阅读说明：本技术 医用生物凝胶止血敷料的制备方法 (Preparation method of medical biogel hemostatic dressing ) 是由 杨鑫 于 2019-10-11 设计创作，主要内容包括：本发明公开一种医用生物凝胶止血敷料的制备方法,涉及生物医学领域。本发明先利用蚕茧和聚乙烯醇溶液,制得蚕丝蛋白混合液,然后将细菌纤维素与蚕丝蛋白混合液混合,并加入聚丙烯酰胺,经冻融循环后,制得凝胶坯料；再将凝胶坯料用壳聚糖季铵盐处理液处理后,依次与含有海藻酸钠的银氨溶液以及钙离子溶液混合,过滤,冷冻干燥,制得医用生物凝胶止血敷料。本发明制备的医用生物凝胶止血敷料具有优异的抗菌性能,且对血液具有较好的吸附能力,并可使血液快速凝固,同事本发明制备的医用生物凝胶止血敷料具有优异的力学性能。(The invention discloses a preparation method of a medical biogel hemostatic dressing, and relates to the field of biomedicine. Firstly, preparing fibroin mixed solution by using silkworm cocoons and polyvinyl alcohol solution, then mixing bacterial cellulose and the fibroin mixed solution, adding polyacrylamide, and performing freeze-thaw cycle to prepare a gel blank; and treating the gel blank with a chitosan quaternary ammonium salt treatment solution, sequentially mixing with a silver ammonia solution containing sodium alginate and a calcium ion solution, filtering, and freeze-drying to obtain the medical biogel hemostatic dressing. The medical biogel hemostatic dressing prepared by the invention has excellent antibacterial performance, has better adsorption capacity to blood, can quickly coagulate the blood, and has excellent mechanical property.)

1. A preparation method of a medical biogel hemostatic dressing mainly comprises the following preparation steps:

(1) mixing a silkworm cocoon and a 10% sodium carbonate solution according to a mass ratio of 1:8, stirring and reacting for 80min at a temperature of 60 ℃ and a rotating speed of 300r/min, filtering to obtain a degummed silkworm cocoon, crushing the degummed silkworm cocoon in a crusher for 40min to obtain refined silk, mixing the refined silk and a 3% trypsin solution according to a mass ratio of 1:12 in a beaker, stirring and reacting for 50min at a temperature of 36 ℃ and a rotating speed of 300r/min, heating the material in the beaker to 90 ℃, keeping the temperature for 30min, inactivating enzyme to obtain a fibroin dispersion liquid, and mixing the fibroin dispersion liquid and a 10% polyvinyl alcohol solution according to a mass ratio of 3: 1;

(2) mixing bacterial cellulose and the substance obtained in the step (1) according to a mass ratio of 1:20, adding polyacrylamide with the mass of 0.4 time that of the bacterial cellulose, stirring and mixing for 40min at a temperature of 35 ℃ and a rotating speed of 280r/min to obtain a mixed dispersion liquid, freezing the mixed dispersion liquid for 6h at a temperature of-30 ℃, then unfreezing for 6h at room temperature, and performing freeze-thaw cycling for 3 times to obtain a gel blank;

(3) mixing the gel blank obtained in the step (2) with a chitosan quaternary ammonium salt treatment solution according to a mass ratio of 1:8, standing and reacting for 4 hours at the temperature of 40 ℃, filtering to obtain a pretreated gel blank, mixing the pretreated gel blank and a silver ammonia solution with the mass fraction of 10% in a conical flask according to a mass ratio of 1:10, stirring and reacting for 3 hours at the temperature of 45 ℃ and the rotating speed of 180r/min, filtering, and freeze-drying;

(4) mixing the substance obtained in the step (3) with a calcium ion solution according to a mass ratio of 1:10, standing and reacting for 3 hours at the temperature of 35 ℃, filtering, and freeze-drying;

