CN111317857A

Movatterモバイル変換

Info

Publication number: CN111317857A
Application number: CN201811540249.6A
Authority: CN
Inventors: 张立群; 张云飞; 郑静; 宫敏; 谢纹奇; 池骋; 姜奕彤
Original assignee: Beijing University of Chemical Technology
Current assignee: Beijing University of Chemical Technology
Priority date: 2018-12-17
Filing date: 2018-12-17
Publication date: 2020-06-23

Abstract

The invention discloses a degradable and absorbable bone hemostatic material containing chitosan and a preparation method thereof. The bone hemostatic material comprises: two polyethylene glycols and chitosan of different molecular weights; wherein the mass fraction of the chitosan is 1-30%. The preparation method comprises the following steps: stirring and mixing two polyethylene glycols with different molecular weights, and heating to enable the mixture to be in a clear liquid state; slowly adding the chitosan with the dosage, stirring and mixing uniformly, curing and sterilizing to obtain the bone hemostatic material. The bone hemostatic material is white wax, has good kneading plasticity, can perform hemostasis on bone wounds in a physical blocking mode, and contains the chitosan component with good hemostatic and healing promoting performances. Meanwhile, the material can be degraded and absorbed in human body, and has no toxic or harmful effect on human body.

Description

Translated fromChinese

一种含壳聚糖可降解吸收的骨止血材料及其制备方法A kind of chitosan-containing degradable and absorbable bone hemostatic material and preparation method thereof

技术领域technical field

本发明涉及生物医用材料技术领域，进一步地说，是涉及一种含壳聚糖可降解吸收的骨止血材料及其制备方法。The invention relates to the technical field of biomedical materials, and more particularly, to a degradable and absorbable bone hemostatic material containing chitosan and a preparation method thereof.

背景技术Background technique

骨手术创伤往往较大，术中对于松质骨骨面出血、静脉窦出血以及骨髓腔的出血等目前尚无确切的止血手段，术中造成大量失血较为常见，往往需要及时输血补充血容量，但这却加大了患血液性传染病的风险。因此，在对骨骼进行外科手术修复时，采取适当的止血措施成为重中之重，而当今临床上，最常用的骨止血材料是骨蜡(Bone Wax)。Bone surgery trauma is often relatively large, and there is currently no definite hemostasis method for cancellous bone surface bleeding, venous sinus bleeding, and bone marrow cavity bleeding during surgery. Mass blood loss is common during surgery, and blood transfusion is often needed to replenish blood volume. But this increases the risk of blood-borne infections. Therefore, in the surgical repair of bones, taking appropriate hemostasis measures has become the top priority, and in today's clinic, the most commonly used bone hemostatic material is bone wax (Bone Wax).

现在常用的骨蜡止血材料通常是由蜂蜡、石蜡和凡士林等原料按一定质量比混合制备而成，该类产品为白色或淡黄色，具有良好的软化性能，用手搓揉变软后能塑形且无毒。它是通过纯粹的物理方法对骨中渗血的毛细血管进行堵塞，从而达到止血效果，产品本身并无止血和促进骨愈合功能。尽管该类传统型骨蜡在骨止血中应用广泛，且其制备成本较低、工艺相对简单，但由于不可被降解吸收的特性，导致其将作为异体始终存在于人体中进而抑制骨愈合过程，同时还有可能引起慢性炎症和明显的异物反应、降低细菌清除率增加骨感染风险等。The commonly used bone wax hemostatic material is usually prepared by mixing raw materials such as beeswax, paraffin and vaseline in a certain mass ratio. This kind of product is white or light yellow, and has good softening properties. Form and non-toxic. It uses pure physical methods to block the capillaries oozing blood in the bone, so as to achieve the effect of hemostasis. The product itself does not have the function of hemostasis and bone healing. Although this type of traditional bone wax is widely used in bone hemostasis, and its preparation cost is relatively low, the process is relatively simple, but due to the characteristics of non-degradable absorption, it will always exist in the human body as an allogeneic body and thus inhibit the bone healing process. At the same time, it may cause chronic inflammation and obvious foreign body reaction, reduce the bacterial clearance rate and increase the risk of bone infection.

发明内容SUMMARY OF THE INVENTION

为解决现有技术中出现的问题，本发明提供了一种含壳聚糖可降解吸收的骨止血材料及制备方法。用以代替传统骨蜡。In order to solve the problems in the prior art, the present invention provides a chitosan-containing degradable and absorbable bone hemostatic material and a preparation method. Used in place of traditional bone wax.

