CN114540977B - 用于伤口修复的柞蚕丝素蛋白复合纳米纤维 - Google Patents

用于伤口修复的柞蚕丝素蛋白复合纳米纤维 Download PDF

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CN114540977B
CN114540977B CN202210153691.3A CN202210153691A CN114540977B CN 114540977 B CN114540977 B CN 114540977B CN 202210153691 A CN202210153691 A CN 202210153691A CN 114540977 B CN114540977 B CN 114540977B
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silk fibroin
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CN114540977A (zh
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张显华
冯向伟
于宾
刘杰
邹清云
陈莉娜
周金龙
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Henan Institute of Engineering
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Abstract

本发明公开了一种用于伤口修复的柞蚕丝素蛋白复合纳米纤维，本发明的复合纳米纤维是由合适配比的柞蚕丝丝素蛋白、羧甲基壳聚糖、明胶以及抗菌剂通过静电纺丝技术在一定的纺丝条件下制备获得；同时柞蚕丝素蛋白溶液中还添加了乙醇，其既保证了复合纳米纤维膜中柞蚕丝素由无归卷曲结构形成β‑折叠结构，也解决了现有技术中将纺丝形成的纳米纤维膜浸入乙醇溶液中造成纳米纤维膜黏连以及纳米纤维溶解在乙醇中的问题，同时提高了纳米纤维的机械强度，扩大了纳米纤维膜的应用范围；获得的纳米纤维在创口修复方面具有良好的应用前景。

