CN108057132A - 一种表面截留明胶的条带化聚丙交酯-己内酯薄膜及其制备方法 - Google Patents

一种表面截留明胶的条带化聚丙交酯-己内酯薄膜及其制备方法 Download PDF

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CN108057132A
CN108057132A CN201810078510.9A CN201810078510A CN108057132A CN 108057132 A CN108057132 A CN 108057132A CN 201810078510 A CN201810078510 A CN 201810078510A CN 108057132 A CN108057132 A CN 108057132A
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高长有
张德腾
许圣均
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Zhejiang University ZJU
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Abstract

本发明涉及一种表面截留明胶的条带化聚丙交酯‑己内酯(PLCL)薄膜及其制备方法,该薄膜同时结合了取向结构的物理拓扑形貌和生物活性大分子明胶,可促进雪旺细胞快速定向迁移。其制备方法为:通过浇筑成膜法制备聚丙交酯‑己内酯聚合物薄膜;利用聚二甲基硅氧烷(PDMS)模板热压法在上述聚合物薄膜表面引入条带微图案;将处理后的薄膜在丙酮/水混合液中溶胀后迅速转移到明胶水溶液中,明胶被截留到条带化聚丙交酯‑己内酯薄膜表面。本发明首次利用一种简单易行的溶胀‑截留法改性条带化聚丙交酯‑己内酯聚酯薄膜。该薄膜可用于调控细胞的微环境,适用于神经、血管、肌腱、尿道等组织的再生和功能重建,在组织工程与再生医学领域具有良好的应用前景。

