CN104540531A - 细胞因子吸附片及其制备方法以及利用其的血液过滤器 - Google Patents

细胞因子吸附片及其制备方法以及利用其的血液过滤器 Download PDF

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CN104540531A
CN104540531A CN201380032360.3A CN201380032360A CN104540531A CN 104540531 A CN104540531 A CN 104540531A CN 201380032360 A CN201380032360 A CN 201380032360A CN 104540531 A CN104540531 A CN 104540531A
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cytokine
mentioned
fiber net
nanometer fiber
suction sheet
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黄俊植
徐尚哲
金灿
李承勋
金庆洙
金喜赞
李晶灿
徐佶葰
权云龙
金景洙
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Seoul National University Industry Foundation
SNU R&DB Foundation
Amogreentech Co Ltd
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Seoul National University Industry Foundation
Amogreentech Co Ltd
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Abstract

本发明的细胞因子吸附片包含将纺丝溶液进行电纺丝而成的纳米纤维网,上述纺丝溶液由能够吸附细胞因子的吸附物质和能够进行电纺丝的高分子物质混合而成,由此能够防止吸附物质因血液而被溶出的现象。

Description

细胞因子吸附片及其制备方法以及利用其的血液过滤器
技术领域
本发明涉及能够有效吸附并去除血液中所含有的促炎性细胞因子(Pro-inflammatory cytokine),来治疗败血症的细胞因子吸附片及其制备方法以及利用上述细胞因子吸附片的血液过滤器。
背景技术
众所周知,败血症(sepsis)是为了对抗细菌、病毒、寄生虫或真菌类等排出的毒素而生产的细胞因子(cytokine)的生产过剩而使生物体内部的免疫反应系统崩溃,从而发生的疾病。
败血症的临床症状如下:病原性微生物的感染对宿主的免疫系统、凝固系统、神经激素系统等各种功能系统产生影响,导致以与此相关的全身反应出现,因而呈现复杂且多样的临床表现。并且,若症状严重,则会患上多器官功能障碍综合征,导致死亡,是重症监护室患者死亡的主要原因,且死亡率仍上升20%。
当前,作为治疗败血症的方法,使用通过早期抗生素给药及输液疗法、血液增效剂给药的早期目标导向治疗(Early goal directed therapy),但至今,利用免疫功能调节的有效的治疗方法尚未获得成果。
并且,在用于吸附并去除促炎性细胞因子的血液过滤器相关研究中报告了将多粘菌素-B(EP 424698,Therapeutic Apheresis and Dialysis(2003)7(1):108,polyethyleneimine(PEI)(Artificial Organs(1993)17(9):775,Artificial Organs(1996)),polyvinylpyrrolidone(PVP)(Critical Care Medicine(2004)32(3):801)等涂敷于粒状活性炭(Biomaterials(2006)27:5286,5755,珠(beads)、纤维状底物(substrate)等,来进行处理的方法等。
