CN107757019A - 定向导流非织造材料及其制作方法、使用装置、使用方法 - Google Patents
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Abstract
本发明提供了一种定向导流非织造材料,包括自上而下依次设置的疏水层、混合层和亲水层;所述的疏水层由疏水性熔喷纤维构成,所述的亲水层由壳聚糖亲水膜构成,所述的混合层由壳聚糖亲水膜和疏水性熔喷纤维共同构成。本发明提供了定向导流非织造材料的制作方法、使用装置及使用方法。本发明通过熔喷和涂层技术制备定向导流非织造材料,除了能使细胞依靠自身重力与液体快速分离,避免施加的外力对细胞造成的伤害,对细胞无污染,同时,细胞还能够轻易的从材料表面脱落下来,以便细胞后续的使用。
Description
技术领域
本发明属于生物用非织造材料技术领域,特别涉及一种用于快速收集和释放细胞的定向导流非织造材料。
背景技术
细胞体外培养技术在细胞治疗、支架移植、组织工程和再生药物等方面有广泛的应用。为了能够更有效的利用细胞,对细胞进行分析和使用,经常需要收集和释放细胞。例如,从小鼠体内提取组织细胞,进行体外细胞培养,或者将培养好的细胞种植到支架材料上。
目前常规的方法是利用离心力从细胞悬浮液中收集细胞,离心后细胞沉淀,倾倒上清液,然后对细胞重悬。但在离心处理期间,高速离心力会造成细胞活性的降低,甚至导致细胞的破裂死亡。同时,离心处理的过程会增加细胞受外界感染的风险。当对细胞进行重悬时,细胞容易粘附在一起,不易释放,其吹打力也会造成细胞损伤。
当前已有一些收集细胞的相关专利,如专利公开号为CN105358957A的中国发明专利公开了一种从生物流体的样本收集细胞的装置,包括两个腔室,多孔膜,吸收体及弹性部件,使细胞沉积在多孔膜的上游表面,流体则被抽吸通过多孔膜且由吸收体吸收。因此,细胞不易从多孔膜上释放,同时,抽吸作用也会对细胞造成一定的破坏。再如专利公开号为CN104923091A的中国发明专利公开了一种白细胞过滤膜,其特征在于,在聚对苯二甲酸丁二醇酯非织造织物基材上进行接枝改性。该发明虽然能对白细胞进行过滤与分离,但是改性后的材料亲水性使得细胞容易粘附在材料表面上不易释放,因此不适用于细胞的收集与释放。
因此,为了简化收集和释放细胞的过程,且把对细胞造成的损伤最小化,需要一种无需施加任何外力,能从细胞悬浮液中轻易收集细胞并将其释放的材料。
发明内容
本发明要解决的技术问题是提供一种无需施加任何外力,就能从细胞悬浮液中轻易收集细胞并将其释放的材料,以简化收集和释放细胞的过程,且把对细胞造成的损伤最小化。
为了解决上述技术问题,本发明的技术方案是提供一种定向导流非织造材料,其特征在于:包括自上而下依次设置的疏水层、混合层和亲水层;所述的疏水层由疏水性熔喷纤维构成,所述的亲水层由壳聚糖亲水膜构成,所述的混合层由壳聚糖亲水膜和疏水性熔喷纤维共同构成。
优选地,所述定向导流非织造材料在厚度方向上形成梯度润湿性,亲水层的润湿性最佳,疏水层的润湿性最小,从亲水层到疏水层的润湿性逐渐降低。
优选地,所述疏水性熔喷纤维采用熔喷非织造技术制备,所述壳聚糖亲水膜采用涂层技术制备。
本发明还提供了上述定向导流非织造材料的制备方法,其特征在于,步骤为:
步骤1:采用聚丙烯或聚酯切片为原料,通过熔喷非织造技术制成纤维直径为1~3μm、面密度为20~25g/m2的熔喷非织造材料;
步骤2:将熔喷非织造材料进行单面等离子体处理;
步骤3:将等离子体处理后的熔喷非织造材料处理面用壳聚糖醋酸溶液进行涂层;
步骤4:将涂层后的熔喷非织造材料用交联剂蒸汽交联;
步骤5:将蒸汽交联后的熔喷非织造材料进行洗涤,再放入真空烘箱干燥。
优选地,所述步骤1中,熔喷参数如下:喷丝孔吐出量为0.12~0.05g/(孔·min),热空气速度为400~600m/s,滚筒接收距离为100mm。
优选地,所述步骤2中,等离子体处理参数如下:放电功率为10W,放电时间为10s,真空度为10%。
优选地,所述步骤2中,熔喷非织造材料的上表面未处理、下表面进行离子体处理,下表面的润湿性由疏水变成亲水。
