CN106120327A - 一种具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法 - Google Patents
一种具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法 Download PDFInfo
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Abstract
本发明属于医用材料技术领域,具体涉及一种具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法。本发明以醋酸纤维素、醋酸丁酸纤维素、乙基纤维素等纤维素衍生物为原料,通过静电纺丝工艺,制得纤维素衍生物的纳米纤维薄膜。再经过水解、氧化等过程,得到氧化纤维素纳米纤维薄膜。本发明具有工艺简单,反应温和,安全环保的特点。通过本发明方法制得的氧化纤维素纳米纤维膜片,具有良好的生物相容性和可降解性,并能有效促进伤口中血小板的聚集,实现快速止血。
Description
技术领域
本发明属于医用材料技术领域,具体涉及一种具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法。
背景技术
氧化纤维素也称C-6羧基纤维素,是纤维素C-6位伯羟基被选择性氧化的衍生物。氧化纤维素具有优良的生物相容性、生物安全性和生物可降解性,在医疗行业中可作为止血纱布、手术缝合线等使用,目前临床上最广泛应用的此类产品是由美国强生公司生产的速即纱系列可吸收止血产品。
纤维素的氧化体系主要有两种:NO2氧化体系和TEMPO氧化体系。NO2氧化体系最早可追溯至1942年,该体系对纤维素伯羟基的选择性氧化程度较高。经过几十年的优化和改进,NO2氧化体系现在已具有较高的可控性、均一性和选择性。但作为一种传统的氧化体系,该方法存在废气回收困难、NO2价格昂贵的缺点。TEMPO(2,2,6,6-四甲基哌啶-1-氮氧化物)氧化体系是一种新型的纤维素氧化体系,该体系对伯羟基的选择性高,反应过程简单,反应条件温和,近年来用含TEMPO的共氧化体系对多糖类高分子进行选择性氧化已成为研究热点。
目前的氧化纤维素类止血产品大多为传统工艺纺制而成的纱布,但随着近年来静电纺丝技术的发展,纳米纤维表现出了传统纤维不具有的优异性能,如高比表面积、高孔隙率、小孔径等。然而由于纤维素分子内和分子间强烈的氢键作用导致纤维素难以被常规溶剂溶解,因此很难用常规方法直接用纤维素进行静电纺丝。本发明以纤维素衍生物为原料进行静电纺丝,具有原料易得、溶解方便、纺丝过程简便的特点。
在目前已有的相关文献专利中,通常是将纤维素用TEMPO氧化体系氧化之后用胶原蛋白(CN103333356A)、海藻酸盐(CN104013991A)等进行改性,以提高材料的生物相容性或加快止血速率,但其所用的原料一般为传统的再生纤维素纱布,其网孔较大,对血小板的聚集效果有限。专利CN103520763A将材料加工成了纳米纤维毡,但其氧化方法仍为传统的NO2氧化体系,具有原料昂贵、废气回收困难等不足。本发明结合了静电纺纳米纤维技术以及新型的TEMPO共氧化体系,目前尚未有以该技术路线制备氧化纤维素纳米纤维止血材料的相关研究报道。
发明内容
本发明的目的在于解决现有的氧化纤维素止血材料存在的止血速率慢的问题,而提供一种具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法。
本发明提出的一种具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法,具体步骤如下:
(1)将纤维素衍生物溶于易挥发极性溶剂中,配制成纺丝溶液,通过静电纺丝方法,制得纤维素衍生物的纳米纤维薄膜;
(2)将所得纤维素衍生物的纳米纤维薄膜浸入到碱性溶液中进行水解,得到纤维素纳米纤维薄膜;
