CN112121065A - Medical styptic powder and application thereof - Google Patents
Medical styptic powder and application thereofDownload PDFInfo
- Publication number
- CN112121065A CN112121065ACN202011071241.7ACN202011071241ACN112121065ACN 112121065 ACN112121065 ACN 112121065ACN 202011071241 ACN202011071241 ACN 202011071241ACN 112121065 ACN112121065 ACN 112121065A
- Authority
- CN
- China
- Prior art keywords
- powder
- medical hemostatic
- mesh
- giant salamander
- hemostatic powder
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/65—Amphibians, e.g. toads, frogs, salamanders or newts
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/0005—Ingredients of undetermined constitution or reaction products thereof
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0015—Medicaments; Biocides
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0057—Ingredients of undetermined constitution or reaction products thereof
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/0066—Medicaments; Biocides
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/008—Hydrogels or hydrocolloids
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/30—Compounds of undetermined constitution extracted from natural sources, e.g. Aloe Vera
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/418—Agents promoting blood coagulation, blood-clotting agents, embolising agents
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/04—Materials for stopping bleeding
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Surgery (AREA)
- Dispersion Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Diabetes (AREA)
- Hematology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Materials For Medical Uses (AREA)
Abstract
Translated fromChinese
Description
Translated fromChinese技术领域technical field
本发明属于生物材料领域，涉及医用止血粉，具体涉及一种利用天然成分为原料制备的医用止血粉及其用途。The invention belongs to the field of biological materials and relates to medical hemostatic powder, in particular to a medical hemostatic powder prepared by using natural ingredients as raw materials and uses thereof.
背景技术Background technique
健康的成人体内大约有3.8升到5.6升的血液。若失血量超过全身血液总量的15％，会导致脉搏加快，产生晕眩等不良反应；若失血量超过总量的40％，由于血压过低无法回流充满心房，会出现“心室性心博过速”症状，该症状通常会导致死亡。无法控制的失血已经成为威胁受伤者生命和导致死亡的主要原因，可见对于创伤导致失血者的局部快速止血非常重要。A healthy adult has approximately 3.8 to 5.6 liters of blood. If the blood loss exceeds 15% of the total blood in the whole body, it will cause the pulse to speed up, resulting in dizziness and other adverse reactions; if the blood loss exceeds 40% of the total blood pressure, the blood pressure is too low to return to the atrium, which will cause "ventricular heartbeat". Overspeed" symptoms, which often lead to death. Uncontrollable blood loss has become the main cause of life-threatening injury and death. It can be seen that local and rapid hemostasis of trauma-induced blood loss is very important.
现有技术提供了多种止血方法，比如压迫止血，手术电刀及止血材料等。The prior art provides a variety of hemostasis methods, such as compression hemostasis, electrocautery and hemostatic materials.
止血粉是一种常见的医学材料，主要用于快速撒在面积较大或不规则的伤口上起到快速止血的作用，相比敷贴和包扎等更加简便、快捷。止血粉的应用场景包括日常突发性事故的急救止血，医院对病患手术过程中的创伤止血，战争情况下中对受伤战士的现场救护止血，等。快速有效的止血材料可大幅度降低伤员的死亡率。因此，开发快速、安全、有效的新型止血材料成为医学材料领域的重要研究课题。Hemostatic powder is a common medical material, which is mainly used to quickly stop bleeding on large or irregular wounds. It is simpler and faster than dressing and bandaging. The application scenarios of hemostatic powder include emergency hemostasis in daily sudden accidents, wound hemostasis during hospital operations on patients, on-site rescue and hemostasis of wounded soldiers in war situations, etc. Rapid and effective hemostatic materials can greatly reduce the mortality rate of the wounded. Therefore, the development of fast, safe and effective new hemostatic materials has become an important research topic in the field of medical materials.
现有技术的止血粉，大多是利用材料的吸附性能，吸收出血中的水分，加速出血中的血小板与红细胞结合，起到快速止血的效果。例如采用中医药磨成粉末，或传统的医用纤维粉碎成粉末，或者采用微孔淀粉等。或利用具有吸水和发热效应的材料，通过提高施用处的温度来加速血液中的蛋白质凝固，例如采用特殊工艺制备的沸石粉，兼有吸收出血中的水分和发热的功效。还有将各种多聚糖材料制成微孔颗粒，其微孔起到吸水效果，同时可将血液中的凝血因子、血小板、纤维蛋白、红细胞等聚集在颗粒表面，起到快速止血的效果。Most of the hemostatic powders in the prior art use the adsorption properties of materials to absorb water in bleeding, accelerate the combination of platelets and red blood cells in bleeding, and achieve the effect of rapid hemostasis. For example, traditional Chinese medicine is ground into powder, or traditional medical fiber is pulverized into powder, or microporous starch is used. Or use materials with water absorption and heating effects to accelerate the coagulation of proteins in the blood by increasing the temperature of the application site, such as zeolite powder prepared by a special process, which has both the functions of absorbing water in the blood and heating. In addition, various polysaccharide materials are made into microporous particles, and the micropores can absorb water, and at the same time, coagulation factors, platelets, fibrin, red blood cells, etc. in the blood can be aggregated on the surface of the particles, which can quickly stop bleeding. .
现有技术的主要缺点在于，采用非生物兼容性的材料，可能导致伤口黏连，并且材料无法被人体吸收，需要进行愈后处理否可有可能导致各种副作用。而采用生物兼容性材料制成多孔颗粒的工艺复杂，成本居高不下。The main disadvantage of the prior art is that the use of non-biocompatible materials may lead to wound adhesion, and the materials cannot be absorbed by the human body, and the need for post-healing treatment may lead to various side effects. However, the process of using biocompatible materials to make porous particles is complicated and the cost is high.
大鲵(Andrias davidianus Blanchard)为大型两栖纲、有尾目、隐鳃鲵科，俗名“娃娃鱼”，属国家二级保护动物。大鲵在遇到外部刺激时，身体表层会分泌出一种粘液。目前有研究成果显示可以利用大鲵皮肤粘液制备黏合剂或止血剂。中国发明《一种大鲵粘液用于止血的作用》采用大鲵粘液提取物作为止血剂，制备工艺较为复杂，且大鼠断尾实验表明需要约80秒才能达到止血效果，说明止血效果欠佳。中国发明《大鲵粘液在制备止血材料中的应用》主要是利用大鲵粘液干粉的粘合性能对于浅表型外伤进行止血，而且对于粘合之后的长期止血效果以及伤口感染效果未做评价。不排除粘合之后发生内出血或者愈合过程发生炎症的可能性。Giant salamander (Andrias davidianus Blanchard) is a large amphibian, caudate, cryptogill family, commonly known as "Salamander", and is a national second-class protected animal. When giant salamanders encounter external stimuli, they secrete a kind of mucus on the surface of their bodies. At present, research results have shown that adhesives or hemostatic agents can be prepared from giant salamander skin mucus. The Chinese invention "A kind of giant salamander mucus for hemostasis" uses giant salamander mucus extract as a hemostatic agent, the preparation process is relatively complicated, and the rat tail docking experiment shows that it takes about 80 seconds to achieve the hemostatic effect, indicating that the hemostatic effect is not good. The Chinese invention "Application of giant salamander mucus in the preparation of hemostatic materials" mainly uses the adhesive properties of giant salamander mucus dry powder to stop bleeding in superficial trauma, and has not evaluated the long-term hemostatic effect and wound infection effect after bonding. The possibility of internal bleeding after bonding or inflammation of the healing process cannot be ruled out.
以上显示大鲵皮肤粘液制备的医用止血粉仍有缺陷亟待解决。The above shows that the medical hemostatic powder prepared from giant salamander skin mucus still has defects that need to be solved urgently.
发明内容SUMMARY OF THE INVENTION
综合上述现有技术的缺陷，本发明提供一种医用止血粉及其止血之用途，其中的组份含有大鲵皮肤粘液成分，可有效止血并兼具多种促进伤口修复的益处，以解决前述的技术问题。In view of the above-mentioned defects of the prior art, the present invention provides a medical hemostatic powder and its application for hemostasis. technical problem.
本发明提出的医用止血粉，利用大鲵皮肤粘液具有优良黏合性能和多种促进止血、伤口愈合、组织修复的特点，使制备的医用止血粉可达到快速止血的效果。本发明的医用止血粉具备较好的安全性、较好的生物兼容性、可降解、促再生效果，同时具有抗菌效果，材料来源广泛，制备工艺简单，是满足需求的理想材料。The medical hemostatic powder provided by the present invention utilizes the salamander skin mucus to have excellent adhesion properties and various characteristics of promoting hemostasis, wound healing and tissue repair, so that the prepared medical hemostatic powder can achieve the effect of rapid hemostasis. The medical hemostatic powder of the present invention has better safety, better biocompatibility, degradability, and regeneration-promoting effects, as well as antibacterial effects, wide material sources, simple preparation process, and is an ideal material to meet requirements.
为解决原始采集到的大鲵皮肤粘液为胶冻状、不便于保存、临床使用非常不便、难于彻底杀菌等缺陷，本发明提供了一种医用止血粉，其中包含了经过灭菌的大鲵皮肤黏液干粉；此外，已灭菌的大鲵皮肤黏液干粉具备特定的颗粒尺寸及吸水性，以达到最佳的止血效果。In order to solve the defects that the originally collected giant salamander skin mucus is jelly-like, inconvenient to store, very inconvenient for clinical use, and difficult to completely sterilize, the present invention provides a medical hemostatic powder, which contains sterilized giant salamander skin mucus dry powder. ; In addition, the sterilized giant salamander skin mucus dry powder has a specific particle size and water absorption to achieve the best hemostatic effect.
根据上述目的，本发明提供一种医用止血粉，其中包含已灭菌的大鲵皮肤黏液干粉，其颗粒尺寸为粒径小于4目和大于300目；且大鲵皮肤黏液干粉具备吸水性，于吸收水性溶液后形成水凝胶。According to the above purpose, the present invention provides a kind of medical hemostatic powder, which comprises sterilized giant salamander skin mucus dry powder, the particle size of which is smaller than 4 meshes and larger than 300 meshes; A hydrogel is formed after the solution.
根据本发明优选实施方式，前述医用止血粉包含已灭菌的大鲵皮肤黏液干粉，其优选的颗粒尺寸为-4目至-100目。According to a preferred embodiment of the present invention, the aforementioned medical hemostatic powder comprises sterilized giant salamander skin mucus dry powder, and its preferred particle size is -4 mesh to -100 mesh.
根据本发明优选实施方式，前述医用止血粉吸收的水性溶液选自蒸馏水、生理缓冲液、血液、血浆、血细胞制剂、组织液、富血小板血浆、富血小板血浆纤维蛋白或上述任意组合。According to a preferred embodiment of the present invention, the aqueous solution absorbed by the medical hemostatic powder is selected from distilled water, physiological buffer, blood, plasma, blood cell preparation, tissue fluid, platelet-rich plasma, platelet-rich plasma fibrin or any combination of the above.
根据本发明优选实施方式，前述医用止血粉包含的大鲵皮肤黏液干粉和水性溶液的重量份数比例为1：1至1：6。According to a preferred embodiment of the present invention, the weight ratio of the giant salamander skin mucus dry powder and the aqueous solution contained in the aforementioned medical hemostatic powder is 1:1 to 1:6.
根据上述目的，本发明进一步提供前述医用止血粉的使用方法，将医用止血粉应用在伤口上成为敷料。According to the above purpose, the present invention further provides a method for using the aforementioned medical hemostatic powder, which is applied to a wound to become a dressing.
根据本发明优选实施方式，前述的止血是施用医用止血粉至伤口，从而使医用止血粉在伤口与水性溶液混合并形成水凝胶，成为敷料。According to a preferred embodiment of the present invention, the aforesaid hemostasis is to apply medical hemostatic powder to the wound, so that the medical hemostatic powder is mixed with the aqueous solution in the wound to form a hydrogel to become a dressing.
