CN102526796A - Absorbable hemostatic material prepared based on rat tail collagen and preparation method thereof - Google Patents

Abstract

The invention relates to an absorbable hemostatic material prepared based on rat tail collagen and a preparation method thereof. The components and the weight ratio of the components are 1-10 percent of rat tail collagen, 0.2-5 percent of polyvinyl alcohol and 0.2-2 percent of chitosan, and the balance of water, and the components are prepared by adopting a freeze drying process. The hemostatic material prepared by the invention is mainly prepared from rat tail collagen, and has the defects of non-absorbability, easy causing organism allergy, poor hemostatic effect and the like compared with the traditional hemostatic material; the hemostatic effect is fast, and the hydrophilic property is strong; simultaneously, the aggregation of the platelets can be activated, and once the platelets are aggregated, thrombus can be formed, so that the plasma is promoted to agglomerate to stop blood flow. Meanwhile, a specific area on the surface of the collagen can be combined with glycoprotein similar to fibronectin existing on the surface of cells, and the effect of preventing intestinal adhesion can be achieved.

Description

Translated fromChinese

一种基于鼠尾胶原蛋白制备的可吸收止血材料及其制备方法Absorbable hemostatic material prepared based on rat tail collagen and preparation method thereof

【技术领域】【Technical field】

本发明涉及一种基于鼠尾胶原蛋白制备的产品，可吸收止血材料及其制备方法。The invention relates to a product prepared based on rat tail collagen, an absorbable hemostatic material and a preparation method thereof.

【背景技术】【Background technique】

胶原蛋白是脊椎动物和无脊椎动物支持结构的主要组成。在人的身体中这是皮肤、筋、软骨、骨骼及结缔组织的最主要的蛋白质。胶原蛋白占人体蛋白质总量的三分之一，不论来源于哪一种动物或哪一种组织的胶原都有许多共同特性。氨基酸组成中约有三分之一为甘氨酸，有脯氨酸和羟脯氨酸。根据其遗传分子结构遗传基因的差别，胶原蛋白分为20几个亚型。但最为常见的为I、II、III型胶原蛋白，即间质胶原蛋白。其中I型最为丰富且品质优良。I型胶原蛋白分布非常广泛，是主纤维束的主要成分，给结缔组织以强度；是最丰富的胶原蛋白类型，尤其是在皮肤真皮、骨、筋和腱中。Collagen is a major component of support structures in vertebrates and invertebrates. In the human body it is the most important protein of skin, tendon, cartilage, bone and connective tissue. Collagen accounts for one-third of the total protein in the human body. No matter which animal or tissue it comes from, collagen has many common characteristics. About one-third of the amino acid composition is glycine, and there are proline and hydroxyproline. Collagen is divided into more than 20 subtypes according to the difference of its genetic molecular structure and genetic gene. But the most common ones are type I, II, and III collagen, that is, interstitial collagen. Among them, type I is the most abundant and of good quality. Type I collagen is very widely distributed and is the main component of the main fiber bundles that give strength to connective tissue; it is the most abundant type of collagen, especially in the dermis of the skin, bones, tendons and tendons.

I型胶原蛋白是由两条α1链和一条α2链组成。以共价键交联互相缠绕构成了螺旋形高分子，长约300nm，直径1.4-1.5nm，由约1050个氨基酸构成，分子量约300kD。由于其N端及C端的非胶原性肽(尾肽)部分桥键比较多，加上其螺旋形态结构，故刚性强、溶解度低，给I型胶原蛋白的提取带来一定难度。Type I collagen is composed of two α1 chains and one α2 chain. The helical macromolecule is formed by cross-linking and intertwining with covalent bonds, with a length of about 300nm and a diameter of 1.4-1.5nm. It is composed of about 1050 amino acids and has a molecular weight of about 300kD. Because of its N-terminal and C-terminal non-collagenous peptide (tail peptide) part of the bridge more, coupled with its helical structure, so rigidity, low solubility, brings certain difficulties to the extraction of type I collagen.