优选地,所述步骤3中,涂层参数如下:刮刀速度为15~20mm/min,刮刀高度为0.1mm,往复次数为5~10次。
本发明还提供了上述的定向导流非织造材料的使用装置,其特征在于:包括
用于接收和引导收集释放前的细胞悬浮液的上管状套筒接头;
用于引导和收集分离后的不含细胞的废液的下管状套筒接头;
上管状套筒接头与下管状套筒接头相对设置,环形橡胶垫圈设于下管状套筒接头上,用于快速收集和释放细胞的定向导流非织造材料设于上管状套筒接头与环形橡胶垫圈之间,上管状套筒接头、所述定向导流非织造材料、环形橡胶垫圈、下管状套筒接头由上至下依次设置好后通过卫生级卡箍箍紧。
本发明还提供了上述的定向导流非织造材料的使用方法,其特征在于,步骤为:
步骤一、材料的安装
将用于快速收集和释放细胞的定向导流非织造材料裁剪成相应大小的形状,使其疏水层朝上,亲水层朝下,放置在环形橡胶垫圈的上方;上管状套筒接头置于定向导流非织造材料的上方,下管状套筒接头置于环形橡胶垫圈的下方;用卫生级卡箍箍紧上管状套筒接头和下管状套筒接头,使定向导流非织造材料牢牢地固定在装置中;
步骤二、细胞的收集
将安装好材料的装置灭菌处理后放入无菌的操作台内,将准备好的细胞悬浮液直接从上管状套筒接头倒入;等待后,溶液与细胞分离完毕,细胞被定向导流非织造材料截留在其上表面,液体则通过定向导流非织造材料,顺着下管状套筒接头流出;
步骤三、细胞的释放
将卫生级卡箍取下,拿走上管状套筒接头,用镊子夹取定向导流非织造材料的边缘部分,将其放入新的溶液中,用镊子摇晃定向导流非织造材料,然后取出定向导流非织造材料,得到释放过后的新的细胞悬浮液;
步骤四、重复使用
将步骤三用过的定向导流非织造材料和所述使用装置一同放入蒸汽灭菌锅内进行灭菌处理;然后重复步骤一到步骤三,进行快速收集和释放细胞的重复操作。
相比现有技术,本发明具有如下有益效果:
1)定向导流非织造材料能仅依靠细胞自身重力与液体进行快速收集,收集时间短,也避免了细胞因受到外界的作用力而损伤甚至死亡,增加细胞的活率;
2)定向导流非织造材料的上表面疏水,与细胞接触时,细胞不易粘附在其表面上,有利于细胞快速轻易的释放,释放时不直接接触细胞,不会对细胞造成破坏;
3)收集细胞和释放细胞的操作过程简单方便,且在无菌条件下操作,避免了离心等收集方式使得外界环境对细胞造成的污染风险;
4)定向导流非织造材料经过灭菌处理后也可以重复使用,节约成本。
附图说明
图1为定向导流非织造材料的结构示意图;
图2为用定向导流非织造材料快速收集和释放细胞的装置示意图;
图3为定向导流非织造材料使用过程示意图。
具体实施方式
下面结合具体实施例,进一步阐述本发明。
实施例1
定向导流非织造材料的结构如图1所示,包括自上而下依次设置的疏水层1、混合层2和亲水层3,所述疏水层1是由熔喷纤维4构成,所述混合层2是由熔喷纤维4和壳聚糖亲水膜5共同构成,所述亲水层3是由壳聚糖亲水膜5构成。
其中,疏水层1和混合层2的纤维部分是采用聚对苯二甲酸丁二醇酯PBT聚合物切片,通过熔喷非织造技术制备而成。喷丝孔吐出量为0.012g/(孔·min),喷丝孔个数为168个,PBT熔融指数为15.4g/10min,热空气速度为600m/s,滚筒接收距离为100mm,制成的熔喷非织造材料平均纤维直径为1.5μm,面密度为20g/m2。较小的纤维直径得到的熔喷非织造材料的平均孔径较小,使得细胞能够截留在熔喷非织造材料的表面,确保细胞的收集率。
亲水层3和混合层2的膜部分是通过将壳聚糖粉末溶解在醋酸溶液中,在一定温度下充分搅拌溶解,得到壳聚糖醋酸溶液;然后利用涂层技术将该溶液均匀地涂在熔喷非织造材料的下表面。醋酸浓度为2wt%,壳聚糖粉末添加量为0.04g/ml,溶解温度为50℃,刮刀速度为20mm/min,刮刀高度为0.1mm,往复次数为5次。
通过熔喷和涂层技术来制备定向导流非织造材料,材料朝上部分未处理面疏水,材料朝下部分涂层面亲水,混合层的润湿性介于两者之间。在厚度方向上呈现从上至下润湿性逐渐增加的趋势。
实施例2