Description

用于伤口修复的柞蚕丝素蛋白复合纳米纤维
技术领域
本发明属于复合纳米纤维技术领域，具体涉及一种用于伤口修复的柞蚕丝素蛋白复合纳米纤维。
背景技术
我国的柞蚕丝产量丰富，柞蚕丝蛋白作为一种天然的高分子蛋白，其含有Arg-Gly-Asp（RGD）序列，而RGD序列具有优异的细胞粘附性，可促进细胞增殖，在创口修复医用材料领域具有广泛的应用价值；羧甲基壳聚糖是一种重要的水溶性壳聚糖衍生物，具有促进创面愈合、止血、抑制瘢痕、镇痛和抑菌作用，且其具备良好的生物相容性和生物降解性，在生物医用材料领域广泛应用；明胶作为一种多肽混合物，其具有良好的生物相容性、可降解性和水溶性；可广泛应用于医用止血材料，但单纯的明胶水溶液在常温下呈胶体状态，难以通过静电纺丝技术制备获得纳米纤维；同时，采用柞蚕丝素蛋白、羧甲基壳聚糖、明胶为原料经静电纺丝技术制备获得用于伤口修复的纳米纤维尚未见报道，并且，现有技术中以柞蚕丝素蛋白作为原料制备纳米纤维时，为了使柞蚕丝素蛋白纳米纤维形成β-折叠结构，常将获得的纳米纤维浸泡于乙醇溶液中，但是此种方法不可避免会造成纳米纤维的溶解和黏连而影响获得的纳米纤维的性能。
发明内容
针对现有技术存在的不足，本发明的目的在于提供一种用于伤口修复的柞蚕丝素蛋白复合纳米纤维。
为实现上述目的，本发明的技术方案如下：
一种用于伤口修复的柞蚕丝素蛋白复合纳米纤维，其制备方法具体包括以下步骤：
（1）再生柞蚕丝丝素溶液的制备：将柞蚕丝置于Na2CO3溶液中重复脱胶后，烘干剪碎后，利用硫氰酸锂溶液溶解柞蚕丝素，而后经透析浓缩后，向浓缩液中添加乙醇，获得再生柞蚕丝丝素溶液A；
（2）向再生柞蚕丝丝素溶液A中加入羧甲基壳聚糖，在常温下搅拌溶解获得溶液B；
（3）向溶液B中加入明胶搅拌溶解后获得溶液C；
（4）向溶液C中添加抗菌剂，进行超声分散30-60min，获得溶液D；
（5）将溶液D采用静电纺丝设备进行纺丝，即可获得功能性柞蚕丝素蛋白纳米纤维。
优选地，步骤（1）中Na2CO3溶液的质量浓度为0.5%；所述的脱胶重复次数为3次；
优选地，步骤（1）中添加乙醇的量占浓缩液的重量比为1wt%-5wt%；所述的乙醇为无水乙醇；
优选地，步骤（4）中所述的抗菌剂为纳米银、纳米铜、纳米氧化锌中的一种或多种；
优选地，所述的溶液D中再生柞蚕丝丝素的质量浓度为6%-30%；羧甲基壳聚糖的质量浓度为1-10%；明胶的质量浓度为1-5%；所述抗菌剂的添加量占纺丝液的重量百分比为：0.01-0.6wt%；
优选地，步骤（5）中所述的纺丝条件为：纺丝温度50-60℃；纺丝电压7kV-40kV；纺丝距离3cm-25cm；纺丝速度0.1mL/h-2.5mL/h。
有益效果
本发明公开了一种柞蚕丝丝素蛋白复合纳米纤维，其是由合适配比的柞蚕丝丝素蛋白、羧甲基壳聚糖、明胶以及抗菌剂通过静电纺丝技术制备获得；柞蚕丝丝素具有促进细胞粘附的作用；羧甲基壳聚糖作为一种重要的水溶性壳聚糖衍生物，具有抗菌消炎、促进创面愈合、止血、抑制瘢痕、镇痛等作用；明胶具有良好的吸液性和滋润皮肤的作用；纳米抗菌剂与羧甲基壳聚糖具有协同抗菌作用，上述原料以适宜配比混合反应后可通过静电纺丝制备获得纳米纤维膜，在创口修复方面具有良好的应用前景。
本发明在复合纳米纤维原料配比合理的条件下，在获得的柞蚕丝素溶液中还添加了乙醇，其既保证了复合纳米纤维膜中柞蚕丝素由无归卷曲结构形成β-折叠结构，也解决了现有技术中将纺丝形成的纳米纤维膜浸入乙醇溶液中造成纳米纤维膜黏连以及纳米纤维溶解在乙醇中的问题，同时提高了纳米纤维的机械强度，扩大了纳米纤维膜的应用范围。
附图说明
图1是本发明实施例2制备获得的纳米纤维的扫描电镜图；
图2是本发明对比例1制备获得的纳米纤维的扫描电镜图；
图3是本发明对比例2制备获得的纳米纤维的扫描电镜图。
具体实施方式
下面结合附图和具体实施例对本发明的技术方案进行详细说明，本发明实验所应用的原料如无特殊说明均可从市面上购买获得；
实施例1
一种用于伤口修复的柞蚕丝素蛋白复合纳米纤维，其制备方法具体包括以下步骤：
（1）再生柞蚕丝丝素溶液的制备：将柞蚕丝置于0.5%的Na2CO3溶液中重复脱胶3次后，烘干剪碎后，利用硫氰酸锂溶液溶解柞蚕丝素，而后经透析浓缩后，向浓缩液中添加无水乙醇，获得再生柞蚕丝丝素溶液A；
（2）向再生柞蚕丝丝素溶液A中加入羧甲基壳聚糖，在常温下搅拌溶解获得溶液B；
（3）向溶液B中加入明胶搅拌溶解后获得溶液C；
（4）向溶液C中添加纳米银，进行超声分散30-60min，获得溶液D；