将这种吸附物质涂敷于底物的情况下,所涂敷的吸附物质具有因血液而被溶出的可能性,当向生物体内流出时,呈现强的肾毒性(renal toxicity)或细胞毒性(cytotoxicity),而对于这些的临床功效,尚未发表明确的研究结果。
并且,用作底物的珠或纤维状底物的情况下,由于比表面积的限制,无法提供与血液的充分的接触面积,活性炭的情况下,因涂敷吸附物质而使细孔被堵塞,导致存在无法提供充分的表面积的缺点。
现有的血液过滤器如韩国登录特许公报10-1151139(2012年05月22日)中所公开,包含数均直径在1nm以上且500nm以下,大于500nm且1μm以下的直径范围的短纤维的纤维比率以重量换算时为3%以下的由聚酯或聚酰胺形成的纳米纤维分散体,来过滤或吸附血液中的成分,从而去除白细胞或毒素、蛋白质等。
但是,现有的血液过滤器的材质由聚酯或聚酰胺形成,从而借助纳米纤维的气孔来过滤或吸附血液中的成分,因而存在无法吸附促炎性细胞因子等特殊成分的问题。
并且,现有的血液过滤器在需要吸附细胞因子等特殊成分的情况下,通过在过滤器的表面涂敷吸附物质来使用,而这种情况下,如上所述,存在具有涂敷于过滤器的表面的吸附物质因血液而被溶出的可能性的问题。
发明内容
技术问题
本发明用于改善这种现有技术的问题,本发明的目的在于,提供通过电纺丝方法来形成为纳米纤维网,从而能够增大与细胞因子的接触面积,由此能够提高吸附性能的细胞因子吸附片及其制备方法。
本发明的再一目的在于,提供通过电纺丝方法来形成为纳米纤维网,从而能够自动地调节薄片的厚度,进而能够制备成更加薄,从而将过滤器小型化,并提高吸附性能的细胞因子吸附片及其制备方法。
本发明的另一目的在于,提供通过将细胞因子吸附物质和能够纺丝的高分子物质混合,来以电纺丝方法制备,因而能够尽可能抑制吸附物质因血液而被溶出的现象的细胞因子吸附片及其制备方法。
本发明的还一目的在于,提供设有能够有效吸附并去除细胞因子的纳米纤维网形态的细胞因子吸附片的血液过滤器。
本发明所要解决的问题并不局限于以上所提及的技术问题,未提及的其他技术问题能够由本发明所属技术领域的普通技术人员根据以下纳米纤维网明确地理解。
解决问题的手段
为了达成上述目的,本发明的细胞因子吸附片的特征在于,包含将纺丝溶液进行电纺丝而成的纳米纤维网,上述纺丝溶液由能够吸附细胞因子的吸附物质和能够进行电纺丝的高分子物质混合而成。
本发明的特征在于,纳米纤维网通过上述纺丝溶液的电纺丝来累积纳米纤维,由具有多个气孔的形态形成,上述纳米纤维的直径在100nm~800nm范围内,气孔的平均大小在0.1μm~10μm范围内。
本发明的特征在于,吸附物质使用选自多粘菌素-B(polymyxin-B)、聚乙烯亚胺(PEI)、聚乙烯吡咯烷酮(PVP)、聚苯乙烯-二乙烯基苯(PS-DVB,polystyrene-divinylbenzene)中的一种或两种以上的混合物。
本发明的特征在于,高分子物质使用将聚偏氟乙烯(PVdF,polyvinylidenefluoride)、聚甲基丙烯酸甲酯(PMMA,poly-methylmethacrylate)、聚丙烯腈(PAN,polyacrylonitrile)、聚氨酯(PU,poly-urethane)、聚醚砜(PES,polyethersulfone)、聚酰胺酸(PAA,polyamicacid)、聚乙烯醇(PVA,polyvinylachol)、聚环氧乙烷(PEO,polyethyleneoxide)、聚乳酸(PLA,polylacticacid)、聚乙醇酸(PGA,polyglycolic acid)、聚乳酸-聚乙醇酸类的高分子物质中的一种或它们的两种以上混合的高分子物质。