本发明的目的之一是提供一种含壳聚糖可降解吸收的骨止血材料。One of the objectives of the present invention is to provide a degradable and absorbable bone hemostatic material containing chitosan.

包括：include:

两种不同分子量的聚乙二醇和壳聚糖；Two different molecular weight polyethylene glycol and chitosan;

其中壳聚糖的质量分数为1％～30％；优选为10％～20％；The mass fraction of chitosan is 1%-30%; preferably 10%-20%;

其中，优选：Among them, preferably:

所述壳聚糖的脱乙酰度为50％～95％、分子量为5000至100000。The deacetylation degree of the chitosan is 50%-95%, and the molecular weight is 5,000-100,000.

所述聚乙二醇分子量为200至20000。The polyethylene glycol has a molecular weight of 200 to 20,000.

两种不同分子量的聚乙二醇为低分子量聚乙二醇和高分子量聚乙二醇；Two kinds of polyethylene glycols with different molecular weights are low molecular weight polyethylene glycol and high molecular weight polyethylene glycol;

两种不同分子量的聚乙二醇中，低分子量聚乙二醇的质量百分比为10％～50％，高分子量聚乙二醇的质量百分比为50％～90％。Among the two polyethylene glycols with different molecular weights, the mass percentage of low molecular weight polyethylene glycol is 10%-50%, and the mass percentage of high molecular weight polyethylene glycol is 50%-90%.

低分子量聚乙二醇的分子量为200～600；The molecular weight of low molecular weight polyethylene glycol is 200-600;

高分子量聚乙二醇的分子量为800～20000；The molecular weight of high molecular weight polyethylene glycol is 800-20000;

聚乙二醇(PEG)是一种无毒、无刺激性、具有良好水溶性的聚合物，依相对分子质量不同而性质不同，从无色无臭黏稠液体至蜡状固体。聚乙二醇和聚乙二醇脂肪酸酯在化妆品工业和制药工业中的应用很广泛。由于聚乙二醇兼有很多优良的性质：水溶性、不挥发性、生理惰性、温和性、润滑性和使皮肤润湿、柔软、有愉快用后感等。可选取不同相对分子质量级分的聚乙二醇改变制品的粘度、吸湿性和组织结构。聚乙二醇广泛用于多种药物制剂，如注射剂、局部用制剂、眼用制剂、口服和直肠用制剂。而氧化纤维素具有一定的止血功能，常用于外科手术中不能缝合或结扎的中度流血。Polyethylene glycol (PEG) is a non-toxic, non-irritating, and water-soluble polymer with different properties depending on the relative molecular mass, ranging from colorless, odorless, viscous liquid to waxy solid. Polyethylene glycol and polyethylene glycol fatty acid esters are widely used in the cosmetic and pharmaceutical industries. Because polyethylene glycol has many excellent properties: water-solubility, non-volatile, physiological inertness, mildness, lubricity, and skin moisturizing, softness, pleasant after-feel, etc. Different relative molecular weight fractions of polyethylene glycol can be selected to change the viscosity, hygroscopicity and organizational structure of the product. Polyethylene glycol is widely used in various pharmaceutical formulations such as injections, topical formulations, ophthalmic formulations, oral and rectal formulations. Oxidized cellulose has a certain hemostatic function, and is often used for moderate bleeding that cannot be sutured or ligated in surgical operations.

本发明的目的之二是提供高一种含壳聚糖可降解吸收的骨止血材料的制备方法。The second purpose of the present invention is to provide a method for preparing a chitosan-containing degradable and absorbable bone hemostatic material.

包括：include:

将两种不同分子量的聚乙二醇搅拌混合，加热使混合物呈现清亮液体状；缓慢加入所述用量的壳聚糖，搅拌混合均匀，固化，消毒制得所述骨止血材料。The two kinds of polyethylene glycols with different molecular weights are stirred and mixed, heated to make the mixture appear clear and liquid; the amount of chitosan is slowly added, stirred and mixed evenly, solidified, and sterilized to obtain the bone hemostatic material.

