CN113952114B - 具有图案化可释放气体的多层纳米纤维敷料及其制备方法 - Google Patents
具有图案化可释放气体的多层纳米纤维敷料及其制备方法 Download PDFInfo
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- CN113952114B CN113952114B CN202111122402.5A CN202111122402A CN113952114B CN 113952114 B CN113952114 B CN 113952114B CN 202111122402 A CN202111122402 A CN 202111122402A CN 113952114 B CN113952114 B CN 113952114B
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Abstract
本发明涉及生物材料和界面化学技术领域,尤其是涉及具有图案化可释放气体的多层纳米纤维敷料及其制备方法。所述多层纳米纤维伤口敷料包括一层或多层具有孔隙形貌可控的纳米纤维海绵以及一层或多层具有图案化的纳米纤维膜;所述图案化是指在一层或多层纳米纤维膜上的刻印多边形图案,并且通过紫外引发交联得到稳定的多层纳米纤维伤口敷料。不同层的纳米纤维功能性各不一样,结合气体治疗使敷料整体可以同时具有止血,抗菌,抗氧化,促进伤口愈合的功能,且具有良好的弹性性能。
Description
技术领域
本发明涉及生物材料和界面化学技术领域,尤其是涉及具有图案化可释放气体的多层纳米纤维敷料及其制备方法。
背景技术
慢性伤口的愈合给医疗保健***带来了巨大的负担。愈合过程由几个重叠的阶段组成,包括止血、炎症、增殖和重塑。炎症的延长被认为是慢性伤口延迟愈合的重要因素。为了度过炎症期,必须充分考虑慢性伤口的细菌负荷、组织坏死和水分平衡。
来自慢性伤口的大量渗出液中含有高水平的炎症介质和蛋白酶,这使治疗变得困难,也会使生物活性治疗分子失活和稀释,并破坏细胞外基质和伤口床。传统敷料的渗出液管理的依赖于亲水材料对渗出液的吸收和保留,如水凝胶、亲水纳米纤维、纤维素海绵等。然而,亲水性传统敷料与伤口长时间紧密接触,敷料中残留的过多渗出液会反向渗出到伤口周围,使伤口过度水化,损伤伤口周围组织,从而延迟愈合。为了克服这种困境,必须使用外科清创术,这可能会导致继发性创伤和剧烈疼。因此,非常需要具有高效渗出液管理能力的伤口敷料,以建立改善的伤口床环境。此外,利用多余渗出液触发治疗性气体分子的释放并且协同多功能生物活性分子的递送,有望在平衡伤口微环境的同时促进伤口愈合。
发明内容
针对上述技术问题,本发明的第一个目的在于提供一种具有表面图案化的具有气体释放能力的多层纳米纤维伤口敷料;本发明的第二个目的在于提供上述多层纳米纤维伤口敷料的制备方法,该方法简单易操作,塑形方便,制备得到的伤口敷料具有多层结构且不同层的纳米纤维材料物理性能或化学性质是不同的,可以大幅度减少伤口床体液的浸渍现象,此外,可以根据伤口情况经行抗炎抗菌处理。
本发明是通过以下技术方案实现的:
具有图案化可释放气体的多层纳米纤维敷料的制备方法,所述方法包括:
(1)制备具有触发气体释放能力的立体多级孔结构的纳米纤维海绵:
以壳聚糖、聚乙二醇、海藻酸盐、丝素蛋白、透明质酸钠或胶原蛋白中的至少两种为原料与沸石混合,进行静电纺丝,得到纳米纤维膜;
