CN115245709A - Filter material for selectively removing leukocytes and simultaneously retaining platelets and preparation method thereof - Google Patents
Filter material for selectively removing leukocytes and simultaneously retaining platelets and preparation method thereof Download PDFInfo
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D39/00—Filtering material for liquid or gaseous fluids
- B01D39/14—Other self-supporting filtering material ; Other filtering material
- B01D39/16—Other self-supporting filtering material ; Other filtering material of organic material, e.g. synthetic fibres
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/02—Blood transfusion apparatus
- A61M1/0281—Apparatus for treatment of blood or blood constituents prior to transfusion, e.g. washing, filtering or thawing
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Abstract
本发明属于血液产品白细胞过滤技术领域,具体涉及一种选择性去除白细胞同时保留血小板的滤材及其制备方法。本发明的滤材是是将如下重量份的原料进行交联在无纺布上得到的:四臂聚乙二醇胺化合物4‑10份、交联剂1‑4份;其中,所述四臂聚乙二醇胺化合物是式Ⅰ所示的化合物或其盐。本发明可实现对全血产品中白细胞的过滤,能够简化血液产品的加工流程、降低加工成本,具有很好的应用前景。
The invention belongs to the technical field of leukocyte filtration of blood products, and in particular relates to a filter material for selectively removing leukocytes while retaining platelets and a preparation method thereof. The filter material of the present invention is obtained by cross-linking the following raw materials by weight on the non-woven fabric: 4-10 parts of a four-arm polyethylene glycol amine compound and 1-4 parts of a cross-linking agent; The arm polyethylene glycol amine compound is a compound of formula I or a salt thereof. The invention can realize the filtration of leukocytes in the whole blood product, can simplify the processing flow of the blood product, reduce the processing cost, and has a good application prospect.
Description
技术领域technical field
本发明属于血液产品白细胞过滤技术领域,具体涉及一种选择性去除白细胞同时保留血小板的滤材及其制备方法。The invention belongs to the technical field of white blood cell filtration of blood products, and in particular relates to a filter material for selectively removing white blood cells while retaining platelets and a preparation method thereof.
背景技术Background technique
输血在临床医学中是一种常见的医疗手段,最主要的目的是帮助一些有疾病的患者,手术中出血或者意外受伤大量出血的伤者恢复健康。而血液产品中白细胞的存在会引起许多相关的输血副反应(如非溶血性发热反应、承认呼吸窘迫正、血小板输注无效等);其次血液中的白细胞还是病毒传播主要媒介物。因此,输血前去除血液产品中的白细胞是非常重要的。Blood transfusion is a common medical method in clinical medicine. The main purpose is to help some patients with diseases, bleeding during surgery or accidental injuries with heavy bleeding to recover. The presence of white blood cells in blood products will cause many related side effects of blood transfusion (such as non-hemolytic fever reaction, positive recognition of respiratory distress, ineffective platelet transfusion, etc.); secondly, white blood cells in blood are also the main vehicle for virus transmission. Therefore, it is very important to remove leukocytes from blood products before transfusion.
白细胞过滤器的发展距今已有半个世纪左右,其制备方法和材料选择也随着科技的进步和发展而不断地革新。第三代白细胞过滤器出现于上世纪90年代,该类过滤器主要是通过在树脂类支撑网中间放置无纺布类材料制备而成。通常先对无纺布材料进行处理以增进其与血液之间的润湿性。由于第三代白细胞过滤器具有制备工艺简单且去白效率高等优势,目前国内外无论血液中心还是医院临床上大都采用这种过滤方法滤除血液中的白细胞,但不同公司和厂家生产的过滤器在滤芯材料的选择和使用上各有不同。It has been about half a century since the development of white blood cell filter, and its preparation method and material selection have been continuously innovated with the progress and development of science and technology. The third-generation leukocyte filter appeared in the 1990s. This type of filter is mainly prepared by placing a non-woven material in the middle of a resin support net. Usually the non-woven material is first treated to improve its wettability with blood. Because the third-generation leukocyte filter has the advantages of simple preparation process and high white-removing efficiency, at present, both blood centers and hospitals at home and abroad mostly use this filtering method to filter white blood cells in the blood, but the filters produced by different companies and manufacturers There are differences in the selection and use of filter material.
