WO2019011333A1

WO2019011333A1 - Haemostatic material, haemostatic fibre membrane, and haemostatic product

Info

Publication number: WO2019011333A1
Application number: PCT/CN2018/095636
Authority: WO
Inventors: 李林静; 邓坤学; 袁玉宇
Original assignee: 广州迈普再生医学科技股份有限公司
Priority date: 2017-07-14
Filing date: 2018-07-13
Publication date: 2019-01-17

Abstract

Disclosed are a haemostatic material, a haemostatic fibre membrane, and a haemostatic product. The haemostatic material comprises a structural body having an interlaced structure formed by mutually overlapping a plurality of nano short fibres, the nano short fibres being derived from cross-linked nanofibre material, the diameter of the nano short fibres being between 1 nm and 1000 nm; the structural body is: a nanofibre cluster (a), the size of the nanofibre cluster (a) in any direction in three-dimensional space from the start point of the geometric centre thereof being in the range of 5 μm to 500 μm; and/or the size of the nanofibre cluster (a) in the haemostatic material represented by a median diameter D50 is between 100 μm and 500 μm, preferably between 200 μm and 400 μm; the nanofibre cluster (a) has a porous structure, and the length of the nano short fibres in the nanofibre cluster (a) is less than 1000 μm; or a microfibre (b), the diameter of the microfibre (b) being 1 μm to 500 μm and the length thereof being 0.5 mm to 10 mm, and the length of the nano short fibres in the microfibre (b) being less than 10 mm. The haemostatic material of the present application can be highly fluffy, having excellent tissue adhesion properties and evident haemostatic effects.

Description

Hemostatic material and hemostatic fiber membrane and hemostatic product

Technical field

The present disclosure relates to a hemostatic material as well as a hemostatic fibrous membrane and a hemostatic product, and belongs to the field of biomedical materials.

Background technique

Biomedical materials are a kind of high-tech materials developed in the past 30 years. Among them, hemostatic materials have gradually attracted the attention of the medical community with the increase of accidents such as traffic accidents, severe burns and major disasters. With the rapid development of modern science and technology, the research on hemostatic materials has made very rapid progress, various new hemostatic materials have emerged, and the performance has been greatly improved. At present, commonly used local hemostatic materials are fibrin glue, thrombin powder, gelatin sponge, collagen sponge, chitosan sponge, oxidized cellulose, microfiber collagen, alginic acid fiber, zeolite, cyanoacrylate, plant polysaccharide powder. Wait. Biomedical hemostatic materials with exact hemostatic effect, convenient use, good biocompatibility and ability to control the degradation rate have become the main target of attention and research.

Commonly used hemostatic materials include various forms, such as powdered, such as thrombin lyophilized powder, plant polysaccharide powder, zeolite powder, microfiber collagen powder; solution type, such as cyanoacrylate, chitosan solution; It has a liquid type, but it can form a gel or colloid on the wound surface, such as fibrin glue, glutaraldehyde-albumin Bioglue; there are membranes, such as chitosan membrane, polylactic acid membrane; and sponge-like, such as collagen sponge , gelatin sponge, microfiber collagen sponge, fibrin glue and so on. Various forms of hemostatic materials have their own advantages, but also have their own application advantages, mainly based on the type of wound and clinical treatment.

Among them, the powdered hemostatic material has wide application in the field of surgical hemostasis because of its simple operation and no limitation on the wound site. In the prior art, the powdery hemostatic material is mainly a polysaccharide microsphere or a starch granule, and the microporation of the surface of the material is realized by ultrasonic method, moist heat treatment, microwave method, mechanical method or enzyme perforation technology, and the material is improved. The specific surface area and hydrophilic properties act as molecular sieves on the surface of the wound, and the concentration of blood coagulation factors is increased by adsorbing moisture in the blood, thereby accelerating the occurrence of coagulation mechanism, thereby achieving hemostasis. However, the current hemostatic powder has problems of poor adhesion, relatively dense, complicated preparation process, and needs to be prepared in advance, wasting a lot of time for rescue and hemostasis.

In addition, the existing membranous hemostatic materials mainly include two types, one is directly obtained by electrospinning, and the other is obtained by coating a solution or drying in a mold. Generally, the membranous hemostatic materials prepared by the two methods have unsatisfactory hemostatic effects and cannot meet the needs of clinical use. The fiber membrane directly obtained by electrospinning has poor adhesion to the wound surface and adhesion, and the liquid absorption ability is not high, and the hemostatic membrane obtained by directly drying the solution into a membrane has poor gas permeability. The rate of aspiration is also slow, and the effect of rapid and efficient hemostasis cannot be achieved.

Summary of the invention

technical problem

In view of the above, the technical problem to be solved by the present disclosure is to provide a hemostatic material which has a high specific surface area, excellent adhesion properties, and remarkable hemostatic effect.

Further, the hemostatic material of the present disclosure also has high loft or porosity, high water absorption, and good biocompatibility, and can be rapidly degraded and absorbed by the organism.

Further, the present disclosure also provides a hemostatic fibrous membrane which has excellent adhesion properties and remarkable hemostatic effect, and can quickly separate the hemostatic fibrous membrane by tear according to the actual dosage requirement, and has the characteristics of being convenient to use.

solution

The present disclosure firstly provides a hemostatic material comprising a structure having a staggered structure formed by overlapping a plurality of nano-small fibers having a diameter of between 1 nm and 1000 nm; The body is derived from a crosslinked nanofiber material, the structure being a nanofiber cluster (a) or a microfiber (b),

The nanofiber cluster (a) has a dimension in any direction of the three-dimensional space starting from a geometric center thereof in a range of 5 μm to 500 μm; and/or a median diameter D50 of the nanofiber cluster (a) in the hemostatic material The size indicated is between 100 μm and 500 μm, preferably between 200 μm and 400 μm, and the nanofiber cluster (a) has a porous structure, and the length of the nano short fibers in the nanofiber cluster (a) is less than 1000 μm;

The microfibers (b) have a diameter of 1 μm to 500 μm and a length of 0.5 mm to 10 mm, and the length of the nano short fibers in the microfibers (b) is 10 mm or less.

The hemostatic material of the present disclosure, wherein a specific surface area of the hemostatic material ^{^{4m 2 / g ~ 50m 2 /}} g, a water absorption greater than 1500%.

The present disclosure also provides a hemostatic fibrous membrane, the hemostatic fibrous membrane comprising microfibers (b');

The microfibers (b') are derived from a crosslinked nanofiber material, the microfibers (b') having a diameter between 0.1 mm and 1 mm and a length of 20 mm or less;

The microfiber (b') has a staggered structure formed by overlapping a plurality of nano short fibers;

The nano short fibers have a diameter of between 1 nm and 1000 nm and a length of 10 mm or less.

A hemostatic fibrous membrane according to the present disclosure, wherein a surface of the hemostatic fibrous membrane has a plurality of concave portions and/or convex portions.

The hemostatic fiber membrane according to the present disclosure, wherein the hemostatic fibrous membrane has an areal density of 50 g/m ² to 500 g/m ² , preferably 100 g/m ² to 300 g/m ² ; and the specific surface area of the hemostatic fibrous membrane is 5m ² /g～30m ² /g; the hemostatic fiber membrane has a bulkiness of 500 to 5000 cm ³ /g, preferably 1000 to 3000 cm ³ /g; and the hemostatic fiber membrane has more than 1500%, preferably 1700% Water absorption between 2500%.

The present disclosure also provides a method for preparing a hemostatic material according to the present disclosure or a hemostatic fibrous film according to the present disclosure, comprising the steps of:

Electrospinning step: preparing a nanofiber material by electrospinning;

Cross-linking step: crosslinking the nanofiber material in the presence of a crosslinking agent to obtain a crosslinked nanofiber material;

Shearing step: shearing the crosslinked nanofiber material.

The present disclosure further provides a hemostatic article comprising: a hemostatic material according to the present disclosure, or a hemostatic fibrous membrane of the present disclosure, or a hemostatic material or a hemostatic fibrous membrane prepared by the method of the present disclosure.

Beneficial effect

The hemostatic material of the present disclosure can be highly fluffy, has excellent tissue adhesion performance and remarkable hemostatic effect, and promotes the process of mutual fusion with the tissue, and further improves the water absorption capacity and tissue adhesion ability of the material.

Further, the hemostatic material of the present disclosure also has a high specific surface area, good biological properties and remarkable hemostatic effect, can not only be quickly degraded and absorbed by the organism, but also has convenient clinical use, and can be used for hemostasis in wound healing and clinical surgery.

Further, the hemostatic fiber membrane of the present disclosure is simple and convenient to operate during use, and the material can be quickly separated according to actual usage requirements. Moreover, the surface of the hemostatic fibrous membrane may have a microscopic and/or macroscopic relief structure to better form a bond with the tissue surface while also making the specific surface area higher.

Further features and aspects of the present disclosure will become apparent from the following detailed description of exemplary embodiments.

DRAWINGS

The accompanying drawings, which are incorporated in FIG

1 shows a SEM electron micrograph of a nanofiber cluster (a) in a hemostatic product of Example 1 of the present disclosure.

2 is a graph showing a comparison of hemostatic effectiveness of the hemostatic product of the embodiment 1 of the present disclosure and the commercially available product A.

Fig. 3 is a view showing the pathology of degradation of the hemostatic product of the embodiment 1 of the present disclosure in an animal.

Figure 4 shows a pathological map of the degradation of commercially available product A in animals.

Figure 5 shows a SEM electron micrograph of the hemostatic product of Example 7 of the present disclosure.

Figure 6 shows a SEM electron micrograph of a single microfiber (b) or microfiber (b') of the present disclosure.

Fig. 7 is a graph showing a comparative analysis of hemostatic effectiveness of the hemostatic product and the commercially available product 1 of Example 7 of the present disclosure.

Fig. 8 is a view showing the hemostasis and the in vivo degradation of the hemostatic product and the commercially available product 1 in the animal experiment according to Example 7 of the present disclosure.

Figure 9 is a plan view showing a hemostatic product of Example 9 of the present disclosure.

Fig. 10 is a view showing a comparison of hemostatic effectiveness of the hemostatic product of the embodiment 10 of the present disclosure and a commercially available product.

Figure 11 is a graph showing the degradation of the hemostatic product prepared in Example 9 of the present disclosure when implanted in an animal for one week.

Fig. 12 is a view showing the degradation of the hemostatic product prepared in Example 9 of the present disclosure when implanted in an animal for 2 weeks.

Figure 13 is a graph showing the degradation of the hemostatic product prepared in Example 9 of the present disclosure when implanted in an animal for 4 weeks.

Fig. 14 is a view showing the degradation of the membranous hemostatic material of the control group implanted in the animal for one week.

Fig. 15 is a view showing the degradation of the membranous hemostatic material of the control group implanted in the animal for 2 weeks.

Fig. 16 is a view showing the degradation of the membranous hemostatic material of the control group implanted in the animal for 4 weeks.

Detailed ways

Various exemplary embodiments, features, and aspects of the present disclosure are described in detail below with reference to the drawings. The word "exemplary" is used exclusively herein to mean "serving as an example, embodiment, or illustrative." Any embodiment described herein as "exemplary" is not necessarily to be construed as preferred or preferred.

