CN102268753A - Manufacturing method of polymer fiber and wound dressing and obtained wound dressing - Google Patents
Manufacturing method of polymer fiber and wound dressing and obtained wound dressing Download PDFInfo
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Abstract
Description
技术领域 technical field
本发明涉及一种制造高分子纤维的方法以及制造由高分子纤维组成的创伤敷料的方法。本发明也涉及以本发明方法所制造的创伤敷料。The present invention relates to a method for producing polymer fibers and a method for producing wound dressings composed of polymer fibers. The invention also relates to wound dressings produced by the method of the invention.
背景技术 Background technique
目前常用的创伤敷料大都是以水胶、膜或海绵状的结构材料为主。虽然这些敷料已在市场上使用了相当长的一段时间,但是由于例如机械强度、吸液量及吸液速度等问题而造成这些材料在应用上的限制。根据过去学者研究指出,一般的立体空间构型最常见的有海绵状与不织布状,而其中不织布状的通透性以及吸水性优于海绵状。而且,不织布构型有较佳的立体结构,使细胞能有较佳的空间吸附及细胞增生的功能。另外,不织布构型具有良好的柔软性及一定的机械强度,并可通过纺织技术的连续制造方法来缩减其制造成本,制造出物美价廉的医药级敷伤材料。Most of the currently used wound dressings are based on hydrogel, film or sponge-like structural materials. Although these dressings have been used in the market for a considerable period of time, the application of these materials has been limited due to issues such as mechanical strength, liquid absorption capacity and liquid absorption speed. According to the research of past scholars, the most common three-dimensional configurations are sponge and non-woven fabrics, and the permeability and water absorption of non-woven fabrics are better than those of sponges. Moreover, the non-woven configuration has a better three-dimensional structure, so that cells can have better space adsorption and cell proliferation functions. In addition, the non-woven fabric configuration has good softness and certain mechanical strength, and its manufacturing cost can be reduced through the continuous manufacturing method of textile technology, and a medical-grade dressing material with high quality and low price can be produced.
常见的创伤敷料是以天然再生高分子纤维制造。创伤敷料一般是以湿式纺丝成型技术制得,并使用冻结干燥技术来进行纤维干燥。但冻结干燥制造方法既费时又耗能;且在不织布制造方法中,若使用水抄成型技术,需要再一次的费时又耗能的冻干制造方法,而针扎成型技术则需要额外的针扎机设备。因此,以传统方法制造天然再生高分子纤维或不织布纤维作为创伤敷料耗能耗时且需要昂贵的设备。Common wound dressings are made of natural regenerated polymer fibers. Wound dressings are generally made by wet spinning technology, and the fibers are dried using freeze drying technology. However, the freeze-drying manufacturing method is time-consuming and energy-consuming; and in the non-woven fabric manufacturing method, if the hydroforming technology is used, another time-consuming and energy-consuming freeze-drying manufacturing method is required, while the needle-punching technology requires additional needles. machine equipment. Therefore, the traditional method of manufacturing natural regenerated polymer fibers or non-woven fibers as wound dressings is time-consuming and requires expensive equipment.
发明内容 Contents of the invention
为了克服现有技术中制造创伤敷料使用的纤维或不织布的问题,本发明提供了一种制造创伤敷料的方法,其以不同的干燥方式取代冻结干燥,不仅缩短制造方法的时间且减少能量上的消耗。In order to overcome the problems of the fiber or non-woven fabric used in the manufacture of wound dressings in the prior art, the present invention provides a method for manufacturing wound dressings, which replaces freeze-drying with different drying methods, which not only shortens the time of the manufacturing method but also reduces energy consumption. consume.
因此,本发明首先提供一种高分子纤维的制造方法,其特征在于包含下列步骤:Therefore, at first the present invention provides a kind of manufacture method of polymer fiber, it is characterized in that comprising the following steps:
(i)将含有天然高分子材料的水性纺丝原液(aqueous dope)与纺丝成型液湿式纺丝形成湿式高分子原丝;(i) wet-spinning an aqueous spinning dope (aqueous dope) containing a natural polymer material and a spinning forming liquid to form a wet polymer precursor;
(ii)将该湿式高分子原丝浸入柔软剂中获得经柔软剂处理的湿式高分子原丝;(ii) immersing the wet polymer precursor in a softener to obtain a softener-treated wet polymer precursor;
(iii)将该经柔软剂处理的湿式高分子原丝浸入挥发溶剂中以除去该高分子原丝里的水分而形成高分子纤维。(iii) immersing the softener-treated wet polymer precursors in a volatile solvent to remove moisture from the polymer precursors to form polymer fibers.
在一个具体实施例中,该天然高分子材料为几丁聚糖、海藻酸盐或它们的组合。In a specific embodiment, the natural polymer material is chitosan, alginate or a combination thereof.
在一个具体实施例中,该含有天然高分子材料的水性纺丝原液是浓度为3至10重量%(w/w%)的几丁聚糖溶液,且该纺丝成型液含有至少碱性化合物,其浓度为3至10重量%。In a specific embodiment, the aqueous spinning dope containing natural polymer materials is a chitosan solution with a concentration of 3 to 10% by weight (w/w%), and the spinning forming solution contains at least an alkaline compound , and its concentration is 3 to 10% by weight.
在一个具体实施例中,该含有天然高分子材料的水性纺丝原液是浓度为3至10重量%的海藻酸钠溶液;且该纺丝成型液含有二价阳离子,其浓度为3至10重量%。In a specific embodiment, the aqueous spinning stock solution containing natural polymer materials is a sodium alginate solution with a concentration of 3 to 10% by weight; and the spinning forming solution contains divalent cations at a concentration of 3 to 10% by weight %.