虽然,输血用的血液产品需要过滤去除白细胞,但是人们并不希望血液中的红细胞和血小板等其他细胞被滤除。然而,血小板的生理功能为:血管破裂时,血液流体力学发生改变,血小板可快速粘附在血管破裂处,迅速激活并释放其胞内的物质,快速激活凝血系统,形成凝块减少血液的损失。因此,血小板具有易粘附的特性,这导致在过滤白细胞时,血小板也非常容易黏附在滤材上而被除去。Although blood products for transfusion need to be filtered to remove white blood cells, people do not want other cells such as red blood cells and platelets in the blood to be filtered out. However, the physiological function of platelets is: when a blood vessel breaks, the fluid mechanics of the blood changes, platelets can quickly adhere to the broken blood vessel, quickly activate and release their intracellular substances, quickly activate the coagulation system, and form clots to reduce blood loss . Therefore, platelets have the property of easy adhesion, which leads to the fact that platelets are also very easy to adhere to the filter material and be removed when white blood cells are filtered.
因此,开发能够选择性地滤除白细胞而不会滤除血小板的滤材是本领域的重要课题。USP5895575中根据血液中血小板和白细胞的不同性质用水溶性多糖涂层聚醋无纺布和硝基纤维素薄膜得到的滤材可去除白细胞而保留血小板。USP4936998中介绍根据白细胞和血小板的表面电荷密度不同,用HEMA(甲基丙烯酸羟乙酯)和DHEMA共聚物涂层处理聚醋非织造布,使得白细胞吸附在滤材上而血小板通过。EP0559086中介绍了一种白细胞过滤器,制备糖苷甲基丙烯酸羟乙酯,在聚酯纤维表面通过自由基聚合交联糖苷甲基丙烯酸羟乙酯,二氧己烷和HEMA(引发剂偶氮二异丁氰,70℃4小时),该过滤器根据血液中的血小板和白细胞的不同吸附性能对聚醋纤维进行涂层或共聚改性,对于血小板浓缩液(PlateletConcentrates,PC)具有很好的白细胞的去除率和血小板回收率。Therefore, it is an important subject in this field to develop a filter material that can selectively filter out leukocytes without filtering out platelets. In USP5895575, according to the different properties of platelets and leukocytes in blood, the filter material obtained by coating polyester non-woven fabric and nitrocellulose film with water-soluble polysaccharide can remove leukocytes and retain platelets. In USP4936998, according to the different surface charge densities of leukocytes and platelets, polyester nonwovens are treated with HEMA (hydroxyethyl methacrylate) and DHEMA copolymer coating, so that leukocytes are adsorbed on the filter material and platelets pass through. Introduced a kind of leukocyte filter in EP0559086, prepares glycoside hydroxyethyl methacrylate, crosslinks glycoside hydroxyethyl methacrylate, dioxane and HEMA (initiator azobis Isobutylcyanide, 70°C for 4 hours), the filter coats or copolymerizes polyester fibers according to the different adsorption properties of platelets and leukocytes in the blood, and has good leukocytes for platelet concentrates (Platelet Concentrates, PC) The removal rate and platelet recovery rate.
虽然,这些滤材在对浓缩血小板的过滤中取得了很好的效果,能够同时兼有较高的白细胞去除率和血小板回收率。但是这些滤材只能适用于浓缩血小板样品,对于全血样品,由于其中血小板含量相对较低,因而仍然会有较高比例的血小板被滤材滤除,进而导致血小板的回收率无法达到国家相关标准要求。因此,目前血液中心通常将2U(400mL)全血分别制备成红细胞悬液,血浆和浓缩血小板三种成分血,然后分别去除白细胞。而且,2U全血制备的浓缩血小板仍然不能直接过滤,须汇集成一个治疗剂量(12U或者6袋400mL全血制备的PC)后,才能过滤去除白细胞。只有此时,血小板回收率才能够大于85%,符合国家相关标准要求。Although these filter materials have achieved good results in the filtration of concentrated platelets, they can simultaneously have a high leukocyte removal rate and platelet recovery rate. However, these filter materials can only be used to concentrate platelet samples. For whole blood samples, due to the relatively low platelet content, a high proportion of platelets will still be filtered out by the filter materials, resulting in a recovery rate of platelets that cannot reach the national standard. standard requirement. Therefore, at present, the blood center usually prepares 2U (400 mL) whole blood into three components of red blood cell suspension, plasma and concentrated platelets, and then removes white blood cells respectively. Moreover, the concentrated platelets prepared from 2U whole blood still cannot be directly filtered, and must be pooled into a therapeutic dose (12U or 6 bags of PC prepared from 400mL whole blood) before they can be filtered to remove white blood cells. Only at this time, the platelet recovery rate can be greater than 85%, which meets the requirements of relevant national standards.