一种定向导流非织造材料的制备方法,包括如下几个步骤:
步骤一、熔喷非织造材料的制备
由于在加工工程中,聚酯切片易与水发生水解反应,导致分子量下降,影响产品质量,因此,将PBT聚合物切片放在105℃真空烘箱中干燥5h,保证其含水率在100ppm以下。然后将干燥后的PBT聚合物切片在螺杆挤出机中加热熔融后经喷丝口挤出。螺杆挤出机各段的温度为:一区:230℃,二区:260℃,三区:277℃,四区:280℃,五区:280℃,六区:280℃。加热的拉伸空气从模头喷丝孔两侧风道高速吹出,对PBT聚合物熔体细流进行拉伸,冷空气在模头下方一定位置从两侧补入,使纤维冷却结晶。热空气压力为27.58kPa,滚筒接收距离为100mm。得到的熔喷非织造材料面密度为20g/m2,纤维平均直径为1.5μm。
步骤二、等离子体处理
将步骤一制备的熔喷非织造材料置于等离子体真空反应室内,打开气阀,通入氧气。设置真空度为10%,放电功率为10W,放电时间为10s。熔喷非织造材料处理面暴露在气体中,其润湿性从疏水变为亲水,而未处理面表面依旧疏水。
步骤三、涂层
由于等离子体处理具有时效性,因此将熔喷非织造材料的处理面立即用壳聚糖醋酸溶液进行涂层。先将熔喷非织造材料固定在涂层机上,处理面朝上,然后设定好涂层参数:刮刀速度为20mm/min,刮刀高度为0.1mm,往复涂层5次。然后将材料放置4h,去除多余的液体。
步骤四、交联
为使涂层后的壳聚糖稳定存在于熔喷非织造材料中,将步骤三得到的材料四周固定,放置在装有10ml戊二醛交联剂的干燥器内,静置24h。然后取出材料,用清水冲洗掉未交联好的壳聚糖,放入40℃烘箱中干燥,得到最终的定向导流非织造材料。
实施例3
实施例1所述的用于快速收集和释放细胞的定向导流非织造材料的使用装置如图2所示。该装置是由上管状套筒接头6、环形橡胶垫圈8、下管状套筒接头7和一个卫生级卡箍构成。上管状套筒接头6用于接收和引导收集释放前的细胞悬浮液,下管状套筒接头7用于引导和收集分离后的不含细胞的废液,环形橡胶垫圈8和卫生级卡箍是为了确保定向导流非织造材料在使用的过程中固定牢靠,液体不会从边缘漏出,保证细胞收集率。
该装置的使用过程如图3所示,主要包括如下几个步骤:
步骤一、材料的安装
将定向导流非织造材料9裁剪成相应大小的圆形,使其疏水层朝上,亲水层朝下,放置在环形橡胶垫圈8的上方。上管状套筒接头6置于定向导流非织造材料9的上方,下管状套筒接头7置于环形橡胶垫圈8的下方。用卫生级卡箍箍紧上管状套筒接头6和下管状套筒接头7,使定向导流非织造材料9牢牢地固定在装置中。
步骤二、细胞的收集
将安装好材料的装置灭菌处理后放入无菌的操作台内,将准备好的细胞悬浮液直接从上管状套筒接头6倒入。等待18s后,溶液与细胞分离完毕,细胞被定向导流非织造材料9截留在其上表面,液体则通过定向导流非织造材料9,顺着下管状套筒接头7流出。
步骤三、细胞的释放
将卫生级卡箍取下,拿走上管状套筒接头6,用镊子夹取定向导流非织造材料9的边缘部分,将其放入新的溶液中,用镊子轻轻摇晃定向导流非织造材料9。然后取出定向导流非织造材料9,得到释放过后的新的细胞悬浮液。
步骤四、重复使用
将步骤三用过的定向导流非织造材料9和使用装置一同放入高温蒸汽灭菌锅内进行灭菌处理。灭菌温度为120℃,灭菌时间为30min。然后重复步骤一到步骤三,进行快速收集和释放细胞的重复操作。
以上所述,仅为本发明的较佳实施例,并非对本发明任何形式上和实质上的限制,应当指出,对于本技术领域的普通技术人员,在不脱离本发明方法的前提下,还将可以做出若干改进和补充,这些改进和补充也应视为本发明的保护范围。凡熟悉本专业的技术人员,在不脱离本发明的精神和范围的情况下,当可利用以上所揭示的技术内容而做出的些许更动、修饰与演变的等同变化,均为本发明的等效实施例;同时,凡依据本发明的实质技术对上述实施例所作的任何等同变化的更动、修饰与演变,均仍属于本发明的技术方案的范围内。
Claims (10)
