CN108396461B - 一种促愈合纳米纤维膜敷料及其制备方法 - Google Patents
一种促愈合纳米纤维膜敷料及其制备方法 Download PDFInfo
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Abstract
本发明提供了一种促愈合纳米纤维膜敷料及其制备方法，包括以下步骤：（1）聚乳酸端羟基的羧基改性，（2）步骤（1）所得产物的Ag+适配体修饰，（3）静电纺丝液的制备，（4）纳米纤维膜的制备。本发明提供了一种促愈合纳米纤维膜敷料及其制备方法，本发明将聚氨酯的端羟基进行羧基改性，增加聚氨酯表面的羧基数量，再将氨基修饰的Ag+适配体的一条链修饰在聚氨酯上，通过Ag+适配体对Ag+的亲和吸附，使用抗坏血酸作为还原剂原位合成银颗粒，解决了单独添加银颗粒引起银粒子团聚分散不均的问题。本发明制得的纳米纤维膜对创面愈合效果良好，同时具有良好的机械强度，因此本发明制得的纳米纤维膜在创面愈合方面中具有广泛的应用前景。
Description
技术领域
本发明属于医用材料技术领域，具体涉及一种促愈合纳米纤维膜敷料及其制备方法。
背景技术
聚乳酸是一种典型的合成类可完全生物降解材料，由于其具有可靠的生物安全性、生物可降解性、对环境友好、良好的力学性能及易于加工成形等优点，目前已被广泛应用于生物医用高分子、纺织行业、农用地膜和包装等行业。
创面愈合是创伤后机体功能康复的前提，促进创面愈合也是创伤治疗的重要环节，选用适宜的敷料可以加快创面愈合，提高愈合效果，因此研发可促进创面愈合的敷料，一直是临床关注的热点。
通过静电纺丝技术制备的纳米纤维膜在医用材料领域的应用越来越广泛，静电纺丝聚合物的纳米纤维膜具有比表面积大、孔隙率高等特殊的性能，在组织工程支架、皮肤修复和创伤敷料等生物医学领域有广阔的应用前景。然而现有的创面愈合用纳米纤维膜的愈合效果较差，并且纤维膜的机械强度较差。
发明内容
为了解决以上现有技术存在的问题，本发明的目的在于提供一种促愈合纳米纤维膜敷料及其制备方法，从而提高纳米纤维膜的创面愈合效果和力学性能。
为了实现上述目的，本发明提供以下技术方案：
一种促愈合纳米纤维膜敷料的制备方法，包括以下步骤：
（1）聚乳酸端羟基的羧基改性
将聚乳酸和琥珀酸酐进行混合，加入二氯甲烷在搅拌的状态下使得溶解，然后将其加热搅拌至30-40℃，保持该温度继续反应12-24h，反应完成后，用乙酸乙酯进行萃取收集上层有机层，然后用旋转蒸发仪进行旋干，收集产物A，其中聚乳酸和琥珀酸酐的质量比为1:5-10，聚乳酸和琥珀酸酐在反应体系中的质量百分比为15-30%；
（2）步骤（1）所得产物的Ag+适配体修饰
向步骤（1）所得产物A 100份中加入10-20倍重量份的二氯甲烷溶液，在搅拌的条件下加入10-20份的N-羟基琥珀酰亚胺在室温下搅拌反应15-30min，然后加入100-200份的Ag+适配体DNA1 30-50份的碳二亚胺继续反应4-6h，使用截留分子量为1000的超滤管进行超滤，收集截留产物B；
适配体DNA1：5’-SH-AAAAACTCTCTCTCTCTCTCTCTCTC-3’；
（3）静电纺丝液的制备
将步骤（2）制得的产物B 40-60份、聚丁二酸丁二醇酯5-10份、Ag+适配体DNA2 50-100份、AgNO3 10-20份、聚乙烯醇-苯乙烯吡啶盐8-15份、Ⅰ型胶原蛋白6-12份、壳聚糖5-10份、抗坏血酸3-7份、二氯甲烷20-40份进行混合，在25-30℃的温度下搅拌直至溶液变为深红色并不再变化；然后将β-磷酸三钙4-8份、盐酸聚六亚甲基双胍5-10份、聚乙烯吡咯烷酮3-5份、积雪草苷3-6份、多酚物质4-9份继续搅拌1-2h，并进行超声波处理30-60min，制得静电纺丝液；
