CN112121065A

CN112121065A - Medical styptic powder and application thereof

Info

Publication number: CN112121065A
Application number: CN202011071241.7A
Authority: CN
Inventors: 张曦木
Original assignee: Stomatological Hospital of Chongqing Medical University
Current assignee: Stomatological Hospital of Chongqing Medical University
Priority date: 2020-10-09
Filing date: 2020-10-09
Publication date: 2020-12-25

Abstract

The medical hemostatic powder comprises sterilized giant salamander skin mucus dry powder, the particle size of the medical hemostatic powder is 4-300 meshes, the giant salamander skin mucus dry powder has water absorption, hydrogel is formed after the giant salamander skin mucus dry powder absorbs aqueous solution, in addition, the medical hemostatic powder is applied to hemostasis of wounds, the hydrogel is formed after the giant salamander skin mucus dry powder and the aqueous solution are mixed, the wounds are made to stop bleeding, and multiple beneficial effects of promoting tissue repair are achieved.

Description

Medical styptic powder and application thereof

Technical Field

The invention belongs to the field of biological materials, relates to medical styptic powder, and particularly relates to medical styptic powder prepared by using natural components as raw materials and application thereof.

Background

Healthy adults have about 3.8 to 5.6 liters of blood. If the blood loss exceeds 15 percent of the total blood of the whole body, the pulse is accelerated, and the adverse reactions such as dizziness and the like are generated; if the blood loss exceeds 40% of the total amount, the atria cannot be filled with backflow due to hypotension, and "ventricular tachycardia" symptoms occur, which usually lead to death. Uncontrolled blood loss has become a leading cause of life-threatening and death in victims, and it is seen that it is important for trauma-induced local rapid hemostasis in blood-lost persons.

The prior art provides a variety of hemostatic methods, such as compression hemostasis, electrosurgical knife, hemostatic materials, and the like.

The hemostatic powder is a common medical material, is mainly used for quickly scattering wounds with large or irregular areas to play a role in quickly stopping bleeding, and is simpler, more convenient and quicker than application, bandaging and the like. The application scenes of the hemostatic powder comprise emergency hemostasis of daily sudden accidents, wound hemostasis in the operation process of a hospital on a patient, field rescue hemostasis on an injured fighter in the war condition, and the like. The quick and effective hemostatic material can greatly reduce the death rate of the wounded. Therefore, the development of a novel hemostatic material that is fast, safe and effective has become an important research topic in the field of medical materials.

The hemostatic powder in the prior art mostly utilizes the adsorption performance of materials to absorb water in bleeding, accelerates the combination of platelets and red blood cells in the bleeding and plays a role in rapid hemostasis. For example, traditional Chinese medicine is ground into powder, or traditional medical fiber is ground into powder, or microporous starch is adopted. Or the material with the effects of water absorption and heat generation is used for accelerating the protein coagulation in blood by increasing the temperature at the application, for example, the zeolite powder prepared by a special process has the effects of absorbing the water in the bleeding and generating heat. The microporous particles made of various polysaccharide materials have the effects of absorbing water, and simultaneously can gather blood coagulation factors, platelets, fibrin, red blood cells and the like in blood on the surfaces of the particles to achieve the effect of quickly stopping bleeding.

The main disadvantage of the prior art is that the use of non-biocompatible materials may cause the wound to be sticky and the material not to be absorbed by the body, and the need for post-healing treatment may cause various side effects. The process for preparing the porous particles by adopting the biocompatible material is complex, and the cost is high.

Giant salamanders (Andrias davidianus Andrias) are large amphibia, Ceramiales, and Holotrichia, and are commonly named as giant salamanders, belonging to domestic secondary protection animals. When the giant salamander meets external stimulation, mucus is secreted from the surface layer of the body. Research results show that the giant salamander skin mucus can be used for preparing an adhesive or a hemostatic. The invention of China, namely the effect of giant salamander mucus on hemostasis, adopts the giant salamander mucus extract as a hemostatic agent, the preparation process is complex, and the tail breaking experiment of a rat shows that the hemostasis effect can be achieved within about 80 seconds, which indicates that the hemostasis effect is not good enough. The Chinese invention application of giant salamander mucus in preparation of hemostatic materials mainly utilizes the adhesive property of giant salamander mucus dry powder to stanch superficial trauma, and the long-term hemostatic effect and wound infection effect after adhesion are not evaluated. The possibility of internal bleeding after adhesion or inflammation during the healing process is not excluded.

The medical styptic powder prepared from giant salamander skin mucus still has defects to be solved urgently.

Disclosure of Invention

In view of the defects of the prior art, the invention provides medical styptic powder and the application of hemostasis, wherein the components contain giant salamander skin mucus components, so that the medical styptic powder can effectively stop bleeding and has multiple benefits of promoting wound repair, thereby solving the technical problems.

The medical hemostatic powder provided by the invention utilizes the characteristics that giant salamander skin mucus has excellent adhesive performance and various effects of promoting hemostasis, wound healing and tissue repair, so that the prepared medical hemostatic powder can achieve the effect of rapid hemostasis. The medical styptic powder has the advantages of good safety, good biocompatibility, degradability, regeneration promoting effect, antibacterial effect, wide material source and simple preparation process, and is an ideal material for meeting the requirements.

In order to overcome the defects that the originally collected giant salamander skin mucus is jelly-like, inconvenient to store, inconvenient to use clinically, difficult to thoroughly sterilize and the like, the invention provides medical styptic powder which comprises sterilized giant salamander skin mucus dry powder; in addition, the sterilized giant salamander skin mucus dry powder has specific particle size and water absorbability, so that the optimal hemostatic effect is achieved.

According to the above objects, the present invention provides a medical styptic powder, which comprises sterilized giant salamander skin mucus dry powder, wherein the particle size is less than 4 meshes and more than 300 meshes; and the giant salamander skin mucus dry powder has water absorbability, and can form hydrogel after absorbing aqueous solution.

According to a preferred embodiment of the present invention, the medical styptic powder comprises sterilized giant salamander skin mucus dry powder, and the preferred particle size is-4 mesh to-100 mesh.

According to a preferred embodiment of the present invention, the aqueous solution absorbed by the medical hemostatic powder is selected from distilled water, physiological buffer, blood, plasma, blood cell preparation, tissue fluid, platelet rich plasma fibrin or any combination thereof.

According to a preferred embodiment of the invention, the medical styptic powder comprises the dry giant salamander skin mucus powder and the aqueous solution in a weight part ratio of 1: 1 to 1: 6.

according to the purpose, the invention further provides a using method of the medical hemostatic powder, and the medical hemostatic powder is applied to a wound to form a dressing.