一般的生物体中提取的胶原蛋白多为混合型，之前多从牛皮、猪皮、牛腱中提取，但这类胶原蛋白不仅有油脂等其他杂质，也存在着II型及其他一些胶原蛋白亚型；同时，这种畜牧业大量饲养的动物存在着例如疯牛病等一些生物性疾病及病毒感染，如果用此开发医用产品更加存在着品质的不稳定及潜在的风险及危害；再次，人的真皮中主要含有I型和III型，但III型不易大量获得，故应用单一的I型胶原蛋白是解决体外应用的医用产品引起的抗原性问题的关键，这也是I型胶原蛋白抗原性低的原因之一。同时，I型胶原蛋白的三重螺旋的氨基酸结构决定了它的一些特有的性能，如机械性能、促进细胞生长、弱抗原性、可生物降解性和胶原的自缔合性。因为I型胶原蛋白不仅对细胞起到锚定和支持的作用，还为细胞的生长提供适宜的微环境，积极参与了细胞迁移，分化与增殖等细胞行为，可完全被人体吸收，启动凝血因子，对胚胎发育和创伤修复有积极作用。Collagen extracted from general organisms is mostly mixed type. It was extracted from cowhide, pigskin, and beef tendon before, but this type of collagen not only contains oil and other impurities, but also contains type II and other collagen subtypes. At the same time, there are some biological diseases and viral infections such as mad cow disease in the animals raised in large quantities in this kind of animal husbandry. If this kind of medical product is used to develop medical products, there will be more unstable quality and potential risks and hazards; again, human leather Type I and type III are mainly contained in collagen, but type III is not easy to obtain in large quantities, so the application of a single type I collagen is the key to solving the antigenicity problem caused by medical products applied in vitro, which is also the reason for the low antigenicity of type I collagen one. At the same time, the amino acid structure of the triple helix of type I collagen determines some of its unique properties, such as mechanical properties, promotion of cell growth, weak antigenicity, biodegradability and self-association of collagen. Because type I collagen not only anchors and supports cells, but also provides a suitable microenvironment for cell growth, actively participates in cell migration, differentiation and proliferation, and can be completely absorbed by the human body to activate coagulation factors , has a positive effect on embryonic development and wound repair.

随着现代医学的发展，各种医用耗材的更新换代也是日新月异。止血材料是常见的医疗器械产品。但是现今市面上的止血材料不仅存在着容易引起伤口感染的缺点；同时还存在着容易与伤口粘附不易去除，导致伤口溃烂，或者因为粘附导致感染；再次就是止血材料多是通过吸收血液及添加凝血酶等外在因素止血，很少有止血材料是使用能够自身具有止血性质的材料来生产的。现今市面上的可吸收止血材料有氧化纤维素、α-氰基丙烯酸酯类组织胶、医用止血明胶等，但是都有各自的缺点。如氧化纤维素可引起组织周围pH值升高；α-氰基丙烯酸酯类组织胶降解过程中释放出氰和甲醛等有毒物质；医用止血明胶黏附性较差，易脱落，因其吸收血液后膨胀可能压迫伤口周围组织等。With the development of modern medicine, the replacement of various medical consumables is also changing with each passing day. Hemostatic materials are common medical device products. However, the hemostatic materials on the market today not only have the disadvantage of easily causing wound infection; they also have the disadvantage that they are easy to adhere to the wound and are not easy to remove, resulting in wound ulceration, or infection due to adhesion; Adding external factors such as thrombin to stop bleeding, few hemostatic materials are produced using materials that can have hemostatic properties themselves. Absorbable hemostatic materials currently on the market include oxidized cellulose, α-cyanoacrylate tissue glue, medical hemostatic gelatin, etc., but each has its own shortcomings. For example, oxidized cellulose can cause an increase in the pH value around the tissue; α-cyanoacrylate tissue glue releases toxic substances such as cyanide and formaldehyde during the degradation process; medical hemostatic gelatin has poor adhesion and is easy to fall off because it absorbs blood. The swelling may compress the tissue surrounding the wound, etc.