将所述纳米纤维膜经过均质乳化后成为分散的纳米纤维,将分散的纳米纤维按照一定冷冻速度和不同维度的冷冻方向冷冻一定时间,然后真空干燥后获得孔隙形貌可控的具有触发气体释放能力的立体多级孔结构的纳米纤维海绵;将所述纳米纤维海绵放入压力容器中,在一定压力下暴露于干燥的一氧化氮中,将一氧化氮封闭到纳米纤维海绵中,一氧化氮在湿润的环境中才会逸出,在后续制备过程中并不会逸出,后续将制备好的敷料放入干燥皿中即可保证气体在纳米纤维海绵中不逸出。
在上述步骤中,冷冻时间、冷冻速度和方向(包括单向或多向梯度)的不同会导致冰晶的形貌的不同,通过控制冷冻时间、冷冻速度、冷冻速度,根据冰晶的形貌进行重新排列得到具有冰晶形貌的多级孔纳米纤维海绵,真空干燥后即得到孔隙形貌可控的具有触发气体释放能力的立体多级孔结构的纳米纤维海绵。
优选地,将聚乙烯醇溶液和壳聚糖溶液均匀混合得到混合溶液后进行静电纺丝,得到纳米纤维膜;其中,将壳聚糖溶解于乙酸溶液中,得到所述壳聚糖溶液,在所述壳聚糖溶液中壳聚糖的质量浓度为3%-5%,所述壳聚糖的黏度为300-400mPa.s;将聚乙烯醇溶于水中,得到聚乙烯醇溶液,质量浓度为8%-10%;所述聚乙烯醇重均分子量为18~21万;以壳聚糖、聚乙二醇两种为原料与沸石混合得到混合溶液,进行静电纺丝,得到纳米纤维膜;所述混合溶液中,按质量百分含量包括聚乙烯醇溶液70%-80%、壳聚糖溶液20%-30%、沸石质量为1%-3%;
将纳米纤维膜进行均质乳化后的冷冻时间为2-5小时,冷冻速度为-20摄氏度每小时,方向为单向或多向梯度。
(2)在所述纳米纤维海绵上制备具有图案化的纳米纤维膜:将负载药物的聚合物及光引发剂进行溶解,获得静电纺丝溶液,所述负载药物的聚合物与制备所述纳米纤维海绵所采用的聚合物亲水性不同;以所述多级孔结构的纳米纤维海绵为图案模板,在纳米纤维海绵上方再次进行静电纺丝,可得到刻印有多边形图案的纳米纤维膜;
(3)在254nm-365nm波长的紫外线光照1-8小时后,刻印有多边形图案的纳米纤维膜和纳米纤维海绵发生交联,形成具有稳定结构的表面图案化的多层纳米纤维伤口敷料。
进一步地,步骤(1)中,将分散的纳米纤维按照冷冻速度-5~-50℃/min,冷冻方向可为单向或多向维度,冷冻时间6-48小时的条件进行冷冻;
其中,冷冻方向的控制是:将均质乳化后的纳米纤维混合物倒入冷冻铸造的模具中,模具对侧或四周分别有一个或多个冷却元件,结合加热器和热电偶装置控制冷却梯度,实现样品从一侧到对侧的单向冷冻,或者四周到内部多向维度冷冻。
进一步地,步骤(1)中,制备获得的所述纳米纤维海绵具有高弹性,压缩至非压缩状态下的体积的70%后,仍可恢复原状。
进一步地,步骤(1)中,所述纳米纤维海绵中的纳米纤维的直径为512nm-879nm。
进一步地,步骤(1)中,步骤(1)中,静电纺丝的具体技术参数为:电压14-28kv,针头到接收装置的距离8-16cm,沸石的尺寸为50-200nmm;将所述纳米纤维海绵在4-10个大气压下,暴露于干燥的一氧化氮1-10小时。
进一步地,步骤(1)中,是将所述纳米纤维膜剪成方块后浸入无水叔丁醇溶液后经过均质乳化后成为分散的纳米纤维;
所述纳米纤维膜的质量为1-50g,无水叔丁醇溶液的体积为10-1000ml。优选地,将所述纳米纤维膜剪成1×1cm的方块。
进一步地,步骤(2)中,采用的所述聚合物包括聚二甲基硅氧烷、聚氨酯、乳胶中的任意一种或多种;所述负载药物包括抗氧化和抗菌药物中的一种或多种,所述抗氧化和抗菌药物包括姜黄素、百里香酚、硫酸庆大霉素、氨苄西林、阿莫西林中的一种或多种;所述光引发剂为2-羟基-4'-(2-羟基乙氧基)-2-甲基苯丙酮或Irgacure 2959;
聚合物占静电纺丝溶液质量的5wt.%-20wt.%,光引发剂是聚合物质量的0.1wt.%-1wt.%。
进一步地,步骤(2)中,静电纺丝的具体技术参数为:电压20-28kv,针头到接收装置的距离5-13cm。
具有图案化可释放气体的多层纳米纤维敷料,所述多层纳米纤维伤口敷料包括一层或多层负载沸石的具有触发气体释放能力的立体多级孔结构的纳米纤维海绵以及一层或多层图案化的纳米纤维膜;所述图案化是指以特定冷冻模式获得的定制多孔结构的纳米纤维海绵为图案模板,在其上面印刻带有图案的一层或多层纳米纤维膜;
纳米纤维膜上的图案增加了纳米纤维膜和纳米纤维海绵之间的物理缠链,以减少伤口浸渍。
纳米纤维海绵、纳米纤维膜的物理性质和化学性质是不同的;物理性质的不同在于亲水性的不同,基于亲水性不同,渗出液能主动排离伤口部位,被纳米纤维海绵吸收,继而触发一氧化氮治疗性气体分子的释放;化学性质的不同包括负载药物不同或者纤维原料本身的化学性质不同。负载的药物包括姜黄素、百里香酚、或抗氧化或抗菌药物中的一种或多种;所述抗氧化和抗菌药物包括硫酸庆大霉素、氨苄西林、阿莫西林。
进一步地,所述纳米纤维海绵孔径大小为10-80um,孔形貌为蜂窝状、树枝状、层状和柱状中的内任意一种或多种。
本发明的有益技术效果:
本发明提供了一种具有气体释放能力的表面图案化的多层纳米纤维伤口敷料,其具有有效抗菌、抗氧化和渗出液控制特性。在没有外部驱动力的情况下,这种微图案化的纳米纤维气凝胶不仅可以自主智能单向转移液滴,而且具有更快的液体转移能力,并且在液体转移的同时触发一氧化氮治疗性气体的释放(水与沸石具有较强的亲和力,当水与负载有一氧化氮的沸石接触时,水分会争夺原本吸附一氧化氮的沸石孔道,从而触发一氧化氮释放)。这种快速单向吸液复合结构还可以防止已被吸收的渗出液再次浸渍伤口,有利于伤口恢复。
不同的方向和速度的冷冻可得到蜂窝状、树枝状、层状和柱状结构的纳米纤维海绵,利用光交联将图案化的纳米纤维稳定结合在纳米纤维海绵表面上。本发明提供的多层纳米纤维伤口敷料克服了传统纳米纤维在制备过程中的紧密堆积,这种具有可定制化立体多级孔结构的纳米纤维海绵具有增强的机械性能和液体吸收能力,同时保留了纳米纤维固有的柔软质地和细胞外基质样结构。
本发明提供的具有气体释放能力的表面图案化的多层纳米纤维伤口敷料结合具有抗炎、抗氧化和抗菌活性的生物活性分子可达到协同治疗的效果,实现加速通过慢性伤口的炎症期促进伤口愈合。
附图说明
图1为本发明实施例中具有图案化可释放气体的多层纳米纤维敷料的结构示意图。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白,以下结合实施例,对本发明进行进一步详细描述。应当理解,此处所描述的具体实施例仅仅用于解释本发明,并不用于限定本发明。
相反,本发明涵盖任何由权利要求定义的在本发明的精髓和范围上做的替代、修改、等效方法以及方案。进一步,为了使公众对本发明有更好的了解,在下文对本发明的细节描述中,详尽描述了一些特定的细节部分。对本领域技术人员来说没有这些细节部分的描述也可以完全理解本发明。
实施例1
本实施例提供了一种具有图案化可释放气体的多层纳米纤维敷料的制备方法:
(1)制备具有触发气体释放能力的立体多级孔结构的纳米纤维海绵:
(a)将0.2g黏度为400mPa.s的季铵盐壳聚糖加入10mL水中溶解,将1g重均分子量为1万聚乙烯醇溶于10mL水中搅拌溶解。
(b)季铵盐壳聚糖溶液和聚乙烯醇溶液按照重量比3:7混合后加入1%沸石搅拌均匀后,注入注射器中,调节注射器的推进速度为0.3mL/h,针头和接受装置的距离为13cm,二者之间的电压为20KV,电纺3个小时后得到0.1g的平面结构的纳米纤维膜。
(c)将累计得到的0.5g平面结构的纳米纤维膜剪裁为1cm×1cm放入盛有叔丁醇的烧杯中,再将烧杯放入均质乳化仪中,以10000rpm乳化10分钟。
(d)将乳化好的分散纳米纤维溶液放入方形模具中在下方、左方、右方为0摄氏度,上方为-80摄氏度的冷冻铸造装置中冷冻8小时后取出放入冷冻真空干燥机中干燥12小时,得到厚度为10mm,纤维直径为634nm的立体多级孔结构的纳米纤维海绵,纤维海绵在4个大气压下,暴露于干燥的一氧化氮8小时。
(2)在纳米纤维海绵上制备具有图案化的纳米纤维膜:
将0.8g重均分子量为35万聚乳酸和0.001g 2-羟基-4'-(2-羟基乙氧基)-2-甲基苯丙酮溶于10mL六氟异丙醇中搅拌溶解;
将聚乳酸溶液注入注射器中,调节注射器的推进速度为0.2mL/h,针头和的立体多级孔结构的纳米纤维海绵距离为8cm,二者之间的电压为25KV,电纺20分钟;
(3)在365nm紫外光下照射1小时,发生交联,形成具有稳定结构的表面图案化的多层纳米纤维伤口敷料(如图1所示)。
实施例2
本实施例提供了一种具有图案化可释放气体的多层纳米纤维敷料的制备方法:
(1)制备具有触发气体释放能力的立体多级孔结构的纳米纤维海绵:
(a)将0.2g黏度为300mPa.s的季铵盐壳聚糖加入10mL水中溶解,将1g重均分子量为8000聚乙烯醇溶于10mL水中搅拌溶解。
(b)季铵盐壳聚糖溶液和聚乙烯醇溶液按照重量比2:8混合后加入2%沸石搅拌均匀后注入注射器中,调节注射器的推进速度为0.3mL/h,针头和接受装置的距离为13cm,二者之间的电压为22KV,电纺3个小时后得到0.1g的平面结构的纳米纤维膜。
(c)将累计得到的0.5g平面结构的纳米纤维膜剪裁为1cm×1cm放入盛有叔丁醇的烧杯中,再将烧杯放入均质乳化仪中,以13000rpm乳化10分钟。
(d)将乳化好的分散纳米纤维溶液放入方形模具中在上方、下方、左方和右方都为-20摄氏度的冷冻铸造装置中冷冻8小时后取出放入冷冻真空干燥机中干燥12小时,得到厚度为10mm,纤维直径为685nm的立体多级孔结构的纳米纤维海绵,纤维海绵在5个大气压下,暴露于干燥的一氧化氮9小时。
(2)在纳米纤维海绵上制备具有图案化的纳米纤维膜:
将0.8g重均分子量为35万聚乳酸、0.01g硫酸庆大霉素和和0.002g 2-羟基-4'-(2-羟基乙氧基)-2-甲基苯丙酮溶于10mL六氟异丙醇中搅拌溶解。
将上述混合溶液注入注射器中,调节注射器的推进速度为0.2mL/h,针头和的立体多级孔结构的纳米纤维海绵距离为8cm,二者之间的电压为27KV,电纺20分钟;
(3)在254nm紫外光下照射7小时得到具有图案化的多层纳米纤维伤口敷料。
实施例3
本实施例提供了一种具有图案化可释放气体的多层纳米纤维敷料的制备方法:
(1)制备具有立体多级孔结构的纳米纤维海绵:
(a)将0.2g黏度为1000mPa.s的季铵盐壳聚糖加入10mL水中溶解,将1g重均分子量为8000聚环氧己烷溶于10mL水中搅拌溶解。
(b)季铵盐壳聚糖溶液和聚环氧己烷溶液按照重量比2:8混合后加入1.5%沸石搅拌均匀后,调节注射器的推进速度为0.3mL/h,针头和接受装置的距离为13cm,二者之间的电压为24KV,电纺3个小时后得到0.1g的平面结构的纳米纤维膜。
(c)将累计得到的0.8g平面结构的纳米纤维膜剪裁为1cm×1cm放入盛有叔丁醇的烧杯中,再将烧杯放入均质乳化仪中,以13000rpm乳化10分钟。
(d)将乳化好的分散纳米纤维溶液放入方形模具中在上方、下方、左方和右方都为-80摄氏度的冷冻铸造装置冷冻8小时后取出放入冷冻真空干燥机中干燥12小时,得到厚度为10mm,纤维直径为685nm的立体多级孔结构的纳米纤维海绵,纤维海绵在7个大气压下,暴露于干燥的一氧化氮4小时。
(2)在所述纳米纤维海绵上制备具有图案化的纳米纤维膜:
将0.8g重均分子量为35万聚乳酸、0.02g姜黄素和0.001g Irgacure 2959溶于10mL六氟异丙醇中搅拌溶解。
将上述混合溶液注入注射器中,调节注射器的推进速度为0.2mL/h,针头和的立体多级孔结构的纳米纤维海绵距离为8cm,二者之间的电压为26KV,电纺20分钟;
(3)在365nm紫外光下照射2小时得到具有图案化的多层纳米纤维伤口敷料。
实施例4
本实施例提供了一种具有图案化可释放气体的多层纳米纤维敷料的制备方法:
(1)制备具有触发气体释放能力的立体多级孔结构的纳米纤维海绵:
(a)将0.2g黏度为1000mPa.s的季铵盐壳聚糖加入10mL水中溶解,将1g重均分子量为8000聚环氧己烷溶于10mL水中搅拌溶解。
(b)季铵盐壳聚糖溶液和聚环氧己烷溶液按照重量比2:8混合后加入1%沸石搅拌均匀后注入注射器中,调节注射器的推进速度为0.3mL/h,针头和接受装置的距离为13cm,二者之间的电压为24KV,电纺3个小时后得到0.1g的平面结构的纳米纤维膜。
(c)将累计得到的0.8g平面结构的纳米纤维膜剪裁为1cm×1cm放入盛有叔丁醇的烧杯中,再将烧杯放入均质乳化仪中,以13000rpm乳化10分钟。
(d)将乳化好的分散纳米纤维溶液放入方形模具中在右方为-20摄氏度,上方和下方为-60摄氏度的冷冻铸造装置中冷冻8小时后取出放入冷冻真空干燥机中干燥12小时,得到厚度为10mm,纤维直径为685nm的立体多级孔结构的纳米纤维海绵,纤维海绵在6个大气压下,暴露于干燥的一氧化氮9小时。
(2)在所述具有立体多级孔结构的纳米纤维海绵上制备具有图案化的纳米纤维膜:
将0.8g重均分子量为35万聚乳酸、0.01g没食子酸和0.003g Irgacure 2959溶于10mL六氟异丙醇中搅拌溶解。
将上述混合溶液注入注射器中,调节注射器的推进速度为0.2mL/h,针头和的立体多级孔结构的纳米纤维海绵距离为8cm,二者之间的电压为28KV,电纺20分钟;
(3)在365nm紫外光下照射3小时得到具有图案化的多层纳米纤维伤口敷料。
Claims (6)
1.具有图案化可释放气体的多层纳米纤维敷料的制备方法,其特征在于,所述方法包括:
(1)制备具有触发气体释放能力的立体多级孔结构的纳米纤维海绵:
以壳聚糖、聚乙二醇、海藻酸盐、丝素蛋白、透明质酸钠或胶原蛋白中的至少两种为原料与沸石混合,进行静电纺丝,得到纳米纤维膜;
将所述纳米纤维膜经过均质乳化后成为分散的纳米纤维,将分散的纳米纤维按照一定冷冻速度和不同维度的冷冻方向冷冻一定时间,然后真空干燥后获得孔隙形貌可控的具有触发气体释放能力的立体多级孔结构的纳米纤维海绵;制备获得的所述纳米纤维海绵具有高弹性,压缩至非压缩状态下的体积的70%后,仍可恢复原状;
将所述纳米纤维海绵放入压力容器中,在一定压力下暴露于干燥的一氧化氮中,将一氧化氮封闭到纳米纤维海绵中,一氧化氮在湿润的环境中才会逸出,在后续制备过程中并不会逸出;
其中,将分散的纳米纤维按照冷冻速度-5~-50℃/min,冷冻方向可为单向或多向维度,冷冻时间6-48小时的条件进行冷冻;
其中,冷冻方向的控制是:将均质乳化后的纳米纤维混合物倒入冷冻铸造的模具中,模具对侧或四周分别有一个或多个冷却元件,结合加热器和热电偶装置控制冷却梯度,实现样品从一侧到对侧的单向冷冻,或者四周到内部多向维度冷冻;
(2)在所述纳米纤维海绵上制备具有图案化的纳米纤维膜:将负载药物的聚合物及光引发剂进行溶解,获得静电纺丝溶液,所述负载药物的聚合物与制备所述纳米纤维海绵所采用的聚合物亲水性不同;以所述多级孔结构的纳米纤维海绵为图案模板,在纳米纤维海绵上方再次进行静电纺丝,可得到刻印有多边形图案的纳米纤维膜;
(3)在254nm-365nm波长的紫外线光照1-8小时后,刻印有多边形图案的纳米纤维膜和纳米纤维海绵发生交联,形成具有稳定结构的表面图案化的多层纳米纤维伤口敷料;
步骤(1)中,静电纺丝的具体技术参数为:电压14-28kv,针头到接收装置的距离8-16cm,沸石的尺寸为50-200nmm;将所述纳米纤维海绵在4-10个大气压下,暴露于干燥的一氧化氮1-10小时;
步骤(2)中,静电纺丝的具体技术参数为:电压20-28kv,针头到接收装置的距离5-13cm。
2.根据权利要求1所述具有图案化可释放气体的多层纳米纤维敷料的制备方法,其特征在于,步骤(1)中,所述纳米纤维海绵中的纳米纤维的直径为512nm-879 nm。
3.根据权利要求1所述具有图案化可释放气体的多层纳米纤维敷料的制备方法,其特征在于,步骤(1)中,是将所述纳米纤维膜剪成方块后浸入无水叔丁醇溶液后经过均质乳化后成为分散的纳米纤维。
4.根据权利要求1所述具有图案化可释放气体的多层纳米纤维敷料的制备方法,其特征在于,步骤(2)中,采用的所述聚合物包括聚二甲基硅氧烷、聚氨酯、乳胶中的任意一种或多种;所述负载药物包括抗氧化和抗菌药物中的一种或多种,所述抗氧化和抗菌药物包括姜黄素、百里香酚、硫酸庆大霉素、氨苄西林、阿莫西林中的一种或多种;所述光引发剂为2-羟基-4'-(2-羟基乙氧基)-2-甲基苯丙酮或Irgacure 2959;聚合物占静电纺丝溶液质量的5 wt.%-20 wt.%,光引发剂是聚合物质量的0.1 wt.%-1 wt.% 。
5.具有图案化可释放气体的多层纳米纤维敷料,其特征在于,所述多层纳米纤维伤口敷料包括一层或多层负载沸石的具有触发气体释放能力的立体多级孔结构的纳米纤维海绵以及一层或多层图案化的纳米纤维膜;所述图案化是指以特定冷冻模式获得的定制多孔结构的纳米纤维海绵为图案模板,在其上面印刻带有图案的一层或多层纳米纤维膜;
纳米纤维膜上的图案增加了纳米纤维膜和纳米纤维海绵之间的物理缠链,以减少伤口浸渍;
纳米纤维海绵、纳米纤维膜的物理性质和化学性质是不同的;物理性质的不同在于亲水性的不同,基于亲水性不同,渗出液能主动排离伤口部位,被纳米纤维海绵吸收,继而触发一氧化氮治疗性气体分子的释放;化学性质的不同包括负载药物不同或者纤维原料本身的化学性质不同。
6.根据权利要求5所述具有图案化可释放气体的多层纳米纤维敷料,其特征在于,所述纳米纤维海绵孔径大小为10-80um,孔形貌为蜂窝状、树枝状、层状和柱状中的内任意一种或多种。
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