CN110772379A - 一种负载纳米酶的复合纳米纤维膜的制备方法及其创面敷贴 - Google Patents
一种负载纳米酶的复合纳米纤维膜的制备方法及其创面敷贴 Download PDFInfo
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Abstract
本发明公开了一种负载纳米酶的复合纳米纤维膜的制备方法及其创面敷贴,制备方法包括(1)将0.1‑0.5重量份的纳米酶溶解于100重量份的质量分数为5‑15%的聚乙烯醇‑丙烯酰胺接枝共聚物的水溶液中,配制成纺丝溶液;(2)将制得的纺丝溶液采用静电纺丝方法纺成纳米纤维膜;(3)将制得的纳米纤维膜在120‑150℃温度下交联处理15‑30min,即得所述负载纳米酶的复合纳米纤维膜。本申请提供的复合纳米纤维膜及创面敷贴能够防止疮面痂皮形成,不会粘连新生成的肉芽组织,避免换药时引起疼痛;有利于纤维蛋白及坏死组织的溶解,减少更换次数;创造低氧环境,促进毛细血管生成;促进多种生长因子释放并发挥活性;缓解创面疼痛,减少瘢痕形成。
Description
技术领域
本发明涉及一种负载纳米酶的复合纳米纤维膜的制备方法及其创面敷贴,属于合成纤维技术领域。
背景技术
手术、外伤、烧伤、皮炎等均会导致皮肤破坏和缺损,临床上常采用药物加敷料进行皮肤创面护理。传统的敷料一般由棉花、软麻布或亚麻布加工而成,这类敷料存在的问题有:不能保持创面湿润,干性环境易使伤口表面形成结痂,结痂迫使表皮细胞的迁移绕经痂下,延迟创面愈合;敷料纤维易脱落,容易造成异物反应,影响创面愈合;敷料被浸透后屏蔽作用差,病原体易通过敷料到达创面,引起外源性感染;创面肉芽组织易长入敷料的网眼中,创面与敷料粘连,换药时损伤新生的肉芽组织并引起疼痛;使用不方便,换药工作量大等。
为解决传统敷料存在的上述问题,人们研制出了新型敷料,这类新型敷料大部分是通过在创面形成密闭微环境来隔绝外界细菌感染及吸收伤口的渗出液,虽然解决了吸收创面渗液,维持创面干燥,保护创面不受外界细菌感染等问题,但还仅是属于简单的物理保护,未能解决创面愈合慢等问题。
需要说明的是,上述内容属于发明人的技术认知范畴,并不必然构成现有技术。
发明内容
本发明的目的在于解决现有技术所存在的问题,通过提供一种负载纳米酶的复合纳米纤维膜的制备方法及其创面敷贴,可促进伤口愈合,缓解创面疼痛。
一方面,本发明提供了一种负载纳米酶的复合纳米纤维膜的制备方法,所述纳米酶选自四氧化三铁、四氧化三钴、四氧化三锰中的任意一种或多种,所述制备方法包括如下步骤:
(1)将0.1-0.5重量份的纳米酶溶解于100重量份的质量分数为5-15%的聚乙烯醇-丙烯酰胺接枝共聚物的水溶液中,配制成纺丝溶液;
(2)将步骤(1)制得的纺丝溶液采用静电纺丝方法纺成纳米纤维膜;
(3)将步骤(2)制得的纳米纤维膜在120-150℃温度下交联处理15-30min,即得所述负载纳米酶的复合纳米纤维膜。
在优选的实施方式中,所述纳米酶的粒径为5-50nm。
在优选的实施方式中,所述负载纳米酶的复合纳米纤维膜的厚度为0.1-0.2mm。
在优选的实施方式中,所述聚乙烯醇-丙烯酰胺接枝共聚物的接枝率为45-55%。
在优选的实施方式中,所述步骤(1)中纳米酶的溶解方法为:将纳米酶加入到聚乙烯醇-丙烯酰胺接枝共聚物的水溶液中,超声震荡10-15h。
在优选的实施方式中,所述纺丝溶液中,纳米酶与聚乙烯醇-丙烯酰胺接枝共聚物通过静电吸引作用自组装在一起。
在优选的实施方式中,所述静电纺丝方法为无针静电纺丝。
在优选的实施方式中,所述静电纺丝方法的条件为:纺丝电压为3.2-3.5万伏,接收距离为150-180mm,环境温度为15-25℃,环境湿度为25-35%,溶液盒基材走步速度为0.02-0.03m/min。
所述负载纳米酶的复合纳米纤维膜的制备方法得到的复合纳米纤维膜及其在制备创面敷料中的应用。