（5）将溶液D采用静电纺丝设备进行纺丝，即可获得功能性柞蚕丝素蛋白纳米纤维。
其中，步骤（1）中添加无水乙醇的量占浓缩液的重量百分比为1wt%；
步骤（4）中所述的溶液D中再生柞蚕丝丝素的质量浓度为6%；羧甲基壳聚糖的质量浓度为1%；明胶的质量浓度为1%；所述纳米银的添加量占纺丝液的重量百分比为0.01wt%；
步骤（5）中所述的纺丝条件为：纺丝温度为50℃；纺丝电压7kV；纺丝距离3cm；纺丝速度0.1mL/h。
实施例2
一种用于伤口修复的柞蚕丝素蛋白复合纳米纤维，其制备方法具体包括以下步骤：
（1）再生柞蚕丝丝素溶液的制备：将柞蚕丝置于0.5%的Na2CO3溶液中重复脱胶3次后，烘干剪碎后，利用硫氰酸锂溶液溶解柞蚕丝素，而后经透析浓缩后，向浓缩液中添加无水乙醇，获得再生柞蚕丝丝素溶液A；
（2）向再生柞蚕丝丝素溶液A中加入羧甲基壳聚糖，在常温下搅拌溶解获得溶液B；
（3）向溶液B中加入明胶搅拌溶解后获得溶液C；
（4）向溶液C中添加纳米银，进行超声分散30-60min，获得溶液D；
（5）将溶液D采用静电纺丝设备进行纺丝，即可获得功能性柞蚕丝素蛋白纳米纤维。
其中，步骤（1）中添加无水乙醇的量占浓缩液的重量百分比为3wt%；
步骤（4）中所述的溶液D中再生柞蚕丝丝素的质量浓度为18%；羧甲基壳聚糖的质量浓度为5.5%；明胶的质量浓度为3%；所述纳米银的添加量占纺丝液重量百分比为：0.3wt%。
优选地，步骤（5）中所述的纺丝条件为：纺丝温度为55℃；纺丝电压23kV；纺丝距离14cm；纺丝速度1.3mL/h。
实施例3
一种用于伤口修复的柞蚕丝素蛋白复合纳米纤维，其制备方法具体包括以下步骤：
（1）再生柞蚕丝丝素溶液的制备：将柞蚕丝置于0.5%的Na2CO3溶液中重复脱胶3次后，烘干剪碎后，利用硫氰酸锂溶液溶解柞蚕丝素，而后经透析浓缩后，向浓缩液中添加无水乙醇，获得再生柞蚕丝丝素溶液A；
（2）向再生柞蚕丝丝素溶液A中加入羧甲基壳聚糖，在常温下搅拌溶解获得溶液B；
（3）向溶液B中加入明胶搅拌溶解后获得溶液C；
（4）向溶液C中添加纳米银，进行超声分散30-60min，获得溶液D；
（5）将溶液D采用静电纺丝设备进行纺丝，即可获得功能性柞蚕丝素蛋白纳米纤维。
其中，步骤（1）中添加无水乙醇的量占浓缩液的重量百分比为5wt%；
步骤（4）中所述的溶液D中再生柞蚕丝丝素的质量浓度为30%；羧甲基壳聚糖的质量浓度为10%；明胶的质量浓度为5%；所述抗菌剂的添加量占纺丝液的重量比为0.6wt%；
步骤（5）中所述的纺丝条件为：纺丝温度60℃；纺丝电压40kV；纺丝距离25cm；纺丝速度2.5mL/h。
对比例1
一种用于伤口修复的柞蚕丝素蛋白复合纳米纤维，其制备方法具体包括以下步骤：
（1）再生柞蚕丝丝素溶液的制备：将柞蚕丝置于0.5%的Na2CO3溶液中重复脱胶3次后，烘干剪碎后，利用硫氰酸锂溶液溶解柞蚕丝素，而后经透析浓缩后，获得再生柞蚕丝丝素溶液A；
（2）向再生柞蚕丝丝素溶液A中加入羧甲基壳聚糖，在常温下搅拌溶解获得溶液B；
（3）向溶液B中加入明胶搅拌溶解后获得溶液C；
（4）向溶液C中添加纳米银，进行超声分散30-60min，获得溶液D；
（5）将纺丝液D采用静电纺丝设备进行纺丝，获得纳米纤维，再将纳米纤维浸入乙醇溶液中浸泡1h后取出，即可获得功能性柞蚕丝素蛋白纳米纤维。
其中，步骤（4）所述的溶液D中再生柞蚕丝丝素的质量浓度为18%；羧甲基壳聚糖的质量浓度为5.5%；明胶的质量浓度为3%；所述纳米银的添加量占纺丝液的重量百分比为0.3wt%；
步骤（5）中所述的纺丝条件为：纺丝温度为55℃；纺丝电压23kV；纺丝距离14cm；纺丝速度1.3mL/h。
对比例2
对比例2与对比例1获得功能性柞蚕丝素蛋白纳米纤维的步骤基本相同，不同之处在于步骤（5）中将获得的纳米纤维置于乙醇蒸汽中3天后取出获得功能性柞蚕丝素蛋白纳米纤维。
对比例3
对比例3与实施例2制备获得功能性柞蚕丝素蛋白纳米纤维的步骤基本相同，不同之处在于步骤（5）中控制纺丝温度为25℃，而在此温度下，纺丝液不能进行正常纺丝获得纳米纤维。
本发明实施例2制备获得的功能性柞蚕丝素蛋白纳米纤维的扫描电镜图如图1所示，对比例1和对比例2制备获得的功能性柞蚕丝素蛋白纳米纤维的扫描电镜图如图2和图3所示；从图中可以看出，对比例1和2中的纳米纤维出现了黏连的情况。
以上所述，仅为本发明的具体实施方式，但本发明的保护范围并不仅局限于此，本领域普通技术人员对本发明的技术方案所做的其他修改或者等同替换，只要不脱离本发明技术方案的精神和范围，均应涵盖在本发明的权利要求范围当中。