In addition, numerous specific details are set forth in the Detailed Description of the <RTIgt; Those skilled in the art will appreciate that the present disclosure may be practiced without some specific details. In other instances, well-known methods, means, components, and circuits are not described in detail to facilitate the disclosure.

First embodiment

A first embodiment of the present disclosure provides a hemostatic material. The hemostatic material comprises a structure having a staggered structure formed by overlapping a plurality of nano-short fibers having a diameter between 1 nm and 1000 nm; the structure being derived from a cross-linked nanofiber material The structure is a nanofiber cluster (a) or a microfiber (b),

The nanofiber cluster (a) has a dimension in any direction of the three-dimensional space starting from a geometric center thereof in a range of 5 μm to 500 μm; and/or a median diameter D of the nanofiber cluster (a) in the hemostatic material ₅₀ represents a size between 100 μm and 500 μm, preferably between 200 μm and 400 μm, the nanofiber cluster (a) has a porous structure, and the length of the nano short fibers in the nanofiber cluster (a) is less than 1000 μm;

Among them, the crosslinked nanofiber material can be obtained by crosslinking and modifying the nanofiber material.

The structure can be obtained by shearing the crosslinked nanofiber material. The nano short fibers can be formed simultaneously by shearing, so that the structure has a staggered structure formed by overlapping a plurality of nano short fibers.

The nanofiber material of the present embodiment may be derived from a polymer material having biocompatibility and biodegradable absorption, and more preferably a hydrophilic polymer material. For example, the polymer material may include one or a combination of two or more of collagen, chitosan, hyaluronic acid (HA), alginate, cellulose, and derivatives thereof.

The nanofiber material of the present embodiment may be formed by interlacing fiber filaments. Preferred are nanofiber materials prepared using an electrospinning process. The nanofiber material may be a fiber mass, a fiber bundle or a fiber membrane (ie a textile fiber membrane), preferably a fibrous membrane.

The principle of electrospinning is to apply a high voltage to the polymer liquid during electrospinning to introduce a charge into the liquid. When the charge in the liquid accumulates to a certain amount, the liquid forms a Taylor cone in the nozzle, and overcomes the surface tension to form a liquid jet under the action of an applied electric field force, and then the jet is in common with electrostatic repulsion, Coulomb and surface tension. Under the action, the polymer jet moves along an irregular spiral path. The jet is drawn and stretched in a very short time, and as the solvent evaporates or the heat is lost, the polymer jet solidifies to form micro/nanofibers. In the electrospinning process, many parameters affect the final electrospinning fiber. By controlling the process parameters, micro/nano fibers of different sizes, shapes and structures can be prepared.

In the electrospinning process of the present embodiment, the process parameters have an influence on the nanofiber material obtained by electrospinning, and by controlling the process parameters, nanofiber materials having different sizes, shapes and different structures can be prepared. This embodiment has no particular requirements for the manner of electrospinning, and may be an electrospinning method commonly used in the art. Specifically, in the present embodiment, the polymer material is dissolved in a suitable solvent to prepare a spinning dope of the polymer material; then, the spinning dope is spun by electrospinning into a nanofiber material interwoven with fiber filaments. Preferably, the nanofiber material has a porous structure.

The crosslinked nanofiber material of the present embodiment can be obtained by subjecting a nanofiber material to a crosslinking modification treatment. Specifically, the crosslinking modification is a crosslinking modification in the presence of a crosslinking agent, thereby obtaining a crosslinked product having a suitable degree of crosslinking and uniformity.

In the present embodiment, the purpose of the cross-linking modification is to enable the hemostatic material to maintain a large amount of liquid while maintaining the fiber form, and is not quickly dissolved or dispersed by the absorbed body fluid. In addition, the excessively high degree of crosslinking affects the water absorption rate, the flexibility, and the like. In order to obtain a more suitable degree of crosslinking, the mass ratio of the crosslinking agent to the nanofiber material in the present embodiment is 0.01 to 3:1, for example: It may be from 0.01 to 2:1, may be from 0.1 to 1:1, may be from 0.1 to 3:1, and may also be from 0.5 to 2:1.

The nanoshort fiber of the present embodiment has a diameter of 1 nm to 1000 nm, a length of generally 10 mm or less, a length of 8 mm or less, and a length of 5 mm or less or 1000 μm or less. In general, the nanofibers in the narrow sense are nanofibers having a diameter ranging from 1 nm to 100 nm, and the nanofibers in a broad sense also include nanocomposite fibers, that is, conventional fibers in which zero-dimensional or one-dimensional nanomaterials are combined with conventional fibers. For polymer nanofibers, nanofibers in this embodiment are referred to as diameters because they have produced many special properties on the scale of 1000 nm, such as large surface area, easy surface functionalization, and superior mechanical properties. Nanofibers in the range of 1 nm to 1000 nm.

In a specific embodiment of the present embodiment, a hemostatic material (A) is provided. The hemostatic material (A) comprises a nanofiber cluster (a) having a staggered structure formed by overlapping a plurality of nano short fibers; the nanofiber cluster (a) originating from Associated with nanofiber materials.

The nanofiber cluster (a) can be obtained by shearing a crosslinked nanofiber material. The nano short fibers can be formed simultaneously by shearing, so that the nanofiber clusters (a) have a staggered structure formed by overlapping a plurality of nano short fibers.

The hemostatic material (A) is macroscopically powdery or granular, and can be applied to the wound surface by spraying or spraying, and is especially suitable for a cavity-like wound or a narrow wound.

As shown in Fig. 1, in the hemostatic material (A), the size of the nanofiber cluster (a) represented by the median diameter D ₅₀ is between 100 μm and 500 μm, preferably between 200 μm and 400 μm. In the present embodiment, the median diameter D ₅₀ is the particle diameter corresponding to the nanofiber cluster (a) in the hemostatic material (A) when the cumulative particle size distribution percentage reaches 50%.

The hemostatic material (A) has a bulk density of less than 0.06 g/cm ³ , preferably from 0.025 g/cm ³ to 0.05 g/cm ³ . The hemostatic material (A) has a low bulk density, a high bulkiness and an ultra-high specific surface area. The hemostatic material (A) has a porosity of 50% to 90%, preferably 70% to 90%. The hemostatic material (A) has a specific surface area of ^{^{4m 2 / g ~ 50m 2 /}} g, preferably ^{^{10m 2 / g ~ 40m 2 /}} g. The hemostatic material (A) has a water absorption rate of more than 1500%, preferably between 2000% and 3000%, and has a high water absorption property.

As shown in Fig. 1, the nanofiber cluster (a) of the hemostatic material (A) has a porous structure. The nanofiber cluster (a) can be obtained by shearing a crosslinked nanofiber material. The nano short fibers are simultaneously formed by shearing, so that the nanofiber clusters (a) have a staggered structure formed by overlapping a plurality of nano short fibers. Due to the staggered structure formed by overlapping of the plurality of nano short fibers and the small size of the nanofiber cluster (a), the nanofiber cluster (a) is in a fluffy state, thereby increasing the nanofiber cluster (a) Specific surface area.

The nanofiber cluster (a) has a dimension from any geometric direction of the three-dimensional space starting from a geometric center thereof in a range of 5 μm to 500 μm; specifically, the nanofiber cluster (a) is oriented in any direction of the three-dimensional space with its geometric center as a starting point. The size may be between 5 μm and 50 μm, may be between 5 μm and 100 μm, may be between 5 μm and 200 μm, may be between 5 μm and 250 μm, may be between 5 μm and 300 μm, and may be between 5 μm and 350 μm. It may be between 5 μm and 400 μm, may be between 5 μm and 450 μm, and may be between 5 μm and 500 μm.

In the nanofiber cluster (a) of the hemostatic material (A), the length of the nano short fibers is 1000 μm or less. Crosslinking in the crosslinked nanofiber material is crosslinking in the presence of a crosslinking agent, preferably, the mass ratio of the crosslinking agent to the nanofiber material is from 0.01 to 2:1, preferably from 0.1 to 1:1.

Since the hemostatic material (A) of the present embodiment has a small bulk density, a high specific surface area, a high porosity, and a high water absorption rate, the blood in the blood can be quickly absorbed in the bleeding wound, and the red blood cells and blood coagulation in the blood can be further improved. The concentration of factors, etc., accelerates the endogenous coagulation mechanism and improves the hemostatic effect.

The hemostatic material (A) of the present embodiment has excellent tissue adhesion performance and remarkable hemostatic effect, and excellent tissue adhesion can ensure that the material closely adheres to the wound during hemostasis, prevents blood from being washed away, and significantly improves hemostasis. The effect and promote the process of its integration with the organization.

Further, since the hemostatic material (A) has a high specific surface area and a high porosity, the blood in the blood can be quickly absorbed in the blood environment, the concentration of the effective blood coagulation factor in the blood is increased, and the endogenous and exogenous body of the body are activated. The mechanism of hemostasis accelerates the occurrence of blood coagulation, thereby achieving rapid hemostasis.

In another specific embodiment of the present embodiment, a microfibrous hemostatic material is further provided. The microfibrous hemostatic material comprises microfibers (b) having a staggered structure formed by overlapping a plurality of nanoshort fibers; the microfibers (b) are derived from crosslinked nanometers Fiber material.

The microfibers (b) can be obtained by shearing the crosslinked nanofiber material. The nano short fibers can be formed simultaneously by shearing, so that the micro fibers (b) have a staggered structure formed by overlapping a plurality of nano short fibers.

As shown in Fig. 5, the microfibrous hemostatic material of the present embodiment includes microfibers (b). The microfiber-form hemostatic material has a bulk density of less than 0.03 g/cm ³ , preferably 0.01 to 0.025 g/cm ³ . The microfibrous hemostatic material has a low bulk density, a high bulkiness and an ultra-high specific surface area. The specific surface area microfibre material hemostatic state may ^{^{4m 2 / g ~ 50m 2 /}} g, may be ^{^{6m 2 / g ~ 30m 2 /}} g. And the microfibrous hemostatic material has a water absorption ratio of more than 1500%, preferably between 2000% and 2500%, and has high water absorption performance.

Since the microfibrous hemostatic material of the present embodiment has an ultra-high specific surface area and an ultra-high water absorption property, the blood in the blood can be quickly absorbed in the bleeding wound, and the concentration of red blood cells, blood coagulation factors and the like in the blood can be increased, and the endogenous source can be accelerated. Sexual coagulation mechanism to improve hemostasis.

In the microfibrous hemostatic material of the present embodiment, the microfibrous structure is more likely to form a physical compression with a certain strength relative to the powdery microfibrous hemostatic material, and the powdery material is prevented from bleeding more. It is easy to be washed away.

The microfiber (b) of the microfibrous hemostatic material can be obtained by shearing the crosslinked nanofiber material. As shown in FIG. 6, the nano short fibers are simultaneously formed by shearing, so that the microfibers (b) have a staggered structure formed by overlapping a plurality of nano short fibers. Due to the staggered structure formed by overlapping of the plurality of nano short fibers and the small size of the microfibers (b), the microfibrous hemostatic material is in a more fluffy state, thereby further increasing the ratio of the microfibrous hemostatic material. Surface area.