适配体DNA2：5’-CACACACACACACACACACAC-3’；
（4）纳米纤维膜的制备
将步骤（3）制得的静电纺丝液进行静电纺丝，
搅拌均匀后进行静电纺丝，接收纳米丝，制得促愈合纳米纤维膜敷料；静电纺丝工艺参数如下：电压20-40KV，接收距离为5-20cm，纺丝孔内径0.1-1.0mm，纺丝速度为20-60μL/min。
优选的，所述多酚物质为茶多酚、单宁酸、姜黄素的一种或几种组合。
优选的，所述Ag+适配体DNA1和DNA2上生成的银颗粒的大小为5-10nm。
优选的，所述纳米纤维的直径为50-200nm。
优选的，制备静电纺丝液所用的各原料的重量份为：步骤（2）制得的产物B 50份、聚丁二酸丁二醇酯8份、Ag+适配体DNA2 75份、AgNO3 15份、聚乙烯醇-苯乙烯吡啶盐12份、Ⅰ型胶原蛋白9份、壳聚糖8份、抗坏血酸5份、二氯甲烷30份、β-磷酸三钙6份、盐酸聚六亚甲基双胍8份、聚乙烯吡咯烷酮4份、积雪草苷5份、多酚物质7份。
本发明所述的制备方法制得的促愈合纳米纤维膜敷料。
有益效果：本发明提供了一种促愈合纳米纤维膜敷料及其制备方法，本发明将聚氨酯的端羟基进行羧基改性，增加聚氨酯表面的羧基数量，然后再将氨基修饰的Ag+适配体的一条链修饰在聚氨酯上，通过Ag+适配体对Ag+的亲和吸附，再使用抗坏血酸作为还原剂原位合成银颗粒，解决了单独添加银颗粒引起银粒子团聚从而分散不均的问题。负载银粒子的纳米纤维膜对材料的抗菌性具有重要的作用。从测试结果得出，本发明制得的纳米纤维膜对创面愈合效果良好，银粒子在聚氨酯上的原位生长比未在聚氨酯上生长的对创面愈合更好，聚丁二酸丁二醇酯影响了纳米纤维膜的性能，未添加聚丁二酸丁二醇酯的纳米纤维膜对创面的止血效果较差。同时纳米纤维膜拉伸强度不低于17.2MPa，断裂伸长率不低于61.3%，因此表现出良好的机械强度。并且从结果中得出，聚丁二酸丁二醇酯的添加影响了纳米纤维膜的力学性能。因此本发明制得的纳米纤维膜在创面愈合方面中具有广泛的应用前景。
具体实施方式
下面结合具体实施例来进一步描述本发明，但实施例仅是范例性的，并不对本发明的范围构成任何限制。本领域技术人员应该理解的是，在不偏离本发明的精神和范围下可以对本发明技术方案的细节和形式进行修改或替换，但这些修改和替换均落入本发明的保护范围内。
实施例1
一种促愈合纳米纤维膜敷料的制备方法，包括以下步骤：
（1）聚乳酸端羟基的羧基改性
将聚乳酸和琥珀酸酐进行混合，加入二氯甲烷在搅拌的状态下使得溶解，然后将其加热搅拌至30℃，保持该温度继续反应12h，反应完成后，用乙酸乙酯进行萃取收集上层有机层，然后用旋转蒸发仪进行旋干，收集产物A，其中聚乳酸和琥珀酸酐的质量比为1:5，聚乳酸和琥珀酸酐在反应体系中的质量百分比为15%；
（2）步骤（1）所得产物的Ag+适配体修饰
向步骤（1）所得产物A 100份中加入10倍重量份的二氯甲烷溶液，在搅拌的条件下加入10份的N-羟基琥珀酰亚胺在室温下搅拌反应15min，然后加入100份的Ag+适配体DNA130份的碳二亚胺继续反应4h，使用截留分子量为1000的超滤管进行超滤，收集截留产物B；
适配体DNA1：5’-SH-AAAAACTCTCTCTCTCTCTCTCTCTC-3’；
（3）静电纺丝液的制备
将步骤（2）制得的产物B 40份、聚丁二酸丁二醇酯5份、Ag+适配体DNA2 50份、AgNO310份、聚乙烯醇-苯乙烯吡啶盐8份、Ⅰ型胶原蛋白6份、壳聚糖5份、抗坏血酸3份、二氯甲烷20份进行混合，在25℃的温度下搅拌直至溶液变为深红色并不再变化；然后将β-磷酸三钙4份、盐酸聚六亚甲基双胍5份、聚乙烯吡咯烷酮3份、积雪草苷3份、质量比为1:1的茶多酚和单宁酸4份继续搅拌1h，并进行超声波处理30min，制得静电纺丝液；
