CN117398502A - 一种多功能壳聚糖基多孔纳米纤维膜敷料的制备方法 - Google Patents
一种多功能壳聚糖基多孔纳米纤维膜敷料的制备方法 Download PDFInfo
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Abstract
本发明公开了一种多功能壳聚糖基多孔纳米纤维膜敷料的制备方法。具体步骤包括：通过热致相分离技术制备壳聚糖/聚乙烯吡咯烷酮复合纳米纤维膜，后将聚乙烯吡咯烷酮去除得到壳聚糖多孔纳米纤维膜。利用低温等离子辐照壳聚糖多孔纳米纤维膜引入自由基，后将阿司匹林接枝到纤维膜上得到阿司匹林改性壳聚糖多孔纳米纤维膜。在弱碱性条件下将多巴胺聚合负载到阿司匹林改性壳聚糖多孔纳米纤维膜上得到聚多巴胺涂覆阿司匹林改性壳聚糖多孔纳米纤维膜敷料。多功能壳聚糖基多孔纳米纤维膜敷料可在止血和消炎材料中的广泛应用。
Description
技术领域
本发明涉及一种多功能壳聚糖基多孔纳米纤维膜敷料的制备方法，属于天然高分子材料和生物医学材料领域。
背景技术
战争和外科手术过程中的大量出血是导致病人死亡的一个重要原因。除了出血导致的死亡外，创伤感染是初期创伤病人至死的另一个重要原因。创伤感染过程中的细菌感染是最常见的一种感染。细菌感染是致病菌或条件致病菌侵入血循环中生长繁殖，产生毒素和其他代谢产物所引起的急性全身性感染，引起伤口局部化脓感染，甚至败血症、脓毒症等全身感染等。
壳聚糖为甲壳素脱乙酰化后的产物，甲壳素和壳聚糖具有生物降解性、细胞亲和性和生物效应等许多独特的性质，尤其是含有游离氨基的壳聚糖，是天然多糖中唯一的碱性多糖。具有生物降解性、生物相容性、无毒性、抑菌等多种生理功能，广泛应用于抗菌剂、医用敷料、药物缓释材料、医用可吸收材料、组织工程载体材料等众多领域。
阿司匹林又名乙酰水杨酸，是一种白色结晶有机化合物，主要应用于解热镇痛、抗消炎药，抗血小板聚集、退热、治疗风湿痛等。除了以上作用外，阿司匹林能阻止血栓形成，临床上用于预防短暂脑缺血发作、心肌梗死或其他手术后血栓的形成。怎样将壳聚糖止血功能与阿司匹林的抗消炎相互作用，制备多功能纳米纤维膜敷料是将来研究的重点。
发明内容：
为解决上述技术问题，本发明的目的是提出一种烷基化壳聚糖/海藻酸钠复合海绵材料的制备方法及其用途。
本发明是通过以下技术方案实现的：
一种多功能壳聚糖基多孔纳米纤维膜敷料的制备方法，其包括如下步骤：
制备壳聚糖多孔纳米纤维膜；
在氮气的保护下，用低温等离子体对所述壳聚糖多孔纳米纤维膜进行辐照后，浸泡于阿司匹林的乙醇溶液中，在60～80℃下进行反应后，洗去未反应的阿司匹林，得到阿司匹林改性壳聚糖多孔纳米纤维膜；
将所述阿司匹林改性壳聚糖多孔纳米纤维膜浸泡在多巴胺溶液中，调节pH值至8～9，使多巴胺发生沉积聚合，得到聚多巴胺涂覆阿司匹林改性壳聚糖多孔纳米纤维膜，即所述多功能壳聚糖基多孔纳米纤维膜敷料。
作为优选方案，所述壳聚糖多孔纳米纤维膜的制备方法为：
将聚乙烯吡咯烷酮和壳聚糖加入乙醇/水/乙酸混合溶剂中，搅拌溶解，得到反应液；
将所述反应液在-30～0℃下进行淬火后，得到淬火液；
将质量浓度为1～5％的氢氧化钠凝固液倒入所述淬火液中，浸泡4～8h后，调节pH值至中性，进行冷冻干燥，得到所述壳聚糖多孔纳米纤维膜。
作为优选方案，所述聚乙烯吡咯烷酮和壳聚糖的质量比为(1～3)：(2～5)；所述的淬火的时间为100～200min。