According to a preferred embodiment of the present invention, the hemostasis is achieved by applying the medical hemostatic powder to the wound, so that the medical hemostatic powder is mixed with the aqueous solution at the wound and forms hydrogel, and the medical hemostatic powder becomes the dressing.

According to a preferred embodiment of the present invention, the medical hemostatic powder and the aqueous solution are prepared into hydrogel in advance, and then the hydrogel is used as a dressing.

Drawings

FIG. 1 shows a giant salamander (a) described in example 1 of the present invention; obtaining skin mucus (b) on the back of the giant salamander by adopting a mechanical scraping method; and (c) a body type microscopic image of the giant salamander skin mucus dry powder with different particle sizes, which is obtained by the invention.

FIG. 2 is a graph showing the data of the sterilization effect achieved by the sterilization protocols of experiment 1 of example 1 of the present invention (a) and the data of the adhesion strength (b).

FIG. 3 shows SSAD powders (a) of different particle sizes and their sterilized bulk microscope images (b) that are mixed with anticoagulated whole blood and gelatinized as described in example 2 of the present invention.

FIG. 4 shows Scanning Electron Microscope (SEM) images (a) of SSAD powder and medical hemostatic powder (SSAD powder after sterilization) of different particle sizes obtained by different sieving in example 2 of the present invention after hydration into gel; and the pore size distribution (b) of the porous structure formed by mixing the SSAD powder with different mesh numbers and the aqueous solution to form the glue in the embodiment 2 of the invention.

FIG. 5A shows the results of the cell scratching test for cell proliferation and migration promotion by SSAD powder leaching medium in experiment 4 of the present invention.

FIG. 5B shows the statistical analysis of the cell scratch test of the SSAD meal leaching media of experiment 4 of the present invention.

FIG. 5C shows the results of a transwell chamber experiment in which SSAD powder-extracted medium promoted cell proliferation and migration in experiment 4 of the present invention.

FIG. 6 shows the histological analysis of the in vivo degradation of the medical styptic powder of example 5 of the present invention.

Detailed Description

So that the manner in which the above recited features and advantages of the present invention can be understood and attained, a more particular description of the invention, briefly summarized above, may be had by reference to the appended drawings, in which, in order to facilitate understanding of the nature of the invention, features, and advantages thereof, may be had by reference to the appended claims, which are included to illustrate, by way of example, embodiments of the invention. The drawings referred to below are schematic representations, not necessarily drawn to scale, of features of the invention. The description of the embodiments related to the present invention will not be repeated, except for those skilled in the art.

The invention provides medical styptic powder which is mainly prepared from giant salamander skin mucus dry powder.

The originally collected giant salamander skin mucus is jelly-shaped, is difficult to thoroughly sterilize, is inconvenient to store and has certain difficulty in clinical use. In order to solve the defects, the technical scheme adopted by the invention is to provide the medical styptic powder which contains giant salamander skin mucus dry powder, is obtained after quality confirmation and full sterilization and is applied to hemostasis.

The medical styptic powder provided by the invention is not limited herein, except that the giant salamander skin mucus dry powder is used as a necessary component, and is used for preparing a product which is easy to store, transport and clinically apply, and is further matched with other components or raw materials to prepare a medical styptic powder product suitable for use. For example, according to a preferred embodiment of the present invention, the formula of the medical hemostatic powder provided by the present invention may further include other components having hemostatic, anti-inflammatory or tissue growth promoting effects, so as to further improve the clinical performance, and the present invention is not limited thereto.

According to one of the preferred embodiments of the present invention, the preparation method of the medical styptic powder comprises the following steps. Step 1: mucus was obtained from giant salamander skin and freeze-dried. Step 2: grinding and crushing the freeze-dried giant salamander skin mucus, and sieving to obtain SSAD powder with the particle size or particle diameter meeting the requirement, wherein the SSAD powder is used as one of the conditions for ensuring the quality. And step 3: and then sterilizing the SSAD powder to obtain the medical styptic powder, and storing for later use.

When in use, the sterilized medical hemostatic powder and the aqueous solution are mixed according to the ratio of 1: 1 to 1: 6, mixing to form hydrogel. Preferably, the weight ratio of the giant salamander skin mucus dry powder to the aqueous solution in the hydrogel is 1: 2 to 1: 5.

the preferred embodiments are described below with reference to the accompanying drawings. The mucus collection method in the step 1 is performed strictly according to the Chinese animal protection method, and can avoid the permanent disability of the giant salamander without killing the giant salamander. A scraping method or an electrical stimulation method may be employed. Mucus was also collected before sacrifice of commercially bred giant salamanders. The invention is not limited in this regard.

And 2, crushing the collected dried giant salamander skin mucus by a ball mill at a low temperature, and grinding into fine powder. And further sieving the powder by adopting sieves with different meshes to obtain powder with specific particle size (particle size with specific specification), also called SSAD powder, so as to realize the efficacy of the medical styptic powder.

Because the standard of the screening mesh number is slightly different internationally, the screening mesh number is preferably defined by a Taylor standard screening system commonly used in China, the graduation of the screening mesh number is based on the size of a 200-mesh screen hole of 0.074mm, and the detailed calculation mode is the conventional knowledge of the technicians in the field and is not described again. When the standard is used for describing the particle size of the giant salamander skin mucus freeze-dried powder, the plus sign and the minus sign before the mesh number indicate whether the powder can leak through the meshes of the mesh number after grinding or crushing, the minus sign indicates that the powder can leak through the meshes of the mesh number, and the particle size of the powder obtained after sieving is smaller than the mesh size; and the positive number indicates that the powder cannot leak through the mesh of the mesh, i.e. the particle size of the powder obtained after sieving is larger than the mesh size. For example, a particle size of less than 4 mesh refers to a powder that can pass through a 4 mesh screen, i.e., a powder having a maximum particle size of no more than 4.75 mm; a particle size of more than 300 mesh refers to a powder that cannot pass through a 300 mesh sieve, i.e. a powder with a minimum particle size of more than 48 μm.

In order to obtain better medical hemostatic powder performance, the optimal particle size of the giant salamander skin mucus freeze-dried powder is smaller than-4 meshes (the-4 meshes indicate that the number of the screened meshes is larger than 4 meshes and smaller than 8 meshes; the corresponding average particle size is approximately 4207 microns obtained by experiments, and is shown in the following table 2); larger than-100 mesh (said-100 mesh means a mesh number larger than 100 mesh and smaller than 200 mesh the corresponding average particle size was found to be approximately 105 μm by experiment, see table 2 below).