The microfiber (b) in the present embodiment means a fiber having a smaller size. Specifically, the microfibers (b) have a diameter of 1 μm to 500 μm, may be between 1 μm and 400 μm, may be between 1 μm and 300 μm, may be between 1 μm and 200 μm, and may be between 1 μm and 100 μm; 0.5 mm to 10 mm, may be between 0.5 mm and 8 mm, may be between 0.5 mm and 5 mm, and may be between 1 mm and 5 mm. Since the length of the microfiber (b) of the present embodiment is between 0.5 mm and 10 mm, it is easily shaped to be suitable for various wounds for more effective hemostasis, such as clustering, lumps, and the like.

Further, the ratio of the length to the diameter of the microfiber (b) of the present embodiment may be in the range of 1 to 10,000, preferably in the range of 5 to 8,000, or in the range of 8 to 5,000, and in the range of 5 to 8,000. Within the scope of the microfibrous hemostatic material can maintain a high loft and specific surface area, enhance the adhesion strength and hemostatic effect between the wound surface, and facilitate the doctor to directly hold the microfibrous hemostatic material through the forceps To the wound.

In the microfiber (b) of the present disclosure, the length of the nano short fibers is generally 10 mm or less, may be 8 mm or less, and may be 5 mm or less. Crosslinking in the crosslinked nanofiber material is crosslinking in the presence of a crosslinking agent, and the mass ratio of the crosslinking agent to the nanofiber material is from 0.1 to 3:1, preferably from 0.5 to 2:1.

The microfibrous hemostatic material of the present embodiment can be highly fluffy, has excellent tissue adhesion performance and remarkable hemostatic effect, and not only promotes the process of mutual fusion with the tissue, but also further improves the water absorption capacity and tissue adhesion of the material. Ability, and maintains fiber morphology well while a large amount of liquid absorption.

Further, the hemostatic material (for example, the hemostatic material (A) or the microfibrous hemostatic material) of the present embodiment further contains a drug. Preferably, the drug comprises one or a combination of two or more of thrombin, a blood coagulation factor, and a growth factor. By loading the drug, not only can the hemostatic properties of the hemostatic material be improved, but also the properties of promoting rapid wound healing and anti-adhesion.

Second embodiment

A second embodiment of the present disclosure provides a hemostatic fiber membrane. The hemostatic fibrous film comprises microfibers (b') having a staggered structure formed by overlapping a plurality of nano short fibers. The microfibers (b') are derived from a crosslinked nanofiber material.

Among them, the nanofiber material of the present embodiment may be the nanofiber material in the first embodiment. The crosslinked nanofiber material in the present embodiment may be the crosslinked nanofiber material in the first embodiment. The nano short fiber in the present embodiment may be the nano short fiber in the first embodiment.

The hemostatic fibrous film of the present embodiment has a high specific surface area and water absorption, and has excellent tissue adhesion properties and a remarkable hemostatic effect.

The microfiber (b') of the present embodiment can be obtained by shearing a crosslinked nanofiber material. The microfibers (b') have a diameter of between 0.1 mm and 1 mm and a length of 20 mm or less. In the present embodiment, the microfibers (b') have a staggered structure in which a plurality of nano short fibers are overlapped with each other. Due to the staggered structure formed by overlapping of the plurality of nano short fibers and the small size of the microfibers (b'), the hemostatic fiber membrane is in a relatively bulky state, so that the specific surface area of the hemostatic fibrous membrane can be appropriately increased.

In the microfiber (b') of the present embodiment, the length of the nanospun fiber is generally 10 mm or less, and may be 8 mm or less, and may be 5 mm or less. Crosslinking in the crosslinked nanofiber material is crosslinking in the presence of a crosslinking agent, and the mass ratio of the crosslinking agent to the nanofiber material is from 0.1 to 3:1, preferably from 0.5 to 2:1.

As shown in FIG. 9, the hemostatic fibrous film of the present embodiment has a staggered structure formed by overlapping a plurality of microfibers (b'), and has a plurality of concave portions on the surface thereof, so that the surface of the material is rough, and the surface of the tissue is better. The attachment is formed while also making the specific surface area higher than the existing hemostatic fiber membrane.

The hemostatic fibrous film of the present embodiment has a bulkiness of 500 to 5,000 cm ³ /g, preferably 1,000 to 3,000 cm ³ /g. The hemostatic fibrous film has an areal density of 50 g/m ² to 500 g/m ² , preferably 100 g/m ² to 300 g/m ² . The hemostatic fibrous film has a specific surface area of from 5 m ² /g to 30 m ² /g, preferably from 10 m ² /g to 20 m ² /g, and has a high specific surface area. And the hemostatic fibrous film has a water absorption ratio of more than 1500%, preferably between 1700% and 2500%, and has high water absorption performance.

Since the hemostatic fibrous membrane of the present embodiment has the characteristics of high specific surface area, high bulkiness, and high water absorption rate, the blood in the blood can be quickly absorbed in the bleeding wound, and the concentration of red blood cells and blood coagulation factors in the blood can be increased, and the endogenous source can be accelerated. Sexual coagulation mechanism to improve hemostasis.

Further, the hemostatic fibrous film of the present embodiment further contains a drug. Preferably, the drug comprises one or a combination of two or more of thrombin, a blood coagulation factor, and a growth factor. By loading the drug, not only can the hemostatic properties of the material be improved, but also the rapid healing and anti-adhesion properties of the wound can be promoted.

The hemostatic fibrous membrane has a thickness of between 0.05 mm and 2 mm, preferably between 0.3 mm and 1 mm. The hemostatic fiber membrane has a certain thickness and is more likely to form a physical compression with a certain strength, so as to prevent the powdery material from being easily washed away in the case of more bleeding.

In addition, the hemostatic fibrous membrane of the present embodiment is a non-woven hemostatic fibrous membrane, and since the surface thereof has a plurality of concave portions and/or convex portions, the appearance thereof exhibits a certain network structure, and the hemostatic fibrous membrane of the present embodiment may not pass static electricity. The method of spinning is prepared.

In addition, the hemostatic fiber membrane of the present embodiment has a small tear strength, and the material can be quickly torn by hand according to the actual dosage requirement during use to achieve the purpose of hemostasis, and the operation is simple and convenient during use.

The hemostatic fiber membrane of the present embodiment not only promotes the mutual fusion process with the tissue, but also further improves the water absorption capacity and the tissue adhesion ability of the material, and can well maintain the three-dimensional biomimetic structure while a large amount of liquid absorption.

Further, the hemostatic fiber membrane has good biocompatibility and degradability, can not only be quickly degraded and absorbed by the organism, but also has convenient clinical use, and can be used for hemostasis in wound healing and clinical operation.

Third embodiment

A third embodiment of the present disclosure provides a hemostatic material of the first embodiment of the present disclosure or a method of producing the hemostatic fibrous film of the second embodiment, comprising the steps of:

Electrospinning step: preparing the nanofiber material by electrospinning;

Shearing step: shearing the crosslinked nanofiber material.

The term "crosslinking" as used herein has the same or similar meaning as "crosslinking modification". In the process of "crosslinking", some features of "modification" may be attached, and in the present disclosure, for the sake of simplicity, Use "crosslinking" instead of "crosslinking modification".

In the electrospinning step, a fiber raw material is prepared in advance, and the fiber raw material is dissolved in a suitable solvent to prepare a spinning dope of a fiber raw material having a certain concentration. Wherein, the fiber raw material may be the polymer material in the first embodiment. The specific concentration of the solvent to form the solution is not particularly limited as long as it can satisfy the requirements of the subsequent electrospinning process. For example, a suitable solvent may be one or two of trifluoroethanol, hexafluoroisopropanol, trifluoroacetic acid, cyclohexanone, acetone, methyl ethyl ketone, tetrahydrofuran, chloroform, glacial acetic acid, formic acid, propionic acid or water. More than one combination.

The desired nanofiber material can be prepared by adjusting the spinning parameters during the electrospinning process. For example, voltage, extrusion flow rate and electric field receiving distance, spinning environment, and the like. Preferably, the electrospinning process parameter in the embodiment may be: a pressure of 10 to 40 kV, a solution extrusion flow rate of 0.1 to 15 mL/h, an electric field receiving distance of 5 to 30 cm, and a spinning environment relative temperature of 60%. Hereinafter, the ambient temperature is 10 to 40 °C.

In addition, it is conceivable to load the drug in the spinning dope or in the electrospinning process, which may include one or a combination of two of coagulation factors, growth factors and the like. Not only can improve the hemostatic properties of the material, but also promote the rapid healing of the wound, anti-adhesion and other properties.

In the crosslinking step, the crosslinking agent selected includes one or a combination of two or more of carbodiimide, N-hydroxysuccinimide, genipin, and an aldehyde compound.

The cross-linking agent may be selected from carbodiimide and/or N-hydroxysuccinimide in view of the toxicity of the cross-linking agent to the organism and the crosslinking effect. According to the study of the present embodiment, the crosslinking effect of the carbodiimide can be further improved by using N-hydroxysuccinimide. Therefore, the chemical crosslinking agent is preferably a combination of carbodiimide and N-hydroxysuccinimide. More preferably, the mass ratio of the carbodiimide to the N-hydroxysuccinimide is from 1 to 4:1.

In general, the crosslinking modification is carried out in a solution, and the solvent to be subjected to the crosslinking modification treatment in the present embodiment is not particularly limited as long as the requirements for the crosslinking modification reaction can be satisfied. In the present embodiment, the solvent subjected to the crosslinking modification treatment may be a mixed solution of an alcohol and water of different mass ratios. Among them, the alcohol is preferably ethanol, and more preferably, the mass ratio of ethanol to water may be 70% or more.

Further, the crosslinking modification can be controlled by adjusting the reaction conditions of the crosslinking treatment and the amount of the crosslinking agent. For example, the crosslinking treatment temperature, the crosslinking treatment time, the mass ratio of the crosslinking agent to the nanofiber material, the ratio of the mass of the crosslinking agent to the volume of the solvent, and the like. In addition, the degree of cross-linking can be adjusted to meet the requirements of different trauma or clinical surgery on the degradation cycle.

The reaction conditions in the crosslinking step in the present embodiment may be such that the crosslinking treatment temperature is between 1 and 50 ° C, preferably between 4 and 30 ° C. The crosslinking treatment time is between 1 and 72 h, preferably between 6 and 24 h. The mass ratio of the mass of the chemical crosslinking agent to the nanofiber material is 0.01 to 3:1; for example, it may be 0.1 to 3:1, may be 0.5 to 2:1, or may be 0.01 to 2:1. It can be from 0.1 to 1:1. The ratio of the mass of the chemical crosslinking agent to the volume of the solvent is from 0.1 to 10:100, preferably from 1 to 5:100. The degree of crosslinking can be further controlled by controlling the above reaction conditions.

Further, in the present embodiment, the fiber raw material may be subjected to a crosslinking treatment and then treated by an electrospinning technique, preferably by first performing an electrospinning technique, and then subjecting the nanofiber material to a crosslinking treatment.