适配体DNA2：5’-CACACACACACACACACACAC-3’；
（4）纳米纤维膜的制备
将步骤（3）制得的静电纺丝液进行静电纺丝，
搅拌均匀后进行静电纺丝，接收纳米丝，制得促愈合纳米纤维膜敷料；静电纺丝工艺参数如下：电压20KV，接收距离为5cm，纺丝孔内径0.1mm，纺丝速度为20μL/min。
所述Ag+适配体DNA1和DNA2上生成的银颗粒的大小为5-10nm。
本实施例制得的纳米纤维的直径为50nm。
实施例2
一种促愈合纳米纤维膜敷料的制备方法，包括以下步骤：
（1）聚乳酸端羟基的羧基改性
将聚乳酸和琥珀酸酐进行混合，加入二氯甲烷在搅拌的状态下使得溶解，然后将其加热搅拌至35℃，保持该温度继续反应18h，反应完成后，用乙酸乙酯进行萃取收集上层有机层，然后用旋转蒸发仪进行旋干，收集产物A，其中聚乳酸和琥珀酸酐的质量比为1:8，聚乳酸和琥珀酸酐在反应体系中的质量百分比为22%；
（2）步骤（1）所得产物的Ag+适配体修饰
向步骤（1）所得产物A 100份中加入15倍重量份的二氯甲烷溶液，在搅拌的条件下加入15份的N-羟基琥珀酰亚胺在室温下搅拌反应22min，然后加入150份的Ag+适配体DNA130-50份的碳二亚胺继续反应4-6h，使用截留分子量为1000的超滤管进行超滤，收集截留产物B；
适配体DNA1：5’-SH-AAAAACTCTCTCTCTCTCTCTCTCTC-3’；
（3）静电纺丝液的制备
将步骤（2）制得的产物B 50份、聚丁二酸丁二醇酯8份、Ag+适配体DNA2 75份、AgNO315份、聚乙烯醇-苯乙烯吡啶盐12份、Ⅰ型胶原蛋白9份、壳聚糖8份、抗坏血酸5份、二氯甲烷30份进行混合，在28℃的温度下搅拌直至溶液变为深红色并不再变化；然后将β-磷酸三钙6份、盐酸聚六亚甲基双胍8份、聚乙烯吡咯烷酮4份、积雪草苷5份、质量比为1:1:1的茶多酚、单宁酸和姜黄素7份继续搅拌1.5h，并进行超声波处理45min，制得静电纺丝液；
适配体DNA2：5’-CACACACACACACACACACAC-3’；
（4）纳米纤维膜的制备
将步骤（3）制得的静电纺丝液进行静电纺丝，
搅拌均匀后进行静电纺丝，接收纳米丝，制得促愈合纳米纤维膜敷料；静电纺丝工艺参数如下：电压30KV，接收距离为12cm，纺丝孔内径0.5mm，纺丝速度为40μL/min。
所述Ag+适配体DNA1和DNA2上生成的银颗粒的大小为5-10nm。
本实施例制得的纳米纤维的直径为130nm。
实施例3
（1）聚乳酸端羟基的羧基改性
将聚乳酸和琥珀酸酐进行混合，加入二氯甲烷在搅拌的状态下使得溶解，然后将其加热搅拌至32℃，保持该温度继续反应15h，反应完成后，用乙酸乙酯进行萃取收集上层有机层，然后用旋转蒸发仪进行旋干，收集产物A，其中聚乳酸和琥珀酸酐的质量比为1:6，聚乳酸和琥珀酸酐在反应体系中的质量百分比为20%；
（2）步骤（1）所得产物的Ag+适配体修饰
向步骤（1）所得产物A 100份中加入12倍重量份的二氯甲烷溶液，在搅拌的条件下加入12份的N-羟基琥珀酰亚胺在室温下搅拌反应20min，然后加入120份的Ag+适配体DNA135份的碳二亚胺继续反应4.5h，使用截留分子量为1000的超滤管进行超滤，收集截留产物B；
适配体DNA1：5’-SH-AAAAACTCTCTCTCTCTCTCTCTCTC-3’；
（3）静电纺丝液的制备