作为优选方案，所述低温等离子体的辐照功率为100～150W、压强为300～400Pa、辐照时间3～10min。
作为优选方案，所述多巴胺溶液的质量浓度为0.1～0.3％。
一种由前述制备方法得到的多功能壳聚糖基多孔纳米纤维膜敷料。
一种如前述的多功能壳聚糖基多孔纳米纤维膜敷料在止血和消炎材料中的用途。
本发明的基本原理为：
1、通过热致相分离技术制备壳聚糖/聚乙烯吡咯烷酮复合纳米纤维膜，后将聚乙烯吡咯烷酮去除得到壳聚糖多孔纳米纤维膜。
2、利用低温等离子辐照壳聚糖多孔纳米纤维膜引入自由基，后将阿司匹林接枝到纤维膜上得到阿司匹林改性壳聚糖多孔纳米纤维膜。
3、在弱碱性条件下将多巴胺聚合负载到阿司匹林改性壳聚糖多孔纳米纤维膜上得到聚多巴胺涂覆阿司匹林改性壳聚糖多孔纳米纤维膜，即所述的多功能壳聚糖基多孔纳米纤维膜敷料。
与现有技术相比，本发明具有如下的有益效果：
1、利用壳聚糖多孔纳米纤维膜的高孔隙率和大比表面积可快速吸收血浆，使血细胞积聚在表面上，从而促进血液在伤口表面凝结。
2、利用阿司匹林的缓释和聚多巴胺协同消炎作用，进一步促进伤口愈合速度。
附图说明
通过阅读参照以下附图对非限制性实施例所作的详细描述，本发明的其它特征、目的和优点将会变得更明显：
图1为多功能壳聚糖基多孔纳米纤维膜敷料的制备路线图。
具体实施方式
下面结合具体实施例对本发明进行详细说明。以下实施例将有助于本领域的技术人员进一步理解本发明，但不以任何形式限制本发明。应当指出的是，对本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进。这些都属于本发明的保护范围。
实施例1
将0.1g聚乙烯吡咯烷酮(PVP)和0.3g壳聚糖加入10g乙醇/水/乙酸混合溶剂中，磁力搅拌使其完全溶解。将上述溶液放入-20℃超低温冰箱中淬火180min，淬火结束后，将质量浓度为1％ NaOH凝固液从培养皿边缘缓慢倒入培养皿中浸泡6h，浸泡结束后，用蒸馏水冲洗pH至中性后将水倒出，冷冻干燥，得到壳聚糖多孔纳米纤维膜。
在氮气保护条件下，用低温等离子体辐照壳聚糖多孔纳米纤维膜，辐照功率为120W、压强为350Pa、辐照时间4min。辐照结束后将壳聚糖纳米纤维膜浸泡在阿司匹林/乙醇的混合溶液中，在70℃下进行反应3h。反应结束后，用乙醇洗涤多次以保证除去未反应的阿司匹林，真空干燥，得到阿司匹林改性壳聚糖多孔纳米纤维膜。
将0.1g阿司匹林改性壳聚糖多孔纳米纤维膜浸泡在100mL浓度为0.2mg/mL的多巴胺溶液中，用Tris盐缓冲液调节pH值为8.5，浸泡30min后，使多巴胺在多孔纤维表面沉积聚合。然后，用去离子水对纤维膜清洗3遍，真空干燥得到聚多巴胺涂覆阿司匹林改性壳聚糖多孔纳米纤维膜。制备流程如图1所示。
实施例1制备的多功能壳聚糖基多孔纳米纤维膜敷料孔隙率为94.1％，比表面积为6.14m2/g。采用金黄色葡萄球菌(S.aureus)为实验对象，计算培养皿上的菌落形成单位(colony forming units,CFU)，从而计算敷料对细菌的杀菌率，从而考察敷料的抗菌性能。实施例1制备的多功能复合纳米纤维膜敷料的杀菌率为100％。采用凝血指数(BCI)在体外进行评估，通过血红蛋白溶液的吸光度值来判断其血液凝结速率。多功能壳聚糖基多孔纳米纤维膜敷料的BCI为60.4％。
实施例2
