚鼠和5只对照豚鼠分别采用样品浸提液和空白液进行激发贴片试验。在贴片移去后24h和48h记录各部位得分。在试验中，阴性对照组动物和试验组动物记分均为0(无明显改变)，阳性对照组动物记分为2(中度融合性红斑)。说明样品未发现迟发型接触性致敏反应。
实施例2
本实施例的止血敷料的制备方法：
(1)将生物纤维素纤维、四丙基氢氧化铵、硅基化合物与水混合，得到混合液。其中1L水中加入生物纤维素纤维、四丙基氢氧化铵、硅基化合物，该三种物质对应的摩尔比为0.001:0.05:1；其中，生物纤维素纤维是将八叠球菌属与假单胞菌属发酵得到的生物纤维素经12wt％的氢氧化钠水溶液高温蒸煮20min，然后机械均质得到的纳米纤维直径为30nm、长度为40μm的纤维素纤维；硅基化合物是指白炭黑和硅胶，质量比例为1:1。
(2)上述的混合液中加入氢氧化铝，缓慢搅拌6h，得到混合凝胶，其中铝基化 合物与硅基化合物的摩尔比为0.06:1；
(3)上述混合凝胶在高压釜中缓慢加热至175℃，恒温8h，其中升温速率为10℃/min；
(4)上述所得样品用去离子水清洗，在-30℃下冷冻12h，然后真空干燥48h，然后将干燥后的样品放入气氛炉中碳化处理加热至600℃，恒温6h后，样品在气氛炉炉中缓慢冷却至室温，得到沸石分子筛与生物纤维素基纳米碳纤维复合材料；其中，碳化处理是指材料在真空保护下，由室温升温至600℃，其中100～300℃区间升温速率为10℃/min，300～500℃区间升温速率为5℃/min，500～600℃区间升温速率为25℃/min。
(5)将上述复合材料与生物纤维素纤维、壳聚糖水溶液混合，经冷冻干燥、切割、高能射线灭菌、包装，最终得到可以快速止血并且力学性能良好的止血敷料。其中壳聚糖水溶液是由分子量为30万的壳聚糖配制成的，其质量浓度为4wt％。
本实施例的止血敷料拉伸力学强度为1050MPa；孔隙率为80～95％，孔径为50～500μm，吸水率为材料自身重量的200～1000倍。采用兔致命性股动脉出血伤口模型对止血敷料效果进行评估，结果表明，本实施例的止血敷料在1min内的止血率为100％。
实施例3
(1)将生物纤维素纤维、四丙基氢氧化铵、硅基化合物与水混合，得到混合液。其中1L水中加入生物纤维素纤维、四丙基氢氧化铵、硅基化合物，该三种物质对应的摩尔比为0.2:1:1；其中，生物纤维素纤维是将木醋杆菌、无色杆菌属和产碱菌属发酵得到的生物纤维素经14wt％的氢氧化钠水溶液高温蒸煮30min，然后机械均质得到的纳米纤维直径为40nm、长度为50μm的纤维素纤维；硅基化合物是硅酸钠和正硅酸丁酯，质量比例为1:1。
(2)上述的混合液中加入硫酸铝，缓慢搅拌7h，得到混合凝胶，其中铝基化合物与硅基化合物的摩尔比为0.07:1；
(3)上述混合凝胶在高压釜中缓慢加热至170℃，恒温10h，其中升温速率为6℃/min；
(4)上述所得样品用去离子水清洗，在-40℃下冷冻18h，然后真空干燥48h，然后将干燥后的样品放入气氛炉中碳化处理加热至570℃，恒温7h后，样品在气氛炉炉中缓慢冷却至室温，得到沸石分子筛与生物纤维素基纳米碳纤维复合材料；其中，碳化处 理是指材料在氩气保护下，由室温升温至570℃，其中100～300℃区间升温速率为6℃/min，300～500℃区间升温速率为4℃/min，500～570℃区间升温速率为28℃/min。
(5)将上述复合材料与生物纤维素纤维、壳聚糖水溶液混合，经冷冻干燥、切割、高能射线灭菌、包装，最终得到可以快速止血并且力学性能良好的止血敷料。其中壳聚糖水溶液是由分子量为25万的壳聚糖配制成的，其质量浓度为5wt％。
本实施例的止血敷料拉伸力学强度为900MPa；孔隙率为80～95％，孔径为50～500μm，吸水率为材料自身重量的200～1000倍。采用兔致命性股动脉出血伤口模型对止血敷料效果进行评估，结果表明，本实施例的止血敷料在1min内的止血率为100％。
实施例4
(1)将生物纤维素纤维、四丙基氢氧化铵、硅基化合物与水混合，得到混合液。其中1L水中加入生物纤维素纤维、四丙基氢氧化铵、硅基化合物，该三种物质对应的摩尔比为0.005:0.03:1；其中，生物纤维素纤维是将气杆菌属和固氮菌发酵得到的生物纤维素经16wt％的氢氧化钠水溶液高温蒸煮15min，然后机械均质得到的纳米纤维直径为50nm、长度为60μm的纤维素纤维；硅基化合物是指正硅酸甲酯和正硅酸异丙酯，质量比例为1:1。