其中，优选：Among them, preferably:

加热温度为50～60℃，搅拌时间为1～3小时The heating temperature is 50～60℃, and the stirring time is 1～3 hours

本发明具体可采用以下技术方案：The present invention can adopt the following technical scheme specifically:

一种完全可降解吸收且具有优异止血性能的骨止血材料，所述材料包括：A completely degradable and absorbable bone hemostatic material with excellent hemostatic properties, the material includes:

(1)质量比为10/90的两种不分子量的聚乙二醇(PEG)；(1) two kinds of polyethylene glycol (PEG) with no molecular weight whose mass ratio is 10/90;

(2)质量分数为1％～30％的壳聚糖。(2) Chitosan whose mass fraction is 1% to 30%.

优选的，所述聚乙二醇分子量为200至20000。Preferably, the molecular weight of the polyethylene glycol is 200 to 20,000.

优选的，所述壳聚糖为脱乙酰度50％～95％、分子量为5000至100000的壳聚糖。Preferably, the chitosan is chitosan with a degree of deacetylation of 50% to 95% and a molecular weight of 5000 to 100000.

一种完全可降解吸收且具有优异止血性能的骨止血材料的制备方法，其步骤包括：A method for preparing a bone hemostatic material that is completely degradable and absorbable and has excellent hemostatic properties, the steps comprising:

(1)将两种不同分子量的聚乙二醇在机械搅拌条件下加热到50-60℃，搅拌10分钟，使混合物呈现清亮液体状；(1) Heating two kinds of polyethylene glycols with different molecular weights to 50-60°C under mechanical stirring conditions, stirring for 10 minutes, so that the mixture is in a clear liquid state;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入壳聚糖，并搅拌混合均匀，加热温度为50-60℃，搅拌时间为1-3小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan in the well-mixed liquid, and stir and mix well, the heating temperature is 50-60 °C, and the stirring time is 1-3 hours;

(3)将混合均匀的混合物倒入事先准备好的模具或者分装瓶中，常温下放置2小时固化成型；(3) Pour the well-mixed mixture into the prepared mold or sub-bottling, and place it at room temperature for 2 hours to solidify and form;

(4)将固化成型后的产品包装，密封，高压或γ射线灭菌消毒(4) Packaging, sealing, high pressure or gamma ray sterilization of the cured product

本发明的骨止血材料为白色蜡状物，具有良好的揉捏可塑性，可通过物理堵塞的方式对骨创伤进行止血，且包含的壳聚糖成分具有良好的止血和促愈合性能。同时，该材料在人体内可降解吸收，且对人体无毒害作用。The bone hemostatic material of the present invention is a white waxy substance, has good kneading plasticity, can stop bleeding of bone wounds through physical blockage, and contains chitosan components with good hemostasis and healing-promoting properties. At the same time, the material can be degraded and absorbed in the human body, and has no toxic effect on the human body.

附图说明Description of drawings

图1为不同分子量聚乙二醇的熔融温度；Fig. 1 is the melting temperature of different molecular weight polyethylene glycols;

图2为不同分子量聚乙二醇的结晶温度；Fig. 2 is the crystallization temperature of different molecular weight polyethylene glycol;

图3为本发明实施案例6提供的骨止血材料的细胞毒性图。如图，随着时间增加，细胞数量呈现稳定增长趋势，说明本材料具有良好的生物相容性，无细胞毒性。FIG. 3 is a cytotoxicity diagram of the bone hemostatic material provided in Example 6 of the present invention. As shown in the figure, with the increase of time, the number of cells showed a steady growth trend, indicating that the material has good biocompatibility and no cytotoxicity.

具体实施方式Detailed ways

下面结合实施例，进一步说明本发明。Below in conjunction with embodiment, the present invention is further described.

实施例1Example 1

(1)将2g聚乙二醇200(aladdin公司)和18g聚乙二醇10000(aladdin公司)，在机械搅拌条件下加热到60℃，搅拌10分钟，使混合物呈现清亮液体状；(1) 2g polyethylene glycol 200 (aladdin company) and 18g polyethylene glycol 10000 (aladdin company) were heated to 60°C under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear and liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为5％的脱乙酰度95％、分子量为5000的壳聚糖，并搅拌混合均匀，加热温度为60℃，搅拌时间为3小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 5% with a degree of deacetylation of 95% and a molecular weight of 5000 into the uniformly mixed liquid, and stir and mix evenly. The heating temperature is 60 ℃, the stirring time is 3 hours;

(4)将固化成型后的产品包装，密封，高压或γ射线灭菌消毒。(4) Package, seal, and sterilize the cured and formed product by high pressure or gamma rays.