本发明的特征在于,细胞因子吸附片还包括层叠于纳米纤维网的一面的基片;在上述基片形成有能够使血液通过的多个气孔,上述基片使用将无纺布、织造的织物、高分子或金属泡沫(foam)、纸、金属及塑料网中的一种或两种以上复合化的复合体。
本发明的特征在于,血液过滤器包括细胞因子吸附片,上述细胞因子吸附片隔着恒定间隔,缠绕成辊形态,用于形成使血液通过的通道,在上述通道设有间隔片。
本发明的细胞因子吸附片的制备方法的特征在于,包括:将能够吸附细胞因子的吸附物质和能够进行电纺丝的高分子物质溶解于溶剂,来制备纺丝溶液的步骤;以及将上述纺丝溶液进行电纺丝,来形成具有多个气孔的纳米纤维网的步骤。
本发明的特征在于,在形成纳米纤维网的步骤中,若对收集器和纺丝喷嘴之间施加高电压静电力,且由纺丝喷嘴向收集器纺丝纳米纤维,则纳米纤维累积于收集器。
本发明的细胞因子吸附片的制备方法的特征在于,还包括:使纳米纤维网通过加压辊来制备成恒定厚度的步骤;以及在上述纳米纤维网的一面层叠基片的步骤。
本发明的特征在于,基片通过热熔敷、压延、层压、热熔粘合及焊接中的一种方法来粘结于纳米纤维网的一面。
发明的效果
如上所述,本发明的细胞因子吸附片通过电纺丝方法来形成为纳米纤维网,因而具有能够增大与细胞因子的接触面积,以提高吸附性能的优点。
并且,本发明的细胞因子吸附片通过电纺丝方法来形成为纳米纤维网,因而具有能够自动地调节薄片的厚度,进而能够制备成更加薄,从而将过滤器小型化,并提高吸附性能的优点。
并且,本发明的细胞因子吸附片通过将细胞因子吸附物质和能够纺丝的高分子物质混合,来以电纺丝方法制备,因而具有能够尽可能抑制吸附物质因血液而被溶出的现象的优点。
附图说明
图1为根据本发明一实施例制备的细胞因子吸附片的放大照片。
图2为本发明一实施例的细胞因子吸附片制备用电纺丝装置的结构图。
图3为本发明另一实施例的电纺丝装置的结构图。
图4为本发明一实施例的血液过滤器的剖视图。
图5为内置于本发明一实施例的血液过滤器的细胞因子吸附片的立体图。
图6为根据本发明的实施例3制备的细胞因子吸附片的放大照片。
图7为根据本发明的实施例4制备的细胞因子吸附片的放大照片。
具体实施方式
以下,参照附图更加具体说明本发明。在此过程中,为了明确和便于说明,图中所示的结构要素的大小或形状等会有所夸张。并且,考虑本发明的结构及作用而特别定义的术语可根据用户、操作人员的意图或惯例而不同。这种术语应基于本说明书全文内容定义。
图1为本发明一实施例的细胞因子吸附片的放大照片。上述细胞因子吸附片包含将纺丝溶液进行电纺丝而成的直径小于1μm且具有多个气孔的纳米纤维网(nanofiber web)10,上述纺丝溶液由可吸附细胞因子的吸附物质和可进行电纺丝的高分子物质混合而成。
这种细胞因子吸附片通过将可进行电纺丝的高分子物质和可吸附细胞因子的吸附物质混合于溶剂,来制备具有可进行电纺丝的粘度的纺丝溶液,通过将该纺丝溶液进行电纺丝来制备纳米纤维12,并累积该纳米纤维12来形成为具有多个气孔14的纳米纤维网10。
纳米纤维12的直径大致在100nm~800nm范围内,气孔14的平均大小在0.1μm~10μm范围内。在这里,细胞因子吸附性能与吸附片的比表面积以及与血液的接触时间有着密切的关系,因而气孔的平均大小最优选为0.5μm。
并且,细胞因子吸附片的厚度在1μm~150μm范围内,当借助血液过滤器来制备时,若考虑最小的强度及接触面积等,最优选在15μm~20μm范围内。
以纺丝溶液的总含量为基准,高分子物质和吸附物质的含量在5~90重量百分比的范围内,尤其,当考虑纺丝的稳定性、纳米纤维12的强度及气孔14的大小等时,最适合在10~30重量百分比的范围内。
在这里,细胞因子吸附物质使用将选自多粘菌素-B、聚乙烯亚胺、聚乙烯吡咯烷酮、聚苯乙烯-二乙烯基苯(PS-DVB,polystyrene-divinylbenzene)中的一种或两种以上复合化的复合体。当然,除此之外,也可使用能够吸附细胞因子,并溶解于溶剂的任何物质。