将步骤（2）制得的产物B 45份、聚丁二酸丁二醇酯6份、Ag+适配体DNA2 60份、AgNO312份、聚乙烯醇-苯乙烯吡啶盐10份、Ⅰ型胶原蛋白7份、壳聚糖6份、抗坏血酸4份、二氯甲烷25份进行混合，在26℃的温度下搅拌直至溶液变为深红色并不再变化；然后将β-磷酸三钙5份、盐酸聚六亚甲基双胍6份、聚乙烯吡咯烷酮3.5份、积雪草苷4份、质量比为1：1的单宁酸和姜黄素5份继续搅拌1.2h，并进行超声波处理40min，制得静电纺丝液；
适配体DNA2：5’-CACACACACACACACACACAC-3’；
（4）纳米纤维膜的制备
将步骤（3）制得的静电纺丝液进行静电纺丝，
搅拌均匀后进行静电纺丝，接收纳米丝，制得促愈合纳米纤维膜敷料；静电纺丝工艺参数如下：电压25KV，接收距离为10cm，纺丝孔内径0.3mm，纺丝速度为30μL/min。
所述Ag+适配体DNA1和DNA2上生成的银颗粒的大小为5-10nm。
本实施例制得的纳米纤维的直径为80nm。
实施例4
一种促愈合纳米纤维膜敷料的制备方法，包括以下步骤：
（1）聚乳酸端羟基的羧基改性
将聚乳酸和琥珀酸酐进行混合，加入二氯甲烷在搅拌的状态下使得溶解，然后将其加热搅拌至40℃，保持该温度继续反应24h，反应完成后，用乙酸乙酯进行萃取收集上层有机层，然后用旋转蒸发仪进行旋干，收集产物A，其中聚乳酸和琥珀酸酐的质量比为1:10，聚乳酸和琥珀酸酐在反应体系中的质量百分比为30%；
（2）步骤（1）所得产物的Ag+适配体修饰
向步骤（1）所得产物A 100份中加入20倍重量份的二氯甲烷溶液，在搅拌的条件下加入20份的N-羟基琥珀酰亚胺在室温下搅拌反应30min，然后加入200份的Ag+适配体DNA150份的碳二亚胺继续反应6h，使用截留分子量为1000的超滤管进行超滤，收集截留产物B；
适配体DNA1：5’-SH-AAAAACTCTCTCTCTCTCTCTCTCTC-3’；
（3）静电纺丝液的制备
将步骤（2）制得的产物B 60份、聚丁二酸丁二醇酯10份、Ag+适配体DNA2 100份、AgNO3 20份、聚乙烯醇-苯乙烯吡啶盐15份、Ⅰ型胶原蛋白12份、壳聚糖10份、抗坏血酸7份、二氯甲烷40份进行混合，在30℃的温度下搅拌直至溶液变为深红色并不再变化；然后将β-磷酸三钙8份、盐酸聚六亚甲基双胍10份、聚乙烯吡咯烷酮5份、积雪草苷6份、质量比为1:1的茶多酚和姜黄素9份继续搅拌2h，并进行超声波处理60min，制得静电纺丝液；
适配体DNA2：5’-CACACACACACACACACACAC-3’；
（4）纳米纤维膜的制备
将步骤（3）制得的静电纺丝液进行静电纺丝，
搅拌均匀后进行静电纺丝，接收纳米丝，制得促愈合纳米纤维膜敷料；静电纺丝工艺参数如下：电压40KV，接收距离为20cm，纺丝孔内径1.0mm，纺丝速度为60μL/min。
所述Ag+适配体DNA1和DNA2上生成的银颗粒的大小为5-10nm。
本实施例制得的纳米纤维的直径为200nm。
对比例1
对比例1与实施例2的区别在于：对比例1中未添加DNA2。
对比例2
对比例2与实施例2的区别在于：对比例2中DNA1共混到溶液中，未与聚乳酸溶液发生化学交联。
对比例3
对比例3与实施例2的区别在于：对比例3中未添加聚丁二酸丁二醇酯。
将实施例1-4和对比例1-3制备的促愈合纳米纤维膜敷料参照“血余炭纳米纤维膜促进家兔创面愈合的实验研究”（药学实践杂质，2013，31（6），438-458）进行以下性能的测试，表1为家兔背部创面出血模型的止血效果，表2为对创面愈合时间和愈合率的影响，从表1和表2得出，本发明制得的纳米纤维膜对创面愈合效果良好，银粒子在聚氨酯上的原位生长比未在聚氨酯上生长的对创面愈合更好，聚丁二酸丁二醇酯影响了纳米纤维膜的性能，未添加聚丁二酸丁二醇酯的纳米纤维膜对创面的止血效果较差。
表1
表2
