CN108042838A - 一种抗氧化型纳米纤维电纺膜医用敷料的制备方法 - Google Patents
一种抗氧化型纳米纤维电纺膜医用敷料的制备方法 Download PDFInfo
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Abstract
本发明提供一种抗氧化型纳米纤维电纺膜医用敷料的制备方法。所述医用敷料具体制备如下:将三氯甲烷与二甲基甲氨酰混合配置成混合溶剂,并将PCL颗粒溶解于上述混合溶剂中配置成PCL溶液,然后进行静电纺丝得到PCL纳米纤维电纺膜;取胶原蛋白粉末加到乙酸溶液中制成胶状均质溶液A,取Nac溶解于去离子水中得到溶液B,并将溶液A与溶液B混匀超声制成即为载有抗氧化药物Nac的胶原蛋白溶液,将其离心处理去除气泡后用加样枪均匀添加到PCL纳米纤维电纺膜上,依次经过冻干、交联剂溶液浸泡、冷冻干燥得到所述医用敷料。本发明制备的PCL纳米纤维电纺膜具有促进伤口愈合的能力,载有Nac药物的胶原蛋白作为抗氧化药物,有助于加强PCL膜促进愈合的作用。
Description
技术领域
本发明提供一种的医用敷料,具体说是一种抗氧化型纳米纤维电纺膜医用敷料的制备方法。
背景技术
创面愈合困难是整形外科中经常会遇到的问题。临床上对于小型创面处理方式主要通过活力碘消毒后,予以喷涂人表皮生长因子等药物,最后覆盖无菌纱布。对于大型创面则主要采用皮片、皮瓣移植手术治疗后,表面覆盖凡士林纱布,最后覆盖无菌纱布。以上处理方式虽已较为成熟,但仍然存在以下不足:(1)药物难以长期于伤口处保持有效浓度,无法稳定、持续发挥药物作用;(2)医用脱脂纱布的生物相容性稍差,无可降解性,需要以3天/次的频率更换。
发明内容
本发明根据现有技术的不足,提供一种抗氧化型纳米纤维电纺膜医用敷料的制备方法,该方法制备出来的抗氧化型纳米纤维电纺膜材质的医用敷料可以解决目前医用敷料药物作用时间短,药物作用效果差以及敷料自身的生物相容性差,无可降解性等问题。
本发明提供的技术方案:一种抗氧化型纳米纤维电纺膜医用敷料的制备方法,其特征在于具体步骤如下:
(1)将三氯甲烷与二甲基甲氨酰以8.5~9.5:1比例配置成混合溶剂,将聚己内酯(PCL)颗粒作为溶质溶解于上述混合溶剂中,配置成质量分数为20~24%的PCL溶液;
(2)将步骤(1)中制成的PCL溶液进行静电纺丝得到PCL纳米纤维电纺膜,然后经真空干燥处理后置于紫外线下照射2~4h,再采用75%酒精内浸泡2~6h,最后用PBS缓冲液(磷酸盐缓冲液)对材料进行反复冲洗5-6次后置于无菌干燥容器中;
(3)取胶原蛋白粉末,加入到浓度为0.05mmol/L的乙酸溶液中,并在0℃冰水浴下超声8-12min,直至溶解成制备成胶原蛋白质量百分比为2~4%的胶状均质溶液A;取N-乙酰半胱氨酸(Nac)溶解于去离子水中得到浓度为0.5g/L的溶液B;将胶状均质溶液A与溶液B按照质量比为1:0.8~1.2混匀后于0℃冰水浴下超声25-35min至均质胶状溶液,即为载有抗氧化药物Nac的胶原蛋白溶液;
(4)将步骤(3)中制备的载有抗氧化药物Nac的胶原蛋白溶液经过离心处理后去除气泡,并于2~6℃环境下静置30~60min致使其成凝胶状,然后将其用加样枪均匀添加到步骤(2)中的PCL纳米纤维电纺膜上,并置于-20~-26℃环境下冷冻4~6h,之后取出置于冻干机中冻干;
(5)取1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)、N-羟基琥珀酰亚胺(NHS)加入95%乙醇溶液中现场配制交联液,其交联液中EDC的含量为50mmol/L,NHS的含量为12.5mmol/L;然后将步骤(4)中涂有胶原蛋白溶液的膜片材料在室温下浸泡至交联剂溶液中6~8h;
(6)将经过交联剂溶液浸泡后膜片材料用去离子水反复冲洗多次后晾干,再置于-20~-26℃环境下冷冻4~6h后,再通过冻干机冻干,即可得到抗氧化型PCL纳米纤维电纺膜。
本发明较优的技术方案:所述步骤(8)中制备的抗氧化型PCL纳米纤维电纺膜用环氧乙烷37℃条件下进行消毒灭菌处理后储存。
本发明较优的技术方案:所述步骤(2)中的PCL纳米纤维电纺膜在依次经过紫外线照射、酒精浸泡和冲洗后裁剪成3~5cm*3~5cm大小的膜片,分装于无菌干燥的容器中;
本发明较优的技术方案:所述步骤(2)中的静电纺丝的具体参数:流速1.3μl/min,电压6~7kV,距离15cm。
本发明较优的技术方案:所述步骤(3)中的溶液B配制过程中,将溶解温度限定为35-38℃。
本发明较优的技术方案:所述步骤(4)中胶原蛋白溶液的离心条件为:离心转速2000~2500r/min,离心时间5~6min。
本发明较优的技术方案:所述步骤(4)和步骤(6)中的冻干机的工作条件为温度-42~-53℃,压力为5~20Pa,时间为18~24h。
本发明较优的技术方案:所述步骤(6)中经过交联剂溶液浸泡后膜片材料采用去离子水反复冲洗5~6次,每次5~10min,然后取出材料置于滤纸上晾干后。
本发明采用静电纺丝技术制作出PCL(Polycaprolactone,聚己内酯)纳米纤维电纺膜,同时,将PCL纳米纤维电纺膜与载有Nac(N-Acetyl-L-Cysteine,N-乙酰半胱氨酸)抗氧化药物的胶原蛋白进行交联后制成的抗氧化性纳米纤维电纺膜材质的医用敷料,具体具有以下优点:
(1)本发明中的膜是纳米级别的高分子材料可模拟天然的细胞外基质的结构和生物功能,有助于促进伤口愈合;
(2)本发明制备的敷料采用PCL膜,拥有比表面积大、孔隙率高、良好的生物相容性及可降解性等特性;对病人而言舒适度高、异物感小,表面具有纳米级别的细孔,有助于表面皮肤与外界进行液、气交换;
(3)本发明中的抗氧化药物Nac是一种常用的抗氧化剂,它可防止细胞受到体内外氧自由基和各种细胞毒素的损害,促进血管增生并加强血管功能,运用这种载药胶原可加强敷料促进伤口愈合的作用。
(4)本发明将载有抗氧化药物Nac的胶原蛋白与PCL膜进行交联后,可以长时间作用于伤口,加强促进伤口愈合的作用;抗氧化型PCL纳米纤维电纺膜的生物相容性好,具有良好的可降解性,不易诱发伤口的炎症反应。
本发明制成的抗氧化型PCL纳米纤维电纺膜材质的生物敷料,制备过程简单,易于操作,成本低,对仪器要求不高,同时,PCL纳米纤维电纺膜本身具有促进伤口愈合的能力,载有Nac药物的胶原蛋白作为抗氧化药物,有助于加强PCL膜促进愈合的作用。
附图说明:
图1为实施例1中抗氧化型PCL纳米纤维电纺膜的外观示意图;
图2为实施例1中抗氧化性PCL纳米纤维电纺膜的微观结构图;
图3为普通医用敷料和抗氧化型PCL纳米纤维电纺膜的摄水率的对比图;
图4是普通医用敷料和抗氧化型PCL纳米纤维电纺膜各时间点的创面愈合外观图;
图5是普通医用敷料和抗氧化型PCL纳米纤维电纺膜各时间点的创面愈合率对比图。
具体实施方式
下面结合实施例和附图对本发明作进一步说明。其具体实施例如下:
(1)将三氯甲烷(氯仿)与二甲基甲氨酰以9:1比例配置成溶剂,将PCL(Polycaprolactone,聚己内酯)颗粒作为溶质溶解于此溶剂中,配置成质量分数为22%的PCL溶液;
(2)将制成的聚己内酯(PCL)溶液进行静电纺丝(具体参数:流速:1.3μl/min,电压6-7kV,距离15cm),得到PCL纳米纤维电纺膜,经真空干燥处理后,将PCL电纺膜置于紫外线下照射2h、75%酒精内浸泡2h,最后用PBS溶液对材料进行反复冲洗5-6次将PCL电纺膜裁剪成3cm*3cm大小的膜片,分装于无菌干燥的容器中;
(3)取胶原蛋白粉末,加入到浓度为0.05mmol/L的乙酸溶液中,并在0℃冰水浴下超声8-12min,直至溶解成制备成胶原蛋白质量百分比为2%的胶状均质溶液A;取N-乙酰半胱氨酸(Nac)溶解于去离子水中(可用加热至36℃助溶)得到浓度为0.5g/L的溶液B;将胶状均质溶液A与溶液B按照质量比为1:1混匀后于0℃冰水浴下超声30min至均质胶状溶液,即为载有抗氧化药物Nac的胶原蛋白溶液,并将该溶液用离心机2500r/min,离心5min去除气泡,4℃放置半个小时成凝胶状;
(4)将步骤(3)中制备的凝胶状胶原蛋白溶液采用加样枪均匀添加到步骤(2)中的各个PCL电纺膜片上,置于-20℃冰箱内冷冻4h,然后将冷冻后的样品置于冻干机内,调整其温度-42℃,压力为5~20Pa,冻干时间为24小时候取出;
(5)现场配制含有50mmol/L 1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC),12.5mmol/L N-羟基琥珀酰亚胺(NHS)的体积分数为95%乙醇溶液作为交联剂;将冻干后的材料浸泡入上述交联剂溶液中,在室温下(25℃)浸泡6h;
(6)将经过交联剂溶液浸泡后的膜片材料用去离子水反复冲洗材料5-6次,每次5min,将材料置于滤纸上晾干后,置于-20℃冰箱冷冻4h,将冷冻后的样品置于冻干机,调整其温度-42~-53℃,压力为5~20Pa,冻干时间为24小时候取出,即可得到抗氧化型PCL纳米纤维电纺膜,并将其用环氧乙烷37℃条件下进行消毒灭菌处理。
本发明的发明人通过以动物实验为主的一系列实验,证明了本发明制备的抗氧化型PCL纳米纤维电纺膜对于创面是具有促进作用的。其具体以下面几个实验为例,对本发明作进一步说明。
实验1:针对实施例1中制备的抗氧化型PCL纳米纤维电纺膜的大体外观和微观结构进行研究,其中图1为实施例1中抗氧化型PCL纳米纤维电纺膜的外观示意图;图2为实施例1中抗氧化性PCL纳米纤维电纺膜的微观结构图。从图2中可以明显看出,载有Nac抗氧化药物的胶原蛋白与PCL纳米纤维电纺膜形成了类似于“三明治”的结构,这种结构,一方面,有助于Nac药物直接作用于伤口表面,促进药物释放;另一方面,纳米纤维电纺膜作为中层结构可以维持伤口敷料的延伸性与强度,同时具有良好的透气性,有助于伤口与外界气、液进行交换。
实验2:将实施例1中制备的抗氧化型PCL纳米纤维电纺膜与普通敷料置于富含水蒸气的潮湿环境中,记内Δt(单位h)时间内的质量变化量为ΔW(单位mg),材料面积为A(单位cm2)水蒸气透过率(water vapor permeability,WVP)mg·cm-2·h-1=ΔW/(A×Δt)。另外,将所述普通医用敷料和抗氧化型PCL纳米纤维电纺膜浸泡于PBS溶液中,记录样品充分吸水1h、6h、24h、48h后的质量Ms(单位mg),同时记各组材料初始质量为Mi(单位mg),摄水率(water take-up capacity)%=(Ms-Mi)/Mi。
所述为普通医用敷料和抗氧化型PCL纳米纤维电纺膜的水蒸气渗透率的对比如下表所示:
通过对比可看出,抗氧化型纳米纤维电纺膜的水蒸气渗透率高,有利于伤口与外界进行气、液交换。
所述普通医用敷料和抗氧化型PCL纳米纤维电纺膜的摄水率的对比如图3所示。通过对比可以看出,抗氧化型纳米纤维电纺膜的摄水率更高,对于伤口表面的渗出液吸收能力更强,可保持创面的清洁干燥,有助于创面愈合。
实验3:为了验证材料可促进创面愈合的效果,发明人选用了SD大鼠背部创面模型:于SD大鼠背面行2个长径2cm短径1cm的椭圆形创面,分别覆盖普通医用敷料和实施例1中制备的抗氧化型PCL纳米纤维电纺膜,于术后第3、6、9、12、15、18天分别观察拍照,比较各时间点的愈合率,其中,普通医用敷料和抗氧化型PCL纳米纤维电纺膜各时间点的创面愈合情况大致外观如图4所示;两种材料各时间点的创面愈合率的对比如图5所示。
通过以上数据的对比,可以得出结论:抗氧化型PCL纳米纤维电纺膜可以促进创面愈合,且效果优于普通医用敷料。
Claims (8)
1.一种抗氧化型纳米纤维电纺膜医用敷料的制备方法,其特征在于具体步骤如下:
(1)将三氯甲烷与二甲基甲氨酰以8.5~9.5:1比例配置成混合溶剂,将聚己内酯(PCL)颗粒作为溶质溶解于上述混合溶剂中,配置成质量分数为20~24%的PCL溶液;
(2)将步骤(1)中制成的PCL溶液进行静电纺丝得到PCL纳米纤维电纺膜,然后经真空干燥处理后置于紫外线下照射2~4h,再采用75%酒精内浸泡2~6h,最后用PBS缓冲液对材料进行反复冲洗5-6次后置于无菌干燥容器中;
(3)取胶原蛋白粉末,加入到浓度为0.05mmol/L的乙酸溶液中,并在0℃冰水浴下超声8-12min,直至溶解成制备成胶原蛋白质量百分比为2~4%的胶状均质溶液A;取N-乙酰半胱氨酸(Nac)溶解于去离子水中得到浓度为0.5g/L的溶液B;将胶状均质溶液A与溶液B按照质量比为1:0.8~1.2混匀后于0℃冰水浴下超声25-35min至均质胶状溶液,即为载有抗氧化药物Nac的胶原蛋白溶液;
(4)将步骤(3)中制备的载有抗氧化药物Nac的胶原蛋白溶液经过离心处理后去除气泡,并于2~6℃环境下静置30~60min致使其成凝胶状,然后将其用加样枪均匀添加到步骤(2)中的PCL纳米纤维电纺膜上,并置于-20~-26℃环境下冷冻4~6h,之后取出置于冻干机中冻干;
(5)取1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)、N-羟基琥珀酰亚胺(NHS)加入95%乙醇溶液中现场配制交联液,其交联液中EDC的含量为50mmol/L,NHS的含量为12.5mmol/L;然后将步骤(4)中涂有胶原蛋白溶液的膜片材料在室温下浸泡至交联剂溶液中6~8h;
(6)将经过交联剂溶液浸泡后膜片材料用去离子水反复冲洗多次后晾干,再置于-20~-26℃环境下冷冻4~6h后,再通过冻干机冻干,即可得到抗氧化型PCL纳米纤维电纺膜医用敷料。
2.根据权利要求1所述的一种抗氧化型纳米纤维电纺膜医用敷料的制备方法,其特征在于:所述步骤(8)中制备的抗氧化型PCL纳米纤维电纺膜用环氧乙烷37℃条件下进行消毒灭菌处理后储存。
3.根据权利要求1所述的一种抗氧化型纳米纤维电纺膜医用敷料的制备方法,其特征在于:所述步骤(2)中的PCL纳米纤维电纺膜在依次经过紫外线照射、酒精浸泡和冲洗后裁剪成3~5cm*3~5cm大小的膜片,分装于无菌干燥的容器中。
4.根据权利要求1所述的一种抗氧化型纳米纤维电纺膜医用敷料的制备方法,其特征在于:所述步骤(2)中的静电纺丝的具体参数:流速1.3μl/min,电压6~7kV,距离15cm。
5.根据权利要求1所述的一种抗氧化型纳米纤维电纺膜医用敷料的制备方法,其特征在于:所述步骤(3)中的溶液B配制过程中,将溶解温度限定为35-38℃。
6.根据权利要求1所述的一种抗氧化型纳米纤维电纺膜医用敷料的制备方法,其特征在于所述步骤(4)中胶原蛋白溶液的离心条件为:离心转速2000~2500r/min,离心时间5~6min。
7.根据权利要求1所述的一种抗氧化型纳米纤维电纺膜医用敷料的制备方法,其特征在于:所述步骤(4)和步骤(6)中的冻干机的工作条件为温度-42~-53℃,压力为5~20Pa,时间为18~24h。
8.根据权利要求1所述的一种抗氧化型纳米纤维电纺膜医用敷料的制备方法,其特征在于:所述步骤(6)中经过交联剂溶液浸泡后膜片材料采用去离子水反复冲洗5~6次,每次5~10min,然后取出材料置于滤纸上晾干后。
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CN115192763B (zh) * | 2022-07-08 | 2023-12-12 | 重庆科技学院 | 多功能调温创面敷料及其制备方法与应用 |
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