CN107217388B - 抗菌性聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜及制备方法 - Google Patents

抗菌性聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜及制备方法 Download PDF

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CN107217388B
CN107217388B CN201710516225.6A CN201710516225A CN107217388B CN 107217388 B CN107217388 B CN 107217388B CN 201710516225 A CN201710516225 A CN 201710516225A CN 107217388 B CN107217388 B CN 107217388B
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caprolactone
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袁晓燕
李珍光
周芳
周培琼
任丽霞
赵蕴慧
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Tianjin University
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Abstract

本发明公开了一种抗菌性聚(ε‑己内酯)/聚(ε‑己内酯)‑REDV/明胶电纺纤维膜及制备方法；该电纺纤维膜是由直径为200‑1200nm的聚(ε‑己内酯)/聚(ε‑己内酯)‑REDV/明胶纤维和包载在纤维内部的植物源抗菌剂构成的，厚度为50‑150μm。制备方法是将聚(ε‑己内酯)、聚(ε‑己内酯)‑REDV和明胶溶于三氟乙醇中，并滴加冰醋酸至溶液澄清，形成电纺溶液，然后加入植物源抗菌剂丁香酚，使用单道注射泵电纺装置，采用静电纺丝方法得到电纺纤维膜。制得的电纺纤维膜具有加快材料表面内皮化与抗菌的双重功能，在人工血管生物医用材料方面具有应用前景。