迄今为止，还没有十分理想的可吸收止血材料面市。因此开发性能更为优良的相关产品显得很有必要。So far, there is no ideal absorbable hemostatic material available on the market. Therefore, it is necessary to develop related products with better performance.

【发明内容】【Content of invention】

本发明的目的就是为了克服现有制备的止血贴不可吸收、容易引起机体过敏、止血效果差等缺点，提供了一种基于鼠尾胶原蛋白制备的具有人体内可吸收、生物相容性好、止血效果快的止血材料，本发明还提供这种止血材料的制备方法。The purpose of the present invention is to overcome the disadvantages of the existing hemostatic patch, such as non-absorbable, easy to cause body allergy, poor hemostatic effect, etc., to provide a human body absorbable, biocompatible, The hemostatic material with fast hemostatic effect, the invention also provides a preparation method of the hemostatic material.

基于申请号为201210007089.5的发明专利申请“一种制备鼠尾胶原蛋白的方法”所叙述的方法，制备的鼠尾胶原经过SDS-PAGE鉴定后与I型胶原蛋白标准品(SIGMA产品)相比较无杂带，无任何差异，达到了SDS纯度。足以证明此种鼠尾胶原蛋白为单一的I型胶原蛋白。同时，制备所用鼠尾胶原蛋白经检测不存在任何潜在的生物安全问题，制备的胶原蛋白纯度高，品质好。Based on the method described in the invention patent application "a method for preparing rat tail collagen" with the application number 201210007089.5, the prepared rat tail collagen was identified by SDS-PAGE compared with the type I collagen standard (SIGMA product). Miscellaneous bands, without any difference, reached SDS purity. It is sufficient to prove that the rat tail collagen is a single type I collagen. At the same time, the rat tail collagen used in the preparation does not have any potential biological safety problems after testing, and the prepared collagen has high purity and good quality.

基于上述原理及制备方法，本发明模拟细胞外间质的组成特点，研制并推出了胶原蛋白为主要基质的医用生物材料。对胶原蛋白进行相应处理加工，制备成胶原蛋白溶液喷雾剂、胶原蛋白止血材料。Based on the above principle and preparation method, the present invention simulates the composition characteristics of extracellular matrix, develops and releases medical biomaterials with collagen as the main matrix. Collagen is processed accordingly to prepare collagen solution spray and collagen hemostatic material.

本发明制备止血材料的方法包括如下步骤：The method for preparing hemostatic material of the present invention comprises the following steps:

1、选择聚乙烯醇、壳聚糖作为辅料，与鼠尾胶原蛋白联合，共同制成冻干止血材料或止血喷雾剂；各个材料重量配比为鼠尾胶原蛋白∶聚乙烯醇∶壳聚糖为1-10％∶0.2-5％∶0.2-2％，余量的水；1. Select polyvinyl alcohol and chitosan as auxiliary materials, combine with rat tail collagen to make freeze-dried hemostatic material or hemostatic spray; the weight ratio of each material is rat tail collagen: polyvinyl alcohol: chitosan 1-10%: 0.2-5%: 0.2-2%, the rest of the water;

2、将鼠尾胶原蛋白∶聚乙烯醇∶壳聚糖混合后，用无菌水溶解后浇入培养小板，-40-0℃预冻2h-12h，再转入-80℃超低温冰箱急冻4h-12h，再转入真空冷冻干燥机中将复合材料在-4℃下冷冻干燥10-60h，即可以得到复合型冻干止血材料；Co60辐射灭菌，干燥保存；冻干的材料还可直接用于各种伤口的止血材料；2. After mixing rat tail collagen: polyvinyl alcohol: chitosan, dissolve it with sterile water and pour it into a small culture plate, pre-freeze at -40-0°C for 2h-12h, and then transfer to -80°C ultra-low temperature refrigerator for emergency Freeze for 4h-12h, and then transfer to a vacuum freeze dryer to freeze-dry the composite material at -4°C for 10-60h to obtain a composite freeze-dried hemostatic material; Co60 radiation sterilization, dry storage; freeze-dried material Hemostatic materials that can be directly used in various wounds;