在这种细胞因子吸附物质中,可进行电纺丝来形成为纳米纤维网的物质可单独执行电纺丝,但优选地,与医疗用高分子物质混合来实施电纺丝,以提高吸附片的物性。
并且,高分子物质使用可进行电纺丝且能够用于医疗的物质,作为一例,可单独适用聚偏氟乙烯(PVdF,polyvinylidenefluoride)、聚甲基丙烯酸甲酯(PMMA,poly-methylmethacrylate)、聚丙烯腈(PAN,polyacrylonitrile)、聚氨酯(PU,poly-urethane)、聚醚砜(PES,polyethersulfone)、聚酰胺酸(PAA,polyamicacid)、聚乙烯醇(PVA,polyvinylachol)、聚环氧乙烷(PEO,polyethyleneoxide)、聚乳酸(PLA,polylacticacid)、聚乙醇酸(PGA,polyglycolic acid)、聚乳酸-聚乙醇酸类等的高分子物质或者适用将这些复合化的高分子物质。
高分子物质除了罗列于其上的高分子物质之外,作为可进行电纺丝的物质,还可使用合成高分子或天然高分子,只要是对血液不发生异常反应的高分子,那么还可适用任何高分子物质。
溶剂作为得到具有适合将吸附物质和高分子物质进行电纺丝的浓度的恒定粘度的物质,可使用选自N,N-二甲基乙酰胺(DMAc,N,N-Dimethyl acetoamide)、N,N-二甲基甲酰胺(DMF,N,N-Dimethylformamide)、N-甲基-2-吡咯烷酮(NMP,N-methyl-2-pyrrolidinone)、二甲基亚砜(DMSO,dimethyl sulfoxide)、四氢呋喃(THF,tetra-hydrofuran)、碳酸乙烯酯(EC,ethylene carbonate)、碳酸二乙酯(DEC,diethyl carbonate)、碳酸二甲酯(DMC,dimethyl carbonate)、碳酸甲乙酯(EMC,ethyl methyl carbonate)、碳酸丙烯酯(PC,propylene carbonate)、水、乙酸(acetic acid)、甲酸(formic acid)、氯仿(Chloroform)、二氯甲烷(dichloromethane)及丙酮(acetone)中的一种或两种以上的混合物。
细胞因子吸附片通过电纺丝方法来制备,因而厚度根据纺丝溶液的纺丝量而决定。因此,具有容易将细胞因子吸附片的厚度制备成所需的厚度的优点。即,可根据纺丝溶液的纺丝量来调节纳米纤维网的厚度,从而能够以各种厚度制备,尤其,可制备成更薄,从而能够减少制备费用,并将血液过滤器的大小小型化。
像这样,本实施例的细胞因子吸附片能够将厚度制备成更薄,从而当制备血液过滤器时,能够将血液过滤器的尺寸制备成更小,并使吸附性能更优秀。
并且,细胞因子吸附片通过电纺丝方法来形成为累积有纳米纤维12的纳米纤维网10,因而能够将比表面积形成为更宽,从而增大与血液的接触面积,因而能够提高细胞因子吸附性能。
本发明的另一实施例的细胞因子吸附片包括:纳米纤维网,通过将纺丝溶液进行电纺丝而成,其直径小于1μm,且具有多个气孔,上述纺丝溶液由可吸附细胞因子的吸附物质和可进行电纺丝的高分子物质混合而成;以及基片,层叠于纳米纤维网的一面或两面,用于提高纳米纤维网的处理性和物性。
在这里所使用的纳米纤维网10呈与如上所述的纳米纤维网相同的形态,只要是具有可使血液通过的多个气孔,且提高吸附片的处理性或物性,那么基片的材质就可以适用任何材质。作为一例,基片可以为选自由无纺布、织造的织物、高分子泡沫或金属泡沫、纸、金属或塑料网等组成的组中的一种以上。
纳米纤维网和基片可通过热压接、压延(Calendering)、层压(laminating)、热熔粘合焊接、超声波焊接等各种方法来进行贴合,只要是当与血液相接触时不发生副作用的方法,那么就可以适用任何方法。
并且,除了这种贴合方法之外,还可适用通过在基片的表面直接将纳米纤维网进行电纺丝来层叠的方法。