另一方面,本申请还提供了一种创面敷贴,包括依次设置的医用胶带、吸收垫和复合纳米纤维膜。
本申请的有益效果包括但不限于:
(1)本申请得到的纳米纤维膜结构类似于细胞外基质结构,可以为细胞的黏附提供支撑点,便于营养物质和代谢废物的运输,使细胞更加舒展,有利于细胞的生长,促进伤口愈合;纳米纤维膜具有良好的生物相容性、可控的降解性和可吸收性,与创面接触时不易造成过敏及感染;纳米纤维膜具有三维网状结构,透气且孔径小,能够保持创面湿润的微环境,维持创缘到创面中央正常的电势梯度,促使更多生长因子受体与生长因子结合,保持细胞活力,促进修复细胞生长;湿润环境可以加快表皮细胞迁移速度,缩短愈合时间;密闭湿润环境有利于纳米纤维膜上负载的纳米酶发挥酶学清创作用,促进纤维蛋白和坏死组织的溶解,加速创面愈合,刺激细胞增殖,促进生长因子的释放,增强白细胞功能;密闭环境有效隔绝了外界细菌的侵入,防止感染创面细菌传播而造成的感染,低氧或无氧、微酸的愈合环境可以抑制伤口中细菌的生长、促进成纤维细胞的生长、刺激毛细血管增生。
(2)纳米酶具有良好的抗菌功能,且通过自身所具有的氧化还原酶的催化活性可以调节ROS自由基水平,大量ROS自由基进入细菌内可以切断核酸,使蛋白质失活,破坏细胞膜完整性,在生物膜基质中可以降解多种分子包括多糖、蛋白质、胞DNA和脂类,从而达到杀灭多种耐药菌的效果,可以应用于伤口愈合过程中达到抗菌效果防止创面感染;活性ROS与创面炎症过程高度相关,ROS浓度提高可以达到更好的抗菌效果,但ROS浓度过高导致的氧化应激同样会造成正常生物分子的损害,所以要保持体内ROS平衡。纳米酶具有SOD模拟酶和CAT模拟酶催化活性能清除体内高浓度的ROS,来保持体内ROS平衡达到消炎的作用;纳米酶通过化学合成,制备快速简单成本低,稳定性好;通过控制纳米尺度的要素可以调节和优化纳米酶的活性;纳米酶之间可以协同工作增强抗菌效果;功能化修饰和多种纳米效应联用进一步增强其杀菌能力。
综上,本申请提供的复合纳米纤维膜及创面敷贴,纳米酶的负载量高,能够防止疮面痂皮形成,不会粘连新生成的肉芽组织,避免换药时引起疼痛;有利于纤维蛋白及坏死组织的溶解,减少更换次数;创造低氧环境,促进毛细血管生成;促进多种生长因子释放并发挥活性;缓解创面疼痛,减少瘢痕形成。
附图说明
此处所说明的附图用来提供对本申请的进一步理解,构成本申请的一部分,本申请的示意性实施例及其说明用于解释本申请,并不构成对本申请的不当限定。在附图中:
图1为本申请提供的创面敷贴的结构示意图;
图2为本申请实施例1制备的复合纳米纤维不同放大倍数的扫描电镜图;
图3为本申请实施例1制备的复合纳米纤维的直径分布图;
图4本申请实施例1制备的复合纳米纤维的细胞毒性试验结果。
具体实施方式
在以下内容中将会对本发明进行进一步的详细描述。但是需要指出的是,以下的具体实施方式仅仅以示例性的方式给出本发明的具体操作实例,但是本发明的保护范围不仅限于此。本发明的保护范围仅仅由权利要求书所限定。本领域技术人员能够显而易见地想到,可以在本发明权利要求书限定的保护范围之内对本发明所述的实施方式进行各种其它的改良和替换,并且仍然能够实现相同的技术效果,达到本发明的最终技术目的。
在本发明中,室温指实验室内常规的环境温度,随季节和位置变化,通常为25℃。
本申请提供的负载纳米酶的复合纳米纤维膜的制备方法,制备方法采用如下步骤:
(1)将0.1-0.5重量份的纳米酶溶解于100重量份的质量分数为5-15%的聚乙烯醇-丙烯酰胺接枝共聚物的水溶液中,配制成纺丝溶液;
其中,纳米酶选自四氧化三铁、四氧化三钴、四氧化三锰中的任意一种或多种,粒径为5-50nm;聚乙烯醇-丙烯酰胺接枝共聚物的接枝率为45-55%;
进一步的,在纺丝溶液中,纳米酶与聚乙烯醇-丙烯酰胺接枝共聚物中的氨基作用,通过静电吸引作用自组装在一起,增加了四氧化三铁的溶解度及溶液稳定性,纳米酶与聚乙烯醇-丙烯酰胺接枝共聚物的混悬液更稳定,纺出的纤维膜中纳米酶含量高。