实施例2Example 2

(1)将4g聚乙二醇400(aladdin公司)和16g聚乙二醇1500(aladdin公司)，在机械搅拌条件下加热到60℃，搅拌10分钟，使混合物呈现清亮液体状；(1) 4g polyethylene glycol 400 (aladdin company) and 16g polyethylene glycol 1500 (aladdin company) were heated to 60 ℃ under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为1％的脱乙酰度80％、分子量为50000的壳聚糖，并搅拌混合均匀，加热温度为60℃，搅拌时间为3小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 1% with a degree of deacetylation of 80% and a molecular weight of 50,000 into the uniformly mixed liquid, and stir and mix evenly. The heating temperature is 60 ℃, the stirring time is 3 hours;

实施例3Example 3

(1)将10g聚乙二醇200(aladdin公司)和10g聚乙二醇8000(aladdin公司)，在机械搅拌条件下加热60℃，搅拌10分钟，使混合物呈现清亮液体状；(1) 10g polyethylene glycol 200 (aladdin company) and 10g polyethylene glycol 8000 (aladdin company) were heated at 60°C under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear and liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为10％的脱乙酰度80％、分子量为10000的壳聚糖，并搅拌混合均匀，加热温度为60℃，搅拌时间为3小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 10% with a degree of deacetylation of 80% and a molecular weight of 10000 in the well-mixed liquid, and stir and mix evenly. The heating temperature is 60 ℃, the stirring time is 3 hours;

实施例4Example 4

(1)将8g聚乙二醇400(aladdin公司)和12g聚乙二醇8000(aladdin公司)，在机械搅拌条件下加热到60℃，搅拌10分钟，使混合物呈现清亮液体状；(1) 8g polyethylene glycol 400 (aladdin company) and 12g polyethylene glycol 8000 (aladdin company) were heated to 60° C. under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear and liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为5％的脱乙酰度95％、分子量为50000壳聚糖，并搅拌混合均匀，加热温度为60℃，搅拌时间为3小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 5% with a degree of deacetylation of 95% and a molecular weight of 50,000 into the uniformly mixed liquid, and stir and mix evenly. The heating temperature is 60 ° C , the stirring time is 3 hours;

实施例5Example 5

(1)将4g聚乙二醇200(aladdin公司)和16g聚乙二醇1500(aladdin公司)，在机械搅拌条件下加热到60℃，搅拌10分钟，使混合物呈现清亮液体状；(1) 4g polyethylene glycol 200 (aladdin company) and 16g polyethylene glycol 1500 (aladdin company) were heated to 60 ℃ under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为15％的脱乙酰度95％、分子量100000的壳聚糖，并搅拌混合均匀，加热温度为60℃，搅拌时间为3小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 15% with a degree of deacetylation of 95% and a molecular weight of 100,000 into the well-mixed liquid, and stir and mix evenly. The heating temperature is 60°C , the stirring time is 3 hours;

实施例6Example 6

(1)将2g聚乙二醇400(aladdin公司)和18g聚乙二醇1500(aladdin公司)，在机械搅拌条件下加热到60℃，搅拌10分钟，使混合物呈现清亮液体状；(1) 2g polyethylene glycol 400 (aladdin company) and 18g polyethylene glycol 1500 (aladdin company) were heated to 60°C under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear and liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为20％的脱乙酰度95％、分子量100000壳聚糖，并搅拌混合均匀，加热温度为60℃，搅拌时间为3小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 20% with a degree of deacetylation of 95% and a molecular weight of 100,000 into the uniformly mixed liquid, and stir and mix evenly. The heating temperature is 60 ° C, The stirring time is 3 hours;

(4)将固化成型后的产品包装，密封，高压或γ射线灭菌消毒；(4) Packing, sealing, high pressure or gamma ray sterilization of the cured product;

(5)将上述制备骨止血材料，与前成骨细胞MC3T3-E1共培养。不同时间点测试细胞液450nm的吸光度。如图3所示，细胞数量随着时间的延长而增长，表明本发明制备的可吸收骨止血材料具有良好的生物相容性，无细胞毒性。(5) The bone hemostatic material prepared above was co-cultured with the pre-osteoblast MC3T3-E1. The absorbance at 450 nm of the cell fluid was tested at different time points. As shown in FIG. 3 , the number of cells increases with time, indicating that the absorbable bone hemostatic material prepared by the present invention has good biocompatibility and no cytotoxicity.