上述血液样品的处理过程中,受限于血小板回收率的问题,导致处理步骤复杂,使用滤材较多(需对三种成分血分别进行白细胞滤除)。因此,本领域亟需一种针对全血样品的白细胞滤材,能够在对全血样品的过滤中尽可能地滤除白细胞,并保留血小板。In the process of processing the above-mentioned blood samples, limited by the recovery rate of platelets, the processing steps are complicated, and more filter materials are used (leukocytes need to be filtered separately for the three components of blood). Therefore, there is an urgent need in the art for a leukocyte filter material for whole blood samples, which can filter out leukocytes as much as possible and retain platelets during the filtration of whole blood samples.
四臂聚乙二醇及其衍生物是一类由四个聚乙二醇及其衍生物的分子链接枝在同一个季碳上形成的聚合物。现有技术中,这类化合物可通过交联等方式形成凝胶、胶束等,可用于载药(刘东红等,四臂聚乙二醇-聚丙交酯立体复合胶束及其药物传输性能)等用途。而目前,四臂聚乙二醇及其衍生物用于制备血液样品滤材的技术尚未见到有相关报道。Four-arm polyethylene glycol and its derivatives are a class of polymers formed by grafting four molecular chains of polyethylene glycol and its derivatives on the same quaternary carbon. In the prior art, such compounds can form gels, micelles, etc. by means of cross-linking, etc., and can be used for drug loading (Liu Donghong et al., four-armed polyethylene glycol-polylactide stereocomposite micelles and their drug delivery properties) and other purposes. At present, there is no relevant report on the technology of four-armed polyethylene glycol and its derivatives for preparing blood sample filter materials.
发明内容SUMMARY OF THE INVENTION
针对现有技术的问题,本发明提供一种选择性去除白细胞同时保留血小板的滤材及其制备方法,目的在于实现对全血的白细胞过滤。Aiming at the problems in the prior art, the present invention provides a filter material for selectively removing leukocytes while retaining platelets and a preparation method thereof, aiming at realizing leukocyte filtration of whole blood.
一种选择性去除白细胞同时保留血小板的滤材,它是将如下重量份的原料交联在无纺布上得到的:四臂聚乙二醇胺化合物4-10份、交联剂1-4份;A filter material for selectively removing white blood cells while retaining platelets, which is obtained by cross-linking the following raw materials on non-woven fabrics in parts by weight: 4-10 parts of four-arm polyethylene glycol amine compound, 1-4 parts of cross-linking agent share;
其中,所述四臂聚乙二醇胺化合物是式Ⅰ所示的化合物或其盐:Wherein, the four-arm polyethylene glycol amine compound is a compound represented by formula I or a salt thereof:
其中,n1、n2、n3和n4分别独立取自25-75。Wherein, n 1 , n 2 , n 3 and n 4 are independently selected from 25-75.
优选的,所述四臂聚乙二醇胺化合物的数均分子量为5000-20000。Preferably, the number average molecular weight of the four-arm polyethylene glycol amine compound is 5000-20000.
优选的,所述四臂聚乙二醇胺化合物的数均分子量为9000-11000。Preferably, the number average molecular weight of the four-arm polyethylene glycol amine compound is 9000-11000.
优选的,所述交联剂选自二醛类化合物。Preferably, the crosslinking agent is selected from dialdehyde compounds.
优选的,所述二醛类化合物选自戊二醛、丁二醛、己二醛中的一种或两种及以上的组合。Preferably, the dialdehyde compound is selected from one or a combination of two or more of glutaraldehyde, succinic dialdehyde, and adipaldehyde.
优选的,所述无纺布为PBT无纺布、PET无纺布或PP无纺布。Preferably, the non-woven fabric is PBT non-woven fabric, PET non-woven fabric or PP non-woven fabric.
优选的,包括如下步骤:Preferably, the following steps are included:
步骤1,将所述四臂聚乙二醇胺化合物配制成溶液A,交联剂配制成溶液B;Step 1, the four-arm polyethylene glycol amine compound is formulated into solution A, and the cross-linking agent is formulated into solution B;
步骤2,将所述无纺布加入所述溶液A;Step 2, adding the non-woven fabric to the solution A;
步骤3,将所述溶液B加入溶液A,进行交联,即得。
优选的,步骤3中,所述交联的反应条件为25-80℃下反应2-12小时。Preferably, in
本发明还提供上述滤材用于过滤全血中白细胞的用途。The present invention also provides the use of the above filter material for filtering leukocytes in whole blood.