Description

抗菌性聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜 及制备方法
技术领域
本发明涉及一种包载植物源抗菌剂的抗菌性聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜及制备方法，属于生物医用材料领域。
背景技术
近年来，心血管疾病成为造成人类死亡的首要原因。采用静电纺丝成膜技术，通过调整电纺参数，可制得具有纤维直径为几百纳米和较大的比表面积的电纺纤维膜材料，其在结构上可以模拟细胞外基质，有利于细胞的粘附，增殖和迁移，可用于心血管重建。
聚(ε-己内酯)具有良好的生物相容性和生物可降解性及优异的力学性能，为小口径人工血管研究的常用材料。但是，聚(ε-己内酯)本身不利于细胞的粘附并且具有较慢的降解速度，在应用上具有诸多限制(Dash T K,Konkimalla V B.Poly-ε-caprolactonebased formulations for drug delivery and tissue engineering:areview.J.Control.Release,2012,158:15-33)。明胶来源于天然胶原的部分水解，含有整合蛋白结合位点，能够促进细胞的粘附，并具有较快的降解速度，能够弥补聚(ε-己内酯)的缺点(Jiang Y C,Jiang L,Huang A,et al.Electrospun polycaprolactone/gelatincomposites with enhanced cell-matrix interactions as blood vessel endotheliallayer scaffolds.Mat.Sci.Eng.C,2017,71:901-908)。然而，小口径人工血管植入体内，易产生再狭窄和细菌感染，从而导致血管重建失败。人工血管材料表面快速内皮化能够抑制再狭窄的发生，同时赋予材料抗菌性能，则能够预防细菌感染。
精氨酸-谷氨酸-天冬氨酸-缬氨酸(Arg-Glu-Asp-Val，REDV)短肽存在于纤维连接蛋白的III-CS区域，含有该序列的合成肽能够特异性地结合血管内皮细胞，实现快速内皮化(Ren X,Feng Y,Guo J,et al.Surface modification and endothelialization ofbiomaterials as potential scaffolds for vascular tissue engineeringapplications.Chem.Soc.Rev.,2015,44:5680-5742)。从植物中提取的天然酚类化合物丁香酚具有优异的广谱抗菌性能和极低的毒副作用，且不易产生耐药性。本发明将本课题组之前合成的聚(ε-己内酯)-REDV(公开号CN106729976A)与聚(ε-己内酯)和明胶进行共混电纺，同时包载丁香酚，制备具有加快材料表面内皮化和抗菌双重功能的电纺纤维膜材料，该项工作目前未见报道。
发明内容
本发明的目的在于提供一种抗菌性聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜及制备方法。采用静电纺丝成膜技术，使制得的纤维膜具备加快材料表面内皮化和抗菌的双重功能。一方面，利用低分子量聚(ε-己内酯)-REDV在电纺过程中向表面的迁移，实现REDV短肽促进血管内皮细胞粘附和增殖的功能；另一方面，以聚(ε-己内酯)/明胶为载药体系，包载植物源抗菌剂，利用明胶快速的降解速度，实现纤维膜中植物源抗菌剂由快到慢的释放行为，这样使纤维膜既能在植入初期快速抵御细菌感染，又能维持持久的抗菌性能。
为达到上述目的，本发明提供了一种包载植物源抗菌剂的聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜，其特征在于纤维膜是由直径为200-1200nm的聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶纤维和包载在纤维内部的植物源抗菌剂构成的厚度为50-150μm的纤维膜。
所述的包载植物源抗菌剂的聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜，其特征在于所述的电纺纤维中聚(ε-己内酯)的质量分数为20-80wt％，聚(ε-己内酯)-REDV的质量分数为10-50wt％，明胶的质量分数为10-30wt％，而植物源抗菌剂的量占上述聚合物总量的5-50wt％。
所述的包载植物源抗菌剂的聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜的制备方法，其特征在于包括以下步骤：
1)将聚(ε-己内酯)、聚(ε-己内酯)-REDV和明胶溶于三氟乙醇中，室温下搅拌溶解，向上述溶液中滴加冰醋酸并搅拌溶液至均相澄清，然后加入5-50wt％的植物源抗菌剂，混合均匀形成浓度为100-200mg/mL的聚合物/三氟乙醇溶液；
2)将配制好的溶液置于注射器内，使用单道注射泵，在电压为10-15kV，接收距离为15-20cm，流量为0.4-0.8mL/h条件下进行电纺，得到厚度为50-150μm的包载植物源抗菌剂的聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜。