在一个具体实施例中,该柔软剂是浓度为0.1至25重量%的甘油溶液或浓度为0.1至10重量%的聚山梨醇酯溶液。In a specific embodiment, the softening agent is a glycerin solution with a concentration of 0.1 to 25% by weight or a solution of polysorbate with a concentration of 0.1 to 10% by weight.
在一个具体实施例中,该挥发溶剂是低碳数醇、低碳数酮或低碳数醚。In a specific embodiment, the volatile solvent is a lower carbon number alcohol, a lower carbon number ketone or a lower carbon number ether.
本发明也提供一种创伤敷料的制造方法,其特征在于包含下列步骤:The present invention also provides a manufacturing method of a wound dressing, which is characterized in that it comprises the following steps:
(i)以前述方法获得高分子纤维;(i) obtain high molecular fiber with aforementioned method;
(ii)将该高分子纤维形成高分子纤维不织布;以及(ii) forming the polymer fiber into a polymer fiber non-woven fabric; and
(iii)以天然高分子溶液处理该不织布,以在该不织布的高分子纤维上形成有多孔性涂层,从而获得创伤敷料。(iii) treating the non-woven fabric with a natural polymer solution to form a porous coating on the polymer fibers of the non-woven fabric, thereby obtaining a wound dressing.
在一个具体实施例中,以天然高分子溶液处理高分子纤维不织布的步骤包括:将天然高分子喷覆到不织布上形成具有多孔性涂层在高分子纤维上的不织布;以及将具有多孔性涂层的不织布干燥获得创伤敷料。优选的是,该天然高分子是以每分钟10至100毫升的喷覆速率喷覆到不织布上。以及优选的是,具有多孔性涂层的不织布干燥是在35至85℃的温度下进行。In a specific embodiment, the step of treating the polymer fiber non-woven fabric with the natural polymer solution includes: spraying the natural polymer onto the non-woven fabric to form a non-woven fabric with a porous coating on the polymer fiber; The layers of nonwoven fabric are dried to obtain a wound dressing. Preferably, the natural polymer is sprayed onto the non-woven fabric at a spray rate of 10 to 100 milliliters per minute. And preferably, the drying of the nonwoven fabric with the porous coating is carried out at a temperature of 35 to 85°C.
在一个较佳的具体实施例中,该天然高分子溶液选自由海藻酸钠、胶原蛋白、明胶、透明质酸、几丁聚糖及其衍生物和混合物所组成的群组。In a preferred embodiment, the natural polymer solution is selected from the group consisting of sodium alginate, collagen, gelatin, hyaluronic acid, chitosan, derivatives and mixtures thereof.
另一方面,本发明也提供一种创伤敷料,其以根据本发明的方法所制造。具体而言,以本发明方法制造的创伤敷料将含有具有高分子纤维的不织布网体;以及形成于不织布网体中的高分子纤维上的多孔性涂层。In another aspect, the present invention also provides a wound dressing produced by the method according to the present invention. Specifically, the wound dressing manufactured by the method of the present invention will contain a non-woven mesh with polymer fibers; and a porous coating formed on the polymer fibers in the non-woven mesh.
根据本发明方法可不需繁复的干燥步骤即可大量制造创伤敷料。因此,本发明方法和以该方法获得的创伤敷料有极大的经济价值。且根据本发明方法获得的创伤敷料具有改善的伤口愈合以及凝血方面的效果而不会造成动物刺激性。According to the method of the invention, wound dressings can be mass-produced without complicated drying steps. Therefore, the method according to the invention and the wound dressing obtained by this method are of great economic value. And the wound dressing obtained according to the method of the present invention has improved wound healing and blood coagulation effects without causing animal irritation.
附图说明 Description of drawings
图1为以市售敷料与比较实施例敷料的凝血实验结果比较。Figure 1 is a comparison of the coagulation test results of the commercially available dressing and the dressing of the comparative example.
图2为以本发明方法获得的敷料与比较实施例敷料的凝血实验结果比较。Fig. 2 is a comparison of the coagulation test results of the dressing obtained by the method of the present invention and the dressing of the comparative example.
具体实施方式 Detailed ways
一种高分子纤维的制造方法,其特征在于包含下列步骤:A kind of manufacture method of polymer fiber is characterized in that comprising the following steps:
(i)将含有天然高分子材料的水性纺丝原液与纺丝成型液湿式纺丝形成湿式高分子原丝;(i) wet-spinning the water-based spinning stock solution containing the natural polymer material and the spinning forming solution to form a wet polymer precursor;
(ii)将该湿式高分子原丝浸入柔软剂中获得经柔软剂处理的湿式高分子原丝;(ii) immersing the wet polymer precursor in a softener to obtain a softener-treated wet polymer precursor;
(iii)将该经柔软剂处理的湿式高分子原丝浸入挥发溶剂中以除去该高分子原丝里的水分而形成高分子纤维。(iii) immersing the softener-treated wet polymer precursors in a volatile solvent to remove moisture from the polymer precursors to form polymer fibers.
一种创伤敷料的制造方法,其特征在于包含下列步骤:A kind of manufacture method of wound dressing is characterized in that comprising the following steps:
(i)根据权利要求1的方法获得高分子纤维;(i) obtain polymer fiber according to the method for claim 1;
(ii)将该高分子纤维形成高分子纤维不织布;以及(ii) forming the polymer fiber into a polymer fiber non-woven fabric; and
(iii)以天然高分子溶液处理该不织布,以在该不织布的高分子纤维上形成有多孔性涂层,从而获得创伤敷料。(iii) treating the non-woven fabric with a natural polymer solution to form a porous coating on the polymer fibers of the non-woven fabric, thereby obtaining a wound dressing.