本发明中,“%wt”是指重量百分数,“%v”是指体积百分数。In the present invention, "%wt" means percentage by weight, and "%v" means percentage by volume.
本发明通过将四臂聚乙二醇胺化合物与交联剂交联在无纺布表面,形成水凝胶涂层。按照这种方法制备的材料对白细胞具有很好的过滤效果,同时对血小板的过滤效果较弱。特别地,现有技术中,白细胞滤材只有用于对血小板浓度较高的浓缩血小板进行过滤时,才能够达到较好的血小板回收率。而本发明的滤材可直接用于对全血产品的过滤,血小板回收率可达到80%以上,制成的血液产品质量符合《全血及成分血质量要求》(GB18469-2012)。因此,本发明能够简化血液产品的加工步骤,降低加工成本,具有很好的应用前景。In the invention, the hydrogel coating is formed by cross-linking the four-arm polyethylene glycol amine compound and the cross-linking agent on the surface of the non-woven fabric. The material prepared according to this method has a good filtering effect on leukocytes, and at the same time has a weak filtering effect on platelets. In particular, in the prior art, only when the white blood cell filter material is used to filter concentrated platelets with a high platelet concentration can a better recovery rate of platelets be achieved. However, the filter material of the present invention can be directly used for filtering whole blood products, and the recovery rate of platelets can reach more than 80%, and the quality of the prepared blood products meets the "Quality Requirements for Whole Blood and Blood Components" (GB18469-2012). Therefore, the invention can simplify the processing steps of blood products, reduce the processing cost, and has good application prospects.
显然,根据本发明的上述内容,按照本领域的普通技术知识和惯用手段,在不脱离本发明上述基本技术思想前提下,还可以做出其它多种形式的修改、替换或变更。Apparently, according to the above content of the present invention, according to common technical knowledge and conventional means in this field, without departing from the above basic technical idea of the present invention, other various forms of modification, replacement or change can also be made.
以下通过实施例形式的具体实施方式,对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。The above-mentioned content of the present invention will be further described in detail below through specific implementation in the form of examples. However, this should not be construed as limiting the scope of the above-mentioned subject matter of the present invention to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention.
附图说明Description of drawings
图1为滤材的扫描电镜图片,其中,A:原始PBT无纺布;B:实施例1制备的滤材;C:实施例2制备的滤材;D:实施例3制备的滤材;E:实施例4制备的滤材。Fig. 1 is the scanning electron microscope picture of filter material, wherein, A: original PBT non-woven fabric; B: the filter material prepared by embodiment 1; C: the filter material prepared by embodiment 2; D: the filter material prepared by
具体实施方式Detailed ways
以下实施例和实验例中,未特别说明的试剂和材料均为市售品。以下实施例中,四臂聚乙二醇胺化合物用缩写“4armPEG”表示。四臂聚乙二醇胺化合物的结构式为:In the following examples and experimental examples, all reagents and materials not specified are commercially available. In the following examples, the four-arm polyethylene glycol amine compound is represented by the abbreviation "4armPEG". The structural formula of the four-arm polyethylene glycol amine compound is:
其中,n1、n2、n3和n4分别独立取自25-75。Wherein, n 1 , n 2 , n 3 and n 4 are independently selected from 25-75.
实施例1选择性去除白细胞同时保留血小板的滤材Example 1 Filter material for selectively removing leukocytes while retaining platelets
使用电子天平准确称量1.000g 4armPEG(键凯科技,4ARM-PEG-NH2HCl,数均分子量2000Da),溶解在50mL水溶液中,得到2%wt的4armPEG溶液。20g PBT无纺布浸入上述100mL溶液中,加入0.5mL 50%v的戊二醛溶液(含戊二醛0.25g),放入60℃干燥箱中交联3小时,反应结束后,使用蒸馏水清洗PBT无纺布,得到戊二醛交联4armPEG的PBT无纺布膜备用。1.000g of 4armPEG (Jiankai Technology, 4ARM-PEG-NH2HCl, number average molecular weight 2000Da) was accurately weighed using an electronic balance, and dissolved in 50mL of aqueous solution to obtain a 2%wt 4armPEG solution. Immerse 20g of PBT non-woven fabric in the above 100mL solution, add 0.5mL of 50% v glutaraldehyde solution (containing 0.25g of glutaraldehyde), put it in a 60°C drying oven for cross-linking for 3 hours, and wash it with distilled water after the reaction PBT non-woven fabric, obtain the PBT non-woven fabric membrane of glutaraldehyde cross-linked 4armPEG for subsequent use.