(3)将所得纤维素纳米纤维薄膜用TEMPO氧化体系进行氧化反应,再经酸处理、清洗、干燥后,即得到氧化纤维素纳米纤维膜片;其中:所述氧化体系为TEMPO/溴化钠(NaBr)/次氯酸钠(NaClO)共氧化体系或TEMPO/次氯酸钠(NaClO)/亚氯酸钠(NaClO2)共氧化体系;氧化反应中控制纤维素纳米纤维薄膜与TEMPO/溴化钠(NaBr)/次氯酸钠(NaClO)共氧化体系组成的溶液中pH值为10-11,反应温度为0-25℃,反应时间为0.5-2小时;氧化反应中控制纤维素纳米纤维薄膜与TEMPO/次氯酸钠(NaClO)/亚氯酸钠(NaClO2)共氧化体系组成的溶液中pH值为6-7,反应温度为25-80℃,反应时间为0.5-2小时。
本发明中,所述纤维素衍生物为醋酸纤维素(CA)或醋酸丁酸纤维素(CAB)等纤维素酯,或者甲基纤维素(MC)、羧甲基纤维素(CMC)或乙基纤维素(EC)等纤维素醚中的一种或几种。
本发明中,步骤(1)中所述易挥发极性溶剂为N,N-二甲基甲酰胺(DMF),N,N-二甲基乙酰胺(DMAc),二甲基亚砜(DMSO),丙酮,二氯甲烷(DCM),三氯甲烷(TCM),甲醇,乙醇或水中的一种或几种的混合溶剂。
本发明中,步骤(1)中所述纺丝溶液的质量分数为8%-20%。
本发明中,步骤(1)中所述静电纺丝方法中施加的直流高压为5-60kV,基材与电极间的接收距离为50-300mm。
本发明中,步骤(2)中所述碱性溶液的溶质为氢氧化钠或氢氧化钾,溶剂为甲醇、乙醇或水中的一种或几种的混合溶剂。
本发明中,步骤(2)中所述水解反应温度为15-40℃,反应时间为8-48小时。
本发明中,步骤(3)中所述酸处理过程是将氧化后的纤维素纳米纤维膜片在室温条件下浸入pH值为2-3的盐酸溶液中,处理时间为15-30分钟。
本发明中,TEMPO/溴化钠(NaBr)/次氯酸钠(NaClO)共氧化体系中,控制纤维素纳米纤维薄膜与TEMPO的质量比为1:0.01-1:0.05,纤维素纳米纤维薄膜与NaBr的质量比为1:0.1-1:0.5,纤维素纳米纤维薄膜与NaClO的质量比为1:0.5-1:1.5。
本发明中,TEMPO/次氯酸钠(NaClO)/亚氯酸钠(NaClO2)共氧化体系中,控制纤维素纳米纤维膜与TEMPO的质量比为1:0.01-1:0.05,纤维素纳米纤维薄膜与NaClO的质量比为1:0.1-1:0.5,纤维素纳米纤维薄膜与NaClO2的质量比为1:1-1:5。
本发明的有益效果在于:该氧化纤维素纳米纤维膜片以纤维素衍生物为原料,与纤维素相比具有溶解简便、加工方便的特点;膜片由纳米纤维构成,其比表面积远大于传统纱布,能显著促进血小板的聚集,控制大面积出血;氧化过程利用了TEMPO共氧化体系,具有选择性高、反应温和、安全环保的优点。
附图说明
图1.实施例1中醋酸纤维素纳米纤维的扫描电镜图片。其中:(a)为放大倍数5000倍的电镜图片,(b)为放大倍数40000倍的电镜图片。
图2.实施例1中水解后的纤维素纳米纤维的扫描电镜图片。其中:(a)为放大倍数5000倍的电镜图片,(b)为放大倍数40000倍的电镜图片。
图3.实施例1中氧化纤维素纳米纤维的扫描电镜图片。其中:(a)为放大倍数5000倍的电镜图片,(b)为放大倍数40000倍的电镜图片。
图4.实施例4中几种材料的凝血实验对比。其中:(a)为醋酸纤维素纳米纤维,(b)为纤维素纳米纤维,(c)为氧化纤维素纳米纤维,(d)为明胶止血海绵,(e)为普通医用纱布。
具体实施方式
下面通过实施例进一步说明本发明,所举的实施例仅是对本发明产品或方法作概括性例示,有助于更好地理解本发明,但并不会限制本发明范围
实施例1