根据本发明优选实施方式，前述的使用方法是预先使用医用止血粉以及水性溶液制备成水凝胶后，成为敷料。According to a preferred embodiment of the present invention, the aforementioned method of use is to prepare a hydrogel by using medical hemostatic powder and an aqueous solution in advance, and then it becomes a dressing.
附图说明Description of drawings
图1表示本发明实施例1描述的大鲵(a)；采用机械刮擦法获取大鲵背部皮肤黏液(b)；本发明研获的不同粒径的大鲵皮肤黏液干粉的体式显微镜下图像(c)。Fig. 1 shows the giant salamander described in Example 1 of the present invention (a); the back skin mucus (b) of the giant salamander is obtained by mechanical scraping method; the image (c) under the stereomicroscope of the giant salamander skin mucus dry powder of different particle sizes developed by the present invention .
图2表示本发明实施例1的实验1多种灭菌方案所达成的灭菌效果数据图(a)果及黏附强度数据图(b)。Figure 2 shows the sterilization effect data graph (a) and the adhesion strength data graph (b) achieved by various sterilization schemes in Experiment 1 of Example 1 of the present invention.
图3表示本发明实施例2描述的不同粒径SSAD粉(a)及其灭菌后与抗凝全血混合后成胶的体式显微镜下图像(b)。Figure 3 shows the SSAD powder (a) with different particle sizes described in Example 2 of the present invention and the image (b) under a stereomicroscope of the gelatinized SSAD powder after being mixed with anticoagulated whole blood after sterilization.
图4表示本发明实施例2通过不同过筛取得不同颗粒尺寸的SSAD粉以及医用止血粉(灭菌后的SSAD粉)经过水化成胶后的扫描电镜(SEM)图像(a)；以及本发明实施例2不同目数的SSAD粉与水性溶液混合成胶后形成的多孔结构的孔径分布(b)。Fig. 4 shows the scanning electron microscope (SEM) image (a) of SSAD powder with different particle sizes obtained by different sieving and medical hemostatic powder (SSAD powder after sterilization) after hydration into gel in Example 2 of the present invention; and the present invention Example 2 Pore size distribution (b) of the porous structure formed by mixing SSAD powders of different mesh numbers with an aqueous solution to form a gel.
图5A表示本发明实验4中SSAD粉浸提培养基促细胞增殖迁移的细胞划痕实验结果。FIG. 5A shows the results of the cell scratch experiment in which the SSAD powder extraction medium promotes cell proliferation and migration in
图5B表示本发明实验4中SSAD粉浸提培养基的细胞划痕实验的统计分析结果。FIG. 5B shows the statistical analysis results of the cell scratch test of the SSAD powder leaching medium in
图5C表示本发明实验4中SSAD粉浸提培养基促细胞增殖迁移的transwell小室实验结果。Figure 5C shows the results of a transwell chamber experiment in which the SSAD powder extraction medium in
图6表示本发明实施例5医用止血粉在体内降解的组织学分析。Figure 6 shows the histological analysis of the in vivo degradation of the medical hemostatic powder in Example 5 of the present invention.
具体实施方式Detailed ways
为了使本发明的目的、技术特征及优点，能更为相关技术领域人员所了解，并得以实施本发明，在此配合所附的附图、具体阐明本发明的技术特征与实施方式，并列举较佳实施例进一步说明。以下文中所对照的附图，为表达与本发明特征有关的示意，并未亦无需依据实际情形完整绘制。而关于本案实施方式的说明中涉及本领域技术人员所熟知的技术内容，亦不再加以陈述。In order to make the purpose, technical features and advantages of the present invention more understandable to those in the relevant technical fields, and to implement the present invention, the technical features and embodiments of the present invention are described in detail here in conjunction with the attached drawings, and are listed here. The preferred embodiment is further described. The accompanying drawings compared in the following are schematic representations related to the features of the present invention, and are not and do not need to be completely drawn according to the actual situation. The description about the embodiments of the present application involves technical contents well known to those skilled in the art, and will not be described again.
本发明提供一种医用止血粉，医用止血粉主要是用大鲵皮肤粘液干粉制备而成。The invention provides a medical hemostatic powder, which is mainly prepared from dry powder of giant salamander skin mucus.
原始采集到的大鲵皮肤黏液为胶冻状，难于彻底杀菌，且不便于保存，临床使用存在一定的难度。为解决上述缺陷，本发明采用的技术方案是提供含有大鲵皮肤黏液干粉，经过质量确认并充分灭菌后取得医用止血粉，应用于止血。The originally collected giant salamander skin mucus is in a jelly-like shape, which is difficult to sterilize thoroughly and inconvenient to preserve, so there are certain difficulties in clinical use. In order to solve the above-mentioned defects, the technical scheme adopted in the present invention is to provide dry powder containing giant salamander skin mucus, obtain medical hemostatic powder after quality confirmation and sufficient sterilization, and apply it to hemostasis.
根据本发明提出的医用止血粉，除了大鲵皮肤黏液干粉为其必要组份之外，为制作易于保存、运送和临床应用的产品，对于将进一步搭配其他组份或原料，制成适合使用的医用止血粉产品，在此不做限制。具体举例，根据本发明优选实施方式，本发明所提供的医用止血粉的配方中还可包含具有止血、消炎或促进组织生长的其他成分，进一步提升临床应用上的性能，故本发明对此不加限制。According to the medical hemostatic powder proposed by the present invention, in addition to the salamander skin mucus dry powder as its essential components, in order to make a product that is easy to store, transport and clinically use, it is further combined with other components or raw materials to make suitable medical Hemostatic powder products are not limited here. For example, according to the preferred embodiment of the present invention, the formula of the medical hemostatic powder provided by the present invention may also contain other components with hemostasis, anti-inflammatory or tissue growth promoting properties, so as to further improve the performance in clinical application. Add restrictions.
根据本发明的优选实施例之一，医用止血粉的制备方法包含如下步骤。步骤1：从大鲵皮肤取得黏液，将黏液冷冻干燥。步骤2：将冷冻干燥后的大鲵皮肤黏液进行研磨及粉碎，过筛后得到颗粒尺寸或粒径符合要求的SSAD粉，作为品质确保的条件之一。步骤3：接着再对SSAD粉进行灭菌即得医用止血粉，保存备用。According to one of the preferred embodiments of the present invention, the preparation method of medical hemostatic powder comprises the following steps. Step 1: Obtain mucus from giant salamander skin and freeze-dry the mucus. Step 2: Grinding and pulverizing the freeze-dried giant salamander skin mucus, and sieving to obtain SSAD powder with a particle size or particle size that meets the requirements, as one of the conditions for quality assurance. Step 3: Then sterilize the SSAD powder to obtain a medical hemostatic powder, which is stored for future use.
使用时，可将灭菌后的医用止血粉及水性溶液依1：1至1：6的重量配比，混合形成水凝胶状的水凝胶。优选的，水凝胶中所含大鲵皮肤黏液干粉与水性溶液的重量配比为1：2至1：5。When in use, the sterilized medical hemostatic powder and the aqueous solution can be mixed in a weight ratio of 1:1 to 1:6 to form a hydrogel-like hydrogel. Preferably, the weight ratio of the giant salamander skin mucus dry powder and the aqueous solution contained in the hydrogel is 1:2 to 1:5.
以下依据附图说明优选技术方案。前述步骤1之收集黏液方法，应严格按照中国的动物保护法进行，可以无须杀害大鲵亦可避免大鲵的永久残疾。可以采用刮擦法或电刺激法。亦可将商业养殖的大鲵处死之前采集黏液。本发明对此不做限制。Preferred technical solutions are described below with reference to the accompanying drawings. The method of collecting mucus in the aforementioned step 1 should be carried out in strict accordance with China's animal protection law, which can avoid the permanent disability of the giant salamander without killing the giant salamander. Scraping or electrical stimulation can be used. Mucus can also be collected from commercially farmed giant salamanders before they are sacrificed. The present invention does not limit this.
前述步骤2，将收集到的干燥后的大鲵皮肤黏液，在低温状态下经球磨机粉碎后研磨成细粉。并进一步采用不同目数的筛子过筛，得到特定颗粒尺寸(特定规格的粒径)的粉末，又称为SSAD粉，用以实现本发明医用止血粉的效能。In the aforementioned step 2, the collected dried giant salamander skin mucus is ground into a fine powder after being pulverized by a ball mill at a low temperature. And further use sieves with different mesh numbers to sieve to obtain a powder with a specific particle size (particle size of a specific specification), also called SSAD powder, to achieve the efficacy of the medical hemostatic powder of the present invention.
由于国际上对过筛目数的标准略有差异，本发明优选采用中国常用的泰勒标准筛制定义过筛目数，其分度是以200目筛孔尺寸0.074mm为基准，详细计算方式为本领域技术人员的常规知识，不再赘述。利用此标准描述本发明大鲵皮肤黏液冻干粉的颗粒尺寸时，目数前加正负号表示研磨或粉碎后粉末能否漏过该目数的网孔，负数表示粉末能漏过该目数的网孔，表示过筛后取得的粉末颗粒尺寸小于网孔尺寸；而正数表示粉末不能漏过该目数的网孔，即过筛后取得粉末的颗粒尺寸大于网孔尺寸。例如，粒径小于4目指的是可以漏过4目筛子的粉末，即最大粒径不超过4.75mm的粉末；粒径大于300目指的是无法漏过300目筛子的粉末，即最小粒径大于48μm的粉末。Due to the slight difference in the international standards for the number of sieving meshes, the present invention preferably adopts the Taylor standard sieve system commonly used in China to define the sieving mesh number, and its indexing is based on the 200-mesh sieve size of 0.074mm. The detailed calculation method is as follows: The conventional knowledge of those skilled in the art will not be repeated here. When using this standard to describe the particle size of the giant salamander skin mucus freeze-dried powder of the present invention, a plus or minus sign before the mesh number indicates whether the powder can pass through the mesh of the mesh after grinding or pulverization, and a negative number means that the powder can pass through the mesh. The mesh size of , means that the particle size of the powder obtained after sieving is smaller than the mesh size; and a positive number means that the powder cannot pass through the mesh size of the mesh, that is, the particle size of the powder obtained after sieving is larger than the mesh size. For example, a particle size smaller than 4 mesh refers to powder that can pass through a 4-mesh sieve, that is, powder with a maximum particle size of no more than 4.75mm; a particle size larger than 300 mesh refers to powder that cannot pass through a 300-mesh sieve, that is, the minimum particle size is larger than 48μm powder.
为获得较好的医用止血粉性能，本发明大鲵皮肤黏液冻干粉的优选颗粒粒径小于-4目(所述-4目表示过筛目数大于4目小于8目。通过实验取得对应的平均粒径大致为4207μm，见下文表2)；大于-100目(所述-100目表示过筛目数大于100目小于200目。通过实验取得对应的平均粒径大致为105μm，见下文表2)。In order to obtain better medical hemostatic powder performance, the preferred particle size of giant salamander skin mucus freeze-dried powder of the present invention is less than -4 order (the -4 order represents that the sieving number is greater than 4 order and less than 8 order. Obtain the corresponding The average particle size is approximately 4207 μm, see Table 2 below); greater than -100 mesh (the -100 mesh means that the sieving mesh number is greater than 100 mesh and less than 200 mesh. The corresponding average particle size obtained through experiments is approximately 105 μm, see the following table 2).
