CN112915251A - 生物高分子纤维伤口敷料及其制备方法 - Google Patents

生物高分子纤维伤口敷料及其制备方法 Download PDF

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CN112915251A
CN112915251A CN202110112011.9A CN202110112011A CN112915251A CN 112915251 A CN112915251 A CN 112915251A CN 202110112011 A CN202110112011 A CN 202110112011A CN 112915251 A CN112915251 A CN 112915251A
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wound dressing
mxene
biopolymer
fiber wound
biopolymer fiber
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CN112915251B (zh
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邓怡
殷捷
周雄
毛誉蓉
谢克难
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Sichuan University
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Abstract

本发明公开了一种生物高分子纤维伤口敷料及其制备方法,以聚己内酯、左旋聚乳酸或聚乙烯醇作为载体,负载MXene、壳聚糖和抗炎药物,组成具有抗菌抗炎性能的生物高分子纤维伤口敷料。该敷料创新性地将二维材料MXene的光热光动力杀菌作用与生物高分子纤维的细菌吸附作用相结合,最大限度地降低了伤口细菌感染的风险。此外,该敷料引入阿司匹林作为抗炎药物,通过光热控释的方法,显著减轻了光热疗法引发的炎症反应,避免了传统光热疗法在高温治疗时对皮肤的炎症损害,有利于伤口的快速愈合。