进一步的,纳米酶加入到聚乙烯醇-丙烯酰胺接枝共聚物的水溶液中后,超声震荡10-15h。
(2)将步骤(1)制得的纺丝溶液采用静电纺丝方法纺成厚度为0.1-0.2mm的纳米纤维膜;具体的,静电纺丝方法为无针静电纺丝,静电纺丝方法的条件为:纺丝电压为3.2-3.5万伏,接收距离为150-180mm,环境温度为15-25℃,环境湿度为25-35%,溶液盒基材走步速度为0.02-0.03m/min。
(3)将步骤(2)制得的纳米纤维膜在120-150℃温度下交联处理15-30min,即得所述负载纳米酶的复合纳米纤维膜。
如图1所示,本申请还提供了一种创面敷贴,包括依次设置的医用胶带、吸收垫和复合纳米纤维膜。其中,吸水垫为涤纶材料的吸水棉,能够吸收组织渗液保持创面微环境,医用胶带表面涂覆压敏胶,起固定作用。
下面通过具体实施例对本发明的进行详细介绍。如未特殊说明,以下各实施例中,所用的原料均可通过商业途径购得。
实施例1:
(1)选取17-88型医用级聚乙烯醇,配制成质量分数为15%的聚乙烯醇水溶液;
向聚乙烯醇水溶液中加入浓度为0.04mol/L的K2S2O8,室温下搅拌使K2S2O8充分溶解与聚乙烯醇混合均匀得混合溶液,K2S2O8使聚乙烯醇侧羟基上的氢脱落,形成活性中心,在活性中心引发丙烯酰胺的聚合;
向混合溶液中加入质量分数为15%的丙烯酰胺水溶液,在40℃下反应4h得产物溶液,加入的丙烯酰胺与聚乙烯醇的摩尔比6∶1;
反应结束后,将产物溶液用丙酮洗涤,除去未反应的小分子;抽滤后取有机固相,55℃条件下恒温烘干至恒质量;再用二甲基亚砜洗涤,除去有机相中的丙烯酰胺均聚物;抽滤取滤液,再用丙酮对滤液进行洗涤,沉淀出聚乙烯醇-丙烯酰胺接枝共聚物;抽滤,将所得聚乙烯醇-丙烯酰胺接枝共聚物在55℃下恒温烘干。
(2)取100mg粒径为15-25nm的四氧化三铁纳米酶,加入100ml质量分数为15%的聚乙烯醇-丙烯酰胺接枝共聚物水溶液中,超声震荡12h,配制成纺丝溶液;
(3)采用无针静电纺丝方法将纺丝溶液采用静电纺丝方法纺成厚度为0.1-0.2mm的纳米纤维膜,纺丝电压为3.2万伏,接收距离为150mm,环境温度为15℃,环境湿度为25%,溶液盒基材走步速度为0.02m/min;
(4)将制得的复合纳米纤维膜在150℃环境下交联处理25min,调整纤维膜断裂伸长率,获得所需交联度的复合纳米纤维膜。
实施例2:
(1)取100mg粒径为10-25nm的四氧化三铁纳米酶,加入100ml质量分数为12%的聚乙烯醇-丙烯酰胺接枝共聚物水溶液中,超声震荡15h,配制成纺丝溶液;
(2)采用无针静电纺丝方法将纺丝溶液采用静电纺丝方法纺成厚度为0.1-0.2mm的纳米纤维膜,纺丝电压为3.5万伏,接收距离为180mm,环境温度为25℃,环境湿度为35%,溶液盒基材走步速度为0.03m/min;
(3)将制得的复合纳米纤维膜在120℃环境下交联处理15min,获得所需交联度的复合纳米纤维膜。
本实施例与实施例1中聚乙烯醇-丙烯酰胺接枝共聚物的制备方法相同。
实施例3:
(1)取100mg粒径为10-25nm的四氧化三铁纳米酶,加入100ml质量分数为10%的聚乙烯醇-丙烯酰胺接枝共聚物水溶液中,超声震荡10h,配制成纺丝溶液;
(2)采用无针静电纺丝方法将纺丝溶液采用静电纺丝方法纺成厚度为0.1-0.2mm的纳米纤维膜,纺丝电压为3.3万伏,接收距离为155mm,环境温度为20℃,环境湿度为30%,溶液盒基材走步速度为0.03m/min;
(3)将制得的复合纳米纤维膜在130℃环境下交联处理30min,获得所需交联度的复合纳米纤维膜。
本实施例与实施例1中聚乙烯醇-丙烯酰胺接枝共聚物的制备方法相同。
结构观察:
如图2所示,可以观察到实施例1得到的复合纳米纤维膜呈三维网状分布,无明显粘连的现象,有利于物质的传输和扩散,作为创面敷料时具有良好的通透性。