(5) performing index analysis on the substance obtained in the step (4);

mixing chitosan and an acetic acid solution with the mass fraction of 2% according to the mass ratio of 1:10, adjusting the pH value of a mixed solution of the chitosan and the acetic acid solution to 10, collecting precipitates to obtain pretreated chitosan, mixing the pretreated chitosan and isopropanol according to the mass ratio of 1:8, adding a glycidol trimethyl ammonium chloride solution with the mass fraction of 20% and the mass fraction of 4 times that of the pretreated chitosan, stirring for reaction, adding ethanol for precipitation, filtering to obtain filter residues, drying the filter residues at the temperature of 75 ℃ for 3 hours to obtain chitosan quaternary ammonium salt, mixing the chitosan quaternary ammonium salt and water according to the mass ratio of 1:20, adding citric acid with the mass of 0.2 time of that of the chitosan quaternary ammonium salt and triethanolamine with the mass of 0.3 time of that of the chitosan quaternary ammonium salt, and stirring for mixing to obtain the chitosan quaternary ammonium salt treatment solution;

and (4) the calcium ion solution in the step (4) is a calcium chloride solution with the mass fraction of 5%.

Technical Field

The invention relates to the field of biomedicine, in particular to a preparation method of a medical biogel hemostatic dressing.

Background

The medical science indicates that the patient can die within 6-20 minutes if effective emergency measures are not taken due to acute bleeding. Shock occurs when the blood loss of a human body exceeds 20 percent of the whole blood volume; if the blood loss exceeds 40 percent of the total blood volume, death is imminent. According to statistics, the number of traumatic deaths accounts for 10% of the total number of deaths, and the number of deaths caused by massive bleeding accounts for 30% -40%, so early bleeding control is the main method for reducing the mortality rate.

At present, after various artery and vein puncture and catheter extraction operations in hospitals, compression hemostasis is needed to be carried out on wounds. The most widely applied compression hemostasis mode in hospitals is that medical staff adopt gauze to perform compression hemostasis. Medical personnel superimpose gauze over the wound and then use bandages or straps to secure. Such a way of operation has drawbacks: firstly, the bandage or the bandage does not have extensibility and viscosity, and can not automatically give compression force, so that after the bandage or the bandage is fixed, the compression force can be lost if the bandage or the bandage is not tightly bound, and gauze can possibly move if a patient moves; secondly, the gauze has poor blood absorption performance, is easy to infect wounds and realizes the compression of the wounds only by the compression force given by bandages or bands.

Most of the recently developed hemostatic dressings use chitosan, a novel biomaterial, as the hemostatic material. The chitosan has biocompatibility and biodegradability, is rich in resources and easy to obtain, is applied to the hemostatic dressing, and has the functions of resisting bacteria, diminishing inflammation, stopping bleeding, easing pain, promoting wound healing and the like. In order to further enhance the performance of chitosan as a hemostatic dressing, chitosan and other substances are mixed in the prior art to prepare the hemostatic dressing.

Chitosan is a polymer obtained by deacetylating chitin, which is a natural polymer extracted from organisms and mainly exists in cell walls of crustaceans (shrimps and crabs), insect shells or fungi. Chitin is a linear high molecular polysaccharide polymer formed by beta-1, 4 bonding of glucosamine and N-acetylglucosamine. The chitosan has good biocompatibility, bioactivity, no cytotoxicity and biological decomposability with cells, so the chitosan can be applied to biological materials and medical materials. The chitosan has positive electricity in the acid solution, so that the chitosan can attract the blood platelets to aggregate, and further achieve the effect of quickly stopping bleeding.

At present, the dressing made of chitosan non-woven fabric is widely used in clinic, however, the chitosan dressing has the defect of poor stress, and after contacting blood, the chitosan dressing can rapidly form gel and break. Has good hemostatic effect on general bleeding, good biocompatibility and small side effect on human body during wound healing. However, when acute bleeding or surgical bleeding occurs, the hemostasis speed and amount still cannot meet the requirement of emergency treatment for massive and rapid bleeding.

Therefore, it is an important matter to invent a novel hemostatic dressing which can solve the defects of poor hemostatic effect and the like of the existing hemostatic dressing so as to meet the requirements of related industries.