层叠纳米纤维网和基片的细胞因子吸附片执行灭菌处理,而作为灭菌处理方法,只要是环氧乙烷(ethylene oxide)、高温蒸汽、X-射线(x-ray)方法等不对纳米纤维网和细胞因子吸附物质的物性产生影响的方法,那么就可以适用任何方法。
这种细胞因子吸附片通过在纳米纤维网的一面层叠基片,来能够提高处理性和物性。
图2为制备本发明一实施例的细胞因子吸附片的电纺丝装置的结构图。
本发明的电纺丝装置包括:混合槽30(Mixing Tank),用于储存纺丝溶液;多个纺丝喷嘴34,与高电压发生器相连接,并与混合槽30相连接,用于纺丝纳米纤维14;以及收集器36,将从纺丝喷嘴34纺丝的纳米纤维14累积于上述收集器36而制备纳米纤维网10。
在混合槽30设有搅拌器32,上述搅拌器32用于将可吸附细胞因子的吸附物质及可进行电纺丝的高分子物质和溶剂均匀混合,并使高分子物质维持恒定粘度。
对收集器36和纺丝喷嘴34之间施加高电压静电力,且纳米纤维14从纺丝喷嘴34纺丝,从而在收集器36上形成纳米纤维网10。
此时所使用的电压以在0.5kV~100kV范围内可进行纺丝的电压实施,收集器36能够以接地或以(-)极带电的方式被使用。
优选地,收集器36由导电性金属或剥离纸、无纺布等形成。并且,在收集器36可附着捕集装置(suction collector)来使用,用于当纺丝时,使纤维的集束顺畅,优选地,在5~50范围内,调节纺丝喷嘴34到收集器36为止的距离来使用。
就纳米纤维的排出量而言,优选地,当纺丝时,使用定量泵来以每孔0.01~5cc/hole.min的方式进行排出并纺丝,并在纺丝时可调节温度及湿度的混合槽内,在相对湿度为10~90%的环境条件下进行纺丝。
并且,在各纺丝喷嘴34设有空气喷射装置38,向由纺丝喷嘴34纺丝的纤维束14喷射空气,并引导纤维束14使其向收集器36一侧捕集。
在收集器36的后方设有加压辊40,上述加压辊40加压通过电纺丝方法来制备的纳米纤维网10而制备成恒定厚度,并且,设有纳米纤维网辊42,上述纳米纤维网辊42用于缠绕通过加压辊40并被加压的纳米纤维网10。
以下说明利用如此构成的电纺丝装置来制备细胞因子吸附片的制备方法。
首先,准备纺丝溶液。即,使用适当的溶剂,来使可吸附细胞因子的吸附物质和可进行电纺丝的高分子物质以可纺丝的浓度溶解,从而准备纺丝溶液。
纺丝溶液的浓度为当电纺丝时可维持纤维状形态的浓度,而以纺丝溶液的总含量为基准,吸附物质和高分子物质的含量适当在5~90重量百分比的范围内。
在这里,上述比率小于5重量百分比的情况下,由于低于当电纺丝时形成纳米纤维的浓度,导致形成少量(drop),从而大部分情况下无法形成纳米纤维,比率大于90重量百分比的情况下,由于高分子物质的含量过多,导致发生难以进行电纺丝本身的情况。
因此,有必要根据使用的高分子,在可形成纳米纤维的适当的浓度范围内制备纺丝溶液。尤其,将两种以上的高分子混合而纺丝的情况下,高分子和溶剂应具有相容性,且应在不发生相分离等的条件下进行。并且,优选地,就溶剂而言,将一种至两种混合,从而考虑溶剂的挥发,并制备纺丝溶液。
并且,通过对收集器36和纺丝喷嘴34之间施加高电压静电力,来由纺丝喷嘴34将纺丝溶液制备成纳米纤维18,并向收集器36的上面纺丝。那么,纳米纤维18捕集于收集器的表面而形成纳米纤维网10。
此时,通过设置于收集器36的捕集装置,使纺丝时纳米纤维18的集束顺畅,且由设置于纺丝喷嘴34的空气喷射装置38向纳米纤维18喷射空气,使得纳米纤维18可捕集及积累于收集器36的表面,并防止该纳米纤维18飞散。
如此完成的纳米纤维网10在通过加压辊40并被加压成恒定厚度之后,缠绕于纳米纤维网辊42。
并且,为了提高细胞因子吸附片的物性或处理性,在经过如上所述的过程制备的纳米纤维网的一面贴合基片。
此时,优选地,在防止细胞因子吸附物质改性或溶解的温度范围内,将纳米纤维网和基片复合化。
尤其,当将纳米纤维网和基片复合化时,可通过热熔粘合、热板压延、层压、热熔粘合、超声波焊接等方法来维持纳米纤维的结构,并且,例如,使用胶水等的情况下,可使用在与血液的接触中防止溶出的生物相容性胶水,也可使用如上所述的一种方法。