1.一种定向导流非织造材料,其特征在于:包括自上而下依次设置的疏水层(1)、混合层(2)和亲水层(3);所述的疏水层(1)由疏水性熔喷纤维(4)构成,所述的亲水层(3)由壳聚糖亲水膜(5)构成,所述的混合层(2)由壳聚糖亲水膜(5)和疏水性熔喷纤维(4)共同构成。
2.如权利要求1所述的一种定向导流非织造材料,其特征在于:所述定向导流非织造材料在厚度方向上形成梯度润湿性,亲水层(3)的润湿性最佳,疏水层(1)的润湿性最小,从亲水层(3)到疏水层(1)的润湿性逐渐降低。
3.如权利要求1所述的一种定向导流非织造材料,其特征在于:所述疏水性熔喷纤维(4)采用熔喷非织造技术制备,所述壳聚糖亲水膜(5)采用涂层技术制备。
4.一种如权利要求1~3任一项所述的定向导流非织造材料的制备方法,其特征在于,步骤为:
步骤1:采用聚丙烯或聚酯切片为原料,通过熔喷非织造技术制成纤维直径为1~3μm、面密度为20~25g/m2的熔喷非织造材料;
步骤2:将熔喷非织造材料进行单面等离子体处理;
步骤3:将等离子体处理后的熔喷非织造材料处理面用壳聚糖醋酸溶液进行涂层;
步骤4:将涂层后的熔喷非织造材料用交联剂蒸汽交联;
步骤5:将蒸汽交联后的熔喷非织造材料进行洗涤,再放入真空烘箱干燥。
5.如权利要求4所述的一种定向导流非织造材料的制备方法,其特征在于:所述步骤1中,熔喷参数如下:喷丝孔吐出量为0.12~0.05g/(孔·min),热空气速度为400~600m/s,滚筒接收距离为100mm。
6.如权利要求4所述的一种定向导流非织造材料的制备方法,其特征在于:所述步骤2中,等离子体处理参数如下:放电功率为10W,放电时间为10s,真空度为10%。
7.如权利要求4或6所述的一种定向导流非织造材料的制备方法,其特征在于:所述步骤2中,熔喷非织造材料的上表面未处理、下表面进行离子体处理,下表面的润湿性由疏水变成亲水。
8.如权利要求4所述的一种定向导流非织造材料的制备方法,其特征在于:所述步骤3中,涂层参数如下:刮刀速度为15~20mm/min,刮刀高度为0.1mm,往复次数为5~10次。
9.一种如权利要求1~3任一项所述的定向导流非织造材料的使用装置,其特征在于:包括
用于接收和引导收集释放前的细胞悬浮液的上管状套筒接头(6);
用于引导和收集分离后的不含细胞的废液的下管状套筒接头(7);
上管状套筒接头(6)与下管状套筒接头(7)相对设置,环形橡胶垫圈(8)设于下管状套筒接头(7)上,所述定向导流非织造材料(9)设于上管状套筒接头(6)与环形橡胶垫圈(8)之间,上管状套筒接头(6)、定向导流非织造材料(9)、环形橡胶垫圈(8)、下管状套筒接头(7)由上至下依次设置好后通过卫生级卡箍箍紧。
10.一种如权利要求1~3任一项所述的定向导流非织造材料的使用方法,其特征在于,如权利要求9所述的定向导流非织造材料的使用装置,步骤为:
步骤一、材料的安装
将所述定向导流非织造材料(9)裁剪成相应大小的形状,使其疏水层朝上,亲水层朝下,放置在环形橡胶垫圈(8)的上方;上管状套筒接头(6)置于定向导流非织造材料(9)的上方,下管状套筒接头(7)置于环形橡胶垫圈(8)的下方;用卫生级卡箍箍紧上管状套筒接头(6)和下管状套筒接头(7),使定向导流非织造材料(9)牢牢地固定在装置中;
步骤二、细胞的收集
将安装好材料的装置灭菌处理后放入无菌的操作台内,将准备好的细胞悬浮液直接从上管状套筒接头(6)倒入;等待后,溶液与细胞分离完毕,细胞被定向导流非织造材料(9)截留在其上表面,液体则通过定向导流非织造材料(9),顺着下管状套筒接头(7)流出;
步骤三、细胞的释放
将卫生级卡箍取下,拿走上管状套筒接头(6),用镊子夹取定向导流非织造材料(9)的边缘部分,将其放入新的溶液中,用镊子摇晃定向导流非织造材料(9),然后取出定向导流非织造材料(9),得到释放过后的新的细胞悬浮液;
步骤四、重复使用
将步骤三用过的定向导流非织造材料(9)和所述使用装置一同放入蒸汽灭菌锅内进行灭菌处理;然后重复步骤一到步骤三,进行快速收集和释放细胞的重复操作。
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