所述1)中的聚(ε-己内酯)的数均分子量为(5-12)×104，聚(ε-己内酯)-REDV的数均分子量为(1-5)×104，按照公开号CN106729976A专利中的方式合成。
所述1)中的植物源抗菌剂为丁香酚。
本发明的优点在于，通过静电纺丝的方法制备聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜，制备过程简单，所用的聚(ε-己内酯)与明胶具有良好的生物相容性和生物可降解性，包载的植物源抗菌剂廉价易得，毒性低且具有优异的抗菌性能；电纺过程中迁移到表面的REDV短肽能够促进血管内皮细胞的粘附和增殖；聚(ε-己内酯)与明胶不同的降解速度可调控植物源抗菌剂的释放速度，使纤维膜既能在植入初期快速抵御细菌感染，又能维持持久的抗菌性能，还可设计不同的载药量来满足不同的应用需要；因此，制得的纤维膜具有加快材料表面内皮化与抗菌的双重功能，在与心血管疾病相关的生物医用材料中具有广阔的应用前景。
附图说明
图1为实施例1条件下制得的含有丁香酚的聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶抗菌性电纺纤维膜的扫描电子显微镜照片。
图2为实施例1条件下制得的含有丁香酚的聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶抗菌性电纺纤维膜接触培养E.coil和S.aurseus后的平板菌落图。
具体实施方式
下面结合实施案例对本发明的技术方案作进一步阐述，以下实施案例是对本发明的进一步说明，并不限制本发明的适用范围。
实施例1
本实施例电纺纤维中聚(ε-己内酯)、聚(ε-己内酯)-REDV、明胶的质量分数分别为40wt％、30wt％、30wt％，丁香酚占上述聚合物总量的30wt％。
称取300mg聚(ε-己内酯)(Mn＝8×104)，225mg聚(ε-己内酯)-REDV(Mn＝3×104)，225mg明胶溶解于5mL三氟乙醇中，并滴加冰醋酸搅拌至溶液均相澄清，然后加入225mg的丁香酚，混合均匀；将溶液置于注射器中，使用单道注射泵电纺装置，在电压为12kV、流量为0.6mL/h、接收距离为18cm条件下进行电纺，经过8h后收集到纤维直径为500nm-800nm，厚度为50μm的纤维膜
图1显示出得到的电纺纤维形貌良好，无珠丝等缺陷；图2显示出该电纺膜接触培养E.coil和S.aureus后长成的菌落小且稀疏，经计算对E.coil的抑菌率为71.9％，对S.aureus的抑菌率为85.5％；经测试内皮细胞在该电纺膜上培养3、6、9天后的增殖结果为OD460＝0.55、0.74、0.92。
实施例2
本实施例电纺纤维中聚(ε-己内酯)、聚(ε-己内酯)-REDV、明胶的质量分数分别为80wt％、10wt％、10wt％，丁香酚占上述聚合物总量的5wt％。
称取400mg聚(ε-己内酯)(Mn＝12×104)，50mg聚(ε-己内酯)-REDV(Mn＝1×104)，50mg明胶溶解于5mL三氟乙醇中，并滴加冰醋酸搅拌至溶液均相澄清，然后加入25mg的丁香酚，混合均匀；将溶液置于注射器中，使用单道注射泵电纺装置，在电压为10kV、流量为0.4mL/h、接收距离为15cm条件下进行电纺，经过12h后收集到纤维直径为200nm-500nm，厚度为50μm的纤维膜。
实施例3
本实施例电纺纤维中聚(ε-己内酯)、聚(ε-己内酯)-REDV、明胶的质量分数分别为70wt％、20wt％、10wt％，丁香酚占上述聚合物总量的20wt％。
称取700mg聚(ε-己内酯)(Mn＝5×104)，200mg聚(ε-己内酯)-REDV(Mn＝5×104)，100mg明胶溶于10mL三氟乙醇中，并滴加冰醋酸搅拌至溶液均相澄清，然后加入200mg的丁香酚，混合均匀；将溶液置于注射器中，使用单道注射泵电纺装置，在电压为15kV，流量为0.8mL/h，接收距离为20cm条件下进行电纺，经过12h后收集到纤维直径为200nm-500nm，厚度为100μm的纤维膜。
实施例4
本实施例电纺纤维中聚(ε-己内酯)、聚(ε-己内酯)-REDV、明胶的质量分数分别为50wt％、35wt％、15wt％，丁香酚占上述聚合物总量的25wt％。
称取750mg聚(ε-己内酯)(Mn＝8×104)，525mg聚(ε-己内酯)-REDV(Mn＝3×104)，225mg明胶溶于15mL三氟乙醇中，并滴加冰醋酸搅拌至溶液均相澄清，然后加入375mg的丁香酚，混合均匀；将溶液置于注射器中，使用单道注射泵电纺装置，在电压为12kV，流量为0.6mL/h，接收距离为18cm条件下进行电纺，经过24h后收集到纤维直径为200nm-500nm，厚度为150μm的纤维膜。
实施例5
本实施例电纺纤维中聚(ε-己内酯)、聚(ε-己内酯)-REDV、明胶的质量分数分别为35wt％、40wt％、25wt％，丁香酚占上述聚合物总量的15wt％。
称取262.5mg聚(ε-己内酯)(Mn＝12×104)，300mg聚(ε-己内酯)-REDV(Mn＝1×104)，187.5mg明胶溶于5mL三氟乙醇中，并滴加冰醋酸搅拌至溶液均相澄清，然后加入112.5mg的丁香酚，混合均匀；将溶液置于注射器中，使用单道注射泵电纺装置，在电压为10kV，流量为0.8mL/h，接收距离为20cm条件下进行电纺，经过6h后收集到纤维直径为500nm-800nm，厚度为50μm的纤维膜。