实施例2选择性去除白细胞同时保留血小板的滤材Example 2 Filter material for selectively removing leukocytes while retaining platelets
使用电子天平准确称量1.000g 4armPEG(键凯科技,4ARM-PEG-NH2HCl,数均分子量5000Da),溶解在50mL水溶液中,得到2%wt的4armPEG溶液。20g PBT无纺布浸入上述100mL溶液中,加入0.5mL 50%v的戊二醛溶液(含戊二醛0.25g),放入60℃干燥箱中交联3小时,反应结束后,使用蒸馏水清洗PBT无纺布,得到戊二醛交联4armPEG的PBT无纺布膜备用。1.000 g of 4armPEG (Jiankai Technology, 4ARM-PEG-NH2HCl, number average molecular weight 5000Da) was accurately weighed using an electronic balance, and dissolved in 50 mL of aqueous solution to obtain a 2% wt 4armPEG solution. Immerse 20g of PBT non-woven fabric in the above 100mL solution, add 0.5mL of 50% v glutaraldehyde solution (containing 0.25g of glutaraldehyde), put it in a 60°C drying oven for cross-linking for 3 hours, and wash it with distilled water after the reaction PBT non-woven fabric, obtain the PBT non-woven fabric membrane of glutaraldehyde cross-linked 4armPEG for subsequent use.
实施例3选择性去除白细胞同时保留血小板的滤材Example 3 Filter material for selectively removing leukocytes while retaining platelets
使用电子天平准确称量1.000g 4armPEG(键凯科技,4ARM-PEG-NH2HCl,数均分子量10000Da),溶解在50mL水溶液中,得到2%wt的4armPEG溶液。20g PBT无纺布浸入上述100mL溶液中,加入0.5mL 50%v的戊二醛溶液(含戊二醛0.25g),放入60℃干燥箱中交联3小时,反应结束后,使用蒸馏水清洗PBT无纺布,得到戊二醛交联4armPEG的PBT无纺布膜备用。1.000g of 4armPEG (Jiankai Technology, 4ARM-PEG-NH2HCl, number average molecular weight 10000Da) was accurately weighed using an electronic balance, and dissolved in 50mL of aqueous solution to obtain a 2%wt 4armPEG solution. Immerse 20g of PBT non-woven fabric in the above 100mL solution, add 0.5mL of 50% v glutaraldehyde solution (containing 0.25g of glutaraldehyde), put it in a 60°C drying oven for cross-linking for 3 hours, and wash it with distilled water after the reaction PBT non-woven fabric, obtain the PBT non-woven fabric membrane of glutaraldehyde cross-linked 4armPEG for subsequent use.
实施例4选择性去除白细胞同时保留血小板的滤材Example 4 Filter material for selectively removing leukocytes while retaining platelets
使用电子天平准确称量1.000g 4armPEG(键凯科技,4ARM-PEG-NH2HCl,数均分子量20000Da),溶解在50mL水溶液中,得到2%wt的4armPEG溶液。20g PBT无纺布浸入上述100mL溶液中,加入0.5mL 50%v的戊二醛溶液(含戊二醛0.25g),放入60℃干燥箱中交联3小时,反应结束后,使用蒸馏水清洗PBT无纺布,得到戊二醛交联4armPEG的PBT无纺布膜备用。1.000g of 4armPEG (Jiankai Technology, 4ARM-PEG-NH2HCl, number average molecular weight 20000Da) was accurately weighed using an electronic balance, and dissolved in 50mL of aqueous solution to obtain a 2%wt 4armPEG solution. Immerse 20g of PBT non-woven fabric in the above 100mL solution, add 0.5mL of 50% v glutaraldehyde solution (containing 0.25g of glutaraldehyde), put it in a 60°C drying oven for cross-linking for 3 hours, and wash it with distilled water after the reaction PBT non-woven fabric, obtain the PBT non-woven fabric membrane of glutaraldehyde cross-linked 4armPEG for subsequent use.