Disclosure of Invention

The invention aims to provide a medical biogel hemostatic dressing and a preparation method thereof, and aims to solve the problems in the prior art.

In order to achieve the purpose, the invention provides the following technical scheme:

the medical biogel hemostatic dressing is characterized by mainly comprising the following raw material components in parts by weight: 20-30 parts of silk, 6-12 parts of chitosan quaternary ammonium salt, 8-15 parts of bacterial cellulose, 2-4 parts of polyacrylamide and 5-8 parts of sodium alginate.

The medical biogel hemostatic dressing is characterized by further comprising the following raw material components in parts by weight: 12-18 parts of polyvinyl alcohol and 2-4 parts of nano silver.

Preferably, the chitosan quaternary ammonium salt is prepared by treating chitosan with glycidol trimethyl ammonium chloride.

As optimization, the medical biogel hemostatic dressing mainly comprises the following raw materials in parts by weight: 25 parts of silk, 10 parts of chitosan quaternary ammonium salt, 8 parts of bacterial cellulose, 3 parts of polyacrylamide, 8 parts of sodium alginate, 12 parts of polyvinyl alcohol and 4 parts of nano-silver.

As optimization, the preparation method of the medical biogel hemostatic dressing mainly comprises the following preparation steps:

(1) degumming and crushing silkworm cocoons to prepare refined silk, mixing the refined silk with a protease solution, inactivating enzyme at high temperature, adding a polyvinyl alcohol solution, and stirring and mixing to obtain a fibroin mixed solution;

(2) mixing bacterial cellulose and the fibroin mixed solution obtained in the step (1), adding polyacrylamide, stirring and mixing, and performing freeze-thaw circulation to obtain a gel blank;

(3) mixing the gel blank obtained in the step (2) with the chitosan quaternary ammonium salt treatment solution, filtering to obtain a pretreated gel blank, mixing the pretreated gel blank with a silver ammonia solution, adding sodium alginate, stirring and mixing, filtering, and freeze-drying to obtain gel;

(4) mixing the gel obtained in the step (3) with a calcium ion solution, filtering, freezing and drying to obtain the medical biogel hemostatic dressing;

(5) and (4) performing index analysis on the medical biogel hemostatic dressing obtained in the step (4).

As optimization, the preparation method of the medical biogel hemostatic dressing mainly comprises the following preparation steps:

(1) mixing silkworm cocoons with a sodium carbonate solution with the mass fraction of 10% according to the mass ratio of 1:8, stirring for reaction, filtering to obtain degummed silkworm cocoons, crushing the degummed silkworm cocoons in a crusher for 20-40 min to obtain refined silk, mixing the refined silk with a trypsin solution with the mass fraction of 3% according to the mass ratio of 1:12, stirring for reaction, inactivating enzyme at high temperature to obtain a fibroin dispersion liquid, and mixing the fibroin dispersion liquid with a polyvinyl alcohol solution with the mass fraction of 10% according to the mass ratio of 3: 1;

(2) mixing bacterial cellulose and the substance obtained in the step (1) according to a mass ratio of 1: 20-1: 25, adding polyacrylamide with a mass of 0.2-0.5 times that of the bacterial cellulose, stirring and mixing to obtain a mixed dispersion liquid, freezing the mixed dispersion liquid at-40 to-20 ℃ for 6 hours, then thawing at room temperature, and performing freeze-thaw cycling for 3 times to obtain a gel blank;

(3) mixing the substance obtained in the step (2) with a chitosan quaternary ammonium salt treatment solution according to a mass ratio of 1: 6-1: 8, standing for reaction for 3-4 h, filtering to obtain a pretreated gel blank, mixing the pretreated gel blank with a silver ammonia solution with a mass fraction of 10% according to a mass ratio of 1:10, adding sodium alginate with a mass of 0.1-0.5 times that of the pretreated gel blank, stirring for reaction, filtering, and freeze-drying;

(4) mixing the substance obtained in the step (3) with a calcium ion solution according to the mass ratio of 1:10, standing for reaction, filtering, and freeze-drying;

(5) and (4) carrying out index analysis on the substance obtained in the step (4).