In the shearing step of the present embodiment, the shearing process includes a pre-shearing step. In a specific embodiment, for example, when preparing the hemostatic material (A) of the first embodiment, the pre-shearing step is performed on the cross-linked nanofiber material at a rotational speed of 10,000 to 20,000 rpm/min. The preliminary shearing treatment gives a pre-shear. In general, the treatment time of the pre-shearing step is from 10 to 60 min, preferably from 20 to 30 min. By pre-shearing, the cross-linking system can be initially chopped to obtain a uniform sheet-like material.

Further, the shearing process may also include a high speed shearing step. In the high-speed shearing step, in order to form a more uniform form of the finally obtained hemostatic material (A), the rotation speed and time of the high-speed shearing step are critical; specifically, the rotation speed of the high-speed shearing step may be from 30,000 to 50,000 rpm/min. Preferably, it is between 30,000 and 40,000 rpm/min; the time of the high-speed shearing step is from 10 to 30 min, more preferably from 15 to 20 min.

In addition, the method for preparing the hemostatic material (A) of the present embodiment may further include a re-shearing step; the re-shearing step is a shearing after the shearing at a rotational speed of 40,000 to 50,000 rpm/min. The combined nanofiber material (which may be referred to as a shaped body) is subjected to a reshearing treatment. Further, a nanofiber cluster (a) of a suitable size is obtained. Preferably, the time of the reshearing treatment is 1 to 30 min, preferably 10 to 20 min.

In another specific embodiment, for example, when preparing the microfibrous hemostatic material of the first embodiment, the shearing treatment comprises a pre-shearing step. The pre-shearing step is to perform preliminary shearing treatment on the crosslinked nanofiber material at a rotation speed of 5000 to 10000 rpm/min to obtain a pre-shear. In general, the processing time of the pre-shearing step is 5 to 30 mim. By pre-shearing, the cross-linking system can be initially chopped to obtain a uniform sheet-like material.

Further, when preparing the microfibrous hemostatic material, the shearing treatment may also include a high speed shearing step. In the high-speed shearing step, in order to form a relatively uniform morphology of the finally obtained microfibrous hemostatic material, the rotational speed and time of the high-speed shearing step are critical; specifically, the rotational speed of the high-speed shearing step may be between 20,000 and 40,000 rpm/min. Preferably, it is between 25,000 and 30,000 rpm/min; and the time of the high-speed shearing step is from 1 to 10 min, more preferably from 3 to 5 min.

In still another specific embodiment, for example, preparing the hemostatic fibrous membrane of the second embodiment, wherein the hemostatic fibrous membrane comprises microfibers (b'), the shearing step and the hemostatic material are microfibrous hemostasis The shearing step of the material, that is, the hemostatic material including the microfibers (b), may be the same.

When preparing the microfibrous hemostatic material or the hemostatic fiber membrane, if the rotation speed of the high-speed shearing step is less than 20000 rpm/min, and the high-speed shearing step time is less than 1 min, the microfibers are mostly slightly flaky, and sheared. Non-uniform phenomenon; if the rotation speed is greater than 40,000 rpm/min, the time of the high-speed shearing step is more than 10 minutes, which will cause the microfibers to be powdered to some extent, while the powdery microfibers have low bulkiness, compact shape, and The clinical use operation and hemostasis effect can not reach the same level of microfibrous hemostatic material, and it is also difficult to form a nanofiber membrane.

The shearing treatment of the present disclosure is carried out in a state where a flowing gas is introduced. Specifically, the shearing treatment is carried out in a container, and a flowing gas is introduced into the container. The container can be sealed or non-closed. For example, the shearing process of the present embodiment may be processed using a specific shearing machine. The shear can have a container for holding the crosslinked nanofiber material. The crosslinked nanofiber material can be sheared by passing a flowing gas through the vessel. Among the nano short fibers obtained after shearing, at least some of the nano short fibers overlap each other to form nanofiber clusters (a), microfibers (b) or microfibers (b'), and nanofiber clusters (a) and microfibers. (b) or the microfibers (b') can be loosely distributed in the inner space of the container.

Preferably, the shearing machine of the present embodiment can introduce a flowing gas into the container by inflation. It can be continuous inflation or intermittent inflation, or it can be a cyclic inflation method. The flowing gas in the present embodiment may be a gas that forms a convection, or a gas that forms a disturbance.

The material of the container of the present embodiment is preferably a non-metal material, for example, a non-metal material such as plexiglass or tetrafluoroethylene. This is because during the high-speed shearing process, the container is connected to the motor, the metal material is easy to conduct heat, and the high temperature tends to cause the nanofiber cluster (a), or the microfiber (b), or the microfiber (b') to be dissolved and denatured, thereby The extent to which the nanofiber cluster (a), or the microfiber (b), or the microfiber (b') is affected, thereby affecting the properties of the hemostatic material. Therefore, the use of non-metallic materials can avoid affecting the performance of hemostatic materials. Further, in order to facilitate the observation of the shearing effect, a plexiglass material is preferred.

Since the flowing gas is introduced during the shearing process, the shearing is more uniform, and the hemostatic material can be further obtained to obtain a higher bulkiness, thereby further increasing the specific surface area of the hemostatic material.

The sieving step can be carried out in the preparation of the hemostatic material (A) of the first embodiment. In the sieving step in the present embodiment, the sieving treatment is performed by sieving with a mesh of 18 mesh, and the crosslinked nanofiber material (which may be referred to as a molded body) after sieving is collected. Thereby, a nanofiber cluster (a) of a suitable size is obtained, that is, a dimension from any direction of the three-dimensional space starting from the geometric center of the nanofiber cluster (a) is in the range of 5 μm to 500 μm.

In the preparation of the hemostatic fibrous membrane, a molding step is required. In the present embodiment, the microfibers (b') are pressed together using a mold to obtain a molded body. Specifically, the microfibers (b') are placed on the first mold at room temperature, and then covered with the second mold over the microfibers (b'), and pressed for 0.1-3 h, preferably 0.5- 1h, the hemostatic fiber membrane is formed. Preferably, the surface of the first mold and/or the second mold that is in contact with the microfibers (b') has a plurality of recesses and/or protrusions such that the surface of the hemostatic fiber membrane has a plurality of recesses and/or protrusions unit.

Between the crosslinking step and the shearing step, an elution step and/or a freeze-drying step may also be included.

The purpose of elution is to remove unreacted crosslinker. Specifically, the eluting step may include eluting the crosslinked nanofiber material by a concentration gradient method using an eluent at a low temperature of 0 to 20 ° C to remove the unreacted crosslinking agent. The eluent includes a mixture of an alcohol and water, preferably a mixture of ethanol and water, and more preferably, the mass fraction of ethanol in a mixture of ethanol and water is 70% or more. Further, the eluent in the present embodiment may be the same as or different from the solvent used in the crosslinking treatment.

Further, in the present embodiment, it is possible to perform multiple elution using a concentration gradient method using different concentrations of the alcohol-water solution. Preferably, the elution conditions are: an elution temperature of 1 to 20 ° C, preferably 4 to 10 ° C; an elution time of 0.1 to 5 h, preferably 0.5 to 2 h, repeated 3 to 5 times.

The purpose of lyophilization is to remove excess solvent and eluent from the cross-linking process while helping to increase the porosity of the nanofiber material or hemostatic material. The specific step of the freeze-drying is that the eluted nanofiber material is placed in a container and pre-cooled at -20 to -80 ° C for 1 to 3 hours, and then the container is transferred to a freeze dryer for lyophilization and freezing. The dry temperature is -10 to 40 ° C, preferably -10 to 30 ° C; and it is lyophilized at a vacuum of 1 to 100 Pa, preferably 20 to 40 Pa, for 6 to 72 h, preferably 12 to 24 h.

The hemostatic material prepared by the preparation method of the present embodiment has a simpler production process and a lower cost than the commercially available collagen microfiber hemostatic powder, and the selection source of the raw materials is more extensive, and is more in line with industrial production. Claim.

The hemostatic fiber membrane prepared by the preparation method of the present invention has better tissue adhesion and hemostatic effect compared with the membranous hemostatic material in the prior art, and has a simple production process and meets the requirements of industrial production.

Further, the nanofiber cluster (a), the microfiber (b), and the hemostatic fiber membrane obtained after the shearing may be sealed and packaged, and subjected to Co-60 γ ray irradiation sterilization treatment. Specifically, the sealed package requires rapid encapsulation in a dry environment with an ambient humidity of 30% or less; the Co-60 gamma ray irradiation dose is 15 to 30 kGY.

The hemostatic material or the hemostatic fiber membrane prepared by the present disclosure can be used for wounds only when it is taken out from the package, saving valuable rescue time, facilitating and simplifying the operation, and carrying and storing the product more conveniently. For the sake of simplicity.

Fourth embodiment

A fourth embodiment of the present disclosure also provides a hemostatic article comprising: the hemostatic material according to the first embodiment of the present disclosure, or the hemostatic fibrous film of the second embodiment, or the preparation method of the third embodiment of the present disclosure Hemostatic material or hemostatic fiber membrane.

The hemostatic product of the present disclosure can be used for hemostasis and repair in tissue oozing, capillary bleeding, arterial bleeding, oozing of the cavity, and/or for hemostasis and repair of burns, wounds, surgical wounds, and the like. Application prospects. When applying hemostasis and repair during oozing of the cavity, it can be applied to oozing blood in a cavity or the like by means of an auxiliary device, or the doctor can use this product in combination with other commercially available products according to experience, such as Hemostatic sponge, hemostatic gauze and other products to achieve better hemostasis.

In the hemostatic product of the present disclosure, since the hemostatic material or the hemostatic fiber membrane has good adhesive properties, a gel having a good adhesion ability is formed on the wound surface, and good physical sealing is performed to achieve compression and hemostasis. At the same time, the material has a high hydrophilic property due to the selection of a polymer material having an ultrahigh specific surface area and hydrophilicity. In the bleeding wound, it can quickly absorb the water in the blood, thereby providing the concentration of red blood cells and blood coagulation factors in the blood, accelerating the endogenous coagulation mechanism and improving the hemostatic effect.

Example

The embodiments of the present disclosure will be described in detail below with reference to the embodiments, but those skilled in the art will understand that the following examples are only intended to illustrate the disclosure, and are not intended to limit the scope of the disclosure. Those who do not specify the specific conditions in the examples are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are conventional products that can be obtained commercially.

Example 1

(1) Gelatin is dissolved in hexafluoroisopropanol, wherein the gelatin has a mass concentration of 12% (g/mL), and is stirred and dissolved to obtain a uniform polymer solution, which is a spinning dope. The polymer solution was placed in an electrospinning syringe, the rate of the micro syringe pump was adjusted to 6 mL/h, the voltage of the high voltage generator was adjusted to 25 kV, the receiving distance of the receiving device was adjusted to 10 cm, and the relative humidity of the spinning environment was set. 40%, the ambient temperature is 30 ° C, electrospinning, through the high-voltage electrospinning technology, a nanofiber material obtained by interweaving fiber filaments and having a porous structure is prepared.

(2) In a 500 mL reactor, add 225 mL of absolute ethanol solution, add 25 mL of aqueous solution, stir and mix, and then weigh 3.57 g of carbodiimide and 1.43 g of N-hydroxysuccinimide dissolved in the above ethanol at room temperature. In the aqueous solution, 5 g of the nanofiber material was placed in a reactor, and cross-linked modification was carried out at 25 ° C for 12 hours to obtain a crosslinked nanofiber material.