实施例5选择性去除白细胞同时保留血小板的滤材Example 5 Filter material for selectively removing leukocytes while retaining platelets
使用电子天平准确称量1.000g 4armPEG(键凯科技,4ARM-PEG-NH2HCl,数均分子量10000Da),溶解在50mL水溶液中,得到2%wt的4armPEG溶液。20g PBT无纺布浸入上述100mL溶液中,加入2mL 50%v的戊二醛溶液(含戊二醛1.000g),放入60℃干燥箱中交联3小时,反应结束后,使用蒸馏水清洗PBT无纺布,得到戊二醛交联4armPEG的PBT无纺布膜备用。1.000g of 4armPEG (Jiankai Technology, 4ARM-PEG-NH2HCl, number average molecular weight 10000Da) was accurately weighed using an electronic balance, and dissolved in 50mL of aqueous solution to obtain a 2%wt 4armPEG solution. Immerse 20g of PBT non-woven fabric in the above 100mL solution, add 2mL of 50% v glutaraldehyde solution (containing 1.000g of glutaraldehyde), put it in a 60°C drying oven for cross-linking for 3 hours, and wash the PBT with distilled water after the reaction Non-woven fabric, obtain the PBT non-woven fabric membrane of glutaraldehyde cross-linked 4armPEG for subsequent use.
实施例6选择性去除白细胞同时保留血小板的滤材Example 6 Filter material for selectively removing leukocytes while retaining platelets
使用电子天平准确称量2.500g 4armPEG(键凯科技,4ARM-PEG-NH2HCl,数均分子量10000Da),溶解在50mL水溶液中,得到2%wt的4armPEG溶液。20g PBT无纺布浸入上述100mL溶液中,加入2mL 50%v的戊二醛溶液(含戊二醛1.000g),放入60℃干燥箱中交联3小时,反应结束后,使用蒸馏水清洗PBT无纺布,得到戊二醛交联4armPEG的PBT无纺布膜备用。An electronic balance was used to accurately weigh 2.500g 4armPEG (Jiankai Technology, 4ARM-PEG-NH2HCl, number average molecular weight 10000Da), and dissolve it in 50mL aqueous solution to obtain a 2%wt 4armPEG solution. Immerse 20g of PBT non-woven fabric in the above 100mL solution, add 2mL of 50% v glutaraldehyde solution (containing 1.000g of glutaraldehyde), put it in a 60°C drying oven for cross-linking for 3 hours, and wash the PBT with distilled water after the reaction Non-woven fabric, obtain the PBT non-woven fabric membrane of glutaraldehyde cross-linked 4armPEG for subsequent use.
下面通过实验对本发明的技术方案作进一步的说明。The technical scheme of the present invention will be further described through experiments below.
实验例1滤材性能表征实验Experimental example 1 Filter material performance characterization experiment
一、实验方法1. Experimental method
1)表面张力测定:1) Determination of surface tension:
对原始PBT无纺布和实施例1-4制备的滤材进行表面张力测定,对材料亲水性进行表征。分别配制不同浓度的氯化钠(1%-4.5%)及氯化钙(3%-5%)溶液,用液体自动表面张力仪检测溶液的表面张力。用注射器将不同浓度的液体滴在试样表面,以10min内液滴完全铺展的数目来进行衡量。静置10min,在第10min至11min内进行观察。如10min内10滴中至少有9滴浸湿多孔介质,则认为该种表面张力的液体浸湿了介质材料;反之,如10min内10滴中有2滴或更多的液滴未浸湿或不被吸收,则认为该种表面张力的液体不能浸湿该介质。例如,用纯水(其表面张力为72×10-5N/cm)的液滴10滴滴在材料表面上,10min内有9滴浸湿了;而用表面张力75×10-5N/cm的液滴10滴滴在材料表面上,10min内有3滴未浸湿,则该材料的临界表面张力按定义为73.5×10-5N/cm。The surface tension of the original PBT non-woven fabric and the filter material prepared in Examples 1-4 was measured to characterize the hydrophilicity of the material. Sodium chloride (1%-4.5%) and calcium chloride (3%-5%) solutions with different concentrations were prepared respectively, and the surface tension of the solutions was detected with a liquid automatic surface tensiometer. Use a syringe to drop different concentrations of liquid on the surface of the sample, and measure it by the number of drops that are completely spread within 10 minutes. Stand still for 10 minutes, and observe within 10 minutes to 11 minutes. If at least 9 out of 10 drops wet the porous medium within 10 minutes, it is considered that the surface tension liquid wets the medium material; otherwise, if 2 or more drops out of 10 drops within 10 minutes are not wetted or If it is not absorbed, it is considered that the liquid of this surface tension cannot wet the medium. For example, if 10 drops of pure water (whose surface tension is 72×10 -5 N/cm) are dropped on the surface of the material, 9 drops will be wetted within 10 minutes ; If 10 cm of liquid droplets are dropped on the surface of the material and 3 drops are not wetted within 10 minutes, then the critical surface tension of the material is defined as 73.5×10 -5 N/cm.