根据本发明,形成高分子纤维不织布的步骤包括现有技术中已知的由适当长度的纤维制造不织布的任何方法。例如,具有该高分子纤维的不织布可通过梳棉和水抄的方式形成。According to the present invention, the step of forming a nonwoven of polymeric fibers includes any method known in the art for making a nonwoven from fibers of appropriate length. For example, the nonwoven fabric with the polymer fibers can be formed by carding and hydrolaid.
在一个较佳的具体实施例中,形成高分子纤维不织布的步骤包括:将高分子纤维进行梳棉获得高分子纤维棉团。In a preferred specific embodiment, the step of forming the polymer fiber nonwoven fabric includes: carding the polymer fiber to obtain a polymer fiber clump.
在一个较佳的具体实施例中,将高分子纤维均匀分散在卷取罗拉(collecting roller)上接着进行高分子纤维粘合(bonding)。In a preferred embodiment, the polymer fibers are evenly dispersed on a collecting roller, followed by polymer fiber bonding.
在一个较佳的具体实施例中,形成高分子纤维不织布的步骤包括:将高分子纤维剪成短纤维然后铺设在水中形成湿式不织布。In a preferred specific embodiment, the step of forming the polymer fiber non-woven fabric includes: cutting the polymer fiber into short fibers and then laying them in water to form a wet-laid non-woven fabric.
根据本发明,天然高分子材料可为几丁聚糖、海藻酸盐或它们的组合。According to the present invention, the natural polymer material can be chitosan, alginate or a combination thereof.
在本发明较佳的实施例中,含有天然高分子材料的水性纺丝原液是浓度为3至10重量%的几丁聚糖溶液;且该纺丝成型液含有至少碱性化合物,其浓度为3至10重量%。In a preferred embodiment of the present invention, the aqueous spinning stock solution containing the natural polymer material is a chitosan solution with a concentration of 3 to 10% by weight; and the spinning forming solution contains at least an alkaline compound at a concentration of 3 to 10% by weight.
在本发明另一较佳的实施例中,含有天然高分子材料的水性纺丝原液是浓度为3至10重量%的海藻酸钠溶液;且该纺丝成型液含有二价阳离子,其浓度为3至10重量%。In another preferred embodiment of the present invention, the aqueous spinning stock solution containing natural polymer materials is a sodium alginate solution with a concentration of 3 to 10% by weight; and the spinning forming solution contains divalent cations at a concentration of 3 to 10% by weight.
根据本发明,该二价阳离子包括但不限于钙离子。According to the present invention, the divalent cations include but are not limited to calcium ions.
根据本发明,柔软剂可使湿式高分子原丝保持合适的柔软度。According to the present invention, the softening agent can keep the wet-laid polymer precursors with proper softness.
根据本发明,柔软剂包括但不限于甘油、聚山梨醇酯及其衍生物,例如,吐温-20(Tween-20)、吐温-60、吐温-80和斯潘-80(Span-80)。优选的是,柔软剂是浓度为0.1至25重量%的甘油溶液或浓度为0.1至10重量%的聚山梨醇酯溶液。According to the present invention, softening agents include but are not limited to glycerin, polysorbate and derivatives thereof, for example, Tween-20 (Tween-20), Tween-60, Tween-80 and Span-80 (Span-80). 80). Preferably, the softening agent is a solution of glycerol with a concentration of 0.1 to 25% by weight or a solution of polysorbate with a concentration of 0.1 to 10% by weight.
根据本发明,将经柔软剂处理的湿式高分子原丝浸入挥发溶剂中。该挥发溶剂优选为低碳数醇类、低碳数酮类或低碳数醚类。According to the present invention, the wet polymer precursor treated with the softening agent is immersed in a volatile solvent. The volatile solvent is preferably low carbon number alcohols, low carbon number ketones or low carbon number ethers.
低碳数醇类可为甲醇、乙醇、丙醇、丁醇、戊醇、己醇。低碳数酮类可为C1到C6的酮。低碳数醚类可为C2到C6的醚。The low carbon number alcohols can be methanol, ethanol, propanol, butanol, pentanol, hexanol. The low carbon number ketones can be C 1 to C 6 ketones. The lower carbon number ethers can be C 2 to C 6 ethers.
在本发明的一个较佳的具体实施例中,以天然高分子溶液处理该不织布的步骤是为了增加纤维间的粘合(bonding)效果,其包括:In a preferred embodiment of the present invention, the step of treating the non-woven fabric with a natural polymer solution is to increase the bonding effect between fibers, which includes:
将天然高分子溶液喷覆到不织布上形成在高分子纤维上具有多孔性涂层的不织布;以及Spraying the natural polymer solution onto the non-woven fabric to form a non-woven fabric with a porous coating on the polymer fibers; and
将该具有多孔性涂层的不织布干燥,以获得创伤敷料。The nonwoven fabric with the porous coating is dried to obtain a wound dressing.
优选的是,天然高分子溶液是以每分钟10至100毫升的喷覆速率喷覆到不织布上。Preferably, the natural polymer solution is sprayed onto the non-woven fabric at a spray rate of 10 to 100 milliliters per minute.
优选的是,具有多孔性涂层的不织布干燥在35至85℃的温度下进行,以获得创伤敷料。Preferably, drying of the nonwoven with a porous coating is carried out at a temperature of 35 to 85° C. to obtain a wound dressing.