本发明的具体作法上，要取得颗粒尺寸为-4目的粉末，是采用目数4目和8目的筛子二次过筛，取得粒径能漏过4目筛子而不能漏过8目筛子的粉末(换而言之，所得到的颗粒粒径小于4目筛子的筛孔孔径而大于8目筛子的孔径)；-8目表示颗粒粒径小于8目大于14目；-14目表示颗粒粒径小于14目大于20目；-20目表示颗粒粒径小于20目大于60目；-60目表示颗粒粒径小于60目大于100目；-100目表示颗粒粒径小于100目大于200目；-200目表示颗粒粒径小于200目大于300目，-300目表示颗粒粒径小于300目大于400目。In the specific method of the present invention, to obtain a powder with a particle size of -4 mesh, a sieve with a mesh number of 4 mesh and an 8 mesh is used for secondary sieving to obtain a powder whose particle size can pass through the 4 mesh sieve but cannot pass through the 8 mesh sieve. (in other words, the obtained particle size is smaller than the sieve aperture of the 4-mesh sieve and larger than the aperture of the 8-mesh sieve); -8 mesh means that the particle size is smaller than 8 mesh and larger than 14 mesh; -14 mesh means particle size Less than 14 mesh and more than 20 mesh; -20 mesh means the particle size is less than 20 mesh and more than 60 mesh; -60 mesh means that the particle size is less than 60 mesh and more than 100 mesh; -100 mesh means that the particle size is less than 100 mesh and more than 200 mesh; - 200 mesh means that the particle size is smaller than 200 mesh and larger than 300 mesh, and -300 mesh means that the particle size is smaller than 300 mesh and larger than 400 mesh.
完成步骤2之后，可将粉末储存在低于-20℃冰箱中备用。接着，步骤3原则上是对步骤2所得到的粉末进行灭菌与消毒。此步骤是将制品应用至临床使用的重要步骤，攸关临床使用的性能。依据相关法规，未经灭菌消毒的大鲵皮肤黏液相关产品无法直接应用于临床。而且，分泌于活体大鲵体表的黏液，经步骤1所采集时有可能含有对人体有潜在危害的病毒或病菌，病毒或病菌无法经由步骤2的冻干彻底灭活，若直接用于创口表面，将增加伤口感染可能。After completing step 2, the powder can be stored in a refrigerator below -20°C for later use. Next,
此外，大鲵皮肤黏液的主要成分是蛋白、多肽、黏多糖和抗菌肽等活性成分，最优选的灭菌方法既不会破坏和改变生物大分子的结构，也不会导致大鲵皮肤黏液干粉的黏附、止血性能和生物活性降低。因此，根据当前的技术，大鲵皮肤黏液相关产品的消毒灭菌方法包括低温、紫外线、钴射线、消毒剂灭菌法，然而这些消毒灭菌方法应用在本发明中各有一些缺陷，因此，对照之下，优选的，本发明步骤3的灭菌与消毒选用环氧乙烷灭菌法。In addition, the main components of giant salamander skin mucus are active ingredients such as proteins, polypeptides, mucopolysaccharides and antimicrobial peptides. The most preferred sterilization method will neither destroy and change the structure of biological macromolecules, nor cause the adhesion of giant salamander skin mucus dry powder. , hemostatic properties and biological activity decreased. Therefore, according to the current technology, the disinfection and sterilization methods of giant salamander skin mucus related products include low temperature, ultraviolet rays, cobalt rays, and disinfectant sterilization methods. However, these disinfection and sterilization methods are applied in the present invention. Below, preferably, the sterilization and disinfection of
本发明进一步提供医用止血粉的使用方法，应用在伤口的成为敷料。需先强调的是，本发明所指水性溶液并不限于人工配制或外加的溶液，也广泛包含了人体自身的体液，因此，本发明优选的水性溶液包含但不限于：蒸馏水、生理缓冲液、血液、血浆、血细胞制剂、组织液、富血小板血浆、富血小板血浆纤维蛋白，或是将前述项目依使用需求任意组合成特定水性溶液加以应用。The present invention further provides a method for using the medical hemostatic powder, which is used as a dressing for wounds. It should be emphasized first that the aqueous solution referred to in the present invention is not limited to artificially prepared or externally added solutions, but also widely includes the body fluids of the human body itself. Therefore, the preferred aqueous solutions of the present invention include but are not limited to: distilled water, physiological buffer, Blood, plasma, blood cell preparations, tissue fluid, platelet-rich plasma, platelet-rich plasma fibrin, or any combination of the aforementioned items into a specific aqueous solution for application.
依据本发明以大鲵皮肤黏液干粉为医用止血粉主要组份的技术方案，以下述两项方法作为范例，说明本发明医用止血粉的优选使用方法。According to the technical scheme of taking giant salamander skin mucus dry powder as the main component of the medical hemostatic powder of the present invention, the following two methods are taken as examples to illustrate the preferred method of using the medical hemostatic powder of the present invention.
方法一，在伤口或出血部位，直接施用适量的医用止血粉和水性溶液直接形成水凝胶成为敷料，在止血同时亦可以达到较好的伤口黏合封闭效果。其中，在施用水性溶液时，较优选的作法是将伤口或出血部位的出血和体液渗出液的体积一并纳入估算后，再施用适量水性溶液，选择合适的粉水比例再施用，以保持较理想的伤口黏合效果。若伤口或出血部位的出血和体液渗出液的体积与所施用的医用止血粉可以顺利成胶并达到较好的伤口止血或粘合效果，则可不另外加施用水性溶液。The first method is to directly apply an appropriate amount of medical hemostatic powder and aqueous solution to the wound or bleeding site to directly form a hydrogel as a dressing, which can achieve better wound adhesion and sealing effect while hemostasis. Among them, when applying the aqueous solution, it is more preferable to include the volume of bleeding at the wound or bleeding site and the volume of body fluid exudate into the estimation, and then apply an appropriate amount of the aqueous solution, and select an appropriate ratio of powder to water to maintain the Better wound adhesion. If the volume of hemorrhage and body fluid exudate from the wound or bleeding site and the applied medical hemostatic powder can smoothly form a gel and achieve a better wound hemostasis or adhesion effect, the additional application of the aqueous solution is not necessary.
方法二，按照1份医用止血粉，与1～6份重量比例的水性溶液相混合之后，形成任意形状的凝胶，其形状可符合伤口或出血部位的特殊需形态(立体形状、表面积、厚度等)以满足治疗需要。对于出血量较大或组织液渗出较多的伤口或出血部位，选择合适的粉水比例，使凝胶施用于创口后可以吸收伤口或出血部位的组织液进一步溶胀形成水凝胶成为敷料，以起到止血，湿性黏结和封闭创口的效果。Method 2: After mixing 1 part of medical hemostatic powder with 1 to 6 parts by weight of an aqueous solution, a gel of any shape is formed, and its shape can meet the special needs of the wound or bleeding site (three-dimensional shape, surface area, thickness, etc.). etc.) to meet therapeutic needs. For wounds or bleeding sites with a large amount of bleeding or tissue fluid exuding more, choose an appropriate ratio of powder to water, so that the gel can absorb the tissue fluid of the wound or bleeding site after being applied to the wound and further swell to form a hydrogel to become a dressing. To hemostasis, wet adhesion and wound closure effect.
以下列举实施例具体说明本发明之技术方案与对应达成之效果。The following examples are given to specifically illustrate the technical solutions of the present invention and the corresponding effects achieved.
实施例1：制备医用止血粉Example 1: Preparation of medical hemostatic powder
请参考图1，如前述步骤1，取出活体大鲵(如图1a)，采用机械刺激法获取大鲵黏液(如图1b)，并采集大鲵黏液备用。步骤2，将收集到的大鲵黏液制成干粉(如图1c)(白色粉末)。在此过程中，在采集之后尽快将大鲵黏液冻干，从完成采集到开始冻干的时间不超过1小时(h)。冻干的速度设定为每小时下降10～15℃，在4小时内温度降到-20℃。随后使用研磨机(采用上海净信冷冻研磨仪JXFSTPRP-CL)，粉碎得到细粉，利用过筛方式去除粒径不符合要求的细粉，得到特定粉末粒径(优选为范围，不限定为单一数值或平均值)的干粉。在本实施例中，使用目数分别为4、8、14、20、60、100、200、300目的筛子过筛，得到粒径符合要求的干粉。颗粒尺寸为-4目的粉末，是采用目数4目和8目的筛子二次过筛，取得粒径能漏过4目筛子而不能漏过8目筛子的粉末(换而言之，所得到的颗粒粒径小于4目筛子的筛孔孔径而大于8目筛子的孔径)，通常可得到粒径不超过4.75mm且粒径不小于2.36mm的干粉。-8目表示颗粒粒径小于8目大于14目；-14目表示颗粒粒径小于14目大于20目；-20目表示颗粒粒径小于20目大于60目；-60目表示颗粒粒径小于60目大于100目；-100目表示颗粒粒径小于100目大于200目；-200目表示颗粒粒径小于200目大于300目，-300目表示颗粒粒径小于300目大于400目。将干粉灭菌后置于密闭容器内冷藏保存备用，即可作为本实施例之医用止血粉。为便于说明和对比本发明达成的技术效果，以下对于过筛后的大鲵黏液冻干粉，通常简称或标示为SSAD粉。Please refer to Figure 1, as in the previous step 1, take out the giant salamander (Figure 1a), use the mechanical stimulation method to obtain the giant salamander mucus (Figure 1b), and collect the giant salamander mucus for use. Step 2, the collected giant salamander mucus is made into dry powder (as shown in Figure 1c) (white powder). During this process, giant salamander mucus was lyophilized as soon as possible after collection, and the time from the completion of collection to the start of lyophilization did not exceed 1 hour (h). The rate of lyophilization was set to drop 10-15°C per hour, and the temperature dropped to -20°C within 4 hours. Then use a grinder (using Shanghai Jingxin Freezing Mill JXFSTPRP-CL) to pulverize to obtain fine powder, and use a sieving method to remove the fine powder whose particle size does not meet the requirements to obtain a specific powder particle size (preferably a range, not limited to a single particle size). value or average value) of the dry powder. In this embodiment, sieves with mesh numbers of 4, 8, 14, 20, 60, 100, 200, and 300 are used to sieve to obtain dry powder with a particle size that meets the requirements. The particle size is -4-mesh powder, which is sieved twice by using 4-mesh and 8-mesh sieves to obtain powder whose particle size can pass through the 4-mesh sieve but cannot pass through the 8-mesh sieve (in other words, the obtained The particle size is smaller than the sieve aperture of the 4-mesh sieve and larger than the aperture of the 8-mesh sieve), usually a dry powder with a particle size of not more than 4.75mm and a particle size of not less than 2.36mm can be obtained. -8 mesh means that the particle size is smaller than 8 mesh and larger than 14 mesh; -14 mesh means that the particle size is smaller than 14 mesh and larger than 20 mesh; -20 mesh means that the particle size is smaller than 20 mesh and larger than 60 mesh; -60 mesh means that the particle size is smaller than 60 mesh is larger than 100 mesh; -100 mesh means that the particle size is smaller than 100 mesh and larger than 200 mesh; -200 mesh means that the particle size is smaller than 200 mesh and larger than 300 mesh, and -300 mesh means that the particle size is smaller than 300 mesh and larger than 400 mesh. The dry powder is sterilized and then placed in a sealed container for storage in refrigeration, and can be used as the medical hemostatic powder of this embodiment. For the convenience of description and comparison of the technical effects achieved by the present invention, the sieved giant salamander mucus freeze-dried powder is usually abbreviated or marked as SSAD powder below.
以下举例说明不同规格颗粒(粒径)尺寸的SSAD粉制备方式。The following examples illustrate the preparation methods of SSAD powder with different particle size (particle size).
-14目粉末的制备过程为：将研磨机提前开机预冷到-20℃。将冻干的大鲵粘液放入的15ml的研磨罐中，加入6mm研磨珠一粒，在5HZ的频率下运行5秒(s)研磨一次，将研磨后的粉末采用14目和20目的筛子二次过筛后，可获得最大粒径小于14目筛子孔径大于20目筛子孔径的粉末，其颗粒尺寸(粒径)标示为-14目。The preparation process of -14 mesh powder is as follows: start the grinder in advance and pre-cool to -20°C. Put the freeze-dried giant salamander mucus into a 15ml grinding jar, add a 6mm grinding bead, run for 5 seconds (s) at a frequency of 5HZ and grind once, and use the 14-mesh and 20-mesh sieves to grind the ground powder twice. After sieving, powder with a maximum particle size smaller than the 14-mesh sieve aperture and larger than the 20-mesh sieve aperture can be obtained, and the particle size (particle size) is marked as -14 mesh.