Description

生物高分子纤维伤口敷料及其制备方法
技术领域
本发明涉及医用材料领域,尤其涉及一种生物高分子纤维伤口敷料及其制备方法。
背景技术
皮肤作为人体最大的器官,它具有屏障和吸收,分泌,排泄,调节体温,感觉免疫,呼吸,内分泌等重要的生理功能。一旦皮肤出现严重缺损,其引发的感染将严重影响人们的生活和健康。因此,制备出抗感染的伤口敷料对于保护伤口以及抑制伤口感染具有非常重要的意义。
通过静电纺丝技术制备的生物高分子纤维膜由于具有高比表面积、高孔隙率和良好的生物相容性等优点,非常适合作为伤口敷料用来吸收渗出液,止血,透气和保持湿润的伤口环境。研究表明大部分细菌更易于吸附在疏水材料上,因此聚己内酯(PCL)纤维膜作为疏水材料可将伤口周围的致病菌聚集到纤维膜表面,但是生物高分子纤维膜没有良好的抗菌活性,难以对聚集的细菌进行杀灭。为了解决这一问题,传统的治疗方案是将抗生素加入到敷料中对致病菌造成杀伤。但是,抗生素的长期使用将使大量致病菌对抗生素产生耐药性,轻则伤口的恢复效果达不到预期,重则导致患者病情的恶化甚至死亡。随着光学技术飞速发展,诸多光敏材料由于能够实现光热疗法和光动力疗法协同,在生物医学材料方面得到了广泛的应用。但是,光热试剂的加热温度通常在45–50℃范围内,在这种温度下,最常见的细胞1反应是坏死,这会破坏细胞膜的完整性并释放细胞内的组分,进而产生促炎反应。
发明内容
本发明的目的是为了克服在皮肤修复中滥用抗生素引发的耐药性和光热疗法引发的炎症等问题,克服现有伤口敷料容易引发耐药性和光热疗法引发炎症的缺陷,提供一种制备工艺简单,抗菌抗炎性能好的生物高分子纤维伤口敷料及其制备方法。
为达到上述目的,本发明是按照以下技术方案实施的:
本发明以聚己内酯(PCL)、左旋聚乳酸(PLLA)或聚乙烯醇(PVA)作为载体,负载MXene、壳聚糖(CS)和抗炎药物,组成具有抗菌抗炎性能的生物高分子纤维伤口敷料。
本发明的生物高分子纤维伤口敷料制备方法采用以下技术方案:
利用聚己内酯(PCL)、左旋聚乳酸(PLLA)或聚乙烯醇(PVA)作为载体,MXene作为光敏抗菌剂,均匀混合后电纺,得到负载MXene的生物高分子纤维,然后浸滴壳聚糖(CS)溶液于生物高分子纤维表面,最后将生物高分子纤维浸泡在抗炎药物溶液中通过静电吸附作用完成药物负载,得到具有抗菌抗炎性能的生物高分子纤维伤口敷料。
上述方案中,抗炎药物为阿司匹林或布洛芬。
上述方案中,生物高分子纤维伤口敷料制备方法采用以下工艺:
步骤1:MXene的制备
将1g氟化锂(LiF)粉末加入20mL盐酸溶液(9M)搅拌半小时,1g Ti3AlC2缓慢加入上述混合溶液持续搅拌24-36小时,反应温度保持在(35-55℃);所得样品反复离心(3500-5500rpm)洗涤直至上清液PH≈7;之后样品通过超声浴处理1小时,再离心(8000rpm)取上清液,冻干即为MXene。
步骤2:负载MXene的生物高分子纤维的制备
聚己内酯或左旋聚乳酸或聚乙烯醇:MXene按质量比99:1或95:5或95:5溶于六氟异丙醇(HFIP)溶剂(8%w/v),并超声搅拌过夜获得纺丝液,再将纺丝液加入纺丝注射器中进行静电纺丝,最后样品纺完后自然晾干;纺丝使用24G针头,速率为(1-1.5)mL/h,电压为15kV。
步骤3:负载抗炎药物的生物高分子纤维的制备
首先将3%w/v壳聚糖(CS)溶液浸滴在步骤2获得的负载MXene的生物高分子纤维表面,然后将浸滴后的生物高分子纤维置于常温下干燥,再将干燥后的生物高分子纤维浸泡在(300-500)μg/mL阿司匹林(Asp)/布洛芬(Ibu)溶液中,最后晾干获得一种具有抗菌抗炎性能的生物高分子纤维伤口敷料。
本发明的有益效果是:
本发明是一种生物高分子纤维伤口敷料及其制备方法,与现有技术相比,本发明具有如下技术效果:
1、本发明敷料作为疏水材料可将伤口周围的致病菌聚集到纤维膜表面,进而使得纤维膜表面的二维材料MXene不仅可以通过直接接触细菌的方式导致细菌的细胞膜破裂,还可以在近红外激光(NIR)照射下对聚集的致病菌进行光热光动力杀灭。敷料在吸附作用与杀菌作用的协同下最大限度地降低了伤口细菌感染的风险。
2、本发明敷料创新性地将MXene负载在生物高分子纤维上,通过改变MXene与生物高分子的用料比,灵活改变近红外激光照射强度,从而赋予了纤维光热和光动力协同抗菌的作用。
3、本发明敷料引入了阿司匹林、布洛芬作为抗炎药物,通过光热控释的方法,显著减轻了光热疗法引发的炎症反应,避免了传统光热疗法在高温治疗的同时对皮肤的炎症损害。
综上所述,本发明克服了现有伤口敷料容易引发耐药性和光热疗法引发炎症的缺陷,提供的生物高分子纤维伤口敷料及其制备方法,制备工艺简单,抗菌抗炎性能好。
具体实施方式
下面以及具体实施例对本发明作进一步描述,在此发明的示意性实施例以及说明用来解释本发明,但并不作为对本发明的限定。
实施例1
具有抗菌抗炎性能的生物高分子纤维伤口敷料的制备方法具体包括以下步骤:
步骤1:MXene的制备
1g氟化锂(LiF)粉末加入20mL盐酸溶液(9M)搅拌半小时,然后将1g Ti3AlC2缓慢加入上述混合溶液持续搅拌24小时,反应温度保持在45℃。所得样品反复离心(3500rpm)洗涤直至上清液PH≈7。之后样品通过超声浴处理1小时,再离心(8000rpm)取上清液,冻干即为MXene。
步骤2:聚己内酯(PCL)-MXene纤维的制备
PCL:MXene按质量比99:1溶于六氟异丙醇(HFIP)溶剂(8%w/v)并超声搅拌过夜获得纺丝液,再将5mL纺丝液加入纺丝注射器中进行静电纺丝,最后样品纺完后自然晾干。纺丝使用24G针头,速率为1mL/h,电压为15kV。
步骤3:PCL-MXene-CS-Asp纤维的制备