根据本发明,天然高分子溶液含有下列至少一种:海藻酸钠、胶原蛋白、明胶、透明质酸、几丁聚糖及其衍生物和混合物。According to the present invention, the natural polymer solution contains at least one of the following: sodium alginate, collagen, gelatin, hyaluronic acid, chitosan, derivatives and mixtures thereof.
根据上述说明,一种依据本发明方法制得的创伤敷料将含有具有高分子纤维的不织布网体;以及形成于不织布网体中的高分子纤维上的多孔性涂层。According to the above description, a wound dressing prepared according to the method of the present invention will comprise a non-woven mesh with polymer fibers; and a porous coating formed on the polymer fibers in the non-woven mesh.
文中所用的术语“高分子纤维不织布网体”是指由依据本发明的方法所制得的高分子纤维,利用该技术领域中已知的任何用于制备不织布的方法所制得的织品。The term "polymer fiber non-woven fabric network body" used herein refers to a fabric made from polymer fibers produced by the method of the present invention, using any method known in the art for preparing non-woven fabrics.
以下通过本发明实施例进一步阐述本发明为达到发明目的所采取的技术手段以及相关的实验结果。The technical means adopted by the present invention to achieve the purpose of the invention and the related experimental results are further described below through the embodiments of the present invention.
比较实施例:以现有技术制造几丁聚糖复合纤维Comparative Example: Manufacturing Chitosan Composite Fibers with Prior Art
配制至少包含碱性化合物的碱性成型液。配制5重量%的几丁聚糖溶液,并且将其挤出于(extruded into)前述的成型液中成型,进行湿式纺丝,形成湿式几丁聚糖纺丝。湿式几丁聚糖纺丝经冻结干燥获得几丁聚糖纤维。取一定组成量的几丁聚糖纤维,裁切成2至3公分长度,分散于水中,并以高速搅拌分散器分散,然后以水抄方式形成几丁聚糖纤维不织布,最后再以冷冻干燥除去水分。在该比较实施例中,先后使用两次冻结干燥制造方法,各需花费2至3天冻干,制作时间冗长,且冻干机运作时需要大量的用电量。Formulate an alkaline forming liquid containing at least an alkaline compound. A 5% by weight chitosan solution was prepared and extruded into the above-mentioned molding solution for wet spinning to form wet chitosan spinning. Wet chitosan spinning was followed by freeze-drying to obtain chitosan fibers. Take a certain amount of chitosan fibers, cut them into 2 to 3 cm lengths, disperse them in water, and disperse them with a high-speed stirring disperser, then form chitosan fiber non-woven fabrics by water scooping, and finally freeze-dry Remove moisture. In this comparative example, two freeze-drying manufacturing methods were used successively, each of which took 2 to 3 days for freeze-drying, the manufacturing time was lengthy, and a large amount of electricity was required for the operation of the freeze-drying machine.
实施例1:以本发明方法制造几丁聚糖复合纤维Embodiment 1: manufacture chitosan composite fiber with the method of the present invention
配制5重量%的几丁聚糖(分子量约300k道尔顿)溶液。配制至少包含5重量%的氢氧化钠的碱性成型液。将几丁聚糖溶液挤出于前述的成型液成型,进行湿式纺丝,形成湿式几丁聚糖纺丝。将获得的湿式几丁聚糖纺丝浸泡于2体积%的吐温-20中5分钟,接着分别浸泡于50体积%、60体积%、70体积%的乙醇各5分钟进行溶剂置换后,以压吸机将多余溶剂排除,然后以60℃烘箱干燥2小时获得几丁聚糖复合纤维。全部的制作时程为4小时。A 5% by weight solution of chitosan (molecular weight about 300k Dalton) was prepared. An alkaline forming liquid containing at least 5% by weight of sodium hydroxide is prepared. The chitosan solution is extruded into the above-mentioned forming liquid for molding, and wet spinning is performed to form wet chitosan spinning. Soak the obtained wet chitosan spinning in 2 vol% Tween-20 for 5 minutes, then immerse in 50 vol%, 60 vol%, and 70 vol% ethanol for 5 minutes each for solvent replacement, then The excess solvent was removed by a suction machine, and then dried in an oven at 60° C. for 2 hours to obtain chitosan composite fibers. The total production time is 4 hours.
实施例2:以本发明方法制造海藻酸钠复合纤维Embodiment 2: manufacture sodium alginate composite fiber with the inventive method
配制5重量%的海藻酸钠溶液。配制含有5%的氯化钙的成型液。将海藻酸钠溶液挤出于前述的成型液成型,进行湿式纺丝,形成湿式海藻酸钠纺丝。将获得的湿式海藻酸钠纺丝浸泡于2体积%的吐温-20中5分钟,接着分别浸泡于50体积%、60体积%、70体积%的乙醇中各5分钟进行溶剂置换后,以压吸机将多余溶剂排除,然后以60℃烘箱干燥2小时获得海藻酸钠复合纤维。全部的制作时程为4小时。A 5% by weight sodium alginate solution was prepared. Prepare a molding solution containing 5% calcium chloride. The sodium alginate solution is extruded into the above-mentioned forming solution for molding, and wet spinning is performed to form wet sodium alginate spinning. Soak the obtained wet sodium alginate spinning in 2 vol% Tween-20 for 5 minutes, then soak in 50 vol%, 60 vol%, and 70 vol% ethanol for 5 minutes each for solvent replacement, then use The excess solvent was removed by a suction machine, and then dried in an oven at 60° C. for 2 hours to obtain sodium alginate composite fibers. The total production time is 4 hours.