-300目粉末的制备过程为：将研磨机提前开机预冷到-20℃，将冻干的大鲵粘液放入15ml的研磨罐中，加入6mm研磨珠一粒，在65HZ的频率下按照运行10s中断10s的设定研磨三次，将研磨后的粉末采用300目和400目的筛子二次过筛后，可获得最大粒径小于300目筛子孔径大于400目孔径的粉末，其颗粒尺寸(粒径)标示为-300目。The preparation process of -300 mesh powder is as follows: start the grinder in advance and pre-cool to -20°C, put the freeze-dried giant salamander mucus into a 15ml grinding jar, add a 6mm grinding bead, and run for 10s at a frequency of 65HZ. Interrupt the set grinding for 10s three times. After the ground powder is sieved twice with 300-mesh and 400-mesh sieves, the powder with the maximum particle size smaller than the 300-mesh sieve aperture and larger than the 400-mesh aperture can be obtained. Its particle size (particle size) Marked as -300 mesh.
其余不同粒径的大鲵粘液粉末可以通过调节研磨机的频率、研磨时间和研磨次数，以及更换不同目数的筛子得到。通常而言，如需得到粒径更小的粉末，需要采用的研磨频率越高，研磨时间次数越多，筛子目数越多，于此不再赘述。The other giant salamander mucus powders with different particle sizes can be obtained by adjusting the frequency, grinding time and grinding times of the grinding machine, and changing sieves with different mesh numbers. Generally speaking, in order to obtain powder with a smaller particle size, the higher the grinding frequency, the more times of grinding time and the more meshes of the sieve need to be used, which will not be repeated here.
为避免未经灭菌处理的SSAD粉由于长期储存而导致变性和细菌滋生，将干粉置于密闭容器内保存备用。若环境温度较高，应至于冰箱中低温保存。In order to avoid denaturation and bacterial growth of unsterilized SSAD powder due to long-term storage, store the dry powder in an airtight container for later use. If the ambient temperature is high, it should be stored at a low temperature in the refrigerator.
步骤3，采用环氧乙烷对前述干粉进行灭菌。优选的具体作法是将前述得到的SSAD干粉封装入环氧乙烷专用灭菌包装袋中，或装入开口容器并在容器口疏松塞入棉球后封装入环氧乙烷专用灭菌包装袋中。将该灭菌包装袋放入环氧乙烷(EO)灭菌容器，使用环氧乙烷进行灭菌，灭菌时间和温度以不破坏干粉性能为原则，不特地加限制。随后将完成灭菌的干粉放置，待残留的环氧乙烷挥发。最后，参考《GB/T 16886.7-2001“医疗器械生物学评价”》第七部分：“环氧乙烷灭菌残留量评价环氧乙烷的残留”检验环氧乙烷残留率。检验合格后，即完成整体灭菌步骤。
此外，是针对目前常用的不同灭菌方法应用于本发明医用止血粉产品的整体效果进行评估。例如，采用低温、紫外线的灭菌效果有消毒不完全的疑虑，杀菌处理时间较长，且经低温或紫外线消毒的SSAD粉易于发生水解，保存期限较短；而γ射线方法由于涉及放射线，操作较为复杂。因此，本发明优选采用环氧乙烷作为本发明医用止血粉的灭菌方案，详如以下实验说明。In addition, it is to evaluate the overall effect of different currently commonly used sterilization methods applied to the medical hemostatic powder product of the present invention. For example, the sterilization effect of low temperature and ultraviolet rays has the doubt of incomplete disinfection, the sterilization treatment time is long, and the SSAD powder sterilized by low temperature or ultraviolet rays is prone to hydrolysis, and the storage period is short; more complicated. Therefore, the present invention preferably adopts ethylene oxide as the sterilization scheme of the medical hemostatic powder of the present invention, as detailed in the following experimental description.
实验1：不同灭菌方法的评估和效果。Experiment 1: Evaluation and effect of different sterilization methods.
按照步骤1和步骤2所得SSAD粉，在步骤3采用不同消毒灭菌方法。实验组采用环氧乙烷灭菌法，对照组采用低温灭菌(包括-20℃、-50℃、-80℃和液氮)、紫外线和辐照灭菌(钴60γ射线)。According to the SSAD powder obtained in steps 1 and 2, different disinfection and sterilization methods are used in
实验组：针对本发明提出之环氧乙烷灭菌方法，优选参考GB18279-2000“医疗器械环氧乙烷灭菌确认和常规控制”国家标准执行具体灭菌步骤，简述步骤包括：将粉碎后的大鲵皮肤黏液干粉封装入环氧乙烷专用灭菌包装袋中，或装入开口容器后，容器口疏松塞入棉球，进行封装。将该灭菌包装袋放入环氧乙烷(EO)灭菌容器，使用灭菌参数为100％环氧乙烷，灭菌6小时，随后将完成灭菌的粉末室温放置72小时待环氧乙烷解析后备用。灭菌后参考GB/T16886.7-2001“医疗器械生物学评价”第七部分：环氧乙烷灭菌残留量评价环氧乙烷的残留(残留率应≤10ppm)完成检验合格后，即完成整体灭菌步骤。Experimental group: For the ethylene oxide sterilization method proposed by the present invention, it is preferable to refer to GB18279-2000 "Medical Equipment Ethylene Oxide Sterilization Confirmation and Routine Control" national standard to perform specific sterilization steps. After the giant salamander skin mucus dry powder is packaged into a special sterilized packaging bag for ethylene oxide, or after being put into an open container, the container mouth is loosely stuffed into a cotton ball for packaging. Put the sterilized packaging bag into an ethylene oxide (EO) sterilization container, use the sterilization parameter of 100% ethylene oxide, sterilize it for 6 hours, and then place the sterilized powder at room temperature for 72 hours to wait for the sterilization. After the ethane is resolved, it is ready for use. After sterilization, refer to GB/T16886.7-2001 "Biological Evaluation of Medical Devices" Part VII: Evaluation of Ethylene Oxide Sterilization Residues (residual rate should be less than or equal to 10ppm) Complete the overall sterilization step.
对照组：分别采用低温灭菌、紫外线灭菌以及辐照灭菌。其中低温灭菌的温度和冷冻时间分别为：低温灭菌，-20℃，24小时；-50℃，24小时；-80℃，24小时；液氮，24小时。紫外灭菌所使用的紫外线波长为280～400nm，照射时间为24小时。辐照杀菌使用的放射源为钴-60，辐照强度为600～1000千拉德，照射时间为120分钟。Control group: low temperature sterilization, ultraviolet sterilization and irradiation sterilization were used respectively. The temperature and freezing time of low temperature sterilization are respectively: low temperature sterilization, -20°C, 24 hours; -50°C, 24 hours; -80°C, 24 hours; liquid nitrogen, 24 hours. The ultraviolet wavelength used for ultraviolet sterilization is 280-400 nm, and the irradiation time is 24 hours. The radioactive source used in the irradiation sterilization is cobalt-60, the irradiation intensity is 600-1000 kilorad, and the irradiation time is 120 minutes.
灭菌结束后测试灭菌效果：采取平板涂布后菌落计数的方法。取1000倍无菌生理盐水稀释样品，以涂布法接种于平板，经过24小时的培养之后，直接统计平板上的菌落数(CFU)，结果如表1所示。Test the sterilization effect after the sterilization: adopt the method of counting the colonies after the plate coating. Take 1000 times of sterile saline diluted samples, inoculate on the plate by coating method, after 24 hours of culture, directly count the number of colonies (CFU) on the plate, the results are shown in Table 1.
灭菌结束后测试医用止血粉黏附性能：在万能试验机上测试粘接医用止血粉的对于粘结样品的粘接能力(用100N测力传感器以1mm/min的速度将粘结样品加载至完全分离)。每种灭菌方法分别进行5个样品的实验，取其平均值如表1所示。Test the adhesion performance of the medical hemostatic powder after sterilization: test the adhesion of the medical hemostatic powder to the bonded sample on a universal testing machine (use a 100N load cell to load the bonded sample at a speed of 1mm/min until it is completely separated. ). For each sterilization method, five samples were tested, and the average value was taken as shown in Table 1.
表1：不同灭菌方法的灭菌效果及灭菌后黏附强度比较Table 1: Comparison of the sterilization effect of different sterilization methods and the adhesion strength after sterilization
由实验结果可知，环氧乙烷消毒方法和钴射线消毒方法的灭菌效果优于低温灭菌(包括-20℃、-50℃、-80℃和液氮)和紫外线。但是钴射线消毒方法涉及放射性物质的使用，有一定的局限性。It can be seen from the experimental results that the sterilization effect of ethylene oxide disinfection method and cobalt ray disinfection method is better than that of low temperature sterilization (including -20℃, -50℃, -80℃ and liquid nitrogen) and ultraviolet rays. However, the cobalt ray disinfection method involves the use of radioactive substances, which has certain limitations.
依据临床安全性需求，针对上述环氧乙烷的灭菌方法，其灭菌性能与目前其他灭菌方案的比较结果如图2所示。结果显示，环氧乙烷消毒方法和钴射线消毒方法，其灭菌效果优于低温灭菌(包括-20℃、-50℃、-80℃和液氮)和紫外线(如图2a)，而钴射线消毒方法对于黏附性能有较大影响，环氧乙烷消毒方法能兼顾良好的黏附性能(如图2b)。According to the clinical safety requirements, for the above-mentioned ethylene oxide sterilization method, the comparison results of its sterilization performance and other current sterilization schemes are shown in Figure 2. The results show that the sterilization effect of ethylene oxide disinfection method and cobalt ray disinfection method is better than that of low temperature sterilization (including -20℃, -50℃, -80℃ and liquid nitrogen) and ultraviolet rays (as shown in Figure 2a), while The cobalt ray disinfection method has a great influence on the adhesion performance, and the ethylene oxide disinfection method can take into account the good adhesion performance (Figure 2b).
实验1的结果表示，环氧乙烷消毒方法既能达到灭菌效果，又不破坏原始大鲵皮肤粘液干粉中所含活性物质，是首选的消毒方法。The results of experiment 1 show that the ethylene oxide disinfection method can achieve the sterilization effect without destroying the active substances contained in the original giant salamander skin mucus dry powder, and is the preferred disinfection method.
若无特别说明，以下各实施例所使用的干粉，若非特别指明，则均为通过上述步骤1至3所得到的医用止血粉，以便简要说明本发明之技术方案及效果。本发明提供的医用止血粉，主要通过特定颗粒尺寸的大鲵皮肤黏液干粉与水性溶液形成水凝胶的特性，而实现优异的止血效果，如下述实施例说明。Unless otherwise specified, the dry powders used in the following examples are all medical hemostatic powders obtained through the above steps 1 to 3, so as to briefly describe the technical solutions and effects of the present invention. The medical hemostatic powder provided by the present invention mainly realizes the excellent hemostatic effect through the characteristics of forming a hydrogel between the dry powder of salamander skin mucus with a specific particle size and an aqueous solution, as illustrated in the following examples.
实施例2：医用止血粉和水性溶液形成水凝胶。Example 2: Medical hemostatic powder and aqueous solution form hydrogel.
将前述步骤所得到的-8目、-14目、-20目和-60目灭菌后的SSAD粉末做为本实施例之医用止血粉，按照重量比例，按1份医用止血粉混合3份抗凝全血，静置3分钟之后待其凝结成胶。各组医用止血粉及与抗凝全血水化后的体式显微镜图像如图3所示。The -8 mesh, -14 mesh, -20 mesh and -60 mesh sterilized SSAD powders obtained in the preceding steps are used as the medical hemostatic powder of this embodiment, and 3 parts are mixed by 1 part of the medical hemostatic powder according to the weight ratio. Whole blood was anticoagulated and allowed to stand for 3 minutes until it coagulated into a gel. Figure 3 shows the stereomicroscope images of the medical hemostatic powder in each group and after hydration with anticoagulated whole blood.