实施例6
本实施例电纺纤维中聚(ε-己内酯)、聚(ε-己内酯)-REDV、明胶的质量分数分别为45wt％、30wt％、25wt％，丁香酚占上述聚合物总量的35wt％。
称取675mg聚(ε-己内酯)(Mn＝5×104)，450mg聚(ε-己内酯)-REDV(Mn＝5×104)，375mg明胶溶于10mL三氟乙醇中，并滴加冰醋酸搅拌至溶液均相澄清，然后加入525mg的丁香酚，混合均匀；将溶液置于注射器中，使用单道注射泵电纺装置，在电压为15kV，流量为0.4mL/h，接收距离为15cm条件下进行电纺，经过24h后收集到纤维直径为500nm-800nm，厚度为100μm的纤维膜。
实施例7
本实施例电纺纤维中聚(ε-己内酯)、聚(ε-己内酯)-REDV、明胶的质量分数分别为60wt％、25wt％、15wt％，丁香酚占上述聚合物总量的40wt％。
称取1350mg聚(ε-己内酯)(Mn＝8×104)，562.5mg聚(ε-己内酯)-REDV(Mn＝1×104)，337.5mg明胶溶于15mL三氟乙醇中，并滴加冰醋酸搅拌至溶液均相澄清，然后加入900mg的丁香酚，混合均匀；将溶液置于注射器中，使用单道注射泵电纺装置，在电压为15kV，流量为0.6mL/h，接收距离为18cm条件下进行电纺，经过24h后收集到纤维直径为500nm-800nm，厚度为150μm的纤维膜。
实施例8
本实施例电纺纤维中聚(ε-己内酯)、聚(ε-己内酯)-REDV、明胶的质量分数分别为30wt％、45wt％、25wt％，丁香酚占上述聚合物总量的45wt％。
称取300mg聚(ε-己内酯)(Mn＝5×104)，450mg聚(ε-己内酯)-REDV(Mn＝5×104)，250mg明胶溶于5mL三氟乙醇中，并滴加冰醋酸搅拌至溶液均相澄清，然后加入450mg的丁香酚，混合均匀；将溶液置于注射器中，使用单道注射泵电纺装置，在电压为15kV、流量为0.6mL/h、接收距离为15cm条件下进行电纺，经过8h后收集到纤维直径为800nm-1200nm，厚度为50μm的纤维膜。
实施例9
本实施例电纺纤维中聚(ε-己内酯)、聚(ε-己内酯)-REDV、明胶的质量分数分别为65wt％、15wt％、20wt％，丁香酚占上述聚合物总量的50wt％。
称取1300mg聚(ε-己内酯)(Mn＝8×104)，300mg聚(ε-己内酯)-REDV(Mn＝3×104)400mg明胶溶于10mL三氟乙醇中，并滴加冰醋酸搅拌至溶液均相澄清，然后加入1000mg的丁香酚，混合均匀；将溶液置于注射器中，使用单道注射泵电纺装置，在电压为10kV、流量为0.6mL/h、接收距离为15cm条件下进行电纺，经过16h后收集到纤维直径为800nm-1200nm，厚度为100μm的纤维膜。
实施例10
本实施例电纺纤维中聚(ε-己内酯)、聚(ε-己内酯)-REDV、明胶的质量分数分别为20wt％、50wt％、30wt％，丁香酚占上述聚合物总量的10wt％。
称取600mg聚(ε-己内酯)(Mn＝12×104)，1500mg聚(ε-己内酯)-REDV(Mn＝1×104)，900mg明胶溶于15mL三氟乙醇中，并滴加冰醋酸搅拌至溶液均相澄清，然后加入300mg的丁香酚，混合均匀；将溶液置于注射器中，使用单道注射泵电纺装置，在电压为12kV、流量为0.8mL/h，接收距离为18cm条件下进行电纺，经过18h后收集到纤维直径为800nm-1200nm，厚度为150μm的纤维膜。
以上对本发明做了示例性的描述，应该说明的是，在不脱离本发明的核心的情况下，任何简单的变形、修改或者其他本领域技术人员能够不花费创造性劳动的等同替换均落入本发明的保护范围。

Claims (1)

1.一种抗菌性聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜，其特征在于纤维膜是由直径为200-1200nm的聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶纤维和包载在纤维内部的植物源抗菌剂构成的厚度为50-150μm的纤维膜，其中聚(ε-己内酯)的质量分数为20-80wt％，聚(ε-己内酯)-REDV的质量分数为10-50wt％，明胶的质量分数为10-30wt％，而植物源抗菌剂的含量占聚合物总量的5-50wt％；所包载的植物源抗菌剂为丁香酚；抗菌性聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜的制备方法包括以下步骤：
1)将数均分子量为(5-12)×104的聚(ε-己内酯)、数均分子量为(1-5)×104聚(ε-己内酯)-REDV和明胶溶于三氟乙醇中，室温下搅拌溶解，向溶液中滴加冰醋酸并搅拌溶液至均相澄清，然后加入5-50wt％的植物源抗菌剂，混合均匀形成浓度为100-200mg/mL的聚合物/三氟乙醇溶液；
2)将配制好的溶液置于注射器内，使用单道注射泵电纺装置，在电压为10-15kV，接收距离为15-20cm，流量为0.4-0.8mL/h条件下进行电纺，得到厚度为50-150μm的包载植物源抗菌剂的聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜。
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