3、对复合材料进行性状观察，主要包括均匀度、成膜性、脱膜性、柔韧性、抗拉强度、粘附性、吸水性等。均匀度、成膜性、脱膜性反映材料的制备工艺成熟度；柔韧性、抗拉强度、粘附性、吸水性反应材料的止血性能。3. Observe the properties of composite materials, mainly including uniformity, film forming, release, flexibility, tensile strength, adhesion, water absorption, etc. Uniformity, film-forming property, and release property reflect the maturity of the preparation process of the material; flexibility, tensile strength, adhesion, and water absorption reflect the hemostatic performance of the material.

动物试验表明，无论新西兰兔耳缘静脉、耳缘动脉、股动脉、肝脏创伤出血，均与医用棉花、纱布止血存在极为显著性差异。表明本发明的止血材料具有明显的止血作用。Animal experiments have shown that there are extremely significant differences between New Zealand rabbit's ear vein, ear artery, femoral artery, and liver trauma compared with medical cotton and gauze for hemostasis. It shows that the hemostatic material of the present invention has obvious hemostatic effect.

本发明的制备的鼠尾胶原来自于各种品系的大鼠、小鼠鼠尾。The rat tail collagen prepared in the present invention comes from rat tails of various strains and mice.

本发明的止血材料可以按照本技术领域的技术人员熟知的技术加工成创可贴、喷雾剂等剂型。The hemostatic material of the present invention can be processed into dosage forms such as band-aids and sprays according to techniques well known to those skilled in the art.

为了保证制备的鼠尾胶原蛋白的质量，本发明涉及的鼠尾胶原蛋白的提取也可以来自于SPF级(无特定病原体)成年SD大鼠鼠尾。In order to ensure the quality of the prepared rat tail collagen, the rat tail collagen involved in the present invention can also be extracted from the tail of SPF (specific pathogen-free) adult SD rats.

本发明所述壳聚糖为水溶性壳聚糖，其分子量为3000道尔顿-40万道尔顿。The chitosan of the present invention is water-soluble chitosan, and its molecular weight is 3000-400,000 daltons.

可吸收止血材料发明的优点：Advantages of the invention of absorbable hemostatic material:

1、本发明制作的止血材料采用温和的冷冻干燥操作工艺不仅仅提高了生产效率，同时避开了使用其他设备带来的成本上升问题。1. The hemostatic material produced by the present invention adopts a mild freeze-drying operation process, which not only improves the production efficiency, but also avoids the cost increase caused by the use of other equipment.

2、本发明专利生产的止血材料(止血海绵)，其动物实验表明其具有非常好的止血效果。也可应用于内脏出血。具有广阔的应用前景。2. The hemostatic material (hemostatic sponge) produced by the patent of the present invention, its animal experiment shows that it has a very good hemostatic effect. It can also be used for visceral bleeding. have a broad vision of application.

3、本发明制备的止血材料具有可完全被人体吸收，与出血创面一接触，其接触面迅速形成凝胶而粘附在出血创面上，外面则形成连续的压迫干层。启动凝血因子。同时，聚乙烯醇为成膜材料，两种的协同效应形成了一种理想的止血状态和结构。3. The hemostatic material prepared by the present invention can be completely absorbed by the human body. When it comes into contact with the bleeding wound, the contact surface quickly forms a gel and adheres to the bleeding wound, and forms a continuous compressed dry layer on the outside. Activate coagulation factors. At the same time, polyvinyl alcohol is a film-forming material, and the synergistic effect of the two forms an ideal hemostatic state and structure.