实施例3:利用多孔性生物高分子材料制备复合创伤敷料Example 3: Preparation of Composite Wound Dressing Using Porous Biomacromolecule Materials
将由实施例1获得的几丁聚糖复合纤维剪成适当长度的短纤,以梳棉机进行纤维加工,同时在卷取罗拉上对几丁聚糖复合纤维棉网喷覆0.5%的海藻酸钠溶液,单位时间喷覆量为每分钟50毫升,进行纤维粘合,接着干燥形成含有海藻酸钠的多孔性生物高分子材料。进行本步骤时,可将海藻酸钠溶液替换为各种生物高分子溶液,如胶原蛋白、明胶、透明质酸等,以形成各种不同的多孔性生物高分子材料。Cut the chitosan composite fiber obtained in Example 1 into short fibers of appropriate length, and process the fiber with a carding machine, while spraying 0.5% alginic acid on the chitosan composite fiber cotton web on the take-up roller The sodium solution is sprayed at a rate of 50 milliliters per minute per unit time for fiber bonding, followed by drying to form a porous biopolymer material containing sodium alginate. When performing this step, the sodium alginate solution can be replaced with various biopolymer solutions, such as collagen, gelatin, hyaluronic acid, etc., to form various porous biopolymer materials.
然后将获得的多孔性生物高分子材料以压吸机排除多余水分后,以60℃烘箱干燥2小时,最后获得同时具备不织布及多孔性复合型态的创伤敷料。该几丁聚糖复合创伤敷料用于下面的实施例中。Then, the obtained porous biopolymer material was removed with a suction machine to remove excess water, and then dried in an oven at 60° C. for 2 hours, and finally a wound dressing with both non-woven and porous composite forms was obtained. This chitosan composite wound dressing was used in the following examples.
实施例4:复合创伤敷料的基重测试Example 4: Basis Weight Test of Composite Wound Dressings
将获得的几丁聚糖复合创伤敷料裁剪成大小为10cm×10cm的正方形,以电子天平测量重量,记录九个样品的结果取平均值,测量结果如表1所示。经过计算后得出不织布平均基重为0.0146g/cm2,单位换算后基重为146g/m2。The obtained chitosan composite wound dressing was cut into a square with a size of 10cm×10cm, the weight was measured with an electronic balance, and the results of nine samples were recorded and averaged. The measurement results are shown in Table 1. After calculation, the average basis weight of the nonwoven fabric is 0.0146g/cm 2 , and the basis weight after unit conversion is 146g/m 2 .
表1 几丁聚糖复合创伤敷料(100cm2)基重测试结果Table 1 Basis weight test results of chitosan composite wound dressing (100cm 2 )
实施例5:复合创伤敷料的吸液量测试Embodiment 5: liquid absorption test of composite wound dressing
1.实验材料与器材:1. Experimental materials and equipment:
a.以实施例3方法获得的几丁聚糖复合创伤敷料;A. the chitosan composite wound dressing obtained by the method of embodiment 3;
b.去离子水以及不同浓度的生理食盐水。b. Deionized water and different concentrations of physiological saline.
2.实验方法:2. Experimental method:
a.取一定大小的复合创伤敷料先于电子天平上测量重量。a. Take a composite wound dressing of a certain size and measure the weight on an electronic balance.
b.再将测过重量的复合创伤敷料分别浸泡于如下指明的不同浓度的生理食盐水中约8小时。b. Soak the weighted composite wound dressings in physiological saline of different concentrations as indicated below for about 8 hours.
c.取出因浸泡生理食盐水而膨胀的复合创伤敷料并以电子天平测量吸液后重量。c. Take out the composite wound dressing swollen due to immersion in saline, and measure the weight after fluid absorption with an electronic balance.
d.计算浸泡前后的重量差异与吸液程度数值。d. Calculate the weight difference before and after immersion and the value of the degree of liquid absorption.
计算结果显示于表2。由表2可见,在不同浓度的生理食盐水溶液处理下,其吸液率皆大于100%以上,且随着所使用的生理食盐水溶液浓度的增加会使其吸液率下降。The calculation results are shown in Table 2. It can be seen from Table 2 that under the treatment of different concentrations of physiological saline solution, the liquid absorption rate is greater than 100%, and the liquid absorption rate will decrease with the increase of the concentration of the physiological saline solution used.
表2 吸液率测试结果Table 2 Liquid absorption test results
实施例6:复合创伤敷料的凝血测试Example 6: Coagulation Test of Composite Wound Dressings
1.实验材料与器材:1. Experimental materials and equipment:
a.以实施例3方法获得的几丁聚糖复合创伤敷料;A. the chitosan composite wound dressing obtained by the method of embodiment 3;
b.以比较实施例方法获得的几丁聚糖创伤敷料;B. the chitosan wound dressing that obtains with comparative example method;
c.生理食盐水;c. Physiological saline;
d.志愿者全血血液;d. Volunteer whole blood;
e.ELISA(BioAssay Systems QuantiChromTM Hemoglobin Assaykit);e. ELISA (BioAssay Systems QuantiChromTM Hemoglobin Assaykit);
f.市售的创伤敷料(商品名Kaltostat,ConvaTec,USA)作为本实验的控制组。f. Commercially available wound dressing (trade name Kaltostat , ConvaTec, USA) was used as the control group in this experiment.