实施例7Example 7

(1)将2g聚乙二醇600(aladdin公司)和18g聚乙二醇800(aladdin公司)，在机械搅拌条件下加热到60℃，搅拌10分钟，使混合物呈现清亮液体状；(1) 2g polyethylene glycol 600 (aladdin company) and 18g polyethylene glycol 800 (aladdin company) were heated to 60 ℃ under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为30％的脱乙酰度95％、分子量10000壳聚糖，并搅拌混合均匀，加热温度为60℃，搅拌时间为3小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 30% with a degree of deacetylation of 95% and a molecular weight of 10,000 into the uniformly mixed liquid, and stir and mix evenly. The heating temperature is 60 ℃, The stirring time is 3 hours;

实施例8Example 8

(1)将8g聚乙二醇600和(aladdin公司)12g聚乙二醇20000(aladdin公司)，在机械搅拌条件下加热到60℃，搅拌10分钟，使混合物呈现清亮液体状；(1)8g polyethylene glycol 600 and (aladdin company) 12g polyethylene glycol 20000 (aladdin company) were heated to 60°C under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear and liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为5％的脱乙酰度80％、分子量50000壳聚糖，并搅拌混合均匀，加热温度为60℃，搅拌时间为3小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 5% with a degree of deacetylation of 80% and a molecular weight of 50,000 into the uniformly mixed liquid, and stir and mix evenly. The heating temperature is 60 °C, The stirring time is 3 hours;

实施例9Example 9

(1)将10g聚乙二醇600(aladdin公司)和10g聚乙二醇800(aladdin公司)，在机械搅拌条件下加热到60℃，搅拌10分钟，使混合物呈现清亮液体状；(1) 10 g of polyethylene glycol 600 (aladdin company) and 10 g of polyethylene glycol 800 (aladdin company) were heated to 60° C. under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear and liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为15％的脱乙酰度95％、分子量100000壳聚糖，并搅拌混合均匀，加热温度为60℃，搅拌时间为3小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 15% with a degree of deacetylation of 95% and a molecular weight of 100,000 into the uniformly mixed liquid, and stir and mix evenly. The heating temperature is 60 ℃, The stirring time is 3 hours;

实施例10Example 10

(1)将2g聚乙二醇400(aladdin公司)和18g聚乙二醇4000(aladdin公司)，在机械搅拌条件下加热到55℃，搅拌10分钟，使混合物呈现清亮液体状；(1) 2g polyethylene glycol 400 (aladdin company) and 18g polyethylene glycol 4000 (aladdin company) were heated to 55 ℃ under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为10％的脱乙酰度90％、分子量20000壳聚糖，并搅拌混合均匀，加热温度为55℃，搅拌时间为2小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 10% with a degree of deacetylation of 90% and a molecular weight of 20,000 into the uniformly mixed liquid, and stir and mix evenly. The heating temperature is 55 ℃, The stirring time is 2 hours;

实施例11Example 11

(1)将2g聚乙二醇200(aladdin公司)和18g聚乙二醇3000(aladdin公司)，在机械搅拌条件下加热到60℃，搅拌10分钟，使混合物呈现清亮液体状；(1) 2g polyethylene glycol 200 (aladdin company) and 18g polyethylene glycol 3000 (aladdin company) were heated to 60 ℃ under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为5％的脱乙酰度50％、分子量5000壳聚糖，并搅拌混合均匀，加热温度为60℃，搅拌时间为1小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 5% with a degree of deacetylation of 50% and a molecular weight of 5000 in the uniformly mixed liquid, and stir and mix evenly. The heating temperature is 60 ℃, The stirring time is 1 hour;

实施例12Example 12

(1)将2g聚乙二醇400(aladdin公司)和18g聚乙二醇3000(aladdin公司)，在机械搅拌条件下加热到50℃，搅拌10分钟，使混合物呈现清亮液体状；(1) 2g polyethylene glycol 400 (aladdin company) and 18g polyethylene glycol 3000 (aladdin company) were heated to 50°C under mechanical stirring conditions, and stirred for 10 minutes to make the mixture appear clear and liquid;

(2)在保持机械搅拌的加热条件下，在混合均匀的液体中，缓慢加入质量分数为5％的脱乙酰度95％、分子量100000壳聚糖，并搅拌混合均匀，加热温度为50℃，搅拌时间为3小时；(2) Under the heating condition of maintaining mechanical stirring, slowly add chitosan with a mass fraction of 5% with a degree of deacetylation of 95% and a molecular weight of 100,000 into the uniformly mixed liquid, and stir and mix evenly. The heating temperature is 50 ℃, The stirring time is 3 hours;