In the concrete practice of the invention, the powder with the particle size of-4 meshes is obtained by secondary sieving through sieves with 4 meshes and 8 meshes to obtain the powder with the particle size capable of passing through a 4-mesh sieve and not capable of passing through an 8-mesh sieve (in other words, the particle size of the obtained particles is smaller than the mesh size of the 4-mesh sieve and larger than the mesh size of the 8-mesh sieve); -8 mesh means particle size less than 8 mesh and greater than 14 mesh; -14 mesh means particle size less than 14 mesh and greater than 20 mesh; -20 mesh means particle size less than 20 mesh and greater than 60 mesh; -60 mesh means particle size less than 60 mesh and greater than 100 mesh; -100 mesh means particle size less than 100 mesh and greater than 200 mesh; -200 mesh means particle size less than 200 mesh and greater than 300 mesh, -300 mesh means particle size less than 300 mesh and greater than 400 mesh.

After step 2 is completed, the powder can be stored in a below-20 ℃ freezer for use. Next, step 3 is, in principle, sterilization and disinfection of the powder obtained in step 2. This step is an important step in applying the product to clinical use, and is critical to the performance of clinical use. According to relevant regulations, the giant salamander skin mucus related products which are not sterilized and disinfected cannot be directly applied to clinic. Moreover, mucus secreted on the body surface of the live giant salamander may contain viruses or germs which are potentially harmful to the human body when collected in the step 1, the viruses or germs cannot be completely inactivated by the freeze-drying in the step 2, and the possibility of wound infection is increased if the mucus is directly applied to the wound surface.

In addition, the main components of the giant salamander skin mucus are active components such as protein, polypeptide, mucopolysaccharide and antibacterial peptide, and the most preferable sterilization method can not damage and change the structure of biological macromolecules and can not cause the adhesion of the giant salamander skin mucus dry powder, the hemostatic performance and the biological activity to be reduced. Therefore, according to the current technology, the disinfection and sterilization method for giant salamander skin mucus related products comprises low temperature, ultraviolet ray, cobalt ray, disinfectant sterilization method, however, the application of these disinfection and sterilization methods in the present invention has some defects, so in contrast, the ethylene oxide sterilization method is preferably used for the sterilization and sterilization in step 3 of the present invention.

The invention further provides a using method of the medical styptic powder, which is applied to wounds to form a dressing. It is emphasized that the aqueous solutions of the present invention are not limited to artificially prepared or applied solutions, but also broadly include the body fluids of the human body itself, and therefore, preferred aqueous solutions of the present invention include, but are not limited to: distilled water, physiological buffer solution, blood, plasma, blood cell preparation, tissue fluid, platelet rich plasma fibrin, or any combination of the above items according to the use requirements.

According to the technical scheme of the invention, the giant salamander skin mucus dry powder is taken as the main component of the medical styptic powder, and the following two methods are taken as examples to illustrate the preferable use method of the medical styptic powder.

The first method is that the medical hemostatic powder and the aqueous solution with proper amount are directly applied to the wound or the bleeding part to directly form hydrogel to become the dressing, and the better wound adhesion and sealing effect can be achieved while hemostasis is achieved. Wherein, when the aqueous solution is applied, it is preferable to take the volumes of the bleeding and the body fluid exudate at the wound or bleeding part into an estimation, then apply a proper amount of the aqueous solution, select a proper powder-water ratio and apply the solution again, so as to maintain a more ideal wound adhesion effect. If the volume of the bleeding and body fluid exudates of the wound or bleeding part and the applied medical hemostatic powder can be successfully gelatinized and achieve better wound hemostasis or adhesion effect, the application of the aqueous solution is not needed.

And secondly, mixing 1 part of the medical styptic powder with 1-6 parts of aqueous solution by weight to form gel with any shape, wherein the shape can meet the special required shapes (three-dimensional shape, surface area, thickness and the like) of the wound or bleeding part so as to meet the treatment requirement. For wounds or bleeding parts with large bleeding amount or tissue fluid exuding, a proper powder-water ratio is selected, so that the gel can absorb the tissue fluid of the wounds or bleeding parts to further swell to form hydrogel to become a dressing after being applied to the wounds, and the effects of stopping bleeding, wet adhesion and closing the wounds are achieved.

The following examples are provided to illustrate the technical solution of the present invention and the corresponding effects achieved.

Example 1: preparing medical styptic powder

Referring to fig. 1, in step 1, the live giant salamander is taken out (see fig. 1a), the giant salamander mucus is obtained by mechanical stimulation (see fig. 1b), and the giant salamander mucus is collected for use. Step 2, preparing the collected giant salamander mucus into dry powder (as shown in fig. 1c) (white powder). In this process, giant salamander mucus was lyophilized as soon as possible after collection, with a time from completion of collection to initiation of lyophilization of no more than 1 hour (h). The freeze-drying speed is set to be 10-15 ℃ per hour, and the temperature is reduced to-20 ℃ within 4 hours. Then pulverizing with a grinder (JXFSTPRP-CL), and sieving to remove the powder with undesirable particle size to obtain dry powder with specific particle size (preferably, not limited to single value or average value). In this embodiment, the dry powder with the desired particle size is obtained by sieving the powder with sieves with 4, 8, 14, 20, 60, 100, 200 and 300 meshes. The powder having a particle size of-4 mesh is obtained by secondary sieving with 4-mesh and 8-mesh sieves to obtain a powder having a particle size that can pass through the 4-mesh sieve but not the 8-mesh sieve (in other words, the obtained particles have a particle size smaller than the mesh size of the 4-mesh sieve but larger than the mesh size of the 8-mesh sieve), and usually a dry powder having a particle size of not more than 4.75mm and a particle size of not less than 2.36mm can be obtained. -8 mesh means particle size less than 8 mesh and greater than 14 mesh; -14 mesh means particle size less than 14 mesh and greater than 20 mesh; -20 mesh means particle size less than 20 mesh and greater than 60 mesh; -60 mesh means particle size less than 60 mesh and greater than 100 mesh; -100 mesh means particle size less than 100 mesh and greater than 200 mesh; -200 mesh means particle size less than 200 mesh and greater than 300 mesh, -300 mesh means particle size less than 300 mesh and greater than 400 mesh. The dry powder is sterilized and then placed in a closed container for refrigeration and preservation for standby use, and the medical hemostatic powder of the embodiment can be obtained. For convenience of illustration and comparison of the technical effects achieved by the present invention, the sieved giant salamander mucus freeze-dried powder is generally referred to as SSAD powder for short or labeled as SSAD powder.

The following illustrates the preparation of SSAD powder in different particle sizes.

The preparation process of the 14-mesh powder comprises the following steps: the mill was started up in advance and precooled to-20 ℃. Putting the freeze-dried giant salamander mucus into a 15ml grinding tank, adding one 6mm grinding bead, grinding once at the frequency of 5HZ for 5 seconds(s), sieving the ground powder for two times by adopting 14-mesh and 20-mesh sieves to obtain the powder with the maximum particle size of less than 14 meshes and more than 20 meshes, wherein the particle size (particle size) is marked as-14 meshes.