藉由本实施例清楚说明，医用止血粉中含有的多肽交联网络与水性溶液混合后，是膨胀而不是溶解，形成水凝胶体，此过程称为成胶。在成胶过程中，多肽链的氨基酸残基发生构象转变，形成类似水凝胶的黏合剂，酚醛羟基和氨基作为氢键供体向高表面能或亲水界面转变，通过氢键和范德瓦尔斯力促进生物粘附。另外，苯环在接触低表面能或疏水界面时，会通过π-π电子或阳离子-π相互作用与基底形成强烈的相互作用。所述水性溶液可以选自蒸馏水、生理缓冲液、血液、血浆、血细胞制剂、组织液、富血小板血浆、富血小板血浆纤维蛋白或上述任意组合。This example clearly illustrates that after the polypeptide cross-linked network contained in the medical hemostatic powder is mixed with the aqueous solution, it swells rather than dissolves to form a hydrogel, and this process is called gelation. During the gelation process, the amino acid residues of the polypeptide chain undergo a conformational transition to form a hydrogel-like adhesive, and the phenolic hydroxyl and amino groups act as hydrogen bond donors to transition to a high surface energy or hydrophilic interface. Vals force promotes bioadhesion. In addition, the benzene ring forms strong interactions with the substrate through π-π electrons or cation-π interactions when in contact with low surface energy or hydrophobic interfaces. The aqueous solution may be selected from distilled water, physiological buffer, blood, plasma, blood cell preparations, tissue fluid, platelet-rich plasma, platelet-rich plasma fibrin, or any combination of the above.
以扫描电子显微镜(Hitachi，S-3400N II，Japan)分析依据本发明所得到的不同颗粒尺寸(粒径)的医用止血粉的多孔结构，通过扫描电镜分析显示，当医用止血粉与水性溶液混合时会形成三维蜂窝结构，如图4a所示。With scanning electron microscope (Hitachi, S-3400N II, Japan) analyze the porous structure of the medical hemostatic powder of different particle size (particle size) obtained according to the present invention, through scanning electron microscope analysis shows, when the medical hemostatic powder is mixed with the aqueous solution A three-dimensional honeycomb structure is formed, as shown in Figure 4a.
具体针对不同粒径医用止血粉形成的多孔结构，分析多孔结构的孔径和孔隙尺寸的一例结果如图4b所示。Specifically for the porous structure formed by the medical hemostatic powder with different particle sizes, an example of the results of analyzing the pore size and pore size of the porous structure is shown in Figure 4b.
本发明提供的医用止血粉在成胶后具有多孔结构，使其在临床应用时，可以利于组织液中营养物质和代谢产物的交换。The medical hemostatic powder provided by the invention has a porous structure after gelation, so that it can facilitate the exchange of nutrients and metabolites in tissue fluid during clinical application.
值得注意的是，扫描电镜分析显示，三维蜂窝结构的孔洞平均直径随着使用的医用止血粉粒径的减小而相应减小；但孔洞平均直径与医用止血粉和水性溶液的比例(简称水粉比例)的关联性没有统计学意义，表示水粉比例不会显着影响医用止血粉的孔洞平均直径。更具体举例说明，粒径为-14目的医用止血粉水化形成水凝胶的孔洞平均直径为116微米，粒径-60目的孔洞平均孔径为37微米，粒径-300目的孔洞平均孔径为6微米。It is worth noting that the scanning electron microscope analysis shows that the average diameter of the pores of the three-dimensional honeycomb structure decreases with the decrease of the particle size of the medical hemostatic powder used; There was no statistical significance in the correlation between the ratio of gouache and powder, indicating that the ratio of gouache powder did not significantly affect the average diameter of the pores of the medical hemostatic powder. More specifically, the average diameter of the pores of the -14 purpose medical hemostatic powder hydration to form the hydrogel is 116 microns, the average diameter of the pores of the -60 purpose is 37 microns, and the average diameter of the pores of the -300 purpose is 6 microns. microns.
本发明提供的医用止血粉可吸收血液中的水分形成具有多孔结构的水凝胶，血液中的血小板、凝血素和纤维蛋白等大分子物质在水凝胶表面富集，并迅速凝固，达到止血效果。血液中的纤维蛋白可以和止血粉中的蛋白、多肽、黏多糖通过氢键、二硫键等形成稳定的交联网络结构，有利于血凝块更快的形成，同时，形成的血凝块机械强度更高，形成的水凝胶具有黏性，可以黏结封闭伤口，防止进一步出血，并且形成的水凝胶，有一定抑菌和抗感染作用，可以减轻伤口感染风险，在可以体内可完全降解，避免了将水凝胶取出造成伤口的二次伤害。The medical hemostatic powder provided by the invention can absorb the water in the blood to form a hydrogel with a porous structure, and the macromolecular substances such as platelets, thrombin and fibrin in the blood are enriched on the surface of the hydrogel and rapidly coagulate to achieve hemostasis. Effect. Fibrin in blood can form a stable cross-linked network structure with proteins, polypeptides, and mucopolysaccharides in hemostatic powder through hydrogen bonds, disulfide bonds, etc., which is conducive to the faster formation of blood clots. At the same time, the formed blood clots The mechanical strength is higher, and the formed hydrogel is viscous, which can bind and seal the wound to prevent further bleeding, and the formed hydrogel has certain antibacterial and anti-infective effects, which can reduce the risk of wound infection, and can be completely cured in vivo. Degradation, avoiding the secondary injury of the wound caused by taking out the hydrogel.
此外，由本实施例得知，无论水性溶液是何种成分，只要不影响医用止血粉充分成胶，不同重量比例的医用止血粉与水性溶液的比例，均可以形成适合不同止血用途的水凝胶，因此本发明对此不作限制。其中医用止血粉与水性溶液的重量比例在1：1～6具有较好的成胶性能，在1：2～5具有更优的成胶性能。In addition, it is known from this example that no matter what the composition of the aqueous solution is, as long as it does not affect the sufficient gel formation of the medical hemostatic powder, the ratios of the medical hemostatic powder and the aqueous solution in different weight ratios can all form hydrogels suitable for different hemostatic purposes. , so the present invention is not limited thereto. Among them, the weight ratio of the medical hemostatic powder to the aqueous solution is 1:1-6, and the gel-forming property is better, and the gel-forming property is better when the weight ratio is 1:2-5.
实施例3：凝血实验。Example 3: Coagulation assay.
采用根据实施例1制得的不同目数的医用止血粉，通过体外凝血实验和肝脏止血模型，测试医用止血粉粒径对于止血效果的影响。采用市售云南白药和Celox止血粉作为对比。云南白药来源于云南白药集团有限公司(昆明)，Celox止血粉来源于MedtradeProducts Ltd(Electra House,Crewe Business Park,Crewe,CW16GL,UK.)。Using the medical hemostatic powder with different mesh numbers prepared according to Example 1, the influence of the particle size of the medical hemostatic powder on the hemostatic effect was tested by in vitro coagulation experiment and liver hemostasis model. Commercially available Yunnan Baiyao and Celox hemostatic powder were used for comparison. Yunnan Baiyao comes from Yunnan Baiyao Group Co., Ltd. (Kunming), and Celox hemostatic powder comes from Medtrade Products Ltd (Electra House, Crewe Business Park, Crewe, CW16GL, UK.).
分别采用-4目、-8目、-14目、-20目、-60目、-100目、-200目、-300目的医用止血粉。采用电镜拍照并使用Image J软件进行图像分析测得其平均粒径。采用氮气吸附法使用全自动气体置换法真密度仪(Accupyc II 1340,美国)测得前述医用止血粉的固体密度，即医用止血粉在绝对密实状态下的体积，以ρ固表示；同时称取新鲜制备的不同粒径松散医用止血粉，倒入量筒测得其体积后计算不同粒径医用止血粉的表观密度。如下公式I：-4 mesh, -8 mesh, -14 mesh, -20 mesh, -60 mesh, -100 mesh, -200 mesh, -300 mesh medical hemostatic powder were used respectively. The average particle size was measured by taking pictures with electron microscope and using Image J software for image analysis. The solid density of the aforementioned medical hemostatic powder, that is, the volume of the medical hemostatic powder in an absolutely compact state, is expressed as ρsolid by using the nitrogen adsorption method and using an automatic gas replacement method true density meter (Accupyc II 1340, USA). Freshly prepared loose medical hemostatic powders with different particle sizes were poured into a graduated cylinder to measure the volume and the apparent density of the medical hemostatic powders with different particle sizes was calculated. Formula I as follows:
表观密度(ρ表观)＝M/V (I)Apparent density (ρapparent ) = M/V (I)
其中：M为粉末质量，V为量筒测量出的粉体积。Among them: M is the powder mass, V is the powder volume measured by the measuring cylinder.
据此计算不同粒径医用止血粉的空隙率，计算公式如下式II：According to this, the porosity of the medical hemostatic powder with different particle sizes is calculated, and the calculation formula is as follows:
空隙率(％)＝100％×(1-ρ表观/ρ固) (II)Porosity (%)=100%×(1-ρapparent /ρ solid) (II)
不同目数的医用止血粉的平均粒径、固体密度以及空隙率数据如表2。The average particle size, solid density and porosity data of medical hemostatic powder with different mesh numbers are shown in Table 2.
表2：不同目数的医用止血粉数据Table 2: Data of medical hemostatic powder with different mesh numbers
体外凝血实验的方法：分别取不同目数的医用止血粉，以及对照组的云南白药或Celox止血粉50mg置于试管底部。滴加抗凝血剂(含10％(w/v)柠檬酸钠的血浆)100微升，静置。静置后30秒、60秒、120秒分别将试管倒置，视试管的底部朝上口部朝下。未凝结的抗凝血会延着试管壁流下，若抗凝血已经凝结则不会流下。实验结果见表3。Methods of in vitro blood coagulation experiment: respectively take medical hemostatic powder with different mesh numbers, and Yunnan Baiyao or Celox hemostatic powder 50mg in the control group and place them at the bottom of the test tube. 100 microliters of anticoagulant (plasma containing 10% (w/v) sodium citrate) was added dropwise and allowed to stand. After standing for 30 seconds, 60 seconds, and 120 seconds, invert the test tube respectively, depending on the bottom of the test tube facing upward and the mouth downward. Uncoagulated anticoagulant will flow down the wall of the test tube, if the anticoagulant has been clotted will not flow down. The experimental results are shown in Table 3.
表3：体外凝血实验结果Table 3: In vitro coagulation test results
肝脏止血模型的建立方法为：将大鼠固定于手术板上，备皮，消毒，然后行中线开腹暴露肝脏，用手挤压两侧腹，从腹腔中将肝脏挤压出，暴露在止血纱布上。用手术刀沿肝脏行1×1cm的十字切口，导致肝脏出血，等待约3秒后应用不同粒径的前述医用止血粉施用于创口处控制出血。出血量的测量方式为：称量肝脏止血后药剂和血液混合形成的凝集物重量，减去施用于创口的药剂重量，加上从肝脏流出被垫底纱布吸收的血液重量。出血时间的测量方式为：施用药剂后开始计时至肉眼观察到出血停止时停止计时。实验结果见表4。The liver hemostasis model was established as follows: fix the rat on the operating board, prepare the skin, sterilize it, then open the midline to expose the liver, squeeze both sides of the abdomen by hand, extrude the liver from the abdominal cavity, and expose it to hemostasis. on gauze. A 1×1 cm cross incision was made along the liver with a scalpel, causing liver bleeding. After waiting for about 3 seconds, the aforementioned medical hemostatic powder with different particle sizes was applied to the wound to control the bleeding. The amount of bleeding was measured by weighing the weight of the agglutinate formed by mixing the drug and blood after hemostasis of the liver, subtracting the weight of the drug applied to the wound, and adding the weight of blood flowing from the liver and absorbed by the bottom gauze. Bleeding time was measured by starting the timing after the administration of the agent and stopping the timing when the bleeding stopped visually observed. The experimental results are shown in Table 4.