2.实验方法:2. Experimental method:
a.抽取志愿者静脉全血血液,开盖摇晃充氧30分钟;a. Extract venous whole blood from volunteers, open the cover, shake and oxygenate for 30 minutes;
b.将0.1g的几丁聚糖创伤敷料与30μL血液混合,并分别在混合30、60、90及120秒后取出敷料于生理食盐水中摇晃4分钟;b. Mix 0.1 g of chitosan wound dressing with 30 μL of blood, and after mixing for 30, 60, 90 and 120 seconds, take out the dressing and shake it in normal saline for 4 minutes;
c.取出上清液以ELISA测量在540nm的吸光值,参考波长为650nm。c. Take out the supernatant and measure the absorbance value at 540nm by ELISA, the reference wavelength is 650nm.
ELISA实验结果显示于图1和图2。图1显示市售敷料Kaltostat、市售以针轧技术制造的敷料与比较实施例以湿式纺丝制造的创伤敷料的凝血效果比较,比较实施例与市售敷料具有相似的效果。菱形和方形表示市售敷料的结果,三角形表示比较实施例中以湿式纺丝制造的创伤敷料的结果。由图可见,比较实施例使用湿式纺丝获得敷料的凝血效果和市售以针轧技术获得的敷料效果相似。The results of the ELISA experiment are shown in Figure 1 and Figure 2. Figure 1 shows the commercially available dressing Kaltostat 1. Comparing the blood coagulation effect of the commercially available dressing made by needle rolling technology and the wound dressing made by wet spinning in the comparative example, the comparative example and the commercially available dressing have similar effects. The diamonds and squares represent the results for commercially available dressings, and the triangles represent the results for the wound dressings produced by wet spinning in Comparative Examples. It can be seen from the figure that the coagulation effect of the dressing obtained by wet spinning in Comparative Example is similar to that of the commercially available dressing obtained by needle rolling technology.
图2显示使用本发明实施例3敷料与比较实施例敷料的凝血效果比较,菱形表示比较实施例的结果,三角形表示以实施例3的创伤敷料的结果。由图可见,和比较实施例的敷料相比,实施例3的敷料有较好的凝血效果。Fig. 2 shows the comparison of the coagulation effect between the dressing of Example 3 of the present invention and the dressing of Comparative Example, the rhombus indicates the result of Comparative Example, and the triangle indicates the result of the wound dressing of Example 3. It can be seen from the figure that compared with the dressing of Comparative Example, the dressing of Example 3 has a better blood coagulation effect.
实施例7:复合创伤敷料动物促愈测试Embodiment 7: animal healing test of compound wound dressing
1.实验材料与器材:1. Experimental materials and equipment:
a.以实施例3方法获得的几丁聚糖复合创伤敷料;A. the chitosan composite wound dressing obtained by the method of embodiment 3;
b.以比较实施例方法获得的几丁聚糖创伤敷料;B. the chitosan wound dressing that obtains with comparative example method;
c.大白鼠(wistar大白鼠);c. Rats (wistar rats);
d.显微镜。d.Microscope.
2.实验方法:2. Experimental method:
a.以肌肉注射方式对大白鼠施打麻醉剂(0.001mL/g),待其进入深度麻醉后进行实验;a. Apply anesthesia (0.001mL/g) to rats by intramuscular injection, and conduct experiments after they enter deep anesthesia;
b.以电动剃毛刀剃除大白鼠两侧的毛,再用除毛膏彻底除毛;b. Use an electric shaver to shave the hair on both sides of the rat, and then use a hair removal cream to remove the hair completely;
c.用碘酒消毒实验部位,在大白鼠背部手术出两个2cm x 2cm的伤口;c. Disinfect the experimental site with iodine, and perform two 2cm x 2cm wounds on the back of the rat;
d.分别在大白鼠背部伤口覆盖以本发明方法获得的复合创伤敷料与比较实施例的敷料;d. respectively cover the composite wound dressing obtained by the method of the present invention and the dressing of Comparative Example on the back wound of the rat;
e.连续观察伤口愈合情形,进行测量与记录。观察皮肤及伤口愈合过程,分别利用数位相机拍照记录敷料与组织贴合情形、伤口液体分泌量、伤口四周组织感染情形及伤口大小情形。e. Continuously observe the wound healing situation, measure and record. The skin and wound healing process were observed, and digital cameras were used to take pictures to record the adhesion between the dressing and the tissue, the amount of wound fluid secretion, the infection of the tissue around the wound, and the size of the wound.
3.结果:3. Results:
表3显示使用以实施例3方法获得的几丁聚糖复合创伤敷料和比较实施例方法获得的几丁聚糖创伤敷料的伤口部位肉眼观察及伤口面积测量结果。Table 3 shows the results of macroscopic observation of the wound site and measurement of the wound area using the chitosan composite wound dressing obtained by the method of Example 3 and the chitosan wound dressing obtained by the method of Comparative Example.
表3以实施例3与比较实施例获得敷料的伤口愈合情形比较Table 3 compares the wound healing situation that obtains dressing with embodiment 3 and comparative example
使用实施例3与比较实施例获得的敷料实验结果在面积复原情形均为良好。使用比较实施例敷料的伤口于第三周大致上已复原为长条结痂状;使用实施例3敷料的伤口于第三周则复原为圆状伤口。敷料容易拆除,不易沾粘伤口。The dressing experiment results obtained by using Example 3 and Comparative Example are all good in area restoration. The wound using the dressing of the comparative example has almost recovered to a long scab shape in the third week; the wound using the dressing of the embodiment 3 has recovered to a round wound in the third week. The dressing is easy to remove and does not stick to the wound.