The preparation process of the-300-mesh powder comprises the following steps: the method comprises the steps of starting a grinder in advance, pre-cooling to-20 ℃, putting freeze-dried giant salamander mucus into a 15ml grinding tank, adding one 6mm grinding bead, grinding for three times according to the setting of 10s interruption of operation under the frequency of 65HZ, and sieving the ground powder twice by using a 300-mesh and 400-mesh sieve to obtain powder with the maximum particle size of less than 300 meshes and the pore size of more than 400 meshes, wherein the particle size (particle size) is marked as-300 meshes.

The rest giant salamander mucus powder with different particle sizes can be obtained by adjusting the frequency, the grinding time and the grinding times of the grinding machine and replacing sieves with different meshes. Generally, if powder with smaller particle size is needed, the higher the grinding frequency, the more the grinding time, and the more the sieve mesh, which will not be described herein.

In order to avoid degeneration and bacterial breeding caused by long-term storage of the SSAD powder which is not subjected to sterilization treatment, the dry powder is placed in a closed container for storage and standby. If the environmental temperature is higher, the product should be stored in a refrigerator at low temperature.

And 3, sterilizing the dry powder by adopting ethylene oxide. The preferred concrete method is to pack the above obtained SSAD dry powder into a special sterilized packaging bag for ethylene oxide, or to pack the SSAD dry powder into an open container and pack the SSAD dry powder into a special sterilized packaging bag for ethylene oxide after the container opening is loosely stuffed with a cotton ball. The sterilization packaging bag is put into an Ethylene Oxide (EO) sterilization container, and ethylene oxide is used for sterilization, and the sterilization time and temperature are not particularly limited on the principle of not damaging the performance of the dry powder. The sterilized dry powder is then allowed to stand until the residual ethylene oxide has evaporated. Finally, refer to section seven of GB/T16886.7-2001 medical device biological evaluation: "residual amount of ethylene oxide sterilized evaluation of residual amount of ethylene oxide" the residual rate of ethylene oxide was examined. And after the test is qualified, the whole sterilization step is finished.

In addition, the overall effect of the medical styptic powder product is evaluated by aiming at different sterilization methods commonly used at present. For example, the sterilization effect by low temperature and ultraviolet ray is not perfect, the sterilization treatment time is long, and the SSAD powder sterilized by low temperature or ultraviolet ray is easy to hydrolyze, and the storage life is short; the gamma ray method involves radiation, and thus is complicated in operation. Therefore, the present invention preferably adopts ethylene oxide as the sterilization scheme of the medical hemostatic powder of the present invention, and the following experiments are detailed.

Experiment 1: evaluation and effectiveness of different sterilization methods.

According to the SSAD powder obtained in the steps 1 and 2, different disinfection and sterilization methods are adopted in the step 3. The experimental group was sterilized by ethylene oxide, and the control group was sterilized by low temperature (including-20 deg.C, -50 deg.C, -80 deg.C and liquid nitrogen), ultraviolet and irradiation (cobalt 60 gamma rays).

Experimental groups: for the ethylene oxide sterilization method provided by the invention, the specific sterilization steps are preferably executed according to national standard GB18279-2000 'medical instrument ethylene oxide sterilization confirmation and conventional control', and the brief steps comprise: and (3) packaging the ground giant salamander skin mucus dry powder into a special ethylene oxide sterilized packaging bag, or packaging into an open container, loosening the container opening, and filling a cotton ball into the container opening for packaging. The sterilized packaging bag is put into an Ethylene Oxide (EO) sterilized container, the sterilization parameter is 100 percent of ethylene oxide, sterilization is carried out for 6 hours, and then the sterilized powder is placed at room temperature for 72 hours to be used after the ethylene oxide is resolved. Reference GB/T16886.7-2001 "biological evaluation of medical devices" part seven after Sterilization: evaluation of residual amount of ethylene oxide for sterilization, the whole sterilization step is completed after the residual amount of ethylene oxide (residual rate should be less than or equal to 10ppm) is inspected to be qualified.

Control group: low-temperature sterilization, ultraviolet sterilization and irradiation sterilization are respectively adopted. Wherein the temperature and the freezing time of the low-temperature sterilization are respectively as follows: sterilizing at low temperature of-20 ℃ for 24 hours; 24 hours at-50 ℃; 24 hours at-80 ℃; liquid nitrogen for 24 hours. The ultraviolet wavelength used for ultraviolet sterilization is 280-400 nm, and the irradiation time is 24 hours. The radioactive source used for irradiation sterilization is cobalt-60, the irradiation intensity is 600-1000 kilorads, and the irradiation time is 120 minutes.

Testing the sterilization effect after the sterilization is finished: the colony count method after plate coating is adopted. The sample was diluted 1000 times with sterile physiological saline, plated on a plate by the plating method, and the number of Colonies (CFU) on the plate was directly counted after 24 hours of incubation, and the results are shown in table 1.

Testing the adhesion performance of the medical styptic powder after the sterilization is finished: the adhesive ability of the adhesive medical hemostatic powder to the adhesive sample was tested on a universal tester (the adhesive sample was loaded with a 100N load cell at a rate of 1mm/min until completely separated). Experiments were performed with 5 samples for each sterilization method, and the average values are shown in table 1.

Table 1: comparison of the sterilizing effects and adhesion strengths after sterilization of different sterilization methods

As can be seen from the experimental results, the sterilization effects of the ethylene oxide sterilization method and the cobalt ray sterilization method are superior to those of low-temperature sterilization (including-20 ℃, -50 ℃, -80 ℃, and liquid nitrogen) and ultraviolet rays. However, the cobalt ray disinfection method relates to the use of radioactive substances and has certain limitations.

The results of comparing the sterilization performance of the sterilization method for ethylene oxide with other current sterilization schemes according to clinical safety requirements are shown in fig. 2. The results show that the ethylene oxide sterilization method and the cobalt ray sterilization method have sterilization effects superior to those of low-temperature sterilization (including-20 ℃, -50 ℃, -80 ℃, and liquid nitrogen) and ultraviolet rays (as shown in fig. 2a), while the cobalt ray sterilization method has a large influence on the adhesion property, and the ethylene oxide sterilization method can be compatible with good adhesion property (as shown in fig. 2 b).

The result of experiment 1 shows that the ethylene oxide disinfection method can achieve the disinfection effect and does not destroy active substances contained in the original giant salamander skin mucus dry powder, and is the preferred disinfection method.

Unless otherwise stated, the dry powders used in the following examples are all medical hemostatic powders obtained through the above steps 1 to 3, so as to briefly explain the technical solutions and effects of the present invention. The medical styptic powder provided by the invention realizes excellent styptic effect mainly by the characteristic that the dry giant salamander skin mucus powder with specific particle size and the aqueous solution form hydrogel, and is illustrated in the following examples.