表4：不同粒径医用止血粉肝脏止血实验结果Table 4: Results of liver hemostasis experiment of medical hemostatic powder with different particle sizes
实验证明，粒径-4目与-300目的医用止血粉有较好的止血效果，其中以粒径-4目至-200目的医用止血粉具较理想的止血效果；而其中又以-4目至-100目的医用止血粉具更理想的止血效果。Experiments have proved that the medical hemostatic powder with particle size of -4 mesh and -300 mesh has better hemostatic effect, and the medical hemostatic powder with particle size of -4 mesh to -200 mesh has the ideal hemostatic effect; Medical hemostatic powder to -100 mesh has a more ideal hemostatic effect.
实施例4:体内止血实验Example 4: In vivo hemostasis experiment
分别采用肝脏止血模型、断尾止血模型、股动脉止血模型3种不同的模型，比较3种药剂：医用止血粉(20目)、市售止血药物Celox(Medtrade Products Ltd,Electra House,Crewe Business Park,Crewe,CW16GL,UK.)和市售云南白药(云南白药集团有限公司，昆明)的止血效果差异。医用止血粉按照实施例1的方式制备，采用20目的筛子过筛，取得的粉末最大粒径小于20目筛子的孔径。Three different models, namely liver hemostasis model, tail docking hemostasis model, and femoral artery hemostasis model, were used to compare three drugs: medical hemostatic powder (20 meshes), commercially available hemostatic drug Celox (Medtrade Products Ltd, Electra House, Crewe Business Park , Crewe, CW16GL, UK.) and commercially available Yunnan Baiyao (Yunnan Baiyao Group Co., Ltd., Kunming) the difference in hemostatic effect. The medical hemostatic powder was prepared in the manner of Example 1, sieved with a 20-mesh sieve, and the maximum particle size of the obtained powder was smaller than the aperture of the 20-mesh sieve.
肝脏止血模型的建立已于前述实施例3说明，本实施例在大鼠肝脏出血，约3秒钟之后应用前述3种药剂施用于创口以控制出血。记录出血时间和失血量。用治疗前后粉末的重量差来记录失血量。The establishment of the hepatic hemostasis model has been described in the aforementioned Example 3. In this example, the hepatic hemorrhage of the rat was applied after about 3 seconds, and the aforementioned three agents were applied to the wound to control the bleeding. The bleeding time and blood loss were recorded. Blood loss was recorded as the difference in powder weight before and after treatment.
出血量的测量方式为：称量肝脏止血后施用药剂和血液混合的凝集物质重量，减去施用于创口的药剂重量，同时垫底纱布上的血液重量也会被计算。空白对照组不施用药剂。出血时间的测量方式为：出血约3秒之后施用药剂，开始计时至肉眼观察到出血停止时停止计时。每一种药剂重复进行3例实验，取平均值。数据如表5。The amount of bleeding was measured by weighing the agglutinated material mixed with the drug and blood after hemostasis of the liver, subtracting the weight of the drug applied to the wound, and the weight of the blood on the bottom gauze was also calculated. The blank control group was not administered the drug. Bleeding time was measured by administering the agent approximately 3 seconds after bleeding, starting the timer and stopping when the bleeding was visually observed to stop. Three experiments were repeated for each drug, and the average value was taken. The data are shown in Table 5.
表5：三种药剂的止血效果对比Table 5: Comparison of hemostatic effects of the three agents
股动脉止血模型的建立：将大鼠固定于手术板上，备皮，消毒，大鼠股动脉损伤模型，先将大鼠股动脉从周围组织剥离，用21G穿刺针造成创伤出血，然后立即应用3种不同的药剂(－20目医用止血粉、Celox和云南白药)控制出血。记录出血时间和失血量。用治疗前后粉末的重量差来记录失血量。Establishment of femoral artery hemostasis model: fix the rat on the operation board, prepare the skin, sterilize the rat femoral artery injury model, first peel the rat femoral artery from the surrounding tissue, use a 21G puncture needle to cause trauma bleeding, and then apply it immediately 3 different agents (-20 mesh medical hemostatic powder, Celox and Yunnan Baiyao) control bleeding. The bleeding time and blood loss were recorded. Blood loss was recorded as the difference in powder weight before and after treatment.
出血量的测量方式为：称量股动脉止血后施用药剂和血液混合的凝集物质重量，减去应用了药剂重量，同时垫底纱布上的血液重量也会被计算。出血时间的测量方式为：出血之后，从应用药剂止血时开始计时，至肉眼观察出血停止时终止计时。空白组从出血时开始计时至肉眼观察出血停止时终止计时。每一种药剂重复进行3例实验，取平均值。数据如表6。The amount of bleeding was measured by weighing the agglutinated material mixed with the applied drug and blood after femoral artery hemostasis, subtracting the weight of the applied drug, and the weight of the blood on the bottom gauze was also calculated. The measurement method of bleeding time is as follows: after bleeding, start timing from the time when the drug is applied to stop bleeding, and stop timing when the bleeding stops by visual observation. In the blank group, the timing was started from the time of bleeding to the time when the bleeding stopped by visual observation. Three experiments were repeated for each drug, and the average value was taken. The data are shown in Table 6.
表6：三种药剂的止血效果对比Table 6: Comparison of hemostatic effects of the three agents
断尾模型止血模型的建立：先将大鼠固定于手术板上，对于大鼠截尾模型，为了引起出血，用手术剪刀将其尾部长度的50％剪断，置于空气中放血15秒。之后，用样本在轻微压力下覆盖伤口。记录出血时间和失血量。Establishment of hemostasis model of tail docking model: First, fix the rat on the operation board. For the rat tail chopping model, in order to cause bleeding, 50% of the tail length was cut with surgical scissors and placed in the air for 15 seconds to bleed. Afterwards, the wound is covered with the sample under gentle pressure. The bleeding time and blood loss were recorded.
描述出血量的测量方式为，称量止血后施用药剂和血液混合的凝集物质重量，减去应用了的药剂重量。出血时间的测量方式为：出血之后15秒，从应用药剂止血时开始计时，至肉眼观察出血停止时终止计时。空白组从出血后15秒开始计时至肉眼观察出血停止时终止计时。每一种药剂重复进行3例实验，取平均值。数据如表7。A measure to describe the amount of bleeding is to weigh the agglutinating material mixed with the administered agent and blood after hemostasis and subtract the weight of the applied agent. The measurement method of bleeding time is as follows: 15 seconds after bleeding, from the time when the drug is applied to stop the bleeding, to the time when the bleeding stops by visual observation. In the blank group, the timing was started from 15 seconds after the bleeding to the time when the bleeding stopped by visual observation. Three experiments were repeated for each drug, and the average value was taken. The data are shown in Table 7.
表7：三种药剂的止血效果对比Table 7: Comparison of hemostatic effects of the three agents
使用SPSS statistics 25软件(IBM公司，Armonk,NY,USA)进行统计分析，用单因素方差分析计算上述四组之间差异是否有统计学意义。分析结果表明，三种动物止血模型中，-20目医用止血粉，其止血效果明显优于云南白药(P＜0.05)和空白对照组(P＜0.01)，与Celox的止血效果大致相当(P＞0.05，差异无统计学意义)。Statistical analysis was performed using SPSS statistics 25 software (IBM, Armonk, NY, USA), and one-way ANOVA was used to calculate whether the differences between the above four groups were statistically significant. The analysis results showed that among the three animal hemostatic models, the hemostatic effect of -20 mesh medical hemostatic powder was significantly better than that of Yunnan Baiyao (P<0.05) and the blank control group (P<0.01), and was roughly equivalent to the hemostatic effect of Celox (P<0.01). >0.05, the difference was not statistically significant).
除了前述本发明医用止血粉的良好止血性能之外，本发明提供的医用止血粉更具备了许多创伤应用所需的特点，对伤口止血带来更多的效益，进一步以下述实验2至实验6进行说明。In addition to the good hemostatic properties of the aforementioned medical hemostatic powder of the present invention, the medical hemostatic powder provided by the present invention has many characteristics required for wound application, and brings more benefits to wound hemostasis. The following experiments 2 to 6 are further used. Be explained.
实验2：抑菌效果实验。Experiment 2: Antibacterial effect experiment.
配置需氧菌实验所用的SSAD粉浸提液：称取1g无菌SSAD干粉，加入适量LB(营养肉汤培养基)培养基使SSAD干粉充分吸水溶胀后，再继续加入培养基，最终使液体培养基的量达到10mL，在4℃冰箱中放置24小时完成浸提。Configure the SSAD powder extract used in the aerobic bacteria experiment: Weigh 1 g of sterile SSAD dry powder, add an appropriate amount of LB (nutrient broth medium) medium to make the SSAD dry powder fully absorb water and swell, then continue to add the medium, and finally make the liquid The amount of medium reached 10 mL, and the leaching was completed by placing in a refrigerator at 4°C for 24 hours.
配置厌氧菌实验所用的SSAD粉浸提液：称取1g无菌SSAD干粉，加入适量TSB(胰蛋白胨大豆肉汤培养基)培养基使其充分吸水溶胀后，再继续加入培养基，最终使液体培养基的量达到10mL，在4℃冰箱中放置24小时完成浸提。Configure the SSAD powder extract used in the anaerobic bacteria experiment: Weigh 1 g of sterile SSAD dry powder, add an appropriate amount of TSB (tryptone soy broth medium) medium to fully absorb water and swell, and then continue to add the medium. The amount of liquid medium reached 10 mL, and the extraction was completed by placing in a refrigerator at 4°C for 24 hours.
需氧菌选用大肠杆菌和金黄色葡萄球菌，培养方法如下：配置需氧菌实验所用的浸提液，浸提24小时之后，将浸提液以3000r/min离心3分钟，以获得最终浸提液。在50mL离心管中加入SSAD粉浸提液10mL，接种500μL菌液，混合均匀，立刻放入培养箱中。Escherichia coli and Staphylococcus aureus were selected as the aerobic bacteria, and the culture method was as follows: configure the leaching solution used in the aerobic bacteria experiment, and after leaching for 24 hours, centrifuge the leaching solution at 3000 r/min for 3 minutes to obtain the final leaching solution. liquid. Add 10 mL of SSAD powder extract to a 50 mL centrifuge tube, inoculate 500 μL of bacterial solution, mix well, and immediately put it into the incubator.
厌氧菌选用牙龈卟啉单胞菌，培养方法如下：配置厌氧菌实验所用的浸提液，浸提24小时之后，将浸提液以3000r/min离心3分钟，以获得最终浸提液。在50mL离心管中加入SSAD粉浸提液10mL，按照5％浓度羊血，每管中加入500μL羊血，再分别接种同浓度500μL菌液，混合均匀，立刻放入厌氧培养箱中。The anaerobic bacteria were Porphyromonas gingivalis, and the cultivation method was as follows: configure the extract used in the anaerobic bacteria experiment, and after 24 hours of extraction, centrifuge the extract at 3000 r/min for 3 minutes to obtain the final extract . Add 10 mL of SSAD powder extract to a 50 mL centrifuge tube, add 500 μL of sheep blood to each tube according to 5% concentration of sheep blood, and then inoculate 500 μL of bacterial solution with the same concentration respectively, mix well, and immediately put it into an anaerobic incubator.
空白对照组的配置方法如下。需氧菌：离心管中加入LB培养基10mL，接种与实验组同浓度的菌液，作为空白对照组。厌氧菌：离心管中加入TSB培养基10mL，500μL羊血，接种与实验组同浓度的菌液，作为空白对照组。The configuration method of the blank control group is as follows. Aerobic bacteria: add 10 mL of LB medium to the centrifuge tube, and inoculate the bacterial solution with the same concentration as the experimental group as a blank control group. Anaerobic bacteria: add 10 mL of TSB medium and 500 μL of sheep blood to the centrifuge tube, and inoculate the bacterial solution with the same concentration as the experimental group as a blank control group.