(3) Dissolve the ethanol-water solution with 70% ethanol content and pre-cool for 4 hours at a low temperature of 4 °C, then transfer the crosslinked nanofiber material to the above ethanol-water solution with 70% ethanol content. Elution; after 1 h of elution, transfer to a 4 °C ethanol-water solution with 95% ethanol content; after 1 h of elution, transfer to an ethanol-water solution at 70 °C with a mass fraction of 70%. , eluted for 1 h. Elution using a concentration gradient method to remove unreacted carbodiimide and N-hydroxysuccinimide was repeated 3 times.

(4) The eluted nanofiber material was placed in a clean container and pre-cooled at -80 ° C for 3 h, then the container was transferred to a freeze dryer for lyophilization, and the lyophilization temperature was set at 10 ° C for 3 h. Then, it was dried at 20 ° C for 24 h, and the degree of vacuum was set to 30 Pa.

(5) Put the freeze-dried nanofiber material into a high-speed shearing machine, set the rotation speed to 10000 rpm/min, and the treatment time is 30 min, perform preliminary shearing to obtain a small piece of pre-shear; then rotate the speed After setting to 30000 rpm/min, the chip-shaped pre-shear was subjected to high-speed shear treatment for 20 minutes, and then sieved with an 18-mesh sieve to collect the molded body through which the sieve was passed, thereby obtaining a partial nanofiber cluster (a).

(6) A larger cluster that has not passed through the mesh screen is again placed in a high-speed shearing machine, and the rotational speed is set to 40,000 rpm/min; and the treatment time is 10 minutes to obtain a partial nanofiber cluster (a).

(7) The nanofiber clusters (a) obtained after the shearing were sealed and packaged, and subjected to a 25-kGY Co-60 γ-ray irradiation sterilization treatment to obtain a hemostatic product. An electron micrograph of a single nanofiber cluster (a) in the hemostatic product is shown in FIG.

Example 2

(1) Silk Fibroin is dissolved in formic acid, wherein the silk fibroin has a mass concentration of 10% (g/mL), and is stirred and dissolved to obtain a uniform polymer solution, that is, a spinning dope. The polymer solution was placed in an electrospinning syringe, the rate of the micro syringe pump was adjusted to 2 mL/h, the voltage of the high voltage generator was adjusted to 30 kV, the receiving distance of the receiving device was adjusted to 15 cm, and the relative humidity of the spinning environment was set to 30%. Electrostatic spinning was carried out at an ambient temperature of 40 °C. A nanofiber material obtained by interweaving fiber filaments and having a porous structure is prepared by a high-voltage electrospinning technique.

(2) In a 500 mL reactor, add 150 mL of absolute ethanol solution, add 50 mL of aqueous solution and 4 mL of glutaraldehyde solution, stir and mix, and place 2 g of nanofiber material in the reactor, and crosslink modification at 40 °C. After treatment for 24 h, a crosslinked nanofiber material was obtained.

(3) Dissolve the ethanol-water solution with 70% ethanol content and pre-cool for 4 hours at a low temperature of 4 °C, then transfer the crosslinked nanofiber material to the above ethanol-water solution with 70% ethanol content. Elution; after 1 hour of elution, transfer to ethanol solution with a mass fraction of 95% in ethanol at 4 °C; after 1 hour of elution, transfer to a 4 °C ethanol-water solution with 70% ethanol content, wash Take off for 1h. Elution was carried out by a concentration gradient method to remove unreacted glutaraldehyde, which was repeated 3 times.

(4) The eluted nanofiber material was placed in a clean container and pre-cooled at -80 ° C for 3 h, then the container was transferred to a freeze dryer for lyophilization, and the lyophilization temperature was set to -10 ° C for 3 h. Then, it was dried at 20 ° C for 24 h, and the degree of vacuum was set to 20 Pa.

(5) The freeze-dried nanofiber material was placed in a high-speed shearing machine at a rotational speed of 15,000 rpm/min, and the treatment time was 20 min, and preliminary shearing was performed to obtain a small piece of pre-shear. Then, the rotation speed was set to 40,000 rpm/min, and the small-sized pre-shear was subjected to high-speed shear treatment for 30 minutes, and then sieved by an 18-mesh sieve to collect the formed body through the sieve filtration to obtain a partial nanofiber cluster (a). ).

(6) A larger cluster that has not passed through the mesh screen is again placed in a high-speed shearing machine, and the rotational speed is set to 50,000 rpm/min; and the treatment time is 10 min to obtain a partial nanofiber cluster (a).

(7) The nanofiber clusters (a) obtained after the shearing were sealed and packaged, and subjected to a 25-kGY Co-60 γ-ray irradiation sterilization treatment to obtain a hemostatic product.

Example 3

(1) Polyvinyl alcohol (PVA) is dissolved in purified water, wherein the polyvinyl alcohol has a mass concentration of 5% (g/mL), and is stirred and dissolved to obtain a uniform polymer solution, that is, a spinning dope. The polymer solution was placed in an electrospinning syringe, the rate of the micro syringe pump was adjusted to 2 mL/h, the voltage of the high voltage generator was adjusted to 30 kV, the receiving distance of the receiving device was adjusted to 15 cm, and the relative humidity of the spinning environment was set to 30%. Electrostatic spinning was carried out at an ambient temperature of 40 °C. A nanofiber material obtained by interweaving fiber filaments and having a porous structure is prepared by a high-voltage electrospinning technique.

(3) Dissolve the ethanol-water solution with 70% ethanol content and pre-cool for 4 hours at a low temperature of 4 °C, then transfer the crosslinked nanofiber material to the above ethanol-water solution with 70% ethanol content. Elution; after 1 hour of elution, transfer to a 4 °C ethanol-water solution with 95% ethanol content; after 1 hour of elution, transfer to a 4 °C ethanol-water solution with 70% ethanol content, wash Take off for 1h. Elution was carried out by a concentration gradient method to remove unreacted glutaraldehyde, which was repeated 3 times.

(5) placing the freeze-dried nanofiber material in a high-speed shearing machine, setting a rotation speed of 10,000 rpm/min, a treatment time of 25 minutes, and performing preliminary shearing to obtain a small piece of pre-shear;

Then, the rotation speed was set to 50,000 rpm/min, and the small-sized pre-shear was subjected to high-speed shear treatment for 10 minutes, and then sieved with an 18-mesh sieve to collect the formed body through the sieve filtration to obtain a partial nanofiber cluster (a). ).

(6) The larger clusters that have not passed through the mesh screen are again placed in a high-speed shearing machine at a rotational speed of 40,000 rpm/min; and the treatment time is 15 minutes to obtain a partial nanofiber cluster (a).

Example 4

(1) Dissolving carboxymethyl chitosan (CMCH) in a mixed solution of purified water and hexafluoroisopropanol, wherein the mass concentration of carboxymethyl chitosan is 5% (g/mL), stirring and dissolving A homogeneous polymer solution is obtained, which is a spinning dope. The polymer solution was placed in an electrospinning syringe, the rate of the micro syringe pump was adjusted to 2 mL/h, the voltage of the high voltage generator was adjusted to 30 kV, the receiving distance of the receiving device was adjusted to 15 cm, and the relative humidity of the spinning environment was set to 30%. Electrostatic spinning was carried out at an ambient temperature of 40 °C. A nanofiber material obtained by interweaving fiber filaments and having a porous structure is prepared by a high-voltage electrospinning technique.

(5) Put the freeze-dried nanofiber material into a high-speed shearing machine, set the rotation speed to 10000 rpm/min, and the treatment time is 20 min, perform preliminary shearing to obtain a small piece of pre-shear; then rotate the speed After setting to 30000 rpm/min, the chip-shaped pre-shear was subjected to high-speed shear treatment for 20 minutes, and then sieved with an 18-mesh sieve to collect the molded body through which the sieve was passed, thereby obtaining a partial nanofiber cluster (a).

Example 5

(1) Dissolving a hydroxypropyl methylcellulose (HPMC) material in a mixed solution of hexafluoroisopropanol and water, wherein the mass concentration of hydroxypropylmethylcellulose is 10% (g/mL), stirring Dissolved to obtain a uniform polymer solution, which is a spinning dope. At the same time, 20% by mass of fibrinogen (coagulation factor) of hydroxypropylmethylcellulose was added to the above polymer solution for dissolution. The polymer solution in which fibrinogen was dissolved was placed in an electrospinning syringe, the rate of the micro syringe pump was adjusted to 5 mL/h, the voltage of the high voltage generator was adjusted to 38 kV, and the receiving distance of the receiving device was adjusted to 10 cm, and the spinning environment was Electrospinning was carried out with a relative humidity of 30% and an ambient temperature of 40 °C. A nanofiber material composed of a fiber entangled and having a porous structure of a composite coagulation factor is prepared by a high-voltage electrospinning technique.

(2) In a 500 mL reactor, add 160 mL of absolute ethanol solution, then add 40 mL of aqueous solution and 8 mL of glutaraldehyde solution, and adjust the pH to acidity (pH<4), stir and mix, and 6 g of nanofiber material. It was placed in a reactor, cross-linked and modified at 35 ° C, and treated for 48 hours to obtain a crosslinked nanofiber material.

(4) The eluted nanofiber material was placed in a clean container and pre-cooled at -80 ° C for 3 h, and then the container was transferred to a freeze dryer for lyophilization, and the lyophilization temperature was set at 30 ° C for 3 h. It was then dried at 20 ° C for 24 h and the vacuum was set to 40 Pa.

(5) Put the freeze-dried nanofiber material into a high-speed shearing machine, set the rotation speed to 20000 rpm/min, and the treatment time is 10 min, perform preliminary shearing to obtain a small piece of pre-shear; then rotate the speed After setting at 40000 rpm/min, the chip-shaped pre-shear was subjected to high-speed shear treatment for 30 min, and then sieved with an 18-mesh sieve to collect the molded body passed through the sieve to obtain a partial nanofiber cluster (a).

Example 6

(1) Dissolving carboxymethyl chitosan (CMCH) in purified water, wherein the concentration of carboxymethyl chitosan is 5% (g/mL), stirring and dissolving to obtain a uniform polymer solution, that is, spinning Silk stock solution. The polymer solution was placed in an electrospinning syringe, the rate of the micro syringe pump was adjusted to 2 mL/h, the voltage of the high voltage generator was adjusted to 30 kV, the receiving distance of the receiving device was adjusted to 15 cm, and the relative humidity of the spinning environment was set to 30%. Electrostatic spinning was carried out at an ambient temperature of 40 °C. A nanofiber material obtained by interweaving fiber filaments and having a porous structure is prepared by a high-voltage electrospinning technique.

(5) Put the freeze-dried nanofiber material into a high-speed shearing machine, set the rotation speed to 5000 rpm/min, and the treatment time is 10 min, perform preliminary shearing to obtain a sheet-like pre-shear; then set the rotation speed. At 25,000 rpm/min, the treatment time was 10 min, and microfibers (b) were obtained.

(6) The microfibers (b) obtained after the shearing were sealed and packaged, and subjected to a 25-kGY Co-60 γ-ray irradiation sterilization treatment to obtain a hemostatic product in a microfibrous state.