Preferably, the chitosan quaternary ammonium salt treatment solution in the step (3) is prepared by mixing chitosan and an acetic acid solution with the mass fraction of 2% according to the mass ratio of 1:10, adjusting the pH value of the mixed solution of the chitosan and the acetic acid solution to 10, collecting precipitates to obtain pretreated chitosan, mixing the pretreated chitosan and isopropanol according to the mass ratio of 1:8, adding a glycidol trimethyl ammonium chloride solution with the mass fraction of 20% and the mass fraction of 3-6 times that of the pretreated chitosan, stirring for reaction, adding ethanol for precipitation, filtering, drying to obtain chitosan quaternary ammonium salt, mixing the chitosan quaternary ammonium salt with water according to the mass ratio of 1:20, adding citric acid with the mass of 0.1-0.3 time that of the chitosan quaternary ammonium salt and triethanolamine with the mass of 0.2-0.3 time that of the chitosan quaternary ammonium salt, and stirring and mixing to obtain the chitosan quaternary ammonium salt treatment solution.

Preferably, the calcium ion solution in the step (4) is any one of a calcium chloride solution with a mass fraction of 5% or a calcium nitrate solution with a mass fraction of 3%.

Compared with the prior art, the invention has the beneficial effects that:

silk and chitosan quaternary ammonium salt are added when the medical biogel hemostatic dressing is prepared; firstly, silk is added into the medical biogel hemostatic dressing, on one hand, the silk can be matched with added bacterial cellulose to be used as a framework of the medical biogel hemostatic dressing, thereby improving the mechanical property of the medical biogel hemostatic dressing, preventing the medical biogel hemostatic dressing from being broken due to blood sucking and coagulation promotion when in use, and improving the strippability of a product, on the other hand, the surface of the silk is degraded after the added silk is treated by protease, thereby improving the number of active groups on the surface of the silk, and can absorb sufficient calcium ions after being mixed with a calcium ion solution, and the calcium ions can promote blood coagulation, thereby improving the coagulation promotion performance of the product on blood when the medical biogel hemostatic dressing is in use, meanwhile, because the silk is protein fiber, the silk can be decomposed by protease in plasma in the use process of the product, and the wound healing does not improve amino acid, thereby improving the use effect of the product; secondly, add chitosan quaternary ammonium salt in medical biogel hemostatic dressing, chitosan quaternary ammonium salt can adsorb and the silk surface under the effect of electrostatic force, thereby improve the antibacterial property of product, after follow-up and the silver ammonia solution that contains sodium alginate mixes, sodium alginate can be under the effect of electrostatic force, carry out self-assembly on the silk surface, simultaneously, because sodium alginate and the group on silk surface have the reductibility, can reduce silver ammonia solution, form nanometer silver, therefore sodium alginate can be with the firm cladding of nanometer silver on the silk surface in self-assembly process, and then further improve the antibacterial property of product, moreover, because sodium alginate can carry out the crosslinking under calcium ion solution, thereby further improve calcium ion content in the product, and improve the fastness of product, and then improve the effect of procoagulant of product to blood.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

In order to more clearly illustrate the method provided by the present invention, the following examples are provided to illustrate the method of testing the indexes of the medical bio-gel hemostatic dressing prepared in the following examples as follows:

sterilization property: the medical biogel hemostatic dressing obtained in each example and the comparative product are respectively placed in a staphylococcus aureus culture solution and an escherichia coli culture solution with the same concentration, and the sterilization rate after the medical biogel hemostatic dressing and the comparative product are placed for 2 hours is measured.

Blood coagulation property: the medical biogel hemostatic dressings obtained in each example and the comparative example were cut into pieces of 1cm × 1cm × 0.5cm, and placed in beakers containing 10mL of blood, respectively, and the time for adsorbing the whole blood and the time for clotting the blood were measured.