4、本发明制备的止血材料，经过动物实验的验证，在腹腔止血时，还具有明显的防止肠粘连方面的功效。4. The hemostatic material prepared by the present invention, through the verification of animal experiments, also has the obvious effect of preventing intestinal adhesion when hemostasis in the abdominal cavity.

5、有促进受损组织创面愈合的作用。所形成的持续时间较长的稳定凝胶，可保证新生创面组织顺利地长入，同时辅料壳聚糖也具有促进伤口愈合的功效，从而达到创面修复的目的。聚乙烯醇为成膜材料，也可以促进伤口愈合。5. It can promote wound healing of damaged tissue. The stable gel formed with a long duration can ensure the smooth growth of new wound tissue. At the same time, the auxiliary material chitosan also has the effect of promoting wound healing, so as to achieve the purpose of wound repair. Polyvinyl alcohol is a film-forming material that also promotes wound healing.

【具体实施方式】【Detailed ways】

实施例1：止血材料的制备Embodiment 1: Preparation of hemostatic material

1、选择能与鼠尾胶原蛋白联合发挥作用的聚乙烯醇、壳聚糖作为辅料。各个材料重量配比为鼠尾胶原蛋白∶聚乙烯醇∶壳聚糖为1％∶0.2％∶0.2％。余量为水。1. Select polyvinyl alcohol and chitosan that can work together with rat tail collagen as auxiliary materials. The weight ratio of each material is rat tail collagen: polyvinyl alcohol: chitosan is 1%: 0.2%: 0.2%. The balance is water.

2、将混合材料用无菌水溶解后浇入培养小板，-40℃预冻2h，再转入-80℃超低温冰箱急冻4h，再转入真空冷冻干燥机中将复合材料在-4℃下冷冻干燥10h，即可以得到复合型冻干止血材料。Co60辐射灭菌，干燥保存。冻干的材料可直接用于各种伤口的止血。2. Dissolve the mixed material in sterile water and pour it into a small culture plate, pre-freeze at -40°C for 2 hours, then transfer it to a -80°C ultra-low temperature refrigerator for 4 hours, and then transfer it to a vacuum freeze dryer to store the composite material at -4 The composite freeze-dried hemostatic material can be obtained by freeze-drying at ℃ for 10 hours. Co60 radiation sterilization, dry storage. The freeze-dried material can be directly used for hemostasis of various wounds.

实施例2：止血材料的制备Embodiment 2: Preparation of hemostatic material

1、选择能与鼠尾胶原蛋白联合发挥作用的聚乙烯醇、壳聚糖作为辅料。各个材料重量配比为鼠尾胶原蛋白∶聚乙烯醇∶壳聚糖为5％∶2％∶1％，余量为水。1. Select polyvinyl alcohol and chitosan that can work together with rat tail collagen as auxiliary materials. The weight ratio of each material is rat tail collagen: polyvinyl alcohol: chitosan is 5%: 2%: 1%, and the balance is water.

2、将混合材料用无菌水溶解后浇入培养小板，-20℃预冻7h，再转入-80℃超低温冰箱急冻8h，再转入真空冷冻干燥机中将复合材料在-4℃下冷冻干燥30h，即可以得到复合型冻干止血材料。Co60辐射灭菌，干燥保存。冻干的材料可直接用于各种伤口的止血。2. Dissolve the mixed material in sterile water and pour it into a small culture plate, pre-freeze at -20°C for 7 hours, then transfer it to a -80°C ultra-low temperature refrigerator for 8 hours, and then transfer it to a vacuum freeze dryer to store the composite material at -4 The composite freeze-dried hemostatic material can be obtained by freeze-drying at ℃ for 30 hours. Co60 radiation sterilization, dry storage. The freeze-dried material can be directly used for hemostasis of various wounds.