Example 7

(1) The collagen (Collagen) is dissolved in trifluoroethanol, wherein the mass concentration of the collagen is 7% (g/mL), and the mixture is stirred and dissolved to obtain a uniform polymer solution, that is, a spinning dope. The polymer solution was placed in an electrospinning syringe, the rate of the micro syringe pump was adjusted to 5 mL/h, the voltage of the high voltage generator was adjusted to 25 kV, the receiving distance of the receiving device was adjusted to 12 cm, and the relative humidity of the spinning environment was set to 40%, the ambient temperature is 30 ° C, electrospinning, through the high-voltage electrospinning technology, a nanofiber material obtained by interweaving fiber filaments and having a porous structure is prepared.

(5) Put the freeze-dried nanofiber material into a high-speed shearing machine, set the rotation speed to 8000 rpm/min, and the treatment time is 5 min, perform preliminary shearing to obtain a sheet-like pre-shear; then set the rotation speed. At 30,000 rpm/min, the treatment time was 5 min, and microfibers (b) were obtained.

(6) The microfibers (b) obtained after shearing were sealed and packaged, and subjected to a 25-kGY Co-60 γ-ray irradiation sterilization treatment to obtain a hemostatic product in a microfibrous state, as shown in Fig. 5 and Fig. 6.

Example 8

(5) Put the freeze-dried nanofiber material into a high-speed shearing machine, set the rotation speed to 10000 rpm/min, and the treatment time is 5 min, perform preliminary shearing to obtain a sheet-like pre-shear; then set the rotation speed. At 40,000 rpm/min and a treatment time of 1 min, microfibers (b) were obtained.

(6) The microfibers (b) obtained after shearing were sealed and packaged, and subjected to a 25-kGY Co-60 γ-ray irradiation sterilization treatment to obtain a microfiber-formed hemostatic product.

Example 9

(1) Dissolving Silk Fibroin in hexafluoroisopropanol, wherein the silk fibroin has a mass concentration of 10% (g/mL), and is stirred and dissolved to obtain a uniform polymer solution, that is, spinning. Stock solution. The polymer solution was placed in an electrospinning syringe, the rate of the micro syringe pump was adjusted to 6 mL/h, the voltage of the high voltage generator was adjusted to 25 kV, the receiving distance of the receiving device was adjusted to 10 cm, and the relative humidity of the spinning environment was set to 30%. Electrostatic spinning was carried out at an ambient temperature of 35 °C. A nanofiber material obtained by interweaving fiber filaments and having a porous structure is prepared by a high-voltage electrospinning technique.

(2) In a 500 mL reactor, add 200 mL of absolute ethanol solution, add 100 mL of aqueous solution and 1 mL of formaldehyde solution, stir and mix, and place 3 g of nanofiber material in the reactor, and crosslink modification at 40 ° C. 3h, a crosslinked nanofiber material was obtained.

(3) Dissolve the ethanol-water solution with 70% ethanol content and pre-cool for 4 hours at a low temperature of 4 °C, then transfer the crosslinked nanofiber material to the above ethanol-water solution with 70% ethanol content. Elution; after 1 hour of elution, transfer to a 4 °C ethanol-water solution with 95% ethanol content; after 1 hour of elution, transfer to a 4 °C ethanol-water solution with 70% ethanol content, wash Take off for 1h. Elution was carried out by a concentration gradient method to remove unreacted formaldehyde, which was repeated 3 times.

(5) Put the freeze-dried nanofiber material into a high-speed shearing machine, set the rotation speed to 10000 rpm/min, and the treatment time is 5 min, perform preliminary shearing to obtain a small piece of pre-shear; then rotate the speed The setting was 20,000 rpm/min and the treatment time was 5 min to obtain microfibers (b').

(6) The microfibers (b') were uniformly placed in a first mold having a mesh pattern on the surface, and a micro-fiber was covered with a second mold having a mesh structure having a convex portion having a weight of 10 kg ( On b'), press-fit for 1 h to obtain a molded body.

(7) The obtained molded body was cut, sealed, and subjected to a 25-kGY Co-60 γ-ray irradiation sterilization treatment to obtain a hemostatic product, as shown in FIG.

Example 10

(1) Dissolving carboxymethyl starch (CMS) in a mixed solution of water and hexafluoroisopropanol, wherein the mass concentration of carboxymethyl starch is 5% (g/mL), and stirring to obtain a uniform polymer The solution is the spinning dope. The polymer solution is placed in an electrospinning syringe, the rate of the micro syringe pump is adjusted to 2 mL/h, the voltage of the high voltage generator is adjusted to 35 kV, the receiving distance of the receiving device is adjusted to 15 cm, and the relative humidity of the spinning environment is set to 30%, an ambient temperature of 40 ° C, electrospinning, through the high-voltage electrospinning technology, a nanofiber material obtained by interweaving fiber filaments and having a porous structure was prepared.

(2) In a 500 mL reactor, add 100 mL of absolute ethanol solution, add 100 mL of aqueous solution and 5 mL of glutaraldehyde solution, stir and mix, and place 2 g of nanofiber material in the reactor, and crosslink modification at 50 °C. After treatment for 10 h, a crosslinked nanofiber material was obtained.

(5) Put the freeze-dried nanofiber material into a high-speed shearing machine, set the rotation speed to 5000 rpm/min, and the treatment time is 10 min, perform preliminary shearing to obtain a sheet-like pre-shear; then set the rotation speed. At 30,000 rpm/min, the treatment time was 10 min, and microfibers (b') were obtained.

(6) The microfibers (b') were uniformly placed in the first mold with the S-shaped grain, and the second mold covered with the weight of 15 kg of the surface having the convex portion was covered with the microfibers (b' On the test, 0.5 h was pressed to obtain a molded body.

(7) The obtained molded body was cut and packaged, and subjected to a 25-kGY Co-60 γ-ray irradiation sterilization treatment to obtain a hemostatic product.

Example 11

(1) Dissolving a hydroxyethyl cellulose (HEC) material in a mixed solution of hexafluoroisopropanol and water, wherein the hydroxyethyl cellulose has a mass concentration of 7% (g/mL), and is uniformly dissolved by stirring. The polymer solution is the spinning dope. At the same time, 10% by mass of fibrinogen (coagulation factor) of hydroxyethylcellulose was added to the above polymer solution for dissolution. The polymer solution in which fibrinogen was dissolved was placed in an electrospinning syringe, the rate of the micro syringe pump was adjusted to 5 mL/h, the voltage of the high voltage generator was adjusted to 30 kV, and the receiving distance of the receiving device was adjusted to 10 cm, and the spinning environment was Electrospinning was carried out with a relative humidity of 30% and an ambient temperature of 40 °C. A nanofiber material composed of a fiber entangled and having a porous structure of a composite coagulation factor is prepared by a high-voltage electrospinning technique.

(2) In a 500 mL reactor, add 60 mL of absolute ethanol solution, add 140 mL of aqueous solution and 8 mL of glutaraldehyde solution, adjust the pH to acidity (pH<4), stir and mix, and 5g nanofiber material. The mixture was placed in a reactor and cross-linked modified at 40 ° C for 24 hours to obtain a crosslinked nanofiber material.

(5) Put the freeze-dried nanofiber material into a high-speed shearing machine, set the rotation speed to 10000 rpm/min, and the treatment time is 3 min, perform preliminary shearing to obtain a sheet-like pre-shear; then set the rotation speed. At 40,000 rpm/min, the treatment time was 3 min, and microfibers (b') were obtained.

(6) The microfibers (b') were uniformly placed in a first mold having a diamond-shaped grain, and covered with a second mold having a mesh structure having a convex portion having a weight of 8 kg on the microfibers (b' The film was pressed for 2 hours to obtain a molded body.

Example 12

(1) The collagen (Collagen) is dissolved in trifluoroethanol, wherein the mass concentration of the collagen is 8% (g/mL), and the mixture is stirred and dissolved to obtain a uniform polymer solution, that is, a spinning dope. The polymer solution was placed in an electrospinning syringe, the rate of the micro syringe pump was adjusted to 10 mL/h, the voltage of the high voltage generator was adjusted to 28 kV, the receiving distance of the receiving device was adjusted to 8 cm, and the relative humidity of the spinning environment was set to 30%. Electrostatic spinning was carried out at an ambient temperature of 35 °C. A nanofiber material obtained by interweaving fiber filaments and having a porous structure is prepared by a high-voltage electrospinning technique.

(2) In a 500 mL reactor, add 200 mL of absolute ethanol solution, add 100 mL of aqueous solution and 5 mL of Genipin solution (concentration: 2%), stir and mix, and place 3 g of nanofiber material in the reactor at 50 °C. The cross-linking modification was carried out for 5 hours to obtain a crosslinked nanofiber material.

(5) Put the freeze-dried nanofiber material into a high-speed shearing machine, set the rotation speed to 10000 rpm/min, and the treatment time is 2 min, perform preliminary shearing to obtain a small piece of pre-shear; then rotate the speed The setting was 15000 rpm/min and the treatment time was 10 min to obtain microfibers (b').

(6) The microfiber (b') is evenly laid on the first mold whose surface is a flat steel plate, and the second mold having a surface of a weight of 5 kg is covered with the second mold on the microfiber (b'), and pressed. 3h, a molded body was obtained.

Performance Testing

Median particle size D ₅₀ test

Test method: the product to be tested is formulated into a dilute solution of a certain mass concentration, and the solution is dispersed and treated for 10 minutes using an ultrasonic processor (100 W), so that the nanofiber cluster (a) particles are dispersed and uniform, and the solution system reaches a certain stable state. The test was carried out using a Mastersizer 2000 laser particle size analyzer from Malvern, England. Before the measurement, the laser particle size analyzer needs to be preheated for 30 minutes. When measuring, first, the instrument performs the light control, and the control measurement background state is normal. Then, pour the dispersed sample into the sample measuring cell at one time, rinse the residual sample with deionized water and pour it into the sample cell, quickly click “Analyze” to measure the sample and save the data. The test was carried out with the hemostatic products of Examples 1-5, and the test results are shown in Table 1.

Table 1 Test results of the median particle size D ₅₀

样品名称sample name	实施例1Example 1	实施例2Example 2	实施例3Example 3	实施例4Example 4	实施例5Example 5
中位粒径(μm)Median particle size (μm)	260260	480480	350350	300300	200200

As can be seen from Table 1, the nanofiber cluster (a) of the hemostatic material of the present disclosure has a size represented by a median diameter D ₅₀ of between 200 μm and 500 μm.

Specific surface area test

Test method: The product to be tested is placed in the sample tube of the analytical instrument, wherein the analytical instrument is a fast fully automatic specific surface area and pore size analyzer, and the model is American Conta NOVA 4200e. Under a low temperature (liquid nitrogen bath) condition, a certain amount of adsorbate gas (N ₂ ) is introduced into the sample tube, and the adsorption amount of the adsorbed molecule (N ₂ ) of the sample to be tested is determined according to the change of the gas volume before and after the adsorption; The specific surface area of the solid matter is determined by referring to the national standard GB/T24533-2009-gas adsorption BET principle.