如图3所示,可以观察到实施例1得到的复合纳米纤维膜纤维直径呈现正态分布,直径稳定。
细胞毒性实验:
按照表1设定各组别,参照《GB/T16886.5-2017医疗器械生物学评价第5部分:体外细胞毒性试验》中MTT法,对各组对象进行测试。
表1
空白组 | 不含实验样品的细胞培养液 |
实验组 | 实施例1得到的复合纳米纤维膜浸提液 |
阴性对照组 | 聚乙烯膜浸提液 |
阳性对照组 | 10%DMSO(Sigma,批号:RNBC9663)溶液 |
如表2所示,在显微镜下24h观察实验组、阴性对照组绝大部分细胞形态正常,阳性对照组细胞层几乎完全破坏,与空白组相比实验组细胞存活率为73%,说明实施例1得到的复合纳米纤维膜纤维无潜在细胞毒性。
表2
对大鼠皮肤缺损的治疗效果:
根据表3所示,设定空白组和对照组1-4。
表3
采用大鼠全皮层切除的造模方法,使用各组纤维膜对伤口进行处理,每天观察伤口恢复情况,在第6、9、13天拍照及测量创面愈合率。
如图4所示,通过观察动物模型创面恢复情况及计算创面恢复率发现与对照组相比,实施例1得到的复合纳米纤维膜促进创面愈合效果高于其他组,具有明显的促进伤口愈合作用;图中,*表示与空白组有显著性差异,+表示聚乙烯醇接枝物与纳米酶复合组与单纯聚乙烯醇与纳米酶混合具有显著性差异。
上述具体实施方式不能作为对本发明保护范围的限制,对于本技术领域的技术人员来说,对本发明实施方式所做出的任何替代改进或变换均落在本发明的保护范围内。
本发明未详述之处,均为本技术领域技术人员的公知技术。
Claims (10)
1.一种负载纳米酶的复合纳米纤维膜的制备方法,其特征在于,所述纳米酶选自四氧化三铁、四氧化三钴、四氧化三锰中的任意一种或多种,所述制备方法包括如下步骤:
(1)将0.1-0.5重量份的纳米酶溶解于100重量份的质量分数为5-15%的聚乙烯醇-丙烯酰胺接枝共聚物的水溶液中,配制成纺丝溶液;
(2)将步骤(1)制得的纺丝溶液采用静电纺丝方法纺成纳米纤维膜;
(3)将步骤(2)制得的纳米纤维膜在120-150℃温度下交联处理15-30min,即得所述负载纳米酶的复合纳米纤维膜。
2.根据权利要求1所述的负载纳米酶的复合纳米纤维膜的制备方法,其特征在于,所述纳米酶的粒径为5-50nm。
3.根据权利要求1所述的负载纳米酶的复合纳米纤维膜的制备方法,其特征在于,所述负载纳米酶的复合纳米纤维膜的厚度为0.1-0.2mm。
4.根据权利要求1所述的负载纳米酶的复合纳米纤维膜的制备方法,其特征在于,所述聚乙烯醇-丙烯酰胺接枝共聚物的接枝率为45-55%。
5.根据权利要求1所述的负载纳米酶的复合纳米纤维膜的制备方法,其特征在于,所述步骤(1)中纳米酶的溶解方法为:将纳米酶加入到聚乙烯醇-丙烯酰胺接枝共聚物的水溶液中,超声震荡10-15h。
6.根据权利要求1所述的负载纳米酶的复合纳米纤维膜的制备方法,其特征在于,所述纺丝溶液中,纳米酶与聚乙烯醇-丙烯酰胺接枝共聚物通过静电吸引作用自组装在一起。
7.根据权利要求1所述的负载纳米酶的复合纳米纤维膜的制备方法,其特征在于,所述静电纺丝方法为无针静电纺丝。
8.根据权利要求7所述的负载纳米酶的复合纳米纤维膜的制备方法,其特征在于,所述静电纺丝方法的条件为:纺丝电压为3.2-3.5万伏,接收距离为150-180mm,环境温度为15-25℃,环境湿度为25-35%,溶液盒基材走步速度为0.02-0.03m/min。
9.根据权利要求1-8任一项所述的负载纳米酶的复合纳米纤维膜的制备方法得到的复合纳米纤维膜及其在制备创面敷料中的应用。
10.一种创面敷贴,其特征在于,包括依次设置的医用胶带、吸收垫和复合纳米纤维膜,所述复合纳米纤维膜由权利要求1-8任一项所述的制备方法制备得到。
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