实施例3：止血材料的制备Embodiment 3: Preparation of hemostatic material

1、选择能与鼠尾胶原蛋白联合发挥作用的聚乙烯醇、壳聚糖作为辅料。各个材料重量配比为鼠尾胶原蛋白∶聚乙烯醇∶壳聚糖为10％∶5％∶2％，余量为水。1. Select polyvinyl alcohol and chitosan that can work together with rat tail collagen as auxiliary materials. The weight ratio of each material is rat tail collagen: polyvinyl alcohol: chitosan is 10%: 5%: 2%, and the balance is water.

2、将混合材料用无菌水溶解后浇入培养小板，0℃预冻12h，再转入-80℃超低温冰箱急冻12h，再转入真空冷冻干燥机中将复合材料在-4℃下冷冻干燥60h，即可以得到复合型冻干止血材料。Co60辐射灭菌，干燥保存。冻干的材料可直接用于各种伤口的止血。2. Dissolve the mixed material with sterile water and pour it into the culture plate, pre-freeze at 0°C for 12 hours, then transfer to -80°C ultra-low temperature refrigerator for 12 hours, and then transfer the composite material to a vacuum freeze dryer at -4°C After 60 hours of freeze-drying, the composite freeze-dried hemostatic material can be obtained. Co60 radiation sterilization, dry storage. The freeze-dried material can be directly used for hemostasis of various wounds.

实施例4：本发明实施例2的止血效果试验：Embodiment 4: The hemostatic effect test of embodiment 2 of the present invention:

1、以本发明实施例2的止血材料为实验材料，动物实验分别进行了新西兰兔的耳缘静脉出血、耳缘动脉出血、股动脉出血实验，以医用棉花和纱布作为对照。取普通级新西兰兔20只，按体重随机分为对照组、鼠尾胶原止血材料组，10只/组。每只兔分别切断右耳耳缘静脉、左耳耳缘动脉及切开右后肢股动脉3/4，自由出血5s。5s后对照组压敷消毒棉花，鼠尾胶原止血材料组压敷鼠尾胶原止血材料。每隔30s观察伤口的流血情况，并用药棉吸附所出血液。记录止血时间。采用单因素方差分析的方法，实验组所用的复合材料使实验动物的出血时间明显缩短，统计学分析具有显著性差异，是下一步开发成医用止血的好材料。所有结果如表1所示。1. Using the hemostatic material of Example 2 of the present invention as the experimental material, animal experiments were carried out on ear vein bleeding, ear artery bleeding, and femoral artery bleeding in New Zealand rabbits, with medical cotton and gauze as controls. Take 20 ordinary New Zealand rabbits, and divide them randomly into control group and rat tail collagen hemostatic material group according to body weight, 10 rabbits/group. The auricular vein of the right ear, the auricular artery of the left ear and 3/4 of the femoral artery of the right hindlimb were cut off for each rabbit, and the bleeding was free for 5 seconds. After 5 seconds, the control group applied pressure on sterilized cotton, and the rat tail collagen hemostatic material group applied pressure on rat tail collagen hemostatic material. Observe the bleeding of the wound every 30 seconds, and absorb the blood with cotton wool. Record the hemostasis time. Using the method of single-factor analysis of variance, the composite material used in the experimental group can significantly shorten the bleeding time of the experimental animals, and the statistical analysis has a significant difference. It is a good material for medical hemostasis in the next step. All results are shown in Table 1.

表1.止血材料对新西兰兔耳缘静脉、耳缘动脉、股动脉止血时间的影响

Table 1. Effects of hemostatic materials on hemostasis time of marginal ear vein, marginal ear artery and femoral artery of New Zealand rabbits

注：与对照组相比，“△”P＜0.05，“△△”P＜0.01Note: Compared with the control group, "△"P<0.05, "△△"P<0.01

依照本发明专利使用冷冻干燥的制备工艺，所制备的止血材料具有显著性的止血效果，与对照组的医用棉花及纱布相比其止血速度提高1-3倍。According to the preparation process of the patent of the present invention using freeze-drying, the prepared hemostatic material has a significant hemostatic effect, and its hemostatic speed is increased by 1-3 times compared with the medical cotton and gauze of the control group.