细菌检测方法如下。大肠杆菌和金黄色葡萄球菌的检测：吸亮度的测定采用酶标仪(EnSpire，PerkinElmer，新加坡)，操作方法为从第0小时起，每隔2小时吸取各组混合均匀的液体以100μL每孔加入96孔板中，设置三个副孔。在OD＝600的条件下，测定各组的吸亮度。数据如表8和表9。The bacterial detection method is as follows. Detection of Escherichia coli and Staphylococcus aureus: The absorbance was measured using a microplate reader (EnSpire, PerkinElmer, Singapore), and the operation method was from the 0th hour onwards, and every 2 hours, the mixed liquid of each group was drawn with 100 μL per well. Add it to a 96-well plate and set up three sub-wells. Under the condition of OD=600, the absorbance of each group was measured. The data are shown in Table 8 and Table 9.
牙龈卟啉单胞菌的检测：吸亮度的测定采用酶标仪，操作方法为从第0天起，每天吸取各组混合均匀的液体以100μL每孔加入96孔板中，设置五个副孔。在OD＝600的条件下，测定各组的吸亮度。数据如表10。Detection of Porphyromonas gingivalis: The determination of the absorbance was performed using a microplate reader. The operation method was as follows: from the 0th day onwards, the well-mixed liquid of each group was drawn into a 96-well plate with 100 μL per well every day, and five auxiliary wells were set. . Under the condition of OD=600, the absorbance of each group was measured. The data are shown in Table 10.
表8：大肠杆菌抑菌效果对比Table 8: Comparison of bacteriostatic effects of Escherichia coli
表9：金黄色葡萄球菌抑菌效果对比Table 9: Comparison of antibacterial effects of Staphylococcus aureus
表10：牙龈卟啉单胞菌抑菌效果对比Table 10: Comparison of antibacterial effects of Porphyromonas gingivalis
上述实验证明，SSAD粉浸提液对于常见的两种需氧菌(大肠杆菌、金黄色葡萄球菌)具有较为明显的抑制作用，抑菌时间可达到16h。SSAD粉浸提液对于牙龈卟啉单胞菌(厌氧菌)的生长具有明显的抑制作用，抑菌时间可达到5天以上。The above experiments prove that the SSAD powder extract has a relatively obvious inhibitory effect on two common aerobic bacteria (Escherichia coli, Staphylococcus aureus), and the antibacterial time can reach 16h. SSAD powder extract has obvious inhibitory effect on the growth of Porphyromonas gingivalis (anaerobic bacteria), and the antibacterial time can reach more than 5 days.
实验3：抗氧化实验。Experiment 3: Antioxidant experiment.
首先制备SSAD粉浸提液，方法如下：将200mgSSAD干粉放入2mL的去离子水(重量比例约为1：10)浸泡7天，使SSAD干粉中的可溶性物质充分溶解，将液体取上清液得到SSAD粉浸提液。现场配制1mg/mL的DPPH乙醇溶液。First prepare the SSAD powder extract, the method is as follows: put 200mg SSAD dry powder into 2mL of deionized water (weight ratio is about 1:10) and soak for 7 days, so that the soluble substances in the SSAD dry powder are fully dissolved, and the liquid is taken as the supernatant The SSAD powder extract is obtained. Prepare a 1 mg/mL solution of DPPH in ethanol on site.
实验组采用SSAD粉浸提液1mL+DPPH乙醇溶液1mL；对照组采用纯水1mL+DPPH乙醇溶液1mL。用乙醇将分光亮度计调零，并于0.5小时、1小时、2小时分别测量在517nm波长，对照组的吸亮度值为A1，实验组的吸亮度值为A2。自由基清除率的计算公式为如下式III，测定结果如表11。The experimental group was treated with 1 mL of SSAD powder extract + 1 mL of DPPH ethanol solution; the control group was treated with 1 mL of pure water + 1 mL of DPPH ethanol solution. The spectrophotometer was zeroed with ethanol, and the wavelength was measured at 517 nm at 0.5 hours, 1 hour, and 2 hours, respectively. The absorbance value of the control group was A1 , and the absorbance value of the experimental group was A2 . The calculation formula of the free radical scavenging rate is the following formula III, and the measurement results are shown in Table 11.
表11：自由基清除率计算表Table 11: Calculation table of free radical scavenging rate
本实验表明，SSAD粉浸提液可以清除DPPH溶液中大部分自由基，具有抗氧化的功能。This experiment shows that the SSAD powder extract can scavenge most of the free radicals in the DPPH solution and has the function of anti-oxidation.
实验4：促细胞增殖迁移实验。Experiment 4: Promoting cell proliferation and migration experiment.
为模拟体外伤口愈合过程，进行细胞划痕实验和transwell小室实验。To simulate the wound healing process in vitro, cell scratch experiments and transwell chamber experiments were performed.
针对前述两种实验，首先准备完全培养基：取88份DMEM，10份胎牛血清、1份青霉素和1份链霉素，均匀混合，得到完全培养基。接着，按照1mgSSAD干粉添加1mL完全培养基的比例，将SSAD在完全培养基中浸泡7天，使SSAD中的可溶性物质充分溶解，得到浓度为1mg/mL的SSAD粉浸提培养基，并进一步制备5种不同浓度比例的含SSAD粉浸提培养基，浓度分别为0.5mg/mL、0.1mg/mL、0.05mg/mL、0.01mg/mL、0.005mg/mL。For the above two experiments, first prepare complete medium: take 88 parts of DMEM, 10 parts of fetal bovine serum, 1 part of penicillin and 1 part of streptomycin, and mix them evenly to obtain a complete medium. Next, according to the ratio of adding 1 mL of complete medium to 1 mg of SSAD dry powder, SSAD was soaked in the complete medium for 7 days to fully dissolve the soluble substances in SSAD to obtain 1 mg/mL of SSAD powder leaching medium, which was further prepared Five different concentration ratios of SSAD powder-containing extraction medium were used, and the concentrations were 0.5 mg/mL, 0.1 mg/mL, 0.05 mg/mL, 0.01 mg/mL, and 0.005 mg/mL, respectively.
在细胞划痕实验中，分别采用HUVES以及L929两种细胞进行测试，细胞的准备过程为：取对数生长期的脐静脉内皮细胞(HUVES)和成纤维细胞(L929)，胰酶消化得到单细胞悬浊液，细胞计数6×105个/mL；而后将1mL细胞悬浊液接种至6孔板中，再加1mL完全培养基，37℃孵箱中培养，待细胞贴壁且长满6孔板后，用200μL的黄色枪头进行划痕(横、纵向各划一道，使划痕呈“+”，以便后期观察时定位)，划痕后用PBS洗涤2-3遍，洗去漂浮的细胞，最后分别往其中5个小孔中加入前述5种不同浓度的SSAD粉浸提培养基2mL，一个小孔加入不含SSAD粉浸提培养基的完全培养基2mL作为空白对照组。所有的实验样本放入37℃孵箱中培养，分别在0h和24h的时候在显微镜下进行观察拍照，其中以0.1mg/mLSSAD粉浸提培养基的实验结果为代表，呈现如图5A所示，其余浓度则未示出。In the cell scratch experiment, HUVES and L929 cells were used for testing. The preparation process of the cells was as follows: umbilical vein endothelial cells (HUVES) and fibroblasts (L929) in logarithmic growth phase were taken, trypsinized to obtain single cells. Cell suspension, the cell count was 6×105 cells/mL; then 1 mL of cell suspension was inoculated into a 6-well plate, 1 mL of complete medium was added, and cultured in a 37°C incubator until the cells adhered and became confluent After the 6-well plate, scratch with a 200 μL yellow pipette tip (one horizontally and one vertically, so that the scratches are “+” for later observation), and wash with PBS for 2-3 times after scratching, and wash away For the floating cells, 2 mL of the aforementioned 5 different concentrations of SSAD powder extraction medium were added to 5 wells respectively, and 2 mL of complete medium without SSAD powder extraction medium was added to one well as a blank control group. All experimental samples were cultured in a 37°C incubator, and were observed and photographed under a microscope at 0 h and 24 h respectively. The experimental results of 0.1 mg/mL SSAD powder extraction medium were representative, as shown in Figure 5A. , the remaining concentrations are not shown.
利用Image-J软件对所采集图片进行面积计算与分析，通过实验证明，不同浓度的SSAD粉浸提培养基对HUVES以及L929两种细胞的迁移均有一定的促进作用，其中以0.1mg/mL的浓度效果最为显着(如图5B所示)，其细胞生长及迁移明显快于空白对照组，且两者之间的差异具有统计学意义。Image-J software was used to calculate and analyze the area of the collected pictures. It was proved by experiments that different concentrations of SSAD powder extraction medium had a certain promotion effect on the migration of HUVES and L929 cells. The concentration effect of the most significant (as shown in Figure 5B), its cell growth and migration were significantly faster than the blank control group, and the difference between the two was statistically significant.
接着，通过transwell小室实验，进一步验证5个不同浓度的SSAD粉浸提培养基对于HUVES以及L929两种细胞的体外迁移速率的影响，以反映其促进皮肤、黏膜再生的作用。Then, the effect of 5 different concentrations of SSAD powder extraction medium on the in vitro migration rate of HUVES and L929 cells was further verified by transwell experiments, in order to reflect its effect on promoting skin and mucosal regeneration.
分别采用HUVES以及L929两种细胞进行测试，细胞的准备过程为：取对数生长期的HUVES和L929，胰酶消化得到单细胞悬浊液，细胞计数HUVES 2×105个/mL，L929计数3×105个/mL，将transwell小室置于24孔板中，而后将100μL细胞悬浊液接种至transwell上室中，下室中分别加入800μLSSAD粉浸提培养基，37℃孵箱中培养24h后用镊子小心取出小室，吸干上室液体，移到预先加入约800μL甲醇的孔中，室温固定30分钟后取出小室，吸干上室中的固定液，将小室移到预先加入约800μL0.1％结晶紫染液的24孔板中，室温染色15－30分钟，染色完成后用PBS浸泡数次洗去多余染液，最后吸去上室中液体，用湿棉棒小心擦去上室底部膜表面上的细胞；待小室膜干燥后用镊子小心揭下膜，移至载玻片上用封片后，于200倍显微镜下观察，如图5C所示；而后对各组实验组样本随机截取的照片进行细胞计数，每个样本随机选取5个视野计数。图5C所示是呈现前述每个样本的1个视野作为示意；整体计数取得的细胞数量如表12所示，最后对得出细胞数量后进行差异统计分析。HUVES and L929 cells were used for testing. The preparation process of cells was as follows: take HUVES and L929 in logarithmic growth phase, trypsinize to obtain single cell suspension, count HUVES 2×105 cells/mL, and count
表12：transwell小室实验结果Table 12: Results of transwell chamber experiments
实验结果证明，不同浓度的SSAD粉浸提培养基对两种细胞的迁移速率均有不同程度的影响。其中对HUVES而言，0.5mg/mL、0.1mg/mLSSAD粉浸提培养基组和空白对照组之间的差异具有统计学意义；对L929而言，0.1mg/mL、0.05mg/mL、0.01mg/mLSSAD粉浸提培养基组和空白对照组之间的差异具有统计学意义。其中以0.1mg/mLSSAD粉浸提培养基对两种细胞的促进效果最为显着。本实验的实验结果表明，不同浓度的SSAD粉浸提培养基对两种细胞的迁移速率均有不同程度的影响，其中0.1mg/mLSSAD粉浸提培养基的促进效果尤为显着。The experimental results show that different concentrations of SSAD powder extraction medium have different degrees of influence on the migration rates of the two types of cells. Among them, for HUVES, the differences between 0.5mg/mL, 0.1mg/mL SSAD powder extraction medium group and blank control group were statistically significant; for L929, 0.1mg/mL, 0.05mg/mL, 0.01mg/mL The difference between the mg/mL SSAD powder extraction medium group and the blank control group was statistically significant. Among them, the 0.1mg/mL SSAD powder extraction medium had the most significant promoting effect on the two kinds of cells. The experimental results of this experiment showed that different concentrations of SSAD powder extraction medium had different effects on the migration rates of the two types of cells, and the promotion effect of 0.1mg/mL SSAD powder extraction medium was particularly significant.