Claims (5)

1.一种用于伤口修复的柞蚕丝素蛋白复合纳米纤维，其特征在于，制备方法具体包括以下步骤：
步骤(1)，再生柞蚕丝丝素溶液的制备：将柞蚕丝置于Na2CO3溶液中重复脱胶后，烘干剪碎后，利用硫氰酸锂溶液溶解柞蚕丝素，而后经透析浓缩后，向浓缩液中添加乙醇，添加乙醇的量占浓缩液的重量比为1wt％-5wt％，获得再生柞蚕丝丝素溶液A；
步骤(2)，向再生柞蚕丝丝素溶液A中加入羧甲基壳聚糖，在常温下搅拌溶解获得溶液B；步骤(3)，向溶液B中加入明胶搅拌溶解后获得溶液C；
步骤(4)，向溶液C中添加抗菌剂，进行超声分散30-60min，获得溶液D；
步骤(5)，将溶液D采用静电纺丝设备进行纺丝，纺丝条件为：纺丝温度50-60℃；纺丝电压7kV-40kV；纺丝距离3cm-25cm；纺丝速度0.1mL/h-2.5mL/h，即可获得功能性柞蚕丝素蛋白纳米纤维。
2.如权利要求1所述的用于伤口修复的柞蚕丝素蛋白复合纳米纤维，其特征在于，步骤(1)中Na2CO3溶液的质量浓度为0.5％；所述的脱胶重复次数为3次。
3.如权利要求1所述的用于伤口修复的柞蚕丝素蛋白复合纳米纤维，其特征在于，步骤(1)中所述的乙醇为无水乙醇。
4.如权利要求1所述的用于伤口修复的柞蚕丝素蛋白复合纳米纤维，其特征在于，步骤(4)中所述的抗菌剂为纳米银、纳米铜、纳米氧化锌中的一种或多种。
5.如权利要求1所述的用于伤口修复的柞蚕丝素蛋白复合纳米纤维，其特征在于，所述的溶液D中再生柞蚕丝丝素的质量浓度为6％-30％；羧甲基壳聚糖的质量浓度为1-10％；明胶的质量浓度为1-5％；所述抗菌剂的添加量占纺丝液的重量百分比为：0.01-0.6wt％。
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