2、以本发明实施例2的止血材料为实验材料，进行了新西兰兔肝脏创伤模型的止血效果试验，新西兰兔，20只，随机分为对照组，止血材料组，每组10只。麻醉后，固定在实验台，在无菌条件下开腹，暴露肝脏，并在肝表面切除约1cm×1cm的肝组织，造成出血创面。对照组给予消毒海绵止血，止血材料组给予止血材料，敷压伤口，观察止血情况，记录止血时间，测量出血量。术后缝合伤口，动物单笼饲养，自由饮水进食，观察新西兰兔存活情况。于术后第2天、第7天各组取3只新西兰兔，术后第14天和第21天各取3只新西兰兔，剖腹观察腹腔内粘连、腹腔内感染及肝脏愈合情况。2. Using the hemostatic material of Example 2 of the present invention as the experimental material, the hemostatic effect test of the New Zealand rabbit liver trauma model was carried out. Twenty New Zealand rabbits were randomly divided into a control group and a hemostatic material group, 10 in each group. After anesthesia, it was fixed on the laboratory bench, and the liver was exposed by laparotomy under aseptic conditions, and liver tissue of about 1 cm × 1 cm was excised on the surface of the liver to cause a bleeding wound. The control group was given sterile sponge for hemostasis, and the hemostatic material group was given hemostatic material, applied pressure on the wound, observed the hemostatic situation, recorded the hemostasis time, and measured the amount of bleeding. After the operation, the wound was sutured, and the animals were housed in a single cage, with free access to water and food, and the survival of New Zealand rabbits was observed. Three New Zealand rabbits were taken from each group on the 2nd and 7th day after the operation, and 3 New Zealand rabbits were taken from each group on the 14th and 21st days after the operation, and the intra-abdominal adhesion, intra-abdominal infection and liver healing were observed by laparotomy.

结果如下表所示，采用单因素方差分析的方法。与使用医用棉的对照组比较，具有极为显著的差异。The results are shown in the table below, using the method of one-way analysis of variance. Compared with the control group using medical cotton, there is an extremely significant difference.

表2.新西兰兔肝脏创伤模型的止血试验结果表Table 2. Hemostasis test results of New Zealand rabbit liver trauma model

注：与对照组相比，“△”P＜0.05，“△△”P＜0.01Note: Compared with the control group, "△"P<0.05, "△△"P<0.01

观察新西兰兔存活情况。于术后第2天、第7天各组取3只新西兰兔，术后第14天和第21天各取3只新西兰兔，剖腹观察腹腔内粘连、腹腔内感染及肝脏愈合情况。结果如下：术后第2天：止血材料组与创面粘附良好，周围被大网膜包裹并形成胶状物质附着在创口上，未见出血，对照组可见一定程度肿大；术后第7天：止血材料组与创伤口粘附良好，与凝血块在创面上形成一完整覆盖物，伤口周围有一层白色薄膜，伤口恢复良好；术后第14天：止血材料组基本能够被基本吸收，未见粘连，与血凝块在创面上形成一层完整的覆盖物，伤口周围覆盖一层白色的薄膜，创口恢复良好，未见肝脏肿大，对照组的创面组织存在部分粘连，创面恢复缓慢；术后第21天：止血材料组创口恢复良好，未见粘连，止血材料已经被完全吸收，创口处可见一层薄薄的白色组织，肝脏未见异常。Observe the survival of New Zealand rabbits. Three New Zealand rabbits were taken from each group on the 2nd and 7th day after the operation, and 3 New Zealand rabbits were taken from each group on the 14th and 21st days after the operation, and the intra-abdominal adhesion, intra-abdominal infection and liver healing were observed by laparotomy. The results are as follows: on the second day after operation: the hemostatic material group adhered well to the wound surface, and was surrounded by omentum and formed a gelatinous substance attached to the wound. No bleeding was seen, and a certain degree of swelling was seen in the control group; Day: The hemostatic material group adhered well to the wound, and formed a complete cover with the blood clot on the wound surface. There was a white film around the wound, and the wound recovered well; the 14th day after operation: The hemostatic material group could be basically absorbed, No adhesion was seen, and a complete covering was formed with the blood clot on the wound, and a white film was covered around the wound. The wound recovered well, and no liver enlargement was seen. The wound tissue of the control group had some adhesions, and the wound recovered slowly On the 21st day after operation: the wound in the hemostatic material group recovered well, no adhesion was found, the hemostatic material had been completely absorbed, a thin layer of white tissue could be seen at the wound, and no abnormality was found in the liver.