将0.15g聚乙烯吡咯烷酮(PVP)和0.25g壳聚糖加入10g乙醇/水/乙酸混合溶剂中，磁力搅拌使其完全溶解。将上述溶液放入-10℃超低温冰箱中淬火150min，淬火结束后，将质量浓度为1％ NaOH凝固液从培养皿边缘缓慢倒入培养皿中浸泡6h，浸泡结束后，用蒸馏水冲洗pH至中性后将水倒出，冷冻干燥，得到壳聚糖多孔纳米纤维膜。
在氮气保护条件下，用低温等离子体辐照壳聚糖多孔纳米纤维膜，辐照功率为100W、压强为250Pa、辐照时间6min。辐照结束后将壳聚糖纳米纤维膜浸泡在阿司匹林/乙醇的混合溶液中，在70℃下进行反应3h。反应结束后，用乙醇洗涤多次以保证除去未反应的阿司匹林，真空干燥，得到阿司匹林改性壳聚糖多孔纳米纤维膜。
将0.1g阿司匹林改性壳聚糖多孔纳米纤维膜浸泡在100mL浓度为0.15mg/mL的多巴胺溶液中，用Tris盐缓冲液调节pH值为8.5，浸泡30min后，使多巴胺在多孔纤维表面沉积聚合。然后，用去离子水对纤维膜清洗3遍，真空干燥得到聚多巴胺涂覆阿司匹林改性壳聚糖多孔纳米纤维膜
实施例1制备的多功能壳聚糖基多孔纳米纤维膜敷料孔隙率为92.9％，比表面积为6.35m2/g。采用金黄色葡萄球菌(S.aureus)为实验对象，计算培养皿上的菌落形成单位(colony forming units,CFU)，从而计算敷料对细菌的杀菌率，从而考察敷料的抗菌性能。实施例1制备的多功能复合纳米纤维膜敷料的杀菌率为100％。采用凝血指数(BCI)在体外进行评估，通过血红蛋白溶液的吸光度值来判断其血液凝结速率。多功能壳聚糖基多孔纳米纤维膜敷料的BCI为61.9％。
实施例3
将0.15g聚乙烯吡咯烷酮(PVP)和0.3g壳聚糖加入10g乙醇/水/乙酸混合溶剂中，磁力搅拌使其完全溶解。将上述溶液放入-20℃超低温冰箱中淬火150min，淬火结束后，将质量浓度为1％ NaOH凝固液从培养皿边缘缓慢倒入培养皿中浸泡6h，浸泡结束后，用蒸馏水冲洗pH至中性后将水倒出，冷冻干燥，得到壳聚糖多孔纳米纤维膜。
在氮气保护条件下，用低温等离子体辐照壳聚糖多孔纳米纤维膜，辐照功率为130W、压强为300Pa、辐照时间5min。辐照结束后将壳聚糖纳米纤维膜浸泡在阿司匹林/乙醇的混合溶液中，在70℃下进行反应3h。反应结束后，用乙醇洗涤多次以保证除去未反应的阿司匹林，真空干燥，得到阿司匹林改性壳聚糖多孔纳米纤维膜。
将0.1g阿司匹林改性壳聚糖多孔纳米纤维膜浸泡在100mL浓度为0.2mg/mL的多巴胺溶液中，用Tris盐缓冲液调节pH值为8.5，浸泡30min后，使多巴胺在多孔纤维表面沉积聚合。然后，用去离子水对纤维膜清洗3遍，真空干燥得到聚多巴胺涂覆阿司匹林改性壳聚糖多孔纳米纤维膜
实施例1制备的多功能壳聚糖基多孔纳米纤维膜敷料孔隙率为94.2％，比表面积为6.66m2/g。采用金黄色葡萄球菌(S.aureus)为实验对象，计算培养皿上的菌落形成单位(colony forming units,CFU)，从而计算敷料对细菌的杀菌率，从而考察敷料的抗菌性能。实施例1制备的多功能复合纳米纤维膜敷料的杀菌率为100％。采用凝血指数(BCI)在体外进行评估，通过血红蛋白溶液的吸光度值来判断其血液凝结速率。多功能壳聚糖基多孔纳米纤维膜敷料的BCI为63.2％。
对比例1