(2)上述的混合液中加入铝溶胶和仲丁醇铝，质量比为1：1，缓慢搅拌8h，得到混合凝胶，其中铝基化合物与硅基化合物的摩尔比为0.08:1；
(3)上述混合凝胶在高压釜中缓慢加热至170℃，恒温12h，其中升温速率为8℃/min；
(4)上述所得样品用去离子水清洗，在-50℃下冷冻24h，然后真空干燥48h，然后将干燥后的样品放入气氛炉中碳化处理加热至550℃，恒温8h后，样品在气氛炉炉中缓慢冷却至室温，得到沸石分子筛与生物纤维素基纳米碳纤维复合材料；其中，碳化处理是指材料在氮气保护下，由室温升温至550℃，其中100～300℃区间升温速率为10℃/min，300～500℃区间升温速率为3℃/min，500～550℃区间升温速率为26℃/min。
(5)将上述复合材料与生物纤维素纤维、壳聚糖水溶液混合，经冷冻干燥、切割、高能射线灭菌、包装，最终得到可以快速止血并且力学性能良好的止血敷料。其中壳聚糖水溶液是由分子量为20万的壳聚糖配制成的，其质量浓度为6wt％。
本实施例的止血敷料拉伸力学强度为800MPa；孔隙率为80～95％，孔径为50～ 500μm，吸水率为材料自身重量的200～1000倍。采用兔致命性股动脉出血伤口模型对止血敷料效果进行评估，结果表明，本实施例的止血敷料在1min内的止血率为100％。
实施例5
(1)将生物纤维素纤维、四丙基氢氧化铵、硅基化合物与水混合，得到混合液。其中1L水中加入生物纤维素纤维、四丙基氢氧化铵、硅基化合物，该三种物质对应的摩尔比为0.01:0.02:1；其中，生物纤维素纤维是将产碱菌属发酵得到的生物纤维素经18wt％的氢氧化钠水溶液高温蒸煮28min，然后机械均质得到的纳米纤维直径为50nm、长度为70μm的纤维素纤维；硅基化合物是指正硅酸乙酯和正硅酸丙酯，质量比例为1：1。
(2)上述的混合液中加入异丙醇铝，缓慢搅拌5h，得到混合凝胶，其中铝基化合物与硅基化合物的摩尔比为0.09:1；
(3)上述混合凝胶在高压釜中缓慢加热至175℃，恒温24h，其中升温速率为7℃/min；
(4)上述所得样品用去离子水清洗，在-60℃下冷冻24h，然后真空干燥24h，然后将干燥后的样品放入气氛炉中碳化处理加热至600℃，恒温8h后，样品在气氛炉炉中缓慢冷却至室温，得到沸石分子筛与生物纤维素基纳米碳纤维复合材料；其中，碳化处理是指材料在氩气保护下，由室温升温至600℃，其中100～300℃区间升温速率为9℃/min，300～500℃区间升温速率为4℃/min，500～600℃区间升温速率为27℃/min。
(5)将上述复合材料与生物纤维素纤维、壳聚糖水溶液混合，经冷冻干燥、切割、高能射线灭菌、包装，最终得到可以快速止血并且力学性能良好的止血敷料。其中壳聚糖水溶液是由分子量为15万的壳聚糖配制成的，其质量浓度为7wt％。
本实施例的止血敷料拉伸力学强度为850MPa；孔隙率为80～95％，孔径为50～500μm，吸水率为材料自身重量的200～1000倍。采用兔致命性股动脉出血伤口模型对止血敷料效果进行评估，结果表明，本实施例的止血敷料在1min内的止血率为100％。
实施例6
(1)将生物纤维素纤维、四丙基氢氧化铵、硅基化合物与水混合，得到混合液。其中1L水中加入生物纤维素纤维、四丙基氢氧化铵、硅基化合物，该三种物质对应的 摩尔比为0.16:0.5:1；其中，生物纤维素纤维是将木醋杆菌、气杆菌属和固氮菌发酵得到的生物纤维素经20wt％的氢氧化钠水溶液高温蒸煮10min，然后机械均质得到的纳米纤维直径为50nm、长度为100μm的纤维素纤维；硅基化合物是指硅胶、白炭黑、正硅酸甲酯和正硅酸丁酯，比例为1:1:1:1。
(2)上述的混合液中加入偏铝酸钠、氢氧化铝和硫酸铝，质量比例为1:1:1，缓慢搅拌6h，得到混合凝胶，其中铝基化合物与硅基化合物的摩尔比为0.1:1；
(3)上述混合凝胶在高压釜中缓慢加热至170℃，恒温24h，其中升温速率为5℃/min；
(4)上述所得样品用去离子水清洗，在-80℃下冷冻24h，然后真空干燥48h，然后将干燥后的样品放入气氛炉中碳化处理加热至600℃，恒温8h后，样品在气氛炉炉中缓慢冷却至室温，得到沸石分子筛与生物纤维素基纳米碳纤维复合材料；其中，碳化处理是指材料在氮气保护下，由室温升温至600℃，其中100～300℃区间升温速率为7℃/min，300～500℃区间升温速率为3℃/min，600℃区间升温速率为27℃/min。