Example 2: the medical styptic powder and the aqueous solution form hydrogel.

The sterilized SSAD powder of-8 mesh, -14 mesh, -20 mesh and-60 mesh obtained in the above steps is used as the medical hemostatic powder of this example, and 3 parts of anticoagulated whole blood is mixed with 1 part of medical hemostatic powder according to the weight ratio, and after standing for 3 minutes, the mixture is coagulated into gel. Fig. 3 shows the images of the medical hemostatic powder and the hydrated anticoagulated whole blood in each group.

As is clear from this example, the polypeptide cross-linked network contained in the medical hemostatic powder expands rather than dissolves after being mixed with the aqueous solution, forming a hydrogel body, which is called gelling. During the gelling process, the amino acid residues of the polypeptide chain undergo conformational transition to form a hydrogel-like adhesive, phenolic hydroxyl groups and amino groups are converted to high surface energy or hydrophilic interfaces as hydrogen bond donors, and bioadhesion is promoted by hydrogen bonds and van der waals forces. In addition, benzene rings form strong interactions with substrates through pi-pi electron or cation-pi interactions when contacting low surface energy or hydrophobic interfaces. The aqueous solution may be selected from distilled water, physiological buffers, blood, plasma, blood cell preparations, interstitial fluid, platelet rich plasma fibrin or any combination thereof.

The porous structure of the medical hemostatic powder with different particle sizes (particle diameters) obtained according to the present invention was analyzed by scanning electron microscope (Hitachi, S-3400N II, Japan), and it was revealed by the analysis of scanning electron microscope that the medical hemostatic powder forms a three-dimensional honeycomb structure when mixed with an aqueous solution, as shown in fig. 4 a.

Specifically, an example of the results of analyzing the pore diameter and pore size of the porous structure formed by the medical hemostatic powder with different particle sizes is shown in fig. 4 b.

The medical styptic powder provided by the invention has a porous structure after being gelatinized, so that the medical styptic powder can be beneficial to exchange of nutrient substances and metabolites in tissue fluid in clinical application.

It is worth noting that the scanning electron microscope analysis shows that the average diameter of the holes of the three-dimensional honeycomb structure is correspondingly reduced along with the reduction of the particle size of the used medical hemostatic powder; however, the correlation between the average diameter of the pores and the ratio of the medical hemostatic powder to the aqueous solution (referred to as the water-powder ratio for short) has no statistical significance, which means that the water-powder ratio does not significantly affect the average diameter of the pores of the medical hemostatic powder. More specifically, the medical hemostatic powder with the particle size of-14 meshes is hydrated to form hydrogel, the average diameter of the pores with the particle size of-60 meshes is 116 microns, the average pore diameter of the pores with the particle size of-60 meshes is 37 microns, and the average pore diameter of the pores with the particle size of-300 meshes is 6 microns.

The medical hemostatic powder provided by the invention can absorb water in blood to form hydrogel with a porous structure, and macromolecular substances such as platelets, prothrombin, fibrin and the like in the blood are enriched on the surface of the hydrogel and are rapidly solidified to achieve the hemostatic effect. Fibrin in the blood can form stable crosslinked network structure through hydrogen bond, disulfide bond etc. with protein, polypeptide, mucopolysaccharide in the styptic powder, be favorable to the formation of blood clot faster, and simultaneously, the blood clot mechanical strength who forms is higher, the aquogel of formation has the stickness, can bond and seal the wound, prevent further bleeding, and the aquogel of formation has certain antibacterial and anti-infective action, can alleviate wound infection risk, can be in vivo completely degrading, avoided taking out the aquogel and caused the secondary damage of wound.

In addition, it can be seen from the present example that, no matter what kind of components of the aqueous solution is, the medical hemostatic powder can form hydrogel suitable for different hemostatic purposes in different weight ratios of the medical hemostatic powder to the aqueous solution as long as the medical hemostatic powder is not affected to fully gel, and thus the present invention is not limited thereto. Wherein the weight ratio of the medical styptic powder to the aqueous solution is 1: 1-6 has better gelling performance, and the gelling performance is 1: 2-5 has better gelling performance.

Example 3: and (3) blood coagulation experiments.

The medical hemostatic powder with different meshes prepared according to example 1 was used to test the influence of the particle size of the medical hemostatic powder on the hemostatic effect through an in vitro blood coagulation experiment and a liver hemostatic model. The Yunnan white drug powder and the Celox hemostatic powder which are sold in the market are adopted as comparison. Yunnan white drug powder is from Yunnan white drug group Limited (Kunming), and Celox hemostatic powder is from Medtrade Products Ltd (Electrora House, Crewe Business Park, Crewe, CW16GL, UK.).

Respectively adopts medical hemostatic powder of-4 meshes, -8 meshes, -14 meshes, -20 meshes, -60 meshes, -100 meshes, -200 meshes and-300 meshes. The average particle size was determined by taking a photograph using an electron microscope and analyzing the Image using Image J software. Measuring the solid density of the medical hemostatic powder by a nitrogen adsorption method and a full-automatic gas displacement method true density instrument (Accupyc II 1340, USA), namely measuring the volume of the medical hemostatic powder in an absolute compact state by rho_{Fixing device}Represents; meanwhile, the freshly prepared medical loose hemostatic powder with different particle sizes is weighed, poured into a measuring cylinder to measure the volume of the medical loose hemostatic powder, and then the apparent density of the medical hemostatic powder with different particle sizes is calculated. The following formula I:

apparent density (. rho.)_{Apparent appearance})＝M/V (I)

Wherein: m is the powder mass and V is the powder volume measured by the measuring cylinder.

The porosity of the medical hemostatic powder with different particle diameters is calculated according to the formula II:

void ratio (%) < 100% × (1-. rho)_{Apparent appearance}/[ rho ] solid) (II)

The average particle diameter, the solid density and the porosity data of the medical hemostatic powder with different meshes are shown in the table 2.

Table 2: data of different mesh numbers of medical hemostatic powder

Method of in vitro coagulation assay: respectively placing different mesh medical hemostatic powder and Yunnan white drug powder or Celox hemostatic powder of control group at bottom of test tube 50 mg. 100 microliters of anticoagulant (plasma containing 10% (w/v) sodium citrate) is added dropwise and the mixture is allowed to stand. The test tube was inverted 30 seconds, 60 seconds, and 120 seconds after standing, respectively, with the bottom of the test tube facing up and the mouth facing down. Uncoagulated anticoagulated blood flows down along the test tube wall, and does not flow down if the anticoagulated blood is coagulated. The results are shown in Table 3.