2)扫描电镜观察滤材表面形貌2) Scanning electron microscope to observe the surface morphology of the filter material
将原始PBT无纺布和实施例1至实施例4制备的滤材喷金后采用扫描电镜观察滤材的表面形貌。The original PBT non-woven fabric and the filter material prepared in Examples 1 to 4 were sprayed with gold, and the surface morphology of the filter material was observed with a scanning electron microscope.
二、实验结果2. Experimental results
1、亲水性表征结果:在室温下,纯水的表面张力通常为72×10-5N/cm,原始PBT无纺布的表面张力为48×10-5N/cm。戊二醛交联4armPEG后形成的水凝胶涂层覆盖在到PBT无纺布的纤维表面,有助于提高材料的亲水性。由表1中的结果可知,PBT滤材经4arm PEG改性后,表面张力较原始PBT显著提高,且随着4arm PEG的分子量的增加,滤材表面张力增加,材料表面亲水性增加。这有助于提高血液过滤速度。1. Hydrophilic characterization results: at room temperature, the surface tension of pure water is usually 72×10 -5 N/cm, and that of the original PBT non-woven fabric is 48×10 -5 N/cm. The hydrogel coating formed after glutaraldehyde cross-linked 4armPEG covers the fiber surface of the PBT non-woven fabric, which helps to improve the hydrophilicity of the material. It can be seen from the results in Table 1 that the surface tension of the PBT filter material modified by 4arm PEG is significantly higher than that of the original PBT, and with the increase of the molecular weight of 4arm PEG, the surface tension of the filter material increases, and the surface hydrophilicity of the material increases. This helps to increase the rate at which the blood is filtered.
表1原始PBT及改性滤材的表面张力结果Table 1 Surface tension results of original PBT and modified filter material
2、表面形貌表征结果:由图1看出,与原始PBT无纺布相比,4arm PEG改性后的滤材纤维直径增粗,纤维表面有颗粒状附着物,纤维与纤维之间孔隙变窄。说明4arm PEG交联后形成的水凝胶成功覆盖在到PBT无纺布的纤维表面。2. Surface morphology characterization results: It can be seen from Figure 1 that compared with the original PBT non-woven fabric, the fiber diameter of the 4arm PEG modified filter material is thicker, there are granular attachments on the fiber surface, and the pores between the fibers narrowed. It shows that the hydrogel formed after cross-linking of 4arm PEG successfully covers the fiber surface of PBT non-woven fabric.
实验例2全血过滤实验Experimental Example 2 Whole Blood Filtration Experiment
一、实验方法1. Experimental method
将实施例1至实施例4制备的4种滤材分别装入直径为60mm白细胞过滤器的壳后,通过超声焊接过滤器,并连接PVC管路和血袋。每个血袋中装入200mL全血,在一米净压差下血液流过白细胞过滤器,流出的血液通过血袋收集。试验所用全血由德阳中心血站提供。The four kinds of filter materials prepared in Examples 1 to 4 were loaded into the shells of the white blood cell filter with a diameter of 60 mm, and the filters were ultrasonically welded and connected to the PVC pipeline and the blood bag. Each blood bag is filled with 200mL whole blood, and the blood flows through the white blood cell filter under a net pressure difference of one meter, and the outflowing blood is collected through the blood bag. Whole blood used in the experiment was provided by the Deyang Central Blood Bank.
各项参数的检测方法如下:The detection method of each parameter is as follows:
1)血小板回收率计算方法1) Calculation method of platelet recovery rate
血小板回收率(%)=(过滤后血小板浓度×过滤后体积)/(过滤前血小板浓度×过滤前体积)×100Platelet recovery (%) = (platelet concentration after filtration × volume after filtration) / (platelet concentration before filtration × volume before filtration) × 100
通过五分类血细胞计数仪测定血小板过滤前后浓度,通过称重方式获取体积。The concentration of platelets before and after filtration was measured by a five-differential blood cell counter, and the volume was obtained by weighing.