图3为制备本发明另一实施例的细胞因子吸附片的电纺丝装置的结构图。
另一实施例的电纺丝装置包括:混合槽30(Mixing Tank),用于储存纺丝溶液;多个纺丝喷嘴34,与高电压发生器相连接,并与混合槽30相连接,用于纺丝纳米纤维14;收集器36,从纺丝喷嘴34纺丝的纳米纤维14累积于上述收集器36而制备纳米纤维网10;以及基片辊52,配置于收集器36的前方,用于向收集器36供给基片50。
并且,在收集器36的后方设有加压辊40和薄片辊54,上述加压辊40通过加压基片50和纳米纤维网10,来制备成恒定厚度,上述薄片辊54用于缠绕将基片50和纳米纤维网10复合化的细胞因子吸附片。
并且,在纳米纤维网10的两面层叠基片50的结构的情况下,在收集器36的后方还设置一个用于供给基片的基片辊。
观察本实施例的细胞因子吸附片的制备方法如下。
首先,向收集器36供给缠绕于基片辊52的基片50。
并且,通过对收集器36和纺丝喷嘴34之间施加高电压静电力,来由纺丝喷嘴34将纺丝溶液制备成纳米纤维14,并向基片50的表面纺丝。那么,纳米纤维14累积于基片50的表面,从而形成纳米纤维网10。
并且,完成的纳米纤维网10和基片50复合化的吸附片在通过加压辊40并被加压成恒定厚度之后,缠绕于薄片辊42。
图4为本发明的血液过滤器的剖视图,图5为内置于本发明的血液过滤器的细胞因子吸附片的局部立体图。
本发明的血液过滤器包括:壳体70,由圆筒形态形成,设有使血液流入的流入口72和使血液排出的排出口74;以及作为细胞因子吸附片的纳米纤维网10,以隔着恒定间隔的方式缠绕在壳体70的内部。
并且,纳米纤维网10隔着恒定间隔由卷绕成圆状的形态形成,用于确保可使血液通过的通道,在通道设有间隔片76。在这里,作为间隔片76,只要是具有可使血液充分通过的气孔的无纺布等维持通道的间隔并可使血液通过的材质,那么就可以适用任何材质。
在这里,血液过滤器除了这种结构之外,只要是血液可充分与纳米纤维网的表面相接触的结构,那么也可以适用任何结构。
以下,说明本发明的实施例。
(实施例1)
以聚甲基丙烯酸甲酯/聚乙烯亚胺=80/20重量百分比的方式将作为可进行电纺丝的高分子物质的聚甲基丙烯酸甲酯(PMMA,Poly Methyl Methacrylate)和作为细胞因子吸附物质的聚乙烯亚胺(PEI,Polyethyleneimine)混合,并将由此得到的混合物质溶解于作为溶剂的N,N-二甲基乙酰胺,使得上述混合物质达到20重量百分比,从而制备了纺丝溶液。
将上述纺丝溶液移到混合槽中,使施加电压为50kV,纺丝喷嘴和收集器的距离为30cm,每分钟排出量为0.05cc/g.hole,并在30℃温度且相对湿度为60%的纺丝气氛条件下进行电纺丝,从而制备了纳米纤维网。
在150℃温度且5Kgf/cm2条件下,对由此得到的纳米纤维网进行热压接,从而制备了细胞因子吸附片。
(实施例2)
除了以聚甲基丙烯酸甲酯/聚乙烯亚胺=50/50重量百分比的方式混合之外,通过与上述实施例1相同的方法制备了细胞因子吸附片。
图1为以实施例2的方法得到的纳米纤维网的扫描电子显微镜照片,可知纳米纤维的平均直径为500nm,且直径分布为100~700nm。
(实施例3)
使用聚甲基丙烯酸甲酯作为高分子物质,并使用将聚苯乙烯-二乙烯基苯(PS-DVB,Polystyrene-divinylbenzene)和聚乙烯亚胺(聚甲基丙烯酸甲酯/聚乙烯亚胺/聚苯乙烯-二乙烯基苯=45/45/10重量百分比)混合而成的混合物作为细胞因子吸附物质,除此之外,通过与上述实施例1相同的方法制备了细胞因子吸附片。
图6为通过实施例3的方法得到的纳米纤维网的扫描电子显微镜照片,由图6可知,纳米纤维的平均直径为约500nm,且直径分布为100~700nm。
(实施例4)