首先将3w/v%壳聚糖(CS)溶液浸滴在步骤4获得的PCL-MXene纤维表面,然后将浸滴后的纤维置于常温的条件下干燥,再将干燥后的PCL-MXene-CS纤维浸泡在500μg/mL阿司匹林(Asp)溶液中,最后晾干获得一种具有抗菌抗炎性能的生物高分子纤维伤口敷料PCL-MXene-CS-Asp。
实施例2
具有抗菌抗炎性能的生物高分子纤维伤口敷料按实施例1所述的制备方法进行制备,不同的是:
所述步骤1中,混合物在35℃下搅拌反应24小时,所得样品反复离心(4500rpm)洗涤直至上清液PH≈7。
所述步骤2中,左旋聚乳酸(PLLA):MXene按质量比95:5溶于HFIP溶剂(8%w/v)并超声搅拌过夜获得纺丝液。
所述步骤3中,干燥后的PLLA-MXene-CS纤维浸泡在400μg/mL布洛芬溶液中。
实施例3
具有抗菌抗炎性能的生物高分子纤维伤口敷料按实施例1所述的制备方法进行制备,不同的是:
所述步骤1中,混合物在55℃下搅拌反应24小时,所得样品反复离心(5500rpm)洗涤直至上清液PH≈7。
所述步骤2中,聚乙烯醇(PVA):MXene按质量比95:5溶于HFIP溶剂(8%w/v)并超声搅拌过夜获得纺丝液。纺丝使用24G针头,速率为1.5mL/h,电压为15kV。
所述步骤3中,干燥后的PVA-MXene-CS纤维浸泡在300μg/mL阿司匹林(Asp)溶液中。
对比例1
步骤1:聚己内酯(PCL)纤维的制备
PCL溶于HFIP溶剂(8%w/v)并超声搅拌过夜获得纺丝液,再将5mL纺丝液加入纺丝注射器中进行静电纺丝,最后样品纺完后自然晾干。纺丝使用24G针头,速率为1mL/h,电压为15kV。
步骤2:PCL-CS-Asp纤维的制备
首先将3w/v%壳聚糖(CS)溶液浸滴在步骤1获得的PCL纤维表面,然后将浸滴后的纤维置于常温的条件下干燥,再将干燥后的PCL-CS纤维浸泡在500μg/mL阿司匹林(Asp)溶液中,最后晾干获得一种具有抗菌性能的生物高分子纤维伤口敷料PCL-CS-Asp。
1、光热性能分析:
首先,将实施例1制备的PCL-CS、PCL-MXene-CS和实施例2制备的PCL-MXene-CS分别放在48孔板中,其中,以PCL所在实验组为对照组。然后向每个孔内加入500μL的磷酸缓冲盐溶液(PBS),再用近红外激光(808nm,1-2W/cm2)照射每孔至温度波动很小,并每隔15秒利用LIFR红外探测仪捕捉温度变化。
2、药物光热释放分析:
首先,将磷酸缓冲盐溶液(PBS)和含有阿司匹林(Asp)的PBS溶液(100、200、300、400、500μg/mL)进行吸光度(267nm)检测,并且绘制阿司匹林浓度标准曲线。再将实施例1制备的PCL-MXene-CS-Asp和对比例1制备的PCL-Asp、PCL-CS-Asp分别放在48孔板中,每个孔中加入500μL的PBS缓冲液。两个小时后吸取孔中的PBS,再加入新的PBS,进行10分钟的激光照射(808nm,1.5W/cm2)。照射后收集孔中的PBS,再添加新的PBS。以上操作循环两次后,将所得的PBS进行吸光度(267nm)检测,根据测算的阿司匹林浓度标准曲线计算阿司匹林的释放量。
3、抑菌分析:
将实施例1制备的PCL-CS、PCL-MXene、PCL-MXene-CS和对比例1制备的PCL放入48孔板中,再将200μL的大肠杆菌菌液(1×106CFU/mL)分别注入孔板内,然后将各样品对半分为光照组和非光照组,并对光照组进行近红外激光(808nm,1.5W/cm2)照射10分钟,剩下的非光照组放到黑暗之培养10分钟。最后抽取10μL菌液涂覆到LB固体培养基表面,并在37℃的环境下培养24小时后,观察计数培养基上的菌落数,进而分析材料的杀菌效果。
4、动物实验分析:
(1)伤口感染模型的建立:
将小鼠分组编号,并且记录体重(每一只体重测2次,记录2个数值)。小鼠背部脱毛后,用5%的水合氯醛(0.1mL/10g)腹腔注射麻醉,然后用手术剪等在脊柱外2cm处创建直径1cm的全层皮肤伤口(切到浅筋膜即可),再用移液枪滴加10μL的金黄色葡萄球菌菌液(1×108CFU/mL),最后贴无特殊成分创可贴或用无菌纱布包裹以防止小鼠舔舐,抓蹭伤口。
(2)伤口治疗分析:
感染模型建立24小时后记录体重(每一只体重测2次,记录2个数值)。拍摄伤口照片,并用直尺测量伤口的长与宽。依组分别在伤口处放置材料,所有的光照组均接受强度为1.5W/cm2的808nm的近红外光照10min。光照完后使伤口暴露在空气中。之后的五天每一天记录体重(每一只体重测2次,记录2个数值),并且拍摄伤口照片。
结果:
1、光热性能分析:
PCL-MXene-CS伤口敷料在近红外激光(808nm 1.5W/cm2)照射10分钟下能够升温到50℃左右,满足杀菌要求,而对照组PCL-CS敷料并未发产生显著的温度变化。
2、药物光热释放分析:
伤口敷料在10分钟的近红外激光照射(808nm,1.5W/cm2)下,药物释放量明显增多,实现了良好的药物控释作用。
3、抑菌分析:
对比培养24小时后的LB固体培养基上的菌落数,PCL-MXene-CS伤口敷料组在10分钟的近红外激光照射(808nm,1.5W/cm2)下菌落数最少,体现出明显的杀菌效果。
4、动物实验分析:
对于PCL-CS伤口敷料组,小鼠伤口感染后生长较慢;而PCL-MXene-CS-Asp伤口敷料组,通过光热治疗,小鼠伤口恢复迅速,生长较快,表明引入MXene和Asp后的伤口敷料能在体内有效发挥治疗作用。
综上,伤口敷料中MXene的引入有着良好的杀菌作用,而阿司匹林的引入则能够显著降低伤口的炎症反应促进伤口修复,本发明解决了在皮肤修复中滥用抗生素引发的耐药性和光热疗法引发的炎症问题
本发明的技术方案不限于上述具体实施例的限制,凡是根据本发明的技术方案做出的技术变形,均落入本发明的保护范围之内。