Table 3: results of in vitro coagulation experiments

The establishment method of the liver hemostasis model comprises the following steps: rats were fixed on an operating plate, prepared for skin, sterilized, then midline laparotomy to expose the liver, both flanks were squeezed by hand, the liver was squeezed out of the abdominal cavity and exposed on hemostatic gauze. Using a scalpel to perform a 1X 1cm cross incision along the liver to cause the liver to bleed, and waiting for about 3 seconds, applying the medical hemostatic powder with different particle sizes to the wound to control bleeding. The bleeding volume was measured in the following manner: the aggregate formed by the mixture of the agent and blood after hemostasis of the liver was weighed, the weight of the agent applied to the wound was subtracted, plus the weight of the blood flowing from the liver and absorbed by the underlying gauze. The bleeding time was measured as follows: the time was started after the administration of the drug and stopped when bleeding stopped by visual observation. The results are shown in Table 4.

Table 4: liver hemostasis experimental result of medical hemostatic powder with different particle sizes

Experiments prove that the medical hemostatic powder with the grain diameter of-4 meshes and-300 meshes has better hemostatic effect, wherein the medical hemostatic powder with the grain diameter of-4 meshes to-200 meshes has more ideal hemostatic effect; the medical hemostatic powder with-4 meshes to-100 meshes has more ideal hemostatic effect.

Example 4 in vivo hemostasis test

Adopt liver hemostasis model, disconnected tail hemostasis model, 3 kinds of different models of femoral artery hemostasis model respectively, compare 3 medicaments: the hemostatic effect of medical hemostatic powder (20 mesh), commercially available hemostatic drug Celox (Medtrade Products Ltd, Electrora House, Crewe Business Park, Crewe, CW16GL, UK.) and commercially available Yunnan white drug powder (Yunnan white drug group Co., Ltd., Kunming) are different. The medical styptic powder is prepared in the way of example 1, and is sieved by a 20-mesh sieve, and the maximum grain diameter of the obtained powder is smaller than the pore diameter of the 20-mesh sieve.

The establishment of a model of liver hemostasis is described in example 3 above, which applies 3 agents to the wound about 3 seconds after bleeding from rat liver to control bleeding. Bleeding time and blood loss were recorded. The difference in weight of the powder before and after treatment was used to record blood loss.

The bleeding volume was measured in the following manner: the weight of the agglutinated material mixed with the agent applied to the liver after hemostasis and the blood applied to the wound was weighed and subtracted, and the weight of the blood on the gauze pad was calculated. The blank control group was not dosed with drug. The bleeding time was measured as follows: the agent was administered about 3 seconds after bleeding and the timing was started until the cessation of bleeding was visually observed. 3 experiments were repeated for each agent and averaged. The data are shown in Table 5.

Table 5: comparison of hemostatic effects of three agents

Establishing a femoral artery hemostasis model: fixing rat on operation board, preparing skin, sterilizing, and modeling rat femoral artery injury, stripping rat femoral artery from peripheral tissue, using 21G puncture needle to cause wound bleeding, and immediately applying 3 different agents (-20 mesh medical hemostatic powder, Celox and Yunnan white drug powder) to control bleeding. Bleeding time and blood loss were recorded. The difference in weight of the powder before and after treatment was used to record blood loss.

The bleeding volume was measured in the following manner: the amount of agglutinate mixed with the applied agent and blood after femoral artery hemostasis was weighed, and the amount of applied agent was subtracted, and the amount of blood on the underlying gauze was also calculated. The bleeding time was measured as follows: after bleeding, the timing was started from the time of application of the drug to stop bleeding, and was terminated when bleeding was stopped by visual observation. The blank group was timed from bleeding to the end of visual bleeding cessation. 3 experiments were repeated for each agent and averaged. The data are shown in Table 6.

Table 6: comparison of hemostatic effects of three agents

Establishing a tail-breaking model hemostasis model: rats were first fixed on the surgical plate, and in the rat tail-cutting model, 50% of the tail length was cut with surgical scissors and left to bleed in the air for 15 seconds in order to cause bleeding. Thereafter, the wound was covered with the sample under slight pressure. Bleeding time and blood loss were recorded.

The amount of bleeding is measured by weighing the agglutinating substance mixed with the administered agent and blood after hemostasis, and subtracting the applied agent weight. The bleeding time was measured as follows: 15 seconds after bleeding, the time was counted from the time of application of the drug to stop bleeding, and the time was stopped when the bleeding was visually observed to stop. The blank group was timed from 15 seconds after bleeding to the end of visual cessation of bleeding. 3 experiments were repeated for each agent and averaged. The data are shown in Table 7.

Table 7: comparison of hemostatic effects of three agents

Statistical analysis was performed using SPSS statistics 25 software (IBM corporation, Armonk, NY, USA) and one-way anova was used to calculate whether the differences between the four groups were statistically significant. Analysis results show that the hemostatic effect of the-20-mesh medical hemostatic powder in three animal hemostatic models is obviously superior to that of Yunnan white drug powder (P is less than 0.05) and a blank control group (P is less than 0.01), and is approximately equivalent to that of Celox (P is more than 0.05, and the difference has no statistical significance).

In addition to the good hemostatic performance of the medical hemostatic powder of the present invention, the medical hemostatic powder of the present invention has many characteristics required for wound application, and brings more benefits to wound hemostasis, which is further illustrated in experiments 2 to 6 below.

Experiment 2: and (5) bacteriostatic effect experiments.

Preparing an SSAD powder leaching solution for aerobic bacteria experiments: weighing 1g of sterile SSAD dry powder, adding a proper amount of LB (nutrient broth culture medium) culture medium to ensure that the SSAD dry powder fully absorbs water and swells, then continuously adding the culture medium to finally ensure that the amount of the liquid culture medium reaches 10mL, and placing the mixture in a refrigerator at 4 ℃ for 24 hours to complete leaching.

Preparing an SSAD powder leaching solution used for an anaerobic bacteria experiment: weighing 1g of sterile SSAD dry powder, adding a proper amount of TSB (tryptone soya broth) culture medium to fully absorb water and swell, then continuously adding the culture medium to finally make the amount of the liquid culture medium reach 10mL, and placing the mixture in a refrigerator at 4 ℃ for 24 hours to complete leaching.

The aerobic bacteria are selected from escherichia coli and staphylococcus aureus, and the culture method is as follows: preparing a leaching solution for aerobic bacteria experiments, and centrifuging the leaching solution at 3000r/min for 3 minutes after leaching for 24 hours to obtain a final leaching solution. 10mL of SSAD powder leaching liquor is added into a 50mL centrifuge tube, 500 mu L of bacterial liquid is inoculated, the mixture is uniformly mixed, and the mixture is immediately placed into an incubator.