使用聚丙烯腈作为高分子物质,使用聚乙烯亚胺作为细胞因子吸附物质,并以聚丙烯腈/聚乙烯亚胺=40/60重量百分比的方式将聚丙烯腈和聚乙烯亚胺混合,除此之外,通过与上述实施例1相同的方法制备了细胞因子吸附片。
图7为通过实施例4的方法得到的纳米纤维网的扫描电子显微镜照片。如图7所示,可以确认纳米纤维网中纳米纤维均匀形成,且平均直径为约400nm左右。
细胞因子吸附实验
为了确认本发明的细胞因子吸附片的治疗效果,向有效期结束的人类的血浆(新鲜冰冻血浆(fresh frozen human plama))给药重组人细胞因子(Recombinant human cytokine)中的TNF-α、IL-1β、IL-6、IL-8,从而制备试样,并利用生物材料(Biomaterials)((2006)27:5755)的吸附实验方法来实施体外吸附实验如下。
将脂多糖(LPS,Lipopolysaccharide)向动物(狗)(30Kg)注入10分钟,从而诱导败血症,在经过2小时之后,采集血液,并利用离心分离器来仅分离血清之后进行了冷冻保管。
向有效期结束的人类的血浆(新鲜冰冻血浆)投入如上所述的败血症诱导血浆(cytokine spiked plasma),向制成的溶液分别投入利用磷酸盐缓冲液(PBS,Phosphate buffer solution)来进行预处理(预湿(Pre wetting))的吸附材料,并在振荡水浴(Shaking water bath)中在37℃温度且60rpm条件下进行了搅拌。将最初溶液作为比较组(对照组(Control)),经过1小时及2小时之后,采集了各1cc的样品。为了比较吸附材料的吸附量,利用酶联免疫吸附试验(ELISA,enzyme-linked immunosorbent assy)来对所采集的样品进行了定量定性分析,从而得到了结果。
实验结果1
在使吸附片的重量成为0.3g之后,向败血症诱导血浆(cytokinespiked plasma)2cc溶液投入,并实施了吸附,从而比较了随着时间的吸附量,并将由此得到的结果分别示于表1及表2中。
表1
[Table 1]
随着吸附时间的对于TNF-α的吸附量比较
表2
[Table 2]
随着吸附时间的对于IL-6的吸附量比较
如表1所示,可知随着作为吸附物质的聚乙烯亚胺的含量增加,细胞因子的去除率得到提高,但在TNF-α的吸附中,不能说在1小时内,与对照组相比,吸附发生较大的变化。
但是,就对于IL-6的吸附量而言,使用本发明的吸附片的情况下,可以确认呈现与对照组相比,显著得到提高的吸附去除率。并且,实施例4的情况下,可知利用高分子物质与聚丙烯腈混合的情况下,吸附率的提高程度甚微。
实验结果2
在使从实施例2得到的吸附片的重量分别成为0.3g及0.6g之后,向包含败血症诱导血浆(cytokine spiked plasma)的2cc的溶液投入,从而比较了吸附片的吸附量,并将由此得到的结果示于表3和表4中。
表3
[Table 3]
基于吸附片的重量的对于TNF-α的吸附量比较
表4
[Table 4]
基于吸附片的重量的对于IL-6的吸附量比较
如表3及表4所示,在本发明中,随着吸附片中的吸附物质的含量增加,吸附去除率也增加。以1小时的吸附时间为基准,将实施例2的样品(0.3g)的吸附量作为100%的情况下,可知样品(0.6g)的对于TNF-α及IL-6的吸附去除率分别为156.35%、120.58%。
实验结果3
在使吸附片的重量成为0.2g之后,向败血症诱导血浆(cytokinespiked plasma)的10cc的溶液投入,并实施吸附,从而比较了随着时间的吸附量,并将由此得到的结果分别示于表5至表8中。
表5
[Table 5]
随着吸附时间的对于TNF-α的吸附量比较
表6
[Table 6]
随着吸附时间的对于IL-6的吸附量比较
表7
[Table 7]
随着吸附时间的对于IL-1β的吸附量比较
表8
[Table 8]
随着吸附时间的对于IL-8的吸附量比较