表3为纳米纤维膜的拉伸强度和断裂伸长率的测试结果，从表3中得出，本发明制得的纳米纤维膜拉伸强度不低于17.2MPa，断裂伸长率不低于61.3%，因此表现出良好的机械强度。并且从结果中得出，聚丁二酸丁二醇酯的添加影响了纳米纤维膜的力学性能。
表3
序列表
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<211> 21
<212> DNA
<213> 人工合成(2 Ambystoma laterale x Ambystoma jeffersonianum)
<400> 2
cacacacaca cacacacaca c 21
Claims (6)
1. 一种促愈合纳米纤维膜敷料的制备方法，其特征在于，包括以下步骤：
（1）聚乳酸端羟基的羧基改性
将聚乳酸和琥珀酸酐进行混合，加入二氯甲烷在搅拌的状态下使得溶解，然后将其加热搅拌至30-40℃，保持该温度继续反应12-24h，反应完成后，用乙酸乙酯进行萃取收集上层有机层，然后用旋转蒸发仪进行旋干，收集产物A，其中聚乳酸和琥珀酸酐的质量比为1:5-10，聚乳酸和琥珀酸酐在反应体系中的质量百分比为15-30%；
（2）步骤（1）所得产物的Ag+适配体修饰
向步骤（1）所得产物A 100份中加入10-20倍重量份的二氯甲烷溶液，在搅拌的条件下加入10-20份的N-羟基琥珀酰亚胺在室温下搅拌反应15-30min，然后加入100-200份的Ag+适配体DNA1、30-50份的碳二亚胺继续反应4-6h，使用截留分子量为1000的超滤管进行超滤，收集截留产物B；
适配体DNA1：5’-NH2-AAAAACTCTCTCTCTCTCTCTCTCTC-3’；
（3）静电纺丝液的制备
将步骤（2）制得的产物B 40-60份、聚丁二酸丁二醇酯5-10份、Ag+适配体DNA2 50-100份、AgNO3 10-20份、聚乙烯醇-苯乙烯吡啶盐8-15份、Ⅰ型胶原蛋白6-12份、壳聚糖5-10份、抗坏血酸3-7份、二氯甲烷20-40份进行混合，在25-30℃的温度下搅拌直至溶液变为深红色并不再变化；然后将β-磷酸三钙4-8份、盐酸聚六亚甲基双胍5-10份、聚乙烯吡咯烷酮3-5份、积雪草苷3-6份、多酚物质4-9份继续搅拌1-2h，并进行超声波处理30-60min，制得静电纺丝液；
适配体DNA2：5’-CACACACACACACACACACAC-3’；
（4）纳米纤维膜的制备
将步骤（3）制得的静电纺丝液进行静电纺丝，
搅拌均匀后进行静电纺丝，接收纳米丝，制得促愈合纳米纤维膜敷料；静电纺丝工艺参数如下：电压20-40KV，接收距离为5-20cm，纺丝孔内径0.1-1.0mm，纺丝速度为20-60μL/min。
2.根据权利要求1所述的一种促愈合纳米纤维膜敷料的制备方法，其特征在于，所述多酚物质为茶多酚、单宁酸、姜黄素的一种或几种组合。
3.根据权利要求1所述的一种促愈合纳米纤维膜敷料的制备方法，其特征在于，所述Ag+适配体DNA1和DNA2上生成的银颗粒的大小为5-10nm。
4.根据权利要求1所述的一种促愈合纳米纤维膜敷料的制备方法，其特征在于，所述纳米纤维的直径为50-200nm。
5. 根据权利要求1所述的一种促愈合纳米纤维膜敷料的制备方法，其特征在于，制备静电纺丝液所用的各原料的重量份为：步骤（2）制得的产物B 50份、聚丁二酸丁二醇酯8份、Ag+适配体DNA2 75份、AgNO3 15份、聚乙烯醇-苯乙烯吡啶盐12份、Ⅰ型胶原蛋白9份、壳聚糖8份、抗坏血酸5份、二氯甲烷30份、β-磷酸三钙6份、盐酸聚六亚甲基双胍8份、聚乙烯吡咯烷酮4份、积雪草苷5份、多酚物质7份。
6.权利要求1-5任一项所述的制备方法制得的促愈合纳米纤维膜敷料。
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