The anaerobic bacteria are selected from Porphyromonas gingivalis, and the culture method comprises the following steps: preparing a leaching solution for anaerobic bacteria experiment, and centrifuging the leaching solution at 3000r/min for 3 minutes after leaching for 24 hours to obtain a final leaching solution. Adding 10mL of SSAD powder leaching liquor into a 50mL centrifuge tube, adding 500 mu L of sheep blood into each tube according to 5% concentration of sheep blood, respectively inoculating 500 mu L of bacteria liquid with the same concentration, uniformly mixing, and immediately putting into an anaerobic incubator.

The blank control group was prepared as follows. Aerobic bacteria: 10mL of LB medium was added to the centrifuge tube, and a bacterial solution having the same concentration as the experimental group was inoculated as a blank control group. Anaerobic bacteria: 10mL of TSB culture medium and 500 mu L of sheep blood are added into a centrifuge tube, and a bacterial solution with the same concentration as that of the experimental group is inoculated to serve as a blank control group.

The bacteria detection method is as follows. Detection of escherichia coli and staphylococcus aureus: the assay of the absorbance was performed using a microplate reader (EnSpire, PerkinElmer, singapore) by pipetting each set of well-mixed liquids every 2 hours from 0 hour to 100 μ L per well in a 96-well plate, and setting three sub-wells. The absorbance of each group was measured under the condition that OD was 600. The data are shown in tables 8 and 9.

Detection of Porphyromonas gingivalis: the assay of the absorbance was performed by using a microplate reader, and the operation method was that each group of the well-mixed liquids was pipetted every day from day 0, and each 100. mu.L of each well was placed in a 96-well plate, and five wells were set. The absorbance of each group was measured under the condition that OD was 600. The data are shown in Table 10.

Table 8: comparison of bacteriostatic effects of E.coli

Table 9: comparison of bacteriostatic effects of Staphylococcus aureus

Table 10: comparison of bacteriostatic Effect of Porphyromonas gingivalis

The experiment proves that the SSAD powder leaching liquor has a relatively obvious inhibition effect on two common aerobic bacteria (escherichia coli and staphylococcus aureus), and the bacteriostasis time can reach 16 h. The SSAD powder leaching liquor has obvious inhibition effect on the growth of Porphyromonas gingivalis (anaerobe), and the bacteriostasis time can reach more than 5 days.

Experiment 3: and (5) performing an antioxidant experiment.

Firstly, preparing a SSAD powder leaching solution by the following steps: 200mg of the dry powder of the SSAD is put into 2mL of deionized water (the weight ratio is about 1: 10) to be soaked for 7 days, so that soluble substances in the dry powder of the SSAD are fully dissolved, and liquid is taken out to obtain supernatant fluid to obtain the leaching liquor of the SSAD. A1 mg/mL solution of DPPH in ethanol was prepared in situ.

1mL of SSAD powder leaching solution and 1mL of DPPH ethanol solution are adopted in the experimental group; the control group used 1mL of pure water + 1mL of DPPH ethanol solution. The spectrophotometric brightness was set to zero with ethanol and the absorbance value of the control group was A at a wavelength of 517nm measured at 0.5 hour, 1 hour, and 2 hours, respectively₁The absorbance value of the experimental group is A₂. The formula for calculating the radical scavenging rate is shown in the following formula III, and the measurement results are shown in Table 11.

Table 11: free radical clearance calculation table

	Control group (A)₁)	Experimental group (A)₂)	Free radical scavenging rate
				0.5h absorption brightness value	0.461	0.106	77.0％
1h absorption value	0.451	0.099	78.0％
				2h absorption value	0.441	0.082	81.4％

The experiment shows that the SSAD powder leaching liquor can remove most of free radicals in DPPH solution and has an anti-oxidation function.

Experiment 4: cell proliferation and migration promotion experiment.

To simulate the wound healing process in vitro, cell scratch experiments and transwell chamber experiments were performed.

For both of the foregoing experiments, complete media was first prepared: 88 parts of DMEM, 10 parts of fetal bovine serum, 1 part of penicillin and 1 part of streptomycin are uniformly mixed to obtain a complete culture medium. Subsequently, 1mL of complete medium was added to 1mg of SSAD dry powder, and SSAD was immersed in the complete medium for 7 days to dissolve the soluble substances in the SSAD sufficiently, thereby obtaining SSAD powder extract medium with a concentration of 1mg/mL, and 5 SSAD powder-containing extract media with different concentration ratios were further prepared, the concentrations of which were 0.5mg/mL, 0.1mg/mL, 0.05mg/mL, 0.01mg/mL, and 0.005mg/mL, respectively.

In the cell scratch experiment, the test is carried out by respectively adopting HUVES and L929 cells, and the preparation process of the cells is as follows: taking umbilical vein endothelial cells (HUVES) and fibroblasts (L929) in logarithmic growth phase, performing pancreatin digestion to obtain single cell suspension, and counting cells by 6 × 10⁵Per mL; then inoculating 1mL of cell suspension into a 6-well plate, adding 1mL of complete culture medium, culturing in an incubator at 37 ℃, after the cells adhere to the wall and grow over the 6-well plate, scratching (scratching is performed by a 200-L yellow gun head (for later observation and positioning), washing for 2-3 times by PBS after scratching, washing floating cells, finally adding 2mL of the 5 SSAD powder leaching culture media with different concentrations into 5 wells, and adding 2mL of the complete culture medium without the SSAD powder leaching culture medium into one well as serving as a blank control group. All experimental samples were incubated in a 37 ℃ incubator and photographed under a microscope at 0h and 24h, respectively, as represented by the experimental results of 0.1mg/ml SSAD powder extraction medium, as shown in FIG. 5A, with the remaining concentrations not shown.

The area calculation and analysis of the collected pictures are carried out by using Image-J software, and experiments prove that SSAD powder extraction culture media with different concentrations have certain promotion effects on the migration of HUVES and L929 cells, wherein the effect is most obvious at the concentration of 0.1mg/mL (as shown in figure 5B), the cell growth and migration are obviously faster than those of a blank control group, and the difference between the two groups has statistical significance.

Next, the effect of 5 different concentrations of SSAD powder extraction medium on the in vitro migration rate of HUVES and L929 cells was further verified by transwell chamber experiments to reflect its effect on promoting skin and mucosal regeneration.