The specific surface area is calculated by the fact that in the sample placed in a gaseous environment, the surface of the material (the surface area of the outside of the particle and the internal void) will be physically adsorbed at a low temperature. When the adsorption reaches equilibrium, the amount of adsorbed gas at the equilibrium adsorption pressure is measured, and the adsorption amount of the monolayer of the sample is calculated according to the BET equation, thereby calculating the specific surface area of the sample. Among them, the BET equation is:

In the formula:

P——the partial pressure of the adsorbate, in units of Pa;

P0——the saturated vapor pressure of the adsorbent, the unit is Pa;

V——the actual adsorption amount of the sample, the unit is cm ³ ;

Vm - single layer saturated adsorption amount, the unit is cm ³ ;

C - a constant related to the adsorption capacity of the sample.

The test was carried out with the hemostatic products of Examples 1-12, and the test results are shown in Table 2.

Table 2 specific surface area test results

样品名称sample name	实施例1Example 1	实施例2Example 2	实施例3Example 3	实施例4Example 4	实施例5Example 5	实施例6Example 6
比表面积(m ²/g) Specific surface area (m ² /g)	22.18722.187	18.32118.321	15.47715.477	12.38512.385	11.30811.308	10.25110.251
样品名称sample name	实施例7Example 7	实施例8Example 8	实施例9Example 9	实施例10Example 10	实施例11Example 11	实施例12Example 12
比表面积(m ²/g) Specific surface area (m ² /g)	16.31916.319	9.8779.877	15.75415.754	10.85710.857	11.03211.032	9.1229.122

As can be seen from Table 2, the hemostatic product of the present disclosure has a high specific surface area.

Porosity test

The porosity was measured as follows: The porosity of the porous material was determined by a solvent filling method. Since ethanol easily penetrates into the interior of the porous material without causing shrinkage and swelling of the material, ethanol is used as a reagent.

The test method is as follows: a 50 ml small beaker is filled with an anhydrous ethanol solution, and the hemostatic product (mass m ₁ ) dried to a constant weight is weighed in ethanol, and the vacuum is circulated until the hemostatic product no longer has bubble overflow. Weigh the total weight of the beaker containing ethanol and hemostatic product to m ₂ , and then take out the hemostatic product containing ethanol inside, and weigh the remaining beaker and ethanol to m ₃ , and each sample is parallel 3 times. The results are shown in Table 3 below. .

The measured porosity P is:

P=(m ₂ -m ₃ -m ₁ )/(m ₂ -m ₃ )×100%

Wherein: (m _{2 -} m _{3 -} m ₁ ) the mass of ethanol contained in the pores of the blood product;

(m ₂ -m ₃ ) is the total mass of the hemostatic product containing ethanol.

The test was carried out with the hemostatic products of Examples 1-5, and the test results are shown in Table 3.

Table 3 porosity test results

样品名称sample name	实施例1Example 1	实施例2Example 2	实施例3Example 3	实施例4Example 4	实施例5Example 5
孔隙率％Porosity%	9090	8282	7878	6464	7171

As can be seen from Table 3, the hemostatic products of Examples 1-5 of the present disclosure have a high porosity.

Saturated water absorption test

Test method: a certain mass of the sample (m ₁ ) was placed in a petri dish, and 0.9% physiological saline preheated to (37 ± 1) ° C was added, and the mass of the physiological saline was 40 times that of the test material. The dish was transferred to a dry box and kept at (37 ± 1) °C for 30 min. The sample was taken out with tweezers, suspended for 30 s, and m _{2 was} accurately weighed with an electronic balance, and measured in parallel three times. The saturated water absorption (X) is calculated by calculation, and the saturated water absorption (X) is calculated as: X = (m _{2 -} m ₁ ) / m ₁ × 100%.

The hemostatic products of Examples 1-12 and the commercially available products 1-3 (all of which are commercially available as hemostatic materials) were tested. The test results are shown in Table 4, where n is the number of parallel measurements.

Table 4 saturated water absorption test results (n = 3)

样品名称sample name	实施例1Example 1	实施例2Example 2	实施例3Example 3	实施例4Example 4	实施例5Example 5	实施例6Example 6
饱和吸水率％Saturated water absorption%	2812±152812±15	2550±102550±10	2370±82370±8	1951±121951±12	1733±101733±10	2081±102081±10
样品名称sample name	实施例7Example 7	实施例8Example 8	实施例9Example 9	实施例10Example 10	实施例11Example 11	实施例12Example 12
饱和吸水率％Saturated water absorption%	2470±82470±8	1755±121755±12	2150±122150±12	1910±101910±10	1775±101775±10	1585±81585±8
样品名称sample name	市售产品1Commercial products 1	市售产品2Commercial products 2	市售产品3Commercial products 3
饱和吸水率％Saturated water absorption%	1320±101320±10	780±5780±5	600±5600±5

As can be seen from Table 4, the hemostatic product of the present disclosure has a high saturated water absorption rate. Therefore, the hemostatic product of the present disclosure has good water absorption performance, and the saturated water absorption rate of the products of all the examples can be more than 1500%, which is obviously superior to other commercially available products 1-3. Further, the hemostatic product prepared according to Example 7 had a very high saturated water absorption rate of 24.7 times the weight of the material itself. The product prepared according to Example 9 had the highest saturated water absorption and reached 21.5 times the weight of the material itself.

Bulk density test

Test method: the material is freely dropped into a measuring cylinder having a volume of V (cm ³ ) and a mass of m ₁ (g), so that the materials are freely stacked to a predetermined volume, and then the overall mass m ₂ (g) of the measuring cylinder and the material is weighed, Then the bulk density (g/cm ³ ) = (m _{2 -} m ₁ ) / V.

The test was carried out with the hemostatic products of Examples 1-8, and the results are shown in Table 5, where n is the number of parallel tests.

Table 5 bulk density test results (n=3)

样品名称sample name	实施例1Example 1	实施例2Example 2	实施例3Example 3	实施例4Example 4
堆密度(g/cm ³) Bulk density (g/cm ³ )	0.027±0.0010.027±0.001	0.016±0.0020.016±0.002	0.022±0.0020.022±0.002	0.041±0.0030.041±0.003
样品名称sample name	实施例5Example 5	实施例6Example 6	实施例7Example 7	实施例8Example 8
堆密度(g/cm ³) Bulk density (g/cm ³ )	0.033±0.0020.033±0.002	0.021±0.0010.021±0.001	0.015±0.0030.015±0.003	0.011±0.0020.011±0.002

As can be seen from Table 5, the product of the present disclosure is excellent in bulk density and has good bulkiness so that a certain stent structure can be formed on the wound surface. When the hemostatic material quickly absorbs the moisture in the blood, it immediately expands to form a gel, and the wound is physically blocked to stop bleeding. At the same time, the effective concentration of hemostatic components in the plasma can promote the occurrence of coagulation mechanism and play a double hemostasis function.

Thickness and areal density test

The thickness and the areal density test were carried out with the hemostatic products of Examples 9-12, and the test results are shown in Table 6. The test method for the areal density ω is to measure the weight per unit area of a single face while ignoring the thickness of the hemostatic fiber membrane.

Table 6 thickness and surface density test results

测试材料Test material	厚度(mm)Thickness (mm)	面密度(g/m ²) Area density (g/m ² )
实施例9Example 9	0.50.5	189.1189.1
实施例10Example 10	0.30.3	124.8124.8
实施例11Example 11	1.01.0	251.5251.5
实施例12Example 12	0.10.1	75.675.6

As can be seen from Table 6, the present disclosure hemostatic product calculating their thickness of 0.5mm, which is the surface density in the range of ^{^{125g / m 2 -385g / m 2}} , soft texture, a higher degree of bulkiness. Among them, the products of Example 9 to Example 11 have an areal density of 125-210 g/m ² when the thickness thereof is 0.5 mm, and the surface density thereof is smaller, indicating that it is lighter and more fluffy.

Fluffiness test

The bulkiness described in the present disclosure means 1000 times the ratio of the apparent thickness to the areal density of the hemostatic product (hemostatic fiber membrane), that is, the bulkiness B = apparent thickness T _o / areal density ω × 1000.

The bulkiness is expressed in cm ³ /g, the apparent thickness is expressed in mm, and the areal density is expressed in g/m ² . The test method for the apparent thickness T _o is tested by the FAST-1 compressive fabric style meter according to the method of GB/T 7689.1-2001, indicating that the blood product (hemostatic fiber membrane) has a thickness (mm) at a pressure of ² cN/cm ² and The hemostatic product (hemostatic fiber membrane) has a thickness (difference in mm) at a pressure of 100 cN/cm ² . The test method of the areal density ω is to measure the weight per unit area of a single face while ignoring the thickness of the hemostatic product (hemostatic fiber membrane).

Table 7 fluffiness test results

样品名称sample name	实施例9Example 9	实施例10Example 10	实施例11Example 11	实施例12Example 12
蓬松度(cm ³/g) Bulkness (cm ³ /g)	1586.51586.5	1201.91201.9	1988.11988.1	2645.52645.5

As can be seen from Table 7, the hemostatic product (hemostatic fiber membrane) of the present disclosure has a high bulkiness.

Hemostasis test

Test method: rabbit liver oozing model was used to cut abdominal rabbit hair, standard middle laparotomy, free and exposed liver; 10×10×2mm wound was formed in the same part of liver; wound was cleaned with gauze, and hemostasis with the same weight was used. The product covers the wound surface and is covered with a gelatin sponge, pressed for 30 s, the sponge is removed and the wound is oozing. The hemostasis time was recorded and the effectiveness of hemostasis was evaluated.

The products (experimental group) prepared in Example 1, Example 7, and Example 10 of the present disclosure were tested, and the control products (commercial product A, commercial product 1, and commercially available product B) were used as positive controls. The number of parallel groups in the group and the control group was n=10.

As shown in Fig. 2, the hemostasis time of the control product (commercial product A) was 308.8 s; the hemostasis time of the hemostatic product prepared in Example 1 of the present disclosure was 135.5 s; P value = 0.03, P value < 0.05, significant Sexual differences. Thus, the hemostatic time of the products of the present disclosure is significantly less than the hemostatic time of commercially available product A.

As shown in Fig. 7, the hemostatic time of the control product (commercial product 1) was 211.2 s; the hemostasis time of the hemostatic material prepared in Example 7 of the present disclosure was 161.5 s; P value = 0.023, P value < 0.05, significant Sexual differences. Therefore, the hemostatic time of the product of the present disclosure is significantly smaller than the hemostatic time of the commercially available product 1.

As shown in FIG. 10, the hemostatic time of the control product (commercial product B) was 289.7 s; the hemostasis time of the hemostatic product prepared in Example 10 of the present disclosure was 161.2 s; P value = 0.02, P value < 0.05, significant Sexual differences. Therefore, the hemostatic time of the hemostatic product of the present disclosure is significantly less than the hemostasis time of the commercially available product B.

Degradability test

Test method: The product of Example 1 of the present disclosure was used as a test material (experimental group), and the commercially available product A was used as a control material (control group).

Rabbit muscle implantation experiments were used. Cut the abdominal rabbit hair, the standard middle laparotomy, cut the skin and muscles, implant the material, the weight of the material is 0.01g, suture fixation, then suture the skin closed, antibiotic care for three days. After the pre-set observation period (1W (1 week) after surgery, 2W (2 weeks) after surgery, 4W (4 weeks after surgery), the anatomical photo record was taken, and the tissue was sent for pathological analysis. Biocompatibility of the site material.