通常,细胞因子也具有在自然状态下稍微增加或减少的倾向,且在表5和表6的比较组中出现了这种现象,但可知与本发明的实施例相比,细胞因子的吸附去除率相对非常低。尤其,从表6至表8的结果可以确认,与在经过2小时之后,细胞因子的吸附去除率不大于8%的对照组不同,在本发明的实施例中,呈现了以大于60%的水平优秀的吸附去除率。
以上,例举并图示特定的优选实施例来说明了本发明,但本发明并不局限于如上所述的实施例,只要在不脱离本发明的精神的范围内,那么就能够由本发明所属技术领域的普通技术人员进行各种变更和修改。
产业上的可利用性
本发明能够适用于包括细胞因子吸附片的血液过滤器,因而能够有用地适用于败血症的治疗。

Claims (18)

1.一种细胞因子吸附片,其特征在于,包含将纺丝溶液进行电纺丝而成的纳米纤维网,上述纺丝溶液由能够吸附细胞因子的吸附物质和能够进行电纺丝的高分子物质混合而成。
2.根据权利要求1所述的细胞因子吸附片,其特征在于,上述纳米纤维网通过将纺丝溶液进行电纺丝来制备纳米纤维,且累积上述纳米纤维来形成为具有多个气孔的形态。
3.根据权利要求1所述的细胞因子吸附片,其特征在于,形成上述纳米纤维网的纳米纤维的直径在100nm~800nm范围内,气孔的平均大小在0.1μm~10μm范围内。
4.根据权利要求1所述的细胞因子吸附片,其特征在于,上述纳米纤维网的厚度在1μm~150μm范围内。
5.根据权利要求4所述的细胞因子吸附片,其特征在于,上述纳米纤维网的厚度在15μm~20μm范围内。
6.根据权利要求1所述的细胞因子吸附片,其特征在于,上述吸附物质使用选自多粘菌素-B、聚乙烯亚胺、聚乙烯吡咯烷酮、聚苯乙烯-二乙烯基苯中的一种或两种以上的混合物。
7.根据权利要求1所述的细胞因子吸附片,其特征在于,上述高分子物质使用将聚偏氟乙烯、聚甲基丙烯酸甲酯、聚丙烯腈、聚氨酯、聚醚砜、聚酰胺酸、聚乙烯醇、聚环氧乙烷、聚乳酸、聚乙醇酸、聚乳酸-聚乙醇酸类的高分子物质中的一种或两种以上混合的高分子物质。
8.根据权利要求1所述的细胞因子吸附片,其特征在于,还包括层叠于上述纳米纤维网的一面的基片。
9.根据权利要求8所述的细胞因子吸附片,其特征在于,在上述基片形成有能够使血液通过的多个气孔,上述基片为选自无纺布、织造的织物、高分子泡沫、金属泡沫、纸、金属网及塑料网中的一种。
10.一种血液过滤器,其特征在于,内置有权利要求1至9中任一项所述的细胞因子吸附片。
11.根据权利要求10所述的血液过滤器,其特征在于,上述细胞因子吸附片隔着恒定间隔由辊形态形成,用于形成使血液通过的通道,在上述通道设有间隔片。
12.一种细胞因子吸附片的制备方法,其特征在于,包括:
将能够吸附细胞因子的吸附物质和能够进行电纺丝的高分子物质混合溶解于溶剂,来制备纺丝溶液的步骤;以及
将上述纺丝溶液进行电纺丝,来形成具有多个气孔的纳米纤维网的步骤。
13.根据权利要求12所述的细胞因子吸附片的制备方法,其特征在于,以上述纺丝溶液的总含量为基准,上述吸附物质和高分子物质的含量在5~90重量百分比的范围内。
14.根据权利要求12所述的细胞因子吸附片的制备方法,其特征在于,还包括使上述纳米纤维网通过加压辊来制备成恒定厚度的步骤。
15.根据权利要求13所述的细胞因子吸附片的制备方法,其特征在于,还包括在上述纳米纤维网的一面层叠基片的步骤。
16.根据权利要求15所述的细胞因子吸附片的制备方法,其特征在于,上述基片通过热熔敷、压延、层压、热熔粘合及焊接中的一种方法来粘结于纳米纤维网的一面。
17.根据权利要求15所述的细胞因子吸附片的制备方法,其特征在于,还包括对上述基片进行灭菌处理的步骤。
18.一种细胞因子吸附片的制备方法,其特征在于,包括:
将能够吸附细胞因子的吸附物质和能够进行电纺丝的高分子物质溶解于溶剂,来制备纺丝溶液的步骤;
准备基片的步骤;以及
通过在上述基片将上述纺丝溶液进行电纺丝,来将具有多个气孔的纳米纤维网层叠于基片的步骤。
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