采用醋酸纤维素为原料,溶解于DCM/DMF混合溶剂中制成质量分数为11%的溶液,其中DCM与DMF质量比为2:1。将此溶液加入到静电纺丝设备的液槽中,设定直流高压发生器的电压为45kV,电极与基材间接收距离为200mm,可以获得由醋酸纤维素纳米纤维堆积而成的薄膜(结构如图1所示),将此薄膜用去离子水洗涤多次后真空干燥待用。将醋酸纤维素纳米纤维薄膜浸入浓度为0.1mol/L的NaOH/乙醇-水溶液中进行水解,其中乙醇与水的质量比为1:4,反应时间为24小时,反应结束后用去离子水洗涤多次后真空烘干,得到纤维素纳米纤维薄膜(结构如图2所示)。称取0.1gNaBr、0.02gTEMPO,溶于100mL去离子水中,将此溶液置于冰水浴中,保持溶液温度在5℃以下。称取1g纤维素纳米纤维薄膜加入溶液中,在不断搅拌下一次性加入10mLNaClO溶液(有效含量约7.5%),用0.1mol/L NaOH溶液调控溶液pH值在10左右,反应约30分钟后,将膜片取出后放入pH值为2的盐酸中浸泡30分钟,然后用去离子水洗涤多次后真空烘干,得到氧化纤维素纳米纤维膜(结构如图3所示)。
由图1-图3中各材料的扫描电镜图片可以看出,醋酸纤维素纳米纤维表面光滑,纤维直径约800nm;经水解反应后得到的纤维素纳米纤维表面变得粗糙,纤维直径略有减小,约500nm,且纤维之间发生了不同程度的卷曲和粘连;氧化后的纤维素纳米纤维与氧化前在形貌和直径上都没有太大变化。
实施例2
采用醋酸纤维素为原料,溶解于丙酮/DMAc混合溶剂中制成质量分数为12%的溶液,其中丙酮与DMAc质量比为2:1。将此溶液加入到静电纺丝设备的液槽中,设定直流高压发生器的电压为45kV,电极与基材间接收距离为200mm,可以获得由醋酸纤维素纳米纤维堆积而成的薄膜,将此薄膜用去离子水洗涤多次后真空干燥待用。将醋酸纤维素纳米纤维薄膜浸入浓度为0.1mol/L的NaOH/乙醇-水溶液中进行水解,其中乙醇与水的质量比为1:4,反应时间为24小时,反应结束后用去离子水洗涤多次后真空烘干,得到纤维素纳米纤维薄膜。称取0.03g TEMPO、2g NaClO2溶于pH为6.8的磷酸盐缓冲液中,将溶液加热至80℃,然后加入1g纤维素纳米纤维膜和10mL NaClO溶液(有效含量约7.5%),反应30分钟后取出,放入pH为2的盐酸中浸泡30分钟,然后用去离子水洗涤多次后真空烘干,得到氧化纤维素纳米纤维膜。
实施例3
采用乙基纤维素为原料,溶解于乙醇中制成质量分数为15%的溶液。将此溶液加入到静电纺丝设备的液槽中,设定直流高压发生器的电压为45kV,电极与基材间接收距离为200mm,可以获得由乙基纤维素纳米纤维堆积而成的薄膜,将此薄膜用去离子水洗涤多次后真空干燥待用。将乙基纤维素纳米纤维薄膜浸入浓度为0.1mol/L的KOH/乙醇-水溶液中进行水解,其中乙醇与水的质量比为1:4,在50℃反应24小时,反应结束后用去离子水洗涤多次后真空烘干,得到纤维素纳米纤维薄膜。称取0.1g NaBr、0.02g TEMPO,溶于100mL去离子水中,将此溶液置于冰水浴中,保持溶液温度在5℃以下。称取1g纤维素纳米纤维薄膜加入溶液中,在不断搅拌下一次性加入10mL NaClO溶液(有效含量约7.5%),用0.1mol/L NaOH溶液调控溶液pH值在10左右,反应约30分钟后,将膜片取出后放入pH为2的盐酸中浸泡30分钟,然后用去离子水洗涤多次后真空烘干,得到氧化纤维素纳米纤维膜。
实施例4
分别取(a)醋酸纤维素纳米纤维、(b)纤维素纳米纤维、(c)氧化纤维素纳米纤维、(d)明胶止血海绵、(e)医用纱布样品进行凝血实验。
实验方法:将各种材料剪成约1cm×1cm大小平放在表面皿中,取0.1mL柠檬酸盐全血(血液为新鲜兔血,每毫升血液中加入5mg柠檬酸钠)分别滴在每个样品表面,然后再分别滴加10μL 0.2mol/L 氯化钙溶液开始凝血,并在37℃培养箱中放置15分钟。随后将各样品分别放入离心管中,逐滴加入10mL蒸馏水,对比凝血效果。
实验结果说明:氧化纤维素纳米纤维在与血液接触后能迅速形成黑色凝胶状物质,使血液凝固,醋酸纤维素纳米纤维和纤维素纳米纤维均无此效果;与临床上常用的止血材料(明胶止血海绵、医用纱布)相比,氧化纤维素纳米纤维的凝血效果更加明显。
Claims (10)
1.一种具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法,其特征在于具体步骤如下:
(1)将纤维素衍生物溶于易挥发极性溶剂中,配制成纺丝溶液,通过静电纺丝方法,制得纤维素衍生物的纳米纤维薄膜;
(2)将所得纤维素衍生物的纳米纤维薄膜浸入到碱性溶液中进行水解,得到纤维素纳米纤维薄膜;
(3)将所得纤维素纳米纤维薄膜用TEMPO氧化体系进行氧化反应,再经酸处理、清洗、干燥后,即得到氧化纤维素纳米纤维膜片;其中:所述氧化体系为TEMPO/溴化钠(NaBr)/次氯酸钠(NaClO)共氧化体系或TEMPO/次氯酸钠(NaClO)/亚氯酸钠(NaClO2)共氧化体系;氧化反应中控制纤维素纳米纤维薄膜与TEMPO/溴化钠(NaBr)/次氯酸钠(NaClO)共氧化体系组成的溶液中pH值为10-11,反应温度为0-25℃,反应时间为0.5-2小时;氧化反应中控制纤维素纳米纤维薄膜与TEMPO/次氯酸钠(NaClO)/亚氯酸钠(NaClO2)共氧化体系组成的溶液中pH值为6-7,反应温度为25-80℃,反应时间为0.5-2小时。
2.根据权利要求1所述的具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法,其特征在于所述纤维素衍生物为醋酸纤维素、醋酸丁酸纤维素、甲基纤维素、羧甲基纤维素或乙基纤维素中的一种或几种。
3.根据权利要求1所述的具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法,其特征在于步骤(1)中所述易挥发极性溶剂为N,N-二甲基甲酰胺,N,N-二甲基乙酰胺,二甲基亚砜,丙酮,二氯甲烷,三氯甲烷,甲醇,乙醇或水中的一种或几种的混合溶剂。
4.根据权利要求1所述的具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法,其特征在于步骤(1)中所述纺丝溶液的质量分数为8%-20%。
5.根据权利要求1所述的具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法,其特征在于步骤(1)中所述静电纺丝方法中施加的直流高压为5-60kV,基材与电极间的接收距离为50-300mm。
6.根据权利要求1所述的具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法,其特征在于步骤(2)中所述碱性溶液的溶质为氢氧化钠或氢氧化钾,溶剂为甲醇、乙醇或水中的一种或几种的混合溶剂。
7.根据权利要求1所述的具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法,其特征在于步骤(2)中所述水解反应温度为15-40℃,反应时间为8-48小时。
8.根据权利要求1所述的具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法,其特征在于步骤(3)中所述酸处理过程是将氧化后的纤维素纳米纤维膜片在室温条件下浸入pH值为2-3的盐酸溶液中,处理时间为15-30分钟。
9.根据权利要求1所述的具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法,其特征在于TEMPO/溴化钠(NaBr)/次氯酸钠(NaClO)共氧化体系中,控制纤维素纳米纤维薄膜与TEMPO的质量比为1:0.01-1:0.05,纤维素纳米纤维薄膜与NaBr的质量比为1:0.1-1:0.5,纤维素纳米纤维薄膜与NaClO的质量比为1:0.5-1:1.5。
10.根据权利要求1所述的具有快速止血功能的氧化纤维素纳米纤维膜片的制备方法,其特征在于TEMPO/次氯酸钠(NaClO)/亚氯酸钠(NaClO2)共氧化体系中,控制纤维素纳米纤维膜与TEMPO的质量比为1:0.01-1:0.05,纤维素纳米纤维薄膜与NaClO的质量比为1:0.1-1:0.5,纤维素纳米纤维薄膜与NaClO2的质量比为1:1-1:5。
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