与实施例1不同的是，对比例1壳聚糖多孔纳米纤维膜无进行阿司匹林改性，也无进行多巴胺负载，最终只得到壳聚糖多孔纳米纤维膜敷料。对比例1制备的壳聚糖多孔纳米纤维膜敷料孔隙率为87.2％，比表面积为4.88m2/g。对比例1制备多功能壳聚糖基多孔纳米纤维膜敷料的杀菌率为75％。采用凝血指数(BCI)在体外进行评估，通过血红蛋白溶液的吸光度值来判断其血液凝结速率。多功能壳聚糖基多孔纳米纤维膜敷料的BCI为50.6％。
对比例2
与实施例1不同的是，对比例2无多巴胺负载，最终得到阿司匹林改性壳聚糖多孔纳米纤维膜敷料。对比例2制备的阿司匹林改性壳聚糖多孔纳米纤维膜敷料的孔隙率为91.5％，比表面积为5.55m2/g。对比例2制备多功能壳聚糖基多孔纳米纤维膜敷料的杀菌率为100％。采用凝血指数(BCI)在体外进行评估，通过血红蛋白溶液的吸光度值来判断其血液凝结速率。多功能壳聚糖基多孔纳米纤维膜敷料的BCI为59.1％。
对比例3
与实施例1不同的是，对比例3无阿司匹林改性，最终得到聚多巴胺涂覆壳聚糖基多孔纳米纤维膜敷料。对比例3制备的聚多巴胺涂覆壳聚糖基多孔纳米纤维膜敷料的孔隙率为93.1％，比表面积为5.69m2/g。对比例3制备聚多巴胺涂覆壳聚糖基多孔纳米纤维膜敷料的杀菌率为100％。采用凝血指数(BCI)在体外进行评估，通过血红蛋白溶液的吸光度值来判断其血液凝结速率。多功能壳聚糖基多孔纳米纤维膜敷料的BCI为58.4％。
以上对本发明的具体实施例进行了描述。需要理解的是，本发明并不局限于上述特定实施方式，本领域技术人员可以在权利要求的范围内做出各种变形或修改，这并不影响本发明的实质内容。
Claims (7)
1.一种多功能壳聚糖基多孔纳米纤维膜敷料的制备方法，其特征在于，包括如下步骤：
制备壳聚糖多孔纳米纤维膜；
在氮气的保护下，用低温等离子体对所述壳聚糖多孔纳米纤维膜进行辐照后，浸泡于阿司匹林的乙醇溶液中，在60～80℃下进行反应后，洗去未反应的阿司匹林，得到阿司匹林改性壳聚糖多孔纳米纤维膜；
将所述阿司匹林改性壳聚糖多孔纳米纤维膜浸泡在多巴胺溶液中，调节pH值至8～9，使多巴胺发生沉积聚合，得到聚多巴胺涂覆阿司匹林改性壳聚糖多孔纳米纤维膜，即所述多功能壳聚糖基多孔纳米纤维膜敷料。
2.如权利要求1所述的多功能壳聚糖基多孔纳米纤维膜敷料的制备方法，其特征在于，所述壳聚糖多孔纳米纤维膜的制备方法为：
将聚乙烯吡咯烷酮和壳聚糖加入乙醇/水/乙酸混合溶剂中，搅拌溶解，得到反应液；
将所述反应液在-30～0℃下进行淬火后，得到淬火液；
将质量浓度为1～5％的氢氧化钠凝固液倒入所述淬火液中，浸泡4～8h后，调节pH值至中性，进行冷冻干燥，得到所述壳聚糖多孔纳米纤维膜。
3.如权利要求2所述的多功能壳聚糖基多孔纳米纤维膜敷料的制备方法，其特征在于，所述聚乙烯吡咯烷酮和壳聚糖的质量比为(1～3)：(2～5)；所述的淬火的时间为100～200min。
4.如权利要求1所述的多功能壳聚糖基多孔纳米纤维膜敷料的制备方法，其特征在于，所述低温等离子体的辐照功率为100～150W、压强为300～400Pa、辐照时间3～10min。
5.如权利要求1所述的多功能壳聚糖基多孔纳米纤维膜敷料的制备方法，其特征在于，所述多巴胺溶液的质量浓度为0.1～0.3％。
6.一种由权利要求1～5中任意一项所述制备方法得到的多功能壳聚糖基多孔纳米纤维膜敷料。
7.一种如权利要求6所述的多功能壳聚糖基多孔纳米纤维膜敷料在止血和消炎材料中的用途。
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