Claims (7)

1.一种生物高分子纤维伤口敷料,其特征在于:采用聚己内酯、左旋聚乳酸或聚乙烯醇的一种作为载体,负载MXene、壳聚糖和抗炎药物,组成具有抗菌抗炎性能的生物高分子纤维伤口敷料。
2.根据权利要求1所述的生物高分子纤维伤口敷料,其特征在于:所述抗炎药物为阿司匹林或布洛芬的一种。
3.一种生物高分子纤维伤口敷料制备方法,其特征在于,包括以下步骤:
S1:采用聚己内酯、左旋聚乳酸或聚乙烯醇的一种作为载体,MXene作为光敏抗菌剂,均匀混合后电纺,得到负载MXene的生物高分子纤维;
S2:然后浸滴壳聚糖溶液于生物高分子纤维表面;
S3:最后将生物高分子纤维浸泡在抗炎药物溶液中通过静电吸附作用完成药物负载,得到具有抗菌抗炎性能的生物高分子纤维伤口敷料。
4.根据权利要求3所述的生物高分子纤维伤口敷料制备方法,其特征在于:所述抗炎药物为阿司匹林或布洛芬的一种。
5.根据权利要求3所述的生物高分子纤维伤口敷料制备方法,其特征在于:所述MXene的制备方法为:将1g氟化锂粉末加入9M盐酸溶液20mL搅拌半小时,1g Ti3AlC2缓慢加入上述混合溶液持续搅拌24-36小时,反应温度保持在35-55℃;所得样品3500-5500rpm反复离心洗涤直至上清液PH≈7;之后样品通过超声浴处理1小时,再8000rpm离心取上清液,冻干即为MXene。
6.根据权利要求3所述的生物高分子纤维伤口敷料制备方法,其特征在于:所述负载MXene的生物高分子纤维的1制备方法具体为:聚己内酯、左旋聚乳酸或聚乙烯醇中的一种:MXene按质量比99:1或95:5或95:5溶于8%w/v的六氟异丙醇溶剂,并超声搅拌过夜获得纺丝液,再将纺丝液加入纺丝注射器中进行静电纺丝,最后样品纺完后自然晾干;纺丝使用24G针头,速率为1-1.5mL/h,电压为15kV。
7.根据权利要求3所述的生物高分子纤维伤口敷料制备方法,其特征在于:所述具有抗菌抗炎性能的生物高分子纤维的制备方法具体为:首先将3%w/v的壳聚糖溶液浸滴在负载MXene的生物高分子纤维表面,然后将浸滴后的生物高分子纤维置于常温下干燥,再将干燥后的生物高分子纤维浸泡在300-500μg/mL阿司匹林或布洛芬溶液中,最后晾干获得一种具有抗菌抗炎性能的生物高分子纤维伤口敷料。
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