综合以上观察结果，本发明制备的止血材料具有良好的亲水性，止血速度快，与创面具有较强的粘附力，无异常刺激反应，体内吸收速度快，且在腹腔止血时具有防止肠粘连的特殊效果。Based on the above observation results, the hemostatic material prepared by the present invention has good hydrophilicity, fast hemostatic speed, strong adhesion to the wound surface, no abnormal stimulation reaction, fast absorption speed in the body, and has the ability to prevent intestinal bleeding during abdominal hemostasis. Sticky special effects.

本发明的实施例1、3初步试验表明，与实施例2具有相类似的结果。Embodiment 1 of the present invention, 3 preliminary tests show, have similar result with embodiment 2.

本发明不限于上述实施例。The present invention is not limited to the above-described embodiments.

Claims (7)

1. absorbable hemostatic material based on Mus tail collagen protein preparation; The lyophilizing hemostatic material of processing for Mus tail collagen protein, polyvinyl alcohol, chitosan, water; Each material weight proportioning is a Mus tail collagen protein: polyvinyl alcohol: chitosan is 1-10%: 0.2-5%: 0.2-2%, and surplus is a water.

2. the absorbable hemostatic material based on the preparation of Mus tail collagen protein according to claim 1 is characterized in that in the lyophilizing hemostatic material that each material weight proportioning is a Mus tail collagen protein: polyvinyl alcohol: chitosan is 5%: 2%: 1%, the water of surplus.

3. the absorbable hemostatic material based on the preparation of Mus tail collagen protein according to claim 1 and 2 is characterized in that Mus tail collagen protein derives from each strain SFP level rat Mus tail.

4. the absorbable hemostatic material based on the preparation of Mus tail collagen protein according to claim 1 is characterized in that dosage form is an adhesive bandage.

5. the absorbable hemostatic material based on the preparation of Mus tail collagen protein according to claim 1 is characterized in that dosage form is a spray.

6. the absorbable hemostatic preparation methods based on the preparation of Mus tail collagen protein according to claim 1 comprises the steps:

1) take by weighing Mus tail collagen protein, polyvinyl alcohol, chitosan in proportion, each material mixture ratio is a Mus tail collagen protein: polyvinyl alcohol: chitosan is 1-10%: 0.2-5%: 0.2-2%, and surplus is a water;

2) with Mus tail collagen protein: polyvinyl alcohol: after chitosan mixes; With pouring into the cultivation platelet after the sterilized water dissolving;-40-0 ℃ of pre-freeze 2h-12h; Change again that-80 ℃ of ultra cold storage freezers are anxious to freeze 4h-12h over to, change over to again in the vacuum freeze drier composite, promptly obtain the lyophilizing hemostatic material at-4 ℃ of following lyophilization 10-60h; The Co60 radiation sterilization, kept dry.

7. the described absorbable hemostatic material based on the preparation of Mus tail collagen protein of claim 1 prevents the application in the intestinal adhesion in skin injury, viscus hemostasis, abdominal cavity hemostasis.