The pathological results showed that, as shown in Fig. 3, the hemostatic product of the present disclosure had completely degraded at 4 W (4 weeks). As shown in FIG. 4, the commercially available product A can also see that some materials are not completely degraded and absorbed at 4W (4 weeks). Moreover, the tissue validation stimulation of the experimental group was also lower than that of the control group. Thus, the hemostatic products of the present disclosure have good tissue biocompatibility. In addition, it was observed during the course of the experiment that the hemostatic product of the present disclosure has more excellent adhesion properties.

Safety and degradability testing

Test method: The product of Example 7 of the present disclosure was used as a test material (experimental group), and the commercially available product 1 was used as a control material (control group).

A rat liver in situ hemostasis implantation model was used. Prepare the hair in the abdomen of the rat, standard open ventral, free, expose the liver; select the largest piece of liver, and cut a 10 × 10 × 2 mm wound. 0.15g hemostatic product was placed on the surface of the wound, hemostasis was stopped and implanted, suture was closed, and antibiotic treatment was performed for three days. After a pre-set observation period (7 days after surgery, 14 days after surgery, and 28 days after surgery), the photographs were dissected and the tissue was sent for pathological analysis. The pathological analysis was used to evaluate the biocompatibility of the implant site materials. .

The pathological results showed that, as shown in Fig. 8, the hemostatic product of the present disclosure had completely degraded at 28 days. On the other hand, the commercially available product 1 can still see that some materials are not completely degraded and absorbed at 28 days. Moreover, the tissue validation stimulation of the experimental group was also lower than that of the control group. Thus, the hemostatic products of the present disclosure have good tissue biocompatibility. In addition, it was observed during the course of the experiment that the hemostatic product of the present disclosure has more excellent adhesion properties.

Degradation absorption experiment

Twenty healthy rabbits were selected and divided into two groups on average, one of which was the experimental group and the other was the control group. After the rabbit was anesthetized by intravenous injection into the ear, a hemostatic product was implanted in the abdomen of the rabbit, wherein the experimental group was implanted with the hemostatic fiber membrane prepared in Example 9 of the present disclosure, and the control group was implanted with a commercially available similar membrane-like hemostatic product. After suturing and fixation, antibiotics were taken for the first three days after surgery, and anatomical observation was performed after 1, 2, 4, 8 and 12 weeks after implantation. Two animals were randomly selected from each group for anatomy, sampling and fixation for histology. Observed.

As shown in Figures 11-16, the hemostatic product prepared in Example 9 was completely degraded within 2 weeks, and the material of the control group was still mostly present for 1 month. The hemostatic product prepared by the preparation method of the present disclosure is proved to have a rapid degradation absorption effect.

The above-described embodiments of the present disclosure are merely illustrative of the present disclosure, and are not intended to limit the embodiments of the present disclosure. Other variations or modifications of the various forms may be made by those skilled in the art in light of the above description. There is no need and no way to exhaust all of the implementations. Any modifications, equivalent substitutions and improvements made within the spirit and scope of the present disclosure are intended to be included within the scope of the appended claims.

Claims

A hemostatic material, characterized in that the hemostatic material comprises a structure having a staggered structure formed by overlapping a plurality of nano-short fibers, the nano-short fibers having a diameter of between 1 nm and 1000 nm; From the crosslinked nanofiber material, the structure is a nanofiber cluster (a) or a microfiber (b),

The nanofiber cluster (a) has a dimension in any direction of the three-dimensional space starting from a geometric center thereof in a range of 5 μm to 500 μm; and/or a median diameter D50 of the nanofiber cluster (a) in the hemostatic material The size indicated is between 100 μm and 500 μm, preferably between 200 μm and 400 μm, and the nanofiber cluster (a) has a porous structure, and the length of the nano short fibers in the nanofiber cluster (a) is less than 1000 μm;

The microfibers (b) have a diameter of 1 μm to 500 μm and a length of 0.5 mm to 10 mm, and the length of the nano short fibers in the microfibers (b) is 10 mm or less.
The hemostatic material according to claim 1, wherein the specific surface area of the hemostatic material of 4m 2 / g ~ 50m 2 / g, a water absorption greater than 1500%.
The hemostatic material according to claim 1 or 2, wherein the nanofiber material is derived from a polymer material having biocompatibility and biodegradable absorption, the nanofiber material being interwoven by filaments .
The hemostatic material according to any one of claims 1 to 3, characterized in that one of the following conditions is satisfied:

a. the hemostatic material comprises a nanofiber cluster (a), the hemostatic material having a bulk density of less than 0.06 g/cm 3 , preferably from 0.025 g/cm 3 to 0.05 g/cm 3 ; the porosity of the hemostatic material 50% to 90%, preferably 70% to 90%;

b. The hemostatic material comprises microfibers (b) having a bulk density of less than 0.03 g/cm 3 , preferably from 0.01 g/cm 3 to 0.025 g/cm 3 .
The hemostatic material according to any one of claims 1 to 3, characterized in that one of the following conditions is satisfied:

a. The hemostatic material comprises a nanofiber cluster (a), the crosslinking is crosslinking in the presence of a crosslinking agent, preferably, the mass ratio of the crosslinking agent to the nanofiber material is 0.01 to 2 : 1, preferably 0.1 to 1:1;

b. The hemostatic material comprises microfibers (b), the crosslinking being cross-linking in the presence of a crosslinking agent, preferably the mass ratio of the crosslinking agent to the nanofiber material is 0.1 to 3: 1, preferably 0.5 to 2:1.
The hemostatic material according to any one of claims 1 to 5, wherein the ratio of the length to the diameter of the microfiber (b) is in the range of 5 to 8,000.
The hemostatic material according to any one of claims 1 to 6, wherein the hemostatic material further comprises a drug; preferably, the drug comprises one or two of thrombin, a blood coagulation factor, and a growth factor. The combination above.
A hemostatic fibrous membrane, characterized in that the hemostatic fibrous membrane comprises microfibers (b');

The microfibers (b') are derived from a crosslinked nanofiber material, the microfibers (b') having a diameter between 0.1 mm and 1 mm and a length of 20 mm or less;

The microfiber (b') has a staggered structure formed by overlapping a plurality of nano short fibers;

The nano short fibers have a diameter of between 1 nm and 1000 nm and a length of 10 mm or less.
The hemostatic fibrous membrane according to claim 8, wherein the surface of the hemostatic fibrous membrane has a plurality of concave portions and/or convex portions.
The hemostatic fibrous film according to claim 8 or 9, wherein the hemostatic fibrous film has an areal density of 50 g/m 2 to 500 g/m 2 , preferably 100 g/m 2 to 300 g/m 2 ; The hemostatic fibrous membrane has a specific surface area of 5 m 2 /g to 30 m 2 /g, preferably 10 m 2 /g to 20 m 2 /g; and the hemostatic fibrous film has a bulkiness of 500 to 5000 cm 3 /g, preferably 1000 to 3000 cm 3 / g; the hemostatic fibrous membrane has a water absorption of more than 1500%, preferably between 1700% and 2500%.
The hemostatic fibrous membrane according to any one of claims 8 to 10, wherein the hemostatic fibrous membrane has a thickness of between 0.05 mm and 2 mm, preferably between 0.3 mm and 1 mm.
The hemostatic fibrous membrane according to any one of claims 8 to 11, wherein the nanofiber material is derived from a polymer material having biocompatibility and biodegradable absorption, the nanofiber material being composed of fibers Silk interwoven.
The hemostatic fibrous film according to any one of claims 8 to 12, wherein the crosslinking is crosslinking in the presence of a crosslinking agent, preferably, the crosslinking agent and the nanofiber material The mass ratio is from 0.1 to 3:1, preferably from 0.5 to 2:1.
The hemostatic fibrous membrane according to any one of claims 8 to 13, wherein the hemostatic fibrous membrane further comprises a drug; preferably, the drug comprises one of thrombin, a blood coagulation factor and a growth factor or A combination of the two.
A method for preparing a hemostatic material according to any one of claims 1 to 7 or a method for producing a hemostatic fibrous film according to any one of claims 8 to 14, comprising the steps of:

Electrospinning step: preparing a nanofiber material by electrospinning;

Cross-linking step: crosslinking the nanofiber material in the presence of a crosslinking agent to obtain a crosslinked nanofiber material;

Shearing step: shearing the crosslinked nanofiber material.
The method according to claim 15, wherein the crosslinking agent comprises one or more of carbodiimide, N-hydroxysuccinimide, genipin or an aldehyde compound. combination,

Preferably, the crosslinking agent comprises carbodiimide and N-hydroxysuccinimide, and more preferably, the mass ratio of the carbodiimide to N-hydroxysuccinimide is from 1 to 4:1.
The production method according to claim 15 or 16, wherein the shearing treatment is carried out in a state in which a flowing gas is introduced.
The preparation method according to any one of claims 15 to 17, wherein the crosslinking step and the shearing step further comprise: an elution step and/or a freeze-drying step.
The preparation method according to claim 18, wherein the eluting step comprises:

The crosslinked nanofiber material is eluted by a concentration gradient method using an eluent at a low temperature of 0 to 20 ° C to remove unreacted crosslinking agent.
The preparation method according to any one of claims 15 to 19, wherein the preparation method further comprises

Molding step: After the shearing treatment, the mold is pressed using a mold to obtain a molded body.
The preparation method according to any one of claims 15 to 20, wherein the shearing treatment comprises

In the pre-shearing step, the cross-linked nanofiber material is subjected to preliminary shear treatment at a rotation speed of 5000 to 10000 rpm/min to obtain a pre-shear.
The preparation method according to claim 21, wherein the shearing process further comprises

In the high-speed shearing step, the pre-shear is subjected to a high-speed shearing treatment at a rotational speed of 20,000 to 40,000 rpm/min; preferably, the high-speed shearing treatment is performed for 1 to 10 minutes.
The preparation method according to any one of claims 15 to 19, wherein the preparation method further comprises

Screening step: Screening after shearing.
The preparation method according to claim 23, wherein said shearing treatment comprises

a pre-shearing step of preliminary shearing the crosslinked nanofiber material at a rotational speed of 10,000 to 20,000 rpm/min to obtain a pre-shear, and/or

In the high-speed shearing step, the pre-shear is subjected to a high-speed shearing treatment at a rotational speed of 30,000 to 50,000 rpm/min.
The preparation method according to claim 23 or 24, wherein the sieving treatment is performed by sieving with a mesh of 18 mesh, and the sheared crosslinked nanofiber material passed through the sieve is collected.
The preparation method according to claim 25, wherein the preparation method further comprises

The shearing step again: the sheared crosslinked nanofiber material that has not passed through the sieve is subjected to shearing treatment again at a rotation speed of 40,000 to 50,000 rpm/min.
A hemostatic product, comprising: the hemostatic material according to any one of claims 1 to 7, or the hemostatic fiber membrane according to any one of claims 8 to 14, or any one of claims 15-26 The hemostatic material or hemostatic fiber membrane prepared by the preparation method described in the above.