The test is carried out by adopting HUVES and L929 cells respectively, and the preparation process of the cells is as follows: taking HUVES and L929 in logarithmic growth phase, digesting with pancreatin to obtain single cell suspension, and counting HUVES 2 × 10 cells⁵Count per mL, L929 3X 10⁵Placing a transwell chamber in a 24-hole plate, inoculating 100 mu L of cell suspension into an upper chamber of the transwell, adding 800 mu L of LSSAD powder leaching culture medium into a lower chamber respectively, culturing in an incubator at 37 ℃ for 24h, then carefully taking out the chamber by using tweezers, sucking up upper chamber liquid, transferring the upper chamber liquid into a hole in which about 800 mu L of methanol is added in advance, fixing the upper chamber at room temperature for 30 min, then taking out the chamber, sucking up fixing liquid in the upper chamber, transferring the chamber into a 24-hole plate in which about 800 mu L of 0.1% crystal violet dye liquid is added in advance, dyeing at room temperature for 15-30 min, soaking the chamber for several times by using PBS after dyeing is finished, washing off redundant dye liquid, finally sucking out upper chamber liquid, and carefully wiping off cells on the surface of a bottom membrane of the upper chamber by using a wet cotton rod; after the cell membrane was dried, the membrane was carefully removed with tweezers, transferred to a slide and mounted with a mount, and observed under a 200-fold microscope, as shown in FIG. 5C; and then, randomly intercepting photos of samples of each group of experimental groups, counting cells, and randomly selecting 5 fields for counting each sample. FIG. 5C shows, as a schematic, 1 field of view per sample as previously described; the number of cells obtained by the total counting is shown in Table 12, and finally, the number of cells was obtained and then subjected to a statistical analysis of the difference.

Table 12: transwell cell test results

Experimental results prove that SSAD powder extraction culture media with different concentrations have different degrees of influence on the migration rate of two cells. Wherein for HUVES, the difference between 0.5mg/mL, 0.1mg/mLSSAD powder extraction medium group and blank control group is statistically significant; for L929, the differences between the 0.1mg/mL, 0.05mg/mL, 0.01mg/mLSSAD powder extraction medium group and the blank control group were statistically significant. Wherein the promoting effect of 0.1mg/mLSSAD powder leaching culture medium on two kinds of cells is most remarkable. The experimental results of the experiment show that SSAD powder extraction culture media with different concentrations have different degrees of influence on the migration rates of two cells, wherein the promotion effect of 0.1mg/mLSSAD powder extraction culture media is particularly remarkable.

The results of the in vitro cell experiments show that the SSAD powder extraction culture medium has good cell compatibility and no obvious toxicity to cells, and the 0.1mg/mLSSAD powder extraction culture medium can obviously accelerate the migration of HUVES and L929.

Experiment 5: evaluation and effect of the medical styptic powder in vivo degradation.

SD rats are in prone position under general anesthesia with deep inhalation of isoflurane and the back is aseptically prepared for surgery. A skin incision (3 cm) is prepared outside the spinal axis, and is separated from the underlying subcutaneous tissue, so that enough space is provided for implanting the medical hemostatic powder. 100mg of medical hemostatic powder is mixed with 200-. Peripheral tissues and the whole skin are collected for

histological analysis

3, 7 and 14 days after the operation, and the degradation effect of the medical hemostatic powder is evaluated, wherein the degradation effect is shown in fig. 6.

The H & E and Masson staining (shown in fig. 6a and fig. 6b, respectively) showed mild inflammatory reaction after 3, 7 and 14 days of in vivo implantation of the medical hemostatic powder. After 3 days of implantation, a moderate acute inflammatory reaction was observed in the outermost layer of the implanted medical hemostatic powder, with typical inflammatory cells stained dark blue (i.e., cells relatively dark in color as in fig. 6). After 7 days of implantation, the medical styptic powder begins to lose the integrity, is almost filled by invaded inflammatory cells, and almost no fibrous capsule is observed, which indicates that the host has weak reaction to the medical styptic powder. In addition, after 14 days of implantation, there was almost no medical hemostatic powder remaining at the implanted site, and the skin structure was as normal as the blank control, indicating that the medical aqueous hydrogel could be completely degraded in vivo.

The cellular characteristics of the wound healing area were assessed using lymphocyte (CD3) and macrophage (CD68) marker staining, the results of which are shown in fig. 6 c. In FIG. 6c, i, v, ix are partial enlarged views of 3 days after operation in the same field of view; similarly, ii, vi, x represent images 7 days after the operation; iii, vii, xi represent images 14 days after surgery; iv, viii, xii represent images of 21 days after the operation. As can be seen from fig. 6c, on day 3 after implantation, the infiltration rate of lymphocytes around the graft in the medical hemostatic powder treatment group was 0.23 ± 0.06%, and only a small amount of macrophage infiltration was observed; macrophage infiltration reached a maximum (3.21 ± 0.87%) on day 7; the number of lymphocytes and macrophages infiltrated decreased with the passage of time, and almost completely disappeared by day 21, and the above process was quantitatively counted, and the results are shown in fig. 6d and 6e, and the time points are statistically different; the observation result proves that the medical styptic powder has good biocompatibility, can be completely degraded in vivo, has almost no irritation, and has no obvious immunological rejection.

Experiment 6: the main material content of the medical styptic powder.

The main active ingredient of the medical styptic powder is from giant salamander skin mucus dry powder, the content of main substances of the medical styptic powder is detected by adopting a liquid chromatography technology, and more than 87% of the main active ingredient is various amino acids. See table 13.

Table 13: content of main substances in giant salamander skin mucus dry powder

The above description is only the preferred embodiment and experiment of the present invention, and is not intended to limit the scope of the present invention; while the foregoing is directed to embodiments of the present invention, other and further embodiments of the invention may be devised without departing from the basic scope thereof, and the scope thereof is determined by the claims that follow.

Claims

1. The medical styptic powder is characterized by comprising sterilized giant salamander skin mucus dry powder, wherein the particle size of the giant salamander skin mucus dry powder is smaller than 4 meshes and larger than 300 meshes; the giant salamander skin mucus dry powder has water absorbability, and hydrogel is formed after the giant salamander skin mucus dry powder absorbs aqueous solution.

2. The medical styptic powder according to claim 1, wherein the grain size of the giant salamander skin mucus dry powder is from-4 meshes to-100 meshes.

3. The medical styptic powder of claim 1, wherein the aqueous solution is selected from distilled water, physiological buffer, blood, plasma, blood cell preparation, interstitial fluid, platelet rich plasma fibrin, or any combination thereof.

4. The medical styptic powder as claimed in claim 1, wherein the weight part ratio of the giant salamander skin mucus dry powder to the aqueous solution is 1: 1 to 1: 6.

5. a method of using the medical styptic powder as set forth in any one of claims 1 to 4, the method of using comprising direct application and pre-forming post-application.

6. The method of use of claim 5, wherein said directly applying is applying said medical hemostatic powder to a wound, thereby mixing the medical hemostatic powder with said aqueous solution at said wound and forming said hydrogel into a dressing.

7. The use of claim 5, wherein the pre-forming is performed by preparing the hydrogel from the medical hemostatic powder and the aqueous solution, and then adhering the hydrogel to the wound to form a dressing.

8. The medical styptic powder as claimed in claim 1, wherein the sterilized giant salamander skin mucus dry powder is sterilized by ethylene oxide sterilization or irradiation sterilization.