以上体外细胞实验结果均说明，SSAD粉浸提培养基的细胞兼容性较好，对细胞未见明显毒性，且0.1mg/mLSSAD粉浸提培养基能显着加快HUVES以及L929的迁移。The above in vitro cell experiments showed that the SSAD powder extraction medium had good cell compatibility and no obvious toxicity to cells, and 0.1mg/mL SSAD powder extraction medium could significantly accelerate the migration of HUVES and L929.
实验5：医用止血粉在体内降解的评估和效果。Experiment 5: Evaluation and effect of medical hemostatic powder in vivo degradation.
在深吸入异氟醚全身麻醉下，SD大鼠俯卧位，背部无菌准备手术。在脊柱轴外的备皮肤切口(3厘米)，和底层皮下组织分离，为植入医用止血粉提供足够的空间。取100mg医用止血粉，与200-600μL PBS混合后，植入皮下空间，植入后缝合关闭皮肤。术后3、7、14天收集周围组织及全皮肤进行组织学分析，评估医用止血粉的降解效果，其降解效果如图6所示。Under general anesthesia with deep inhalation of isoflurane, SD rats were placed in prone position, and the back was aseptically prepared for surgery. The prepared skin incision (3 cm) outside the spinal axis is separated from the underlying subcutaneous tissue to provide enough space for implanting medical hemostatic powder. Take 100 mg of medical hemostatic powder, mix it with 200-600 μL of PBS, and then implant it into the subcutaneous space, and suture to close the skin after implantation. 3, 7, and 14 days after the operation, the surrounding tissue and the whole skin were collected for histological analysis to evaluate the degradation effect of the medical hemostatic powder. The degradation effect is shown in Figure 6.
医用止血粉在体内植入3、7和14天后，H&E和Masson染色(分别如图6a、图6b所示)显示出轻度炎症反应。植入3天后，在植入的医用止血粉最外层观察到中度急性炎症反应，有典型的炎性细胞染成深蓝色(即如图6中颜色相对较深的细胞)。植入7天后，医用止血粉结构开始失去完整性，几乎被侵入的炎性细胞所填满，几乎没有观察到纤维囊，说明宿主对医用止血粉的反应较弱。此外，植入14天后，植入部位几乎没有医用止血粉残留，皮肤结构与空白对照一样正常，说明医用水性水凝胶在体内可以完全降解。After 3, 7 and 14 days of medical hemostatic powder implantation in vivo, H&E and Masson staining (shown in Figure 6a and Figure 6b, respectively) showed mild inflammatory responses. After 3 days of implantation, a moderate acute inflammatory response was observed in the outermost layer of the implanted medical hemostatic powder, and typical inflammatory cells were stained dark blue (ie, relatively dark cells as shown in Figure 6). After 7 days of implantation, the structure of the medical hemostatic powder began to lose its integrity and was almost filled with invading inflammatory cells, and almost no fibrous capsule was observed, indicating that the host had a weak response to the medical hemostatic powder. In addition, after 14 days of implantation, almost no medical hemostatic powder remained at the implantation site, and the skin structure was as normal as that of the blank control, indicating that the medical aqueous hydrogel can be completely degraded in vivo.
使用淋巴细胞(CD3)和巨噬细胞(CD68)标志物染色，评估创口愈合区域的细胞特征，其结果如图6c所示。在图6c中i、v、ix为术后3天切片同一视野下的局部放大图；同理，ii、vi、x表示术后7天的图像；iii、vii、xi表示术后14天的图像；iv、viii、xii表示术后21天的图像。从图6c可以发现，植入后第3天，在医用止血粉治疗组在移植物周围淋巴细胞浸润率为0.23±0.06％，仅见少量巨噬细胞浸润；巨噬细胞浸润在第7天达到最大值(3.21±0.87％)；随着时间的推移，淋巴细胞和巨噬细胞的浸润的数量都减少，到第21天几乎完全消失，同时，以上过程进行了量化统计，结果如图6d和6e所示，各时间点之间都具备统计上差异；这一观察结果证明医用止血粉具有良好的生物兼容性，在体内可完全降解且几乎无刺激性，未见明显的免疫排斥反应。Using lymphocyte (CD3) and macrophage (CD68) marker staining, the cellular characteristics of the wound healing area were assessed and the results are shown in Figure 6c. In Figure 6c, i, v, and ix are the magnified images of the slices in the same field of view on the 3rd day after operation; similarly, ii, vi, and x represent the images on the 7th day after the operation; iii, vii, and xi represent the images on the 14th day after the operation. Images; iv, viii, xii represent
实验6：医用止血粉主要物质含量。Experiment 6: The main substance content of medical hemostatic powder.
本发明医用止血粉主要活性成分来自大鲵皮肤黏液干粉，采用液相色谱技术对其主要物质含量进行检测，发现其中87％以上为各种氨基酸。见表13。The main active components of the medical hemostatic powder of the present invention come from the dry powder of giant salamander skin mucus, and the content of the main substances is detected by liquid chromatography technology, and it is found that more than 87% of which are various amino acids. See Table 13.
表13：大鲵皮肤黏液干粉主要物质含量Table 13: Contents of main substances in dry powder of giant salamander skin mucus
以上所述仅为本发明之优选实施例及实验，并非用以限定本发明之权利范围；同时以上的描述，对于相关技术领域之专门人士应可明了及实施，因此其他未脱离本发明所揭示之精神下所完成的等效改变或修饰，均应包含在权利要求范围中。The above descriptions are only preferred embodiments and experiments of the present invention, and are not intended to limit the scope of rights of the present invention; at the same time, the above descriptions should be understood and implemented by those skilled in the relevant technical fields, so others do not depart from those disclosed in the present invention. Equivalent changes or modifications accomplished within the spirit of the claims should be included in the scope of the claims.
Claims (8)
Translated fromChinesePriority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011071241.7ACN112121065A (en) | 2020-10-09 | 2020-10-09 | Medical styptic powder and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011071241.7ACN112121065A (en) | 2020-10-09 | 2020-10-09 | Medical styptic powder and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112121065Atrue CN112121065A (en) | 2020-12-25 |
Family
ID=73843929
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011071241.7APendingCN112121065A (en) | 2020-10-09 | 2020-10-09 | Medical styptic powder and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112121065A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114984233A (en)* | 2021-11-05 | 2022-09-02 | 重庆医科大学附属口腔医院 | A local drug delivery system based on salamander skin secretions and its application |
| CN116077731A (en)* | 2021-11-05 | 2023-05-09 | 重庆医科大学附属口腔医院 | Tissue repair material based on giant salamander skin secretion and application thereof |
| CN117797172A (en)* | 2024-02-28 | 2024-04-02 | 合肥启色生物科技有限公司 | Preparation method of anti-inflammatory, hemostatic and healing-promoting combination |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104758978A (en)* | 2015-04-02 | 2015-07-08 | 重庆馗旭生物科技股份有限公司 | Application of giant salamander mucus in preparation of bleeding-stopping material |
| CN106581736A (en)* | 2016-12-28 | 2017-04-26 | 安顺市四海农业科技开发有限公司 | Medical adhesive prepared by adopting skin mucus of giant salamander as raw material and preparation method |
| CN110559469A (en)* | 2019-02-26 | 2019-12-13 | 重庆医科大学附属口腔医院 | Medical adhesive |
| CN110755675A (en)* | 2019-12-10 | 2020-02-07 | 中国人民解放军陆军军医大学第一附属医院 | Composite biological paste capable of rapidly stopping bleeding and preparation method and application thereof |
- 2020
- 2020-10-09CNCN202011071241.7Apatent/CN112121065A/enactivePending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104758978A (en)* | 2015-04-02 | 2015-07-08 | 重庆馗旭生物科技股份有限公司 | Application of giant salamander mucus in preparation of bleeding-stopping material |
| CN106581736A (en)* | 2016-12-28 | 2017-04-26 | 安顺市四海农业科技开发有限公司 | Medical adhesive prepared by adopting skin mucus of giant salamander as raw material and preparation method |
| CN110559469A (en)* | 2019-02-26 | 2019-12-13 | 重庆医科大学附属口腔医院 | Medical adhesive |
| CN110755675A (en)* | 2019-12-10 | 2020-02-07 | 中国人民解放军陆军军医大学第一附属医院 | Composite biological paste capable of rapidly stopping bleeding and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 陈丽萍等: "大鲵黏液低聚糖肽抑菌、抗血小板聚集活性研究", 《亚太传统医药》* |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114984233A (en)* | 2021-11-05 | 2022-09-02 | 重庆医科大学附属口腔医院 | A local drug delivery system based on salamander skin secretions and its application |
| CN116077731A (en)* | 2021-11-05 | 2023-05-09 | 重庆医科大学附属口腔医院 | Tissue repair material based on giant salamander skin secretion and application thereof |
| CN114984233B (en)* | 2021-11-05 | 2023-07-14 | 重庆医科大学附属口腔医院 | A local drug delivery system based on salamander skin secretion and its application |
| CN117797172A (en)* | 2024-02-28 | 2024-04-02 | 合肥启色生物科技有限公司 | Preparation method of anti-inflammatory, hemostatic and healing-promoting combination |
| CN117797172B (en)* | 2024-02-28 | 2024-05-14 | 合肥启色生物科技有限公司 | Preparation method of anti-inflammatory, hemostatic and healing-promoting combination |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112121065A (en) | Medical styptic powder and application thereof | |
| He et al. | Facile preparation of PVA hydrogels with adhesive, self-healing, antimicrobial, and on-demand removable capabilities for rapid hemostasis | |
| JPH02107267A (en) | Composition for wound filling | |
| JP7378486B2 (en) | Medical adhesives, their preparation methods, and their uses | |
| CN112107723B (en) | Medical water-based adhesive and using method thereof | |
| AU2011349659B2 (en) | Hemostatic preparation containing an extract of Golden Moss | |
| RU2699362C2 (en) | Composition based on cerium dioxide nanoparticles and brown algae polysaccharides for treating wounds | |
| CN108404199A (en) | A kind of multiple trauma dressing and preparation method thereof with tissue adhesive property | |
| CN113134113B (en) | Preparation method of absorbable hemostatic fluid gelatin and absorbable hemostatic fluid gelatin | |
| CN111450306A (en) | External application of nano-hydroxyapatite/polydopamine wet adhesion type hemostatic powder and preparation method thereof | |
| WO2020180202A1 (en) | Composition based on cerium dioxide nanoparticles and brown algae polysaccharides for wound treatment | |
| Shi et al. | A multifunctional thermosensitive hydrogel based on phototherapy for promoting the healing of dental extraction wounds | |
| CN112190770A (en) | Degradable tooth extraction wound filling composition and preparation method and application thereof | |
| CN116115820A (en) | Preparation and application of visible light crosslinking composite gel hemostatic dressing | |
| CN115970044A (en) | Double-layer structured chitosan hemostatic sponge and preparation method thereof | |
| CN108355164B (en) | Absorbable platelet-like biomass material and application thereof | |
| CN114796160A (en) | Antibacterial material and application thereof | |
| Dong et al. | Injectable chitosan-based hydrogels harnessing resveratrol micelles for treatment of infected wounds | |
| Raafat et al. | Radiation development and hemostatic performance of innovative hydroxypropyl methyl cellulose-based sponge dressings for controlling severe hemorrhagic wounds | |
| CN114887113B (en) | Preparation method and application of Gelma gel of platelet-supported film-coated cerium oxide | |
| Tuna et al. | Aloehec band-aid and diapers. | |
| CN120285269A (en) | A composite powder with high liquid absorption, tissue adhesion and antibacterial properties and its application | |
| CN119055844A (en) | A zinc-doped bioactive glass composite hydrogel and its preparation method and application in preparing wound repair materials | |
| CN120361298A (en) | Silk fibroin/chitosan/graphene oxide based composite material and preparation method and application thereof | |
| WO2024116120A1 (en) | Matrices for wound healing |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |