CN113293626A - 一种创面敷料及其制备方法和应用 - Google Patents

一种创面敷料及其制备方法和应用 Download PDF

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CN113293626A
CN113293626A CN202110378947.6A CN202110378947A CN113293626A CN 113293626 A CN113293626 A CN 113293626A CN 202110378947 A CN202110378947 A CN 202110378947A CN 113293626 A CN113293626 A CN 113293626A
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wound dressing
layer
fiber
wound
collagen
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蒋朝俊
陈万能
何俊杰
钟永清
钟雄福
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Hubei Sailuo Biomaterials Co ltd
Improve Medical Technology Nanxiong Co ltd
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Hubei Sailuo Biomaterials Co ltd
Improve Medical Technology Nanxiong Co ltd
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Abstract

本发明公开了一种创面敷料及其制备方法和应用，该创面敷料包括纤维支架和裹覆在纤维表面的包覆层，纤维支架由丝素蛋白‑聚己内酯共混液的静电纺丝产物交联而成，其纤维直径为0.1～1μm；包覆层由交替裹覆的壳聚糖层和胶原蛋白层组成，且包覆层的最外层为胶原蛋白层。本发明制备的创面敷料具有较好的力学强度、亲水性和细菌抑制效果，其拉伸强度可达为9.79±0.65MPa，断裂伸长率可达31.94±11.42MPa，水接触角可低至22.28°，该创面敷料孔隙率高，透气性好，吸水率高，可加速创面愈合，使用7天可使创面愈合80％以上。

Description

一种创面敷料及其制备方法和应用
技术领域
本发明涉及创面敷料领域，特别涉及一种创面敷料及其制备方法和应用。
背景技术
皮肤是阻止微生物入侵、维持机体内环境稳定的重要屏障，烧伤(热力、电或化学烧伤)、机械力引起的大片皮肤缺损等皮肤缺损性损伤会引起皮肤屏障功能缺失，运用敷料覆盖创面，临时作为体表保护屏障，促进创面愈合，是临床上治疗的重要措施。消毒纱布等传统的敷料因不能保持创面湿润、易脱落等缺点已不能满足难愈合创面治疗的需求。而天然生物敷料，如自体皮，因来源受限而且后期不能建立有效血供而影响伤口愈合等各种缺陷而仅限一部分的临床运用。
目前，高分子敷料越来越多地应用到创面愈合领域，现有技术中纯丝素敷料韧性差，容易撕裂；丝素/聚已内酯纳米纤维膜的拉力虽远大于纯丝素膜，但丝素/聚已内酯纳米纤维膜抑菌效果差，且亲水性仍不能满足需求，导致丝素/聚已内酯纳米纤维膜的生物相容性差，易从创面表面脱落。
发明内容
本发明将丝素蛋白(SF)与聚己内酯(PCL)共混后经静电纺丝制成纤维支架，再在纤维支架的纤维表面采用静电吸附的方式交替沉积壳聚糖(CS)和胶原蛋白(COL)，最终形成一种亲水性好、强度高、抑菌效果好的创面敷料。
本发明提供的技术方案具体如下：
第一方面，本发明提供一种创面敷料，包括：
纤维支架，其由丝素蛋白-聚己内酯共混液的静电纺丝产物交联而成，其纤维直径为0.1～1μm；和
裹覆在纤维表面的包覆层，其由交替裹覆的壳聚糖层和胶原蛋白层组成，且包覆层的最外层为胶原蛋白层。
作为上述技术方案的优选：按相互邻接的1层壳聚糖层和1层胶原蛋白层为一个双层计，包覆层含6～10个双层。
作为上述技术方案的优选：包覆层含6个双层。
作为上述技术方案的优选：纤维支架的平均纤维直径为519±145nm；创面敷料的平均纤维直径为610±108nm。
作为上述技术方案的优选：纤维支架中丝素蛋白与聚己内酯的重量比为1:1。
第二方面，本发明提供一种创面敷料的制备方法，包括以下步骤：包括以下步骤：对丝素蛋白-聚己内酯共混液进行静电纺丝，将静电纺丝产物浸泡在乙醇中进行交联，交联产物即纤维支架；将纤维支架交替浸泡在电性相反的壳聚糖水溶液和胶原蛋白水溶液中，在纤维表面形成包覆层，即得到创面敷料。
作为上述技术方案的优选：丝素蛋白-聚己内酯共混液中丝素蛋白和聚己内酯的重量比为1:1，溶剂为六氟异丙醇，浓度为7wt％。
作为上述技术方案的优选：包覆层通过如下方法制备：将纤维支架交替放入正电性的壳聚糖水溶液、负电性的胶原蛋白水溶液中浸泡，每次浸泡后用纯水洗去浸泡液，重复若干次，即得到包覆层。
作为上述技术方案的优选：正电性的壳聚糖水溶液为pH＝6.3的壳聚糖水溶液；负电性的胶原蛋白水溶液为pH＝8.0的胶原蛋白水溶液。
第三方面，本发明提供上述创面敷料在制备创面修复材料中的应用。
本发明提供的技术方案带来的有益效果包括：
本发明制备的创面敷料具有较高的力学强度，拉伸强度可达为9.79±0.65MPa，断裂伸长率可达31.94±11.42MPa；亲水性好，水接触角低至22.28°；且抑细菌效果好，孔隙率高，透气性好，吸水性强，可加速创面愈合，使用7天可使创面愈合80％以上。
附图说明
为了更清楚地说明本发明实施例中的技术方案，下面将对实施例描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1为本发明实施例3制备的双层数为0、6、10、10.5的创面敷料的电镜扫描图；其中，图1(a)为SF/PCL的扫描电镜图；图1(b)为(CS/COL)6-SF/PCL的扫描电镜图；图1(c)为(CS/COL)10-SF/PCL的扫描电镜图；图1(d)为(CS/COL)10.5-SF/PCL的扫描电镜图。
图2为本发明实施例3制备的双层数为0、6、10、10.5的创面敷料的纤维直径分布图；其中，图2(a)为SF/PCL的纤维直径分布图；图2(b)为(CS/COL)6-SF/PCL的纤维直径分布图；图2(c)为(CS/COL)10-SF/PCL的纤维直径分布图；图2(d)为(CS/COL)10.5-SF/PCL的纤维直径分布图。
图3为本发明实施例3制备的双层数为0、5.5、6、10、10.5的创面敷料的接触角随时间t的变化图，时间变量为1秒。
图4为大肠杆菌培养48h时的显微镜图；其中，contol代表未使用创面敷料，SF/PCL、(CS/COL)0.5-SF/PCL、(CS/COL)6-SF/PCL、(CS/COL)10-SF/PCL分别代表采用了双层数为0、0.5、6、10的创面敷料。
图5为金黄色葡萄球菌培养48h时的显微镜图；其中，contol代表未使用创面敷料，SF/PCL、(CS/COL)0.5-SF/PCL、(CS/COL)6-SF/PCL、(CS/COL)10-SF/PCL分别代表采用了双层数为0、0.5、6、10、10.5的创面敷料。
图6展示了创面敷料的使用方式；其中，图6(a)展示了小鼠的正方形表皮缺损；图6(b)展示了进行辅助治疗时创面敷料在小鼠伤口上的位置。
图7展示了使用了本发明实施例3制备的创面敷料进行辅助治疗的小鼠在3天和7天时的伤口愈合比例，其中，contol代表未使用创面敷料，SF/PCL、(CS/COL)5.5、(CS/COL)6、(CS/COL)10、(CS/COL)10.5分别代表采用双层数为0、5.5、6、10、10.5的创面敷料进行辅助治疗。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚，下面将结合具体实施例对本发明的技术方案进行清楚、完整地描述，显然，所描述的实施例是本发明的一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动的前提下所获得的所有其他实施例，都属于本发明保护的范围。
第一方面，本发明提供一种创面敷料，包括：
纤维支架，其由丝素蛋白-聚己内酯共混液的静电纺丝产物交联而成，其纤维直径为0.1～1μm；和
裹覆在纤维表面的包覆层，其由交替裹覆的壳聚糖层和胶原蛋白层组成，且包覆层的最外层为胶原蛋白层。
作为上述技术方案的优选：按相互邻接的1层壳聚糖层和1层胶原蛋白层为一个双层计，包覆层含6～10个双层。
作为上述技术方案的优选：包覆层含6个双层。
作为上述技术方案的优选：纤维支架的平均纤维直径为519±145nm；创面敷料的平均纤维直径为610±108nm。
作为上述技术方案的优选：纤维支架中丝素蛋白与聚己内酯的重量比为1:1。
第二方面，本发明提供一种创面敷料的制备方法，包括以下步骤：包括以下步骤：对丝素蛋白-聚己内酯共混液进行静电纺丝，将静电纺丝产物浸泡在乙醇中进行交联，交联产物即纤维支架；将纤维支架交替浸泡在电性相反的壳聚糖水溶液和胶原蛋白水溶液中，在纤维表面形成包覆层，即得到创面敷料。
作为上述技术方案的优选：丝素蛋白-聚己内酯共混液中丝素蛋白和聚己内酯的重量比为1:1，溶剂为六氟异丙醇，浓度为7wt％。
作为上述技术方案的优选：包覆层通过如下方法制备：将纤维支架交替放入正电性的壳聚糖水溶液、负电性的胶原蛋白水溶液中浸泡，每次浸泡后用纯水洗去浸泡液，重复若干次，即得到包覆层。
作为上述技术方案的优选：正电性的壳聚糖水溶液为pH＝6.3的壳聚糖水溶液；负电性的胶原蛋白水溶液为pH＝8.0的胶原蛋白水溶液。
第三方面，本发明提供上述创面敷料在制备创面修复材料中的应用。
以下实施例中所使用的实验方法如无特殊说明，均为常规方法。以下实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。以下实施例中所采用的聚己内酯和壳聚糖均为有机高分子聚合物，购自Sigma-Aldrich公司，所采用的胶原蛋白购自上海源叶生物科技有限公司。
实施例1：制备丝素蛋白-聚己内酯共混液
(1)将家蚕茧壳在碳酸钠溶液中煮沸脱去丝胶蛋白，得到纯净的丝素纤维；
(2)将步骤(1)得到的丝素纤维溶解在9.3mol/L的溴化锂溶液中，得到丝素蛋白粗溶液；
(3)将步骤(2)制备的丝素蛋白粗溶液倒入8000～14000Da的透析袋，在2～8℃条件下透析7天；
(4)取出透析后的丝素蛋白粗溶液，以8000r/min的转速离心，去除离心管底部沉淀的杂质，得到纯净的丝素蛋白溶液；
(5)将步骤(4)得到的丝素蛋白溶液冷冻干燥，得到固体丝素蛋白粉末；
(6)称取1.75g丝素蛋白粉末、1.75g聚己内酯，将二者混合后溶于46.50g六氟异丙醇(HFIP)中，密封后置于磁力搅拌器上，常温搅拌过夜使丝素蛋白完全溶解，制得浓度为7wt％的丝素蛋白-聚己内酯共混液。
实施例2：制备纤维支架
(1)取注射管4支，截去一定长度针头，针尖用砂纸打磨平滑，使针头剩余长度为1.2cm左右；
(2)4支注射管分别抽取实施例1制备的丝素蛋白-聚己内酯共混液8mL，将4支注射管置于推进泵中间位置上，设置推进泵的推进速度为1mL/h，启动推进泵，使注射管有少许液体流出为止；
(2)将铝箔纸平铺于接收辊筒上，且放置于距离针尖12cm处；两针头用铜丝相连后接高压电正极，辊筒接地；
(3)打开辊筒电源开关，使辊筒以500r/min的转速转动；启动推进泵，打开高压电电源开关，设置电压为16kV，点击启动开关，静电纺丝5h，得到静电纺丝产物；
(4)将步骤(3)制备的静电纺丝产物在无水乙醇中浸泡24h，浸泡过程中丝素蛋白和聚己内酯发生交联反应，将交联产物取出晾干，得到纤维支架，命名为SF/PCL。
实施例3：制备创面敷料
(1)配制浸泡液A：浓度为1mg/mL、pH＝6.3的壳聚糖水溶液；配制浸泡液B：浓度为1mg/mL、pH＝8.0的胶原蛋白水溶液；
(2)将实施例2制备的纤维支架SF/PCL依次放入浸泡液A和浸泡液B中各浸泡10分钟，每次浸泡后，将纤维支架SF/PCL在纯水中浸洗3次，并缓慢将纤维支架SF/PCL中的水分挤出，然后再放入下一浸泡液中，完成一次循环后，重新进行下一循环：按浸泡液A、浸泡液B的顺序进行浸泡。浸泡过程中，浸泡液进入纤维支架SF/PCL的孔隙中，在纤维表面沉积，每浸泡1次浸泡液A得到1层壳聚糖层，每浸泡1次浸泡液B得到1层胶原蛋白层，按相互邻接的1层壳聚糖层和1层胶原蛋白层为一个双层计，本实施例按相同的方法分别制备了双层数为0、0.5、1、5.5、6、10、10.5的创面敷料，并按(CS/COL)n-SF/PCL进行命名，其中，下标n代表双层数，双层数为0的创面敷料即为未处理的纤维支架SF/PCL。
性能测试：
1.扫描电镜(SEM)
对实施例3制备的双层数为0、6、10、10.5的创面敷料进行电镜扫描，扫描电镜图如图1所示，纤维的直径分布如图2所示。图1(a)为双层数为0的创面敷料(即SF/PCL)的扫描电镜图，通过图2(a)计算，双层数为0的创面敷料，其平均纤维直径为519±145.7nm；图1(b)为双层数为6的创面敷料(即(CS/COL)6-SF/PCL)的扫描电镜图，通过图2(b)计算，双层数为6的创面敷料，其平均纤维直径为610±108.6nm；图1(c)为双层数为10的创面敷料(即(CS/COL)10-SF/PCL)的扫描电镜图，通过图2(c)计算，双层数为10的创面敷料，其平均纤维直径为673.1±154.2nm；图1(d)为双层数为10.5的创面敷料(即(CS/COL)10.5-SF/PCL)的扫描电镜图，通过图2(d)计算，双层数为10.5的创面敷料，其平均纤维直径为679.5±148.6nm。随着双层数的增加，具有包覆层的纤维直径也随之变粗。
2.亲水性能测试
分别测试实施例3制备的双层数为0、5.5、6、10、10.5的创面敷料的亲水性能，如图3所示，当包覆层最外层为壳聚糖层时，接触角呈增大的趋势，当包覆层最外层为胶原蛋白层时，接触角呈减小的趋势。当包覆层最外层为胶原蛋白层时，(CS/COL)6-SF/PCL与(CS/COL)10-SF/PCL相比，虽然在初始测试时，(CS/COL)6-SF/PCL的接触角大于(CS/COL)10-SF/PCL，但随着时间t的推移(每秒拍照一次)，双层数为6的创面敷料接触角减小明显，亲水性远好于其他双层数的创面敷料。
3.力学性能和透气性测试
分别测试双层数为0、1、5.5、6、10、10.5的创面敷料的力学性能和孔隙率，结果如表1所示：
表1不同双层数的创面敷料的性能测试
Figure BDA0003012063990000091
通过表1可以发现，创面敷料的拉伸强度随着双层数的增加，呈现先增加后降低的趋势，在双层数为6时拉伸强度达到最大值；创面敷料的断裂伸长率则随双层数的增加而增加；孔隙率虽随着双层数的增加而降低，但总体而言，孔隙率较高，创面敷料透气性好、吸水性好。
4.抑菌性实验
分别在双层数为0、0.5、6、10的创面敷料上培养大肠杆菌和金黄色葡萄球菌，培养48h时观察创面敷料上细菌数量，观察图4和图5可以发现，与不具有包覆层的创面敷料相比，具有包覆层的创面敷料能有效抑制大肠杆菌和金黄色葡萄球菌的增长，且随着双层数增加，抑制作用越强。如图5所示，双层数为6的创面敷料的金黄色葡萄球菌菌落总数低至31cfu/g，双层数为10的创面敷料的金黄色葡萄球菌菌落总数低至7cfu/g，抑菌效果非常好。
5.创面愈合实验
如图6(a)所示，在小鼠表皮造边为20mm的正方形表皮缺损，如图6(b)所示，分别用双层数为0、5.5、6、10、10.5的创面敷料对小鼠表皮缺损处进行辅助治疗，在术后3天和7天分别观察小鼠的伤口，并统计伤口愈合比例，结果图7所示；图7显示，使用双层数为6层的创面敷料时，小鼠伤口愈合速度最快，在第7天，伤口愈合比例达80％。
以上所述仅是本发明的具体实施方式，使本领域技术人员能够理解或实现本发明。对这些实施例的多种修改对本领域的技术人员来说将是显而易见的，本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下，在其它实施例中实现。因此，本发明将不会被限制于本文所示的这些实施例，而是要符合与本文所申请的原理和新颖特点相一致的最宽的范围。

Claims (10)

1.一种创面敷料，其特征在于：包括：
纤维支架，其由丝素蛋白-聚己内酯共混液的静电纺丝产物交联而成，其纤维直径为0.1～1μm；和
裹覆在纤维表面的包覆层，其由交替裹覆的壳聚糖层和胶原蛋白层组成，且所述包覆层的最外层为胶原蛋白层。
2.根据权利要求1所述的创面敷料，其特征在于：按相互邻接的1层壳聚糖层和1层胶原蛋白层为一个双层计，所述包覆层含6～10个双层。
3.根据权利要求2所述的创面敷料，其特征在于：所述包覆层含6个双层。
4.根据权利要求1所述的创面敷料，其特征在于：所述纤维支架的平均纤维直径为519±145nm；所述创面敷料的平均纤维直径为610±108nm。
5.根据权利要求1所述的创面敷料，其特征在于：所述纤维支架中丝素蛋白与聚己内酯的重量比为1:1。
6.一种权利要求1～5任一项所述创面敷料的制备方法，其特征在于，包括以下步骤：对丝素蛋白-聚己内酯共混液进行静电纺丝，将静电纺丝产物浸泡在乙醇中进行交联，交联产物即纤维支架；将纤维支架交替浸泡在电性相反的壳聚糖水溶液和胶原蛋白水溶液中，在纤维表面形成包覆层，即得到创面敷料。
7.根据权利要求6所述的创面敷料的制备方法，其特征在于：所述丝素蛋白-聚己内酯共混液中丝素蛋白和聚己内酯的重量比为1:1，溶剂为六氟异丙醇，浓度为7wt％。
8.根据权利要求6所述的创面敷料的制备方法，其特征在于：所述包覆层通过如下方法制备：将纤维支架交替放入正电性的壳聚糖水溶液、负电性的胶原蛋白水溶液中浸泡，每次浸泡后用纯水洗去浸泡液，重复若干次，即得到包覆层。
9.根据权利要求8所述的创面敷料的制备方法，其特征在于：所述正电性的壳聚糖水溶液为pH＝6.3的壳聚糖水溶液；所述负电性的胶原蛋白水溶液为pH＝8.0的胶原蛋白水溶液。
10.权利要求1～5任一项所述的创面敷料在制备创面修复材料中的应用。
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Application publication date: 20210824