(5)将上述复合材料与生物纤维素纤维、壳聚糖水溶液混合，经冷冻干燥、切割、高能射线灭菌、包装，最终得到可以快速止血并且力学性能良好的止血敷料。其中壳聚糖水溶液是由分子量为10万的壳聚糖配制成的，其质量浓度为10wt％。
本实施例的止血敷料拉伸力学强度为750MPa；孔隙率为80～95％，孔径为50～500μm，吸水率为材料自身重量的200～1000倍。采用兔致命性股动脉出血伤口模型对止血敷料效果进行评估，结果表明，本实施例的止血敷料在1min内的止血率为100％。

Claims (15)

  1. 一种生物纤维素纤维，是细菌发酵得到的纤维素经10～20wt％的氢氧化钠水溶液蒸煮处理而得到的，其直径为20～50nm，长度为30～100μm。
  2. 权利要求1所述生物纤维素纤维的制备方法，该方法包括：
    将细菌发酵得到的纤维素经10～20wt％的氢氧化钠水溶液蒸煮10～30min；
    机械均质，以制备得到直径20～50nm、长度30～100μm的纤维。
  3. 权利要求1所述生物纤维素纤维在制备止血敷料中的用途。
  4. 根据权利要求3所述的用途，其中，是将所述生物纤维素纤维作为模板，水热法合成沸石分子筛。
  5. 一种止血敷料的制备方法，该方法包括：
    将权利要求1所述的生物纤维素纤维、四丙基氢氧化铵、硅基化合物与水混合，得到混合液；其中每1L水中加入生物纤维素纤维、四丙基氢氧化铵、硅基化合物对应的摩尔比为(0.001～0.2):(0.01～1):1；
    向上述的混合液中加入铝基化合物，缓慢搅拌5～8h，得到混合凝胶，其中铝基化合物与硅基化合物的摩尔比为(0.05～0.1):1；
    上述混合凝胶缓慢加热至170～175℃，恒温6～24h，得到反应样品；其中升温速率为5～10℃/min；
    上述所得样品用去离子水清洗，干燥，然后加热至550℃～600℃碳化处理，恒温5～8h后，冷却，得到沸石分子筛与生物纤维素基纳米碳纤维复合材料；
    将上述复合材料与生物纤维素纤维、壳聚糖水溶液混合，经冷冻干燥，灭菌，得到止血敷料。
  6. 根据权利要求5所述的方法，其中，所述的硅基化合物包括硅胶、白炭黑、硅酸钠、正硅酸甲酯、正硅酸乙酯、正硅酸丙酯、正硅酸异丙酯和正硅酸丁酯中的任意一种或几种。
  7. 根据权利要求5所述的方法，其中，所述的铝基化合物包括偏铝酸钠、氢氧化铝、硫酸铝、铝溶胶、异丙醇铝和仲丁醇铝中的任意一种或几种。
  8. 根据权利要求5所述的方法，其中，所述的干燥优选为冷冻干燥；所述的冷冻干燥优选是将样品在-20℃～-80℃下冷冻12～24h，然后真空干燥24～48h。
  9. 根据权利要求5所述的方法，其中，所述的碳化处理是指材料在真空、氩气或氮气保护下，由室温升温至550℃～600℃，其中100～300℃区间升温速率为5～ 10℃/min，300～500℃区间升温速率为1～5℃/min，500～600℃区间升温速率优选为30℃/min。
  10. 根据权利要求5所述的方法，其中，所述的冷却是指升温结束后的样品在气氛炉或活化炉中缓慢冷却至室温。
  11. 根据权利要求5所述的方法，其中，复合材料与生物纤维素纤维、壳聚糖水溶液混合时，壳聚糖、沸石分子筛、生物纤维素基碳纤维与生物纤维素纤维的混合质量比为1：(0.01～0.1)：(0.01:0.1)：(0.01～0.1)，更佳的比例为1：(0.04～0.06)：(0.04～0.06)：(0.3～0.6)。
  12. 根据权利要求5所述的方法，其中，所述的壳聚糖水溶液中，壳聚糖质量浓度为3～10wt％，所述的壳聚糖分子量为10～35万。
  13. 一种以生物纤维素纤维制备的止血敷料，其以壳聚糖为多孔材料的基体，其中均匀分布着生物纤维素纤维、纳米碳纤维以及沸石颗粒，该止血敷料的材料孔隙率为80～95％，孔径为50～500μm，吸水率为材料自身重量的200～1000倍，拉伸力学强度为0.5～4GPa。
  14. 根据权利要求13所述的止血敷料，其是按照权利要求5～11任一项所述的方法制备得到的。
  15. 权利要求13或14所述的止血敷料在制备具有止血和/或促进伤口愈合功效的药物中的应用。
PCT/CN2019/085277 2019-04-30 2019-04-30 生物纤维素纤维、包含所述纤维的止血敷料与相关应用 WO2020220300A1 (zh)

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