实施例8:复合型态创伤敷料的细胞毒性测试Example 8: Cytotoxicity test of composite wound dressing
1.实验材料与器材:1. Experimental materials and equipment:
a.以实施例3方法获得的几丁聚糖复合创伤敷料;A. the chitosan composite wound dressing obtained by the method of embodiment 3;
b.L929小鼠结缔组织细胞;b. L929 mouse connective tissue cells;
c.萤光电子显微镜(The Olympus FSX100 Bio Imaging Navigatorall-in-one microscope)。c. Fluorescent electron microscope (The Olympus FSX100 Bio Imaging Navigator all-in-one microscope).
2.实验方法:2. Experimental method:
本测试是依据ANSI/AAMI/ISO10993-5:1999(名称为“医用材料的体外毒性测试”)来进行复合创伤敷料的细胞毒性(cyctotxicity)的评估试验。以培养在培养皿中的L929细胞作为阴性对照组、使L929小鼠细胞用已知有细胞毒性的酚处理作为阳性对照组、以及仅使用PE膜处理作为空白对照组来和以本发明创伤敷料的实验组作比较。This test is based on ANSI/AAMI/ISO10993-5:1999 (titled "in vitro toxicity test of medical materials") to evaluate the cytotoxicity (cyctotxicity) of composite wound dressings. Using L929 cells cultured in a culture dish as a negative control group, making L929 mouse cells treated with known cytotoxic phenol as a positive control group, and using only PE membrane as a blank control group to compare with the wound dressing of the present invention experimental group for comparison.
将阴性对照组、空白对照组、阳性对照组及实验组(实施例3获得的几丁聚糖复合创伤敷料)的细胞收下以台盼蓝(trypan blue)染色并计数后期细胞数目,结果显示于表4。表4结果显示使用实施例3的几丁聚糖复合创伤敷料所得到的细胞数目与阴性对照组和试剂对照组的数目接近,几乎不会使正常小鼠纤维母细胞产生死亡,因此不具有任何细胞毒性。The cells of the negative control group, the blank control group, the positive control group and the experimental group (the chitosan composite wound dressing obtained in Example 3) were collected and stained with trypan blue (trypan blue) and the number of cells in the later stage was counted, the results showed in Table 4. The results in Table 4 show that the number of cells obtained by using the chitosan composite wound dressing of Example 3 is close to the number of the negative control group and the reagent control group, and it will hardly cause the normal mouse fibroblasts to die, so it does not have any Cytotoxicity.
表4 复合创伤敷料的细胞毒性测试结果Table 4 Cytotoxicity test results of composite wound dressings
实施例9:复合创伤敷料皮内刺激测试Embodiment 9: Composite wound dressing intradermal irritation test
1.实验材料与器材:1. Experimental materials and equipment:
a.以实施例3方法获得的几丁聚糖复合创伤敷料;A. the chitosan composite wound dressing obtained by the method of embodiment 3;
b.生理食盐水;b. Physiological saline;
c.雄性纽西兰大白兔。c. Male New Zealand white rabbits.
2.实验方法:2. Experimental method:
a.按照ANSI/AAMI/ISO10993-12:2007(名称为“医疗器材生物性评估-第12部份:样品制备与参考材料”)来制备样品。以生理食盐水及棉籽油作为萃取液,萃取比例为6cm2/mL。将试验物质按比例浸泡于萃取溶剂中,置于旋转速度为100rpm的震荡器上,于37℃温度下萃取72小时。此外,将萃取溶剂在不含试验物质的情形下,以相同条件处理,作为空白对照;a. Prepare samples according to ANSI/AAMI/ISO10993-12:2007 (titled "Biological Evaluation of Medical Devices-Part 12: Sample Preparation and Reference Materials"). Physiological saline and cottonseed oil are used as the extraction solution, and the extraction ratio is 6cm 2 /mL. The test substances were soaked in the extraction solvent in proportion, placed on a shaker with a rotation speed of 100 rpm, and extracted at a temperature of 37°C for 72 hours. In addition, the extraction solvent was treated under the same conditions without the test substance as a blank control;
b.本试验以2只兔子进行测试,试验前18-24小时,以电动剪毛器去除兔子背部被毛,面积大小约15cm x 10cm。试验前,肉眼检查试验兔子背部皮肤,确保无任何损伤;b. In this experiment, 2 rabbits were used for testing. 18-24 hours before the test, the hair on the back of the rabbit was removed with an electric clipper, with an area of about 15cm x 10cm. Before the test, visually inspect the skin on the back of the test rabbit to ensure that there is no damage;
c.试验时,将试验物质生理食盐水萃取液以皮内注射方式分别注射于2只兔子背部左前侧5个部位。同时,将空白对照液皮内注射于试验兔子背部左后侧5个部位作为对照。以相同方式将试验物质棉籽油萃取液和空白对照液皮内注射于试验兔子背部右侧。每个部位注射量为0.2mL;c. During the test, the test substance physiological saline extract was injected intradermally into 5 parts on the left front side of the back of 2 rabbits. At the same time, the blank control solution was intradermally injected into 5 sites on the left rear side of the test rabbit's back as a control. In the same way, the test substance cottonseed oil extract and the blank control solution were intradermally injected on the right side of the back of the test rabbit. The injection volume for each site is 0.2mL;
d.在完成注射后24、48及72小时,根据皮内反应评分系统(如表5)进行观察,记录试验部位和对照部位每个注射点所显现的红斑及水肿情形于表6。d. 24, 48 and 72 hours after the injection, observe according to the intradermal reaction scoring system (as in Table 5), and record the erythema and edema at each injection point of the test site and the control site in Table 6.