2)白细胞去除率计算方法2) Calculation method of white blood cell removal rate
白细胞回收率(%)=(过滤前白细胞浓度×过滤前体积-过滤后剩余白细胞浓度×过滤后体积)/(过滤前白细胞浓度×过滤前体积)×100White blood cell recovery (%) = (leukocyte concentration before filtration × volume before filtration - remaining leukocyte concentration after filtration × volume after filtration) / (leukocyte concentration before filtration × volume before filtration) × 100
通过五分类血细胞计数仪测定过滤前白细胞浓度,通过细胞计数板获取过滤后剩余白细胞浓度,通过称重方式获取体积。The white blood cell concentration before filtration was measured by a five-differential blood cell counter, the remaining white blood cell concentration after filtration was obtained by a cell counting plate, and the volume was obtained by weighing.
3)红细胞回收率计算方法3) Calculation method of red blood cell recovery rate
红细胞回收率(%)=(过滤后红细胞浓度×过滤后体积)/(过滤前红细胞浓度×过滤前体积)×100Red blood cell recovery (%) = (red blood cell concentration after filtration × volume after filtration) / (red blood cell concentration before filtration × volume before filtration) × 100
通过五分类血细胞计数仪测定红细胞过滤前后浓度,通过称重方式获取体积。The concentration of red blood cells before and after filtration was measured by a five-differential blood cell counter, and the volume was obtained by weighing.
4)溶血率计算方法4) Calculation method of hemolysis rate
本实验按照国家医药标准《一次性使用去白细胞滤器》YY-0329-2020中溶血率的测定方法实验。通过用体积分数为的0.9%的氯化钠注射液作为浸提介质制备去白细胞滤器器材或过滤材料的检验液与血液直接接触,测定红细胞释放的血红蛋白量以检测去白细胞滤器或过滤材料体外溶血程度。This experiment was carried out in accordance with the determination method of hemolysis rate in the national medical standard "Disposable Leukocyte Removal Filter" YY-0329-2020. By using 0.9% sodium chloride injection as the leaching medium to prepare the test solution for the leukocyte-removing filter device or filter material and directly contact with the blood, measure the amount of hemoglobin released by red blood cells to detect the hemolysis of the leukocyte-removing filter or filter material in vitro degree.
5)过滤时间5) Filtration time
过滤完200mL全血需要的时间。The time required to filter 200mL of whole blood.
6)血液损失率计算方法6) Calculation method of blood loss rate
血液损失率(%)=(过滤前血液重量-过滤后血液重量)/(过滤前血液重量)×100Blood loss rate (%)=(blood weight before filtration-blood weight after filtration)/(blood weight before filtration)×100
通过称重方式获取血液重量。Obtain blood weight by weighing.
二、实验结果2. Experimental results
全血过滤结果:Whole blood filtration results:
检测过滤前后的血液相关指标,结果见表2。The blood-related indicators before and after filtration were detected, and the results are shown in Table 2.
表2Table 2
从表中各项性能来看,随着4arm PEG分子量的升高,血小板回收率、白细胞去除率、红细胞回收率和血液损失率均表现为先升高后降低;溶血率和过滤时间表现为先降低后升高。即整体来看,滤材的过滤性能随着4arm PEG分子量的升高呈现先增强后减弱的趋势。其中当4arm PEG分子量为10000Da时,制成的滤材具有最佳的过滤性能。过滤后的全血质量符合《全血及成分血质量要求》(GB18469-2012)。From the properties in the table, with the increase of the molecular weight of 4arm PEG, the platelet recovery rate, white blood cell removal rate, red blood cell recovery rate and blood loss rate all showed an increase first and then decreased; the hemolysis rate and filtration time showed a first increase. Decrease and then increase. That is to say, overall, the filtration performance of the filter material shows a trend of first increasing and then weakening with the increase of the molecular weight of 4arm PEG. Among them, when the molecular weight of 4arm PEG is 10000Da, the filter material has the best filtration performance. The quality of filtered whole blood complies with the "Quality Requirements for Whole Blood and Blood Components" (GB18469-2012).
通过上述实施例和实验例可以看到,本发明提供了一种新的白细胞过滤滤材,采用该滤材可直接对全血进行过滤,血小板回收率可达到80%以上,制成的血液产品质量符合《全血及成分血质量要求》(GB18469-2012)。本发明可简化血液产品的加工流程、降低加工成本,具有很好的应用前景。It can be seen from the above examples and experimental examples that the present invention provides a new leukocyte filter material, which can directly filter whole blood, and the recovery rate of platelets can reach more than 80%. The quality complies with the "Quality Requirements for Whole Blood and Blood Components" (GB18469-2012). The invention can simplify the processing flow of the blood product, reduce the processing cost, and has good application prospect.
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