表5 皮内反应评分系统Table 5 Scoring system for intradermal reactions
表6 兔子皮肤刺激反应计分表Table 6 Rabbit skin irritation reaction scoring table
由表6结果可知,根据本发明方法获得的几丁聚糖复合创伤敷料在动物皮肤上没有刺激性反应。From the results in Table 6, it can be seen that the chitosan composite wound dressing obtained according to the method of the present invention has no irritating reaction on animal skin.
实施例10:复合创伤敷料过敏性测试Embodiment 10: Composite wound dressing allergy test
1.实验材料与器材:1. Experimental materials and equipment:
a.以实施例3方法获得的几丁聚糖复合创伤敷料;A. the chitosan composite wound dressing obtained by the method of embodiment 3;
b.生理食盐水;b. Physiological saline;
c.雄性天竺鼠。c. Male guinea pigs.
2.实验方法:2. Experimental method:
a.按照ANSI/AAMI/ISO10993-12:2007来制备样品。以生理食盐水作为萃取液,萃取比例为3cm2/mL。萃取过程将试验物质按比例浸泡于萃取溶剂中,置于旋转速度为100rpm的震荡器上,于37℃温度下萃取72小时。此外,将萃取溶剂在不含试验物质的情形下,以相同条件处理,作为空白对照;a. Prepare samples according to ANSI/AAMI/ISO10993-12:2007. Physiological saline was used as the extraction liquid, and the extraction ratio was 3 cm 2 /mL. Extraction process The test substance was immersed in the extraction solvent in proportion, placed on a shaker with a rotation speed of 100rpm, and extracted at 37°C for 72 hours. In addition, the extraction solvent was treated under the same conditions without the test substance as a blank control;
b.试验前,将15只天竺鼠随机分配至试验组及对照组。试验组和对照组各使用10只和5只天竺鼠进行试验;b. Before the test, 15 guinea pigs were randomly assigned to the test group and the control group. The test group and the control group respectively use 10 guinea pigs and 5 guinea pigs to test;
c.试验前18-24小时,以电动剪毛器剃除所有试验动物肩胛部位的被毛,面积约5cm x 7cm。试验前,肉眼检查试验兔子背部皮肤,确保无任何损伤;c. 18-24 hours before the test, use an electric clipper to shave off the coat of the scapula of all experimental animals, with an area of about 5cm x 7cm. Before the test, visually inspect the skin on the back of the test rabbit to ensure that there is no damage;
d.试验前一天,将制备的萃取液以0.1mL的体积皮内注射于每只动物肩胛两侧剃毛的部位;d. The day before the test, the prepared extract was injected intradermally at the shaved site on both sides of the shoulder blade of each animal with a volume of 0.1 mL;
e.试验第六天再次将试验部位的被毛剃除,并涂抹含10%十二烷基磺酸钠的凡士林;e. On the sixth day of the test, the hair of the test site was shaved again, and vaseline containing 10% sodium dodecylsulfonate was applied;
f.试验第七天,将0.5mL试验物质萃取液或空白对照液滴在大小约2cm x 4cm纱布上,使其充分吸收后,以局部贴肤的方式施加于试验组或对照组动物肩胛剃除被毛的部位,并在48小时后取下皮肤贴片;f. On the seventh day of the test, drop 0.5mL of the test substance extract or the blank control solution on a gauze about 2cm x 4cm in size, make it fully absorbed, and apply it to the scapula of the animals in the test group or the control group in a local way. Remove the hairy area and remove the skin patch after 48 hours;
g.在取下皮肤贴片后,根据表7的评等标准对每只动物试验部位可能出现的任何过敏反应进行观察及评分。g. After removing the skin patch, observe and score any allergic reactions that may occur at the test site of each animal according to the evaluation criteria in Table 7.
表7过敏性试验(Magnusson-Kligman)评等标准Table 7 Allergy Test (Magnusson-Kligman) Rating Criteria
贴片试验反应 评分数值Patch test reaction Score value
无可见的变化 0No
分散或片状的红斑 1Scattered or patchy erythema 1
中度且连成一片的红斑 2Moderate and contiguous erythema 2
严重红斑及水肿 3Severe erythema and edema 3
在皮肤贴片取下后24小时、48小时、72小时的检测中,均无造成天竺鼠皮下刺激反应。也就是说,根据本发明获得的几丁聚糖复合创伤敷料经过过敏性测试后在动物皮肤上无刺激反应发生。In the detection of 24 hours, 48 hours, and 72 hours after the skin patch was removed, there was no subcutaneous irritation in guinea pigs. That is to say, the chitosan composite wound dressing obtained according to the present invention has no irritation reaction on the animal skin after the allergy test.
以上所述仅是本发明的较佳实施例而已,并非对本发明做任何形式上的限制,虽然发明已以较佳实施例揭露如上,然而并非用以限定本发明。本领域技术人员在不脱离本发明的范围内,可利用上述揭示的技术内容作出些许更动或修饰为等同变化的等效实施例,但凡是未脱离本发明技术方案的内容,依据本发明的技术实质对以上实施例所作的任何简单修改、等同变化与修饰,均仍属于本发明技术方案的范围内。The above descriptions are only preferred embodiments of the present invention, and do not limit the present invention in any form. Although the invention has been disclosed as above with preferred embodiments, it is not intended to limit the present invention. Without departing from the scope of the present invention, those skilled in the art can use the technical contents disclosed above to make some changes or modify them into equivalent embodiments with equivalent changes. Technical Essence Any simple modifications, equivalent changes and modifications made to the above embodiments still fall within the scope of the technical solution of the present invention.
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