Description

一种表面截留明胶的条带化聚丙交酯-己内酯薄膜及其制备 方法
技术领域
本发明涉及条带化聚丙交酯-己内酯薄膜及其制备方法,特别涉及一种表面截留明胶的条带化聚丙交酯-己内酯薄膜及其制备方法。
背景技术
周围神经系统具有一定的自修复能力,但对于大尺寸损伤难以实现自我修复。迄今为止,神经自体移植物与人工合成的神经导管都不能达到有效的修复效果,特别是神经功能的恢复。在生物体内,雪旺细胞是神经髓鞘细胞,有序排列形成髓鞘膜包裹在神经纤维外部,使得神经轴突具有一定的取向结构。从仿生设计角度出发,在神经导管内壁同时引入取向的物理拓扑形貌和促进轴突生长的生物活性大分子,实现物理引导和化学信号刺激的协同作用,调控雪旺细胞的定向迁移,可能会更有效地实现神经组织的修复和再生。目前,由于易加工、可降解、力学性能可调、可修饰等优点,人工合成的聚酯材料正越来越多的用于神经组织再生研究。聚丙交酯-己内酯由于力学性能、降解性能可灵活调节,成为组织修复领域的常用材料。但传统聚酯材料由于缺少反应活性位点不易进行改性。
材料表面引入合适的条带微图案可以有效地发挥物理引导作用。通常来讲,材料表面固定生物活性大分子如细胞外基质蛋白、多肽、因子等可以显著促进细胞的粘附、铺展、分化等行为。明胶是胶原蛋白部分水解的产物,含有肽和蛋白质的混合物,可以与细胞表面的整合素受体结合,促进细胞的粘附、铺展和增殖。将生物大分子明胶固定到聚酯材料表面,通常采用化学方法如胺解、碱解、等离子体处理后进一步接枝或物理方法直接吸附、涂覆等。然而,化学方法程序复杂;物理方法修饰到材料表面的明胶不均匀且不稳定,容易脱附。表面截留法是一种简单、方便的物理改性方法,基本原理是将聚酯材料在其良溶剂中适度溶胀,此时大分子链处于舒展状态,随后迅速置于其不良溶剂、生物大分子的良溶剂中进行截留。在此过程中,聚酯材料的溶胀区在生物大分子溶液中发生相分离,生物大分子进入疏松的大分子链间隙与聚酯材料分子链相互缠结,从而实现改性。许多天然生物大分子如壳聚糖和海藻酸盐均可通过这种方法对脂肪族聚酯材料表面进行改性。然而,通过溶胀-截留法在带有拓扑形貌的聚酯条带表面固定生物活性大分子的研究尚未报道。
本发明将聚丙交酯-己内酯聚酯表面条带化,利用溶胀-截留法固定生物大分子明胶,适用于具有取向结构的组织修复领域。
发明内容
本发明的目的是提供一种表面截留明胶的条带化聚丙交酯-己内酯薄膜及其制备方法,该方法通过简单易行的溶胀-截留法来改性条带化聚丙交酯-己内酯聚酯薄膜,该薄膜条带形貌保持良好,同时修饰了生物活性大分子,能够调控细胞雪旺细胞快速定向迁移,在神经、血管、肌腱、尿道等具有取向结构的组织再生领域具有良好的应用前景。
本发明的一种表面截留明胶的条带化聚丙交酯-己内酯薄膜的制备方法,包括以下步骤:
1)将聚丙交酯-己内酯溶于三氯甲烷,浇筑到聚四氟乙烯模具中待溶剂挥发后成膜;
2)将具有条带化表面的聚二甲基硅氧烷模板置于步骤1)制备的聚丙交酯-己内酯薄膜表面,热压,自然冷却到室温后得到表面条带化的聚丙交酯-己内酯薄膜;
3)将步骤2)制备的条带化聚丙交酯-己内酯薄膜浸于丙酮和水的混合液中,溶胀20min后立即转移到明胶水溶液中静置至少2h;
4)将步骤3)制备的聚丙交酯-己内酯薄膜用膜过滤水冲洗多次,40℃烘箱干燥。
所述的步骤1)所用的聚丙交酯-己内酯中丙交酯与己内酯的投料比为75:25,聚丙交酯-己内酯的重均分子量为50kDa。聚丙交酯-己内酯与三氯甲烷溶剂的质量体积比为7%-25%g/mL。
所述的步骤2)中聚二甲基硅氧烷模板的条带化表面为:沟槽与嵴的宽度范围为3-40μm、深度为4-10μm。
所述的步骤2)中热压温度优选为166-180℃。
所述的步骤3)保持在37℃下完成,所述的丙酮与水以体积比为7:3混合。明胶水溶液的浓度通常为0.25-20mg/mL。
本发明的表面截留明胶的条带化聚丙交酯-己内酯薄膜具有良好的生物相容性,同时结合了物理条带和生物活性大分子明胶:条带可以使细胞发生接触引导效应,引导细胞取向、伸长和定向迁移;明胶利于细胞的粘附、铺展、增殖。二者结合,可以有效调控生物材料与细胞接触的微环境,特别在周围神经组织再生中可以避免雪旺细胞的无规迁移,利于神经再生。此外,该薄膜同样适用于带有取向结构的其他组织再生。
附图说明
图1是表面截留明胶的条带化聚丙交酯-己内酯薄膜的制备过程及促进雪旺细胞定向迁移的示意图。
图2是实施例1制备的聚丙交酯-己内酯薄膜表面的扫描电镜照片。
图3是实施例1制备的聚丙交酯-己内酯薄膜表面的台阶仪测试结果。
图4是雪旺细胞在实施例1制备的聚丙交酯-己内酯薄膜表面的迁移轨迹图。
图5是实施例2制备的聚丙交酯-己内酯薄膜表面的扫描电镜照片。
图6是实施例2制备的聚丙交酯-己内酯薄膜表面的台阶仪测试结果。
图7是雪旺细胞在实施例2制备的聚丙交酯-己内酯薄膜表面的迁移轨迹图。
图8是实施例3制备的聚丙交酯-己内酯薄膜表面的扫描电镜照片。
图9是实施例3制备的聚丙交酯-己内酯薄膜表面的台阶仪测试结果。
图10是雪旺细胞在实施例3制备的聚丙交酯-己内酯薄膜表面的迁移轨迹图。
图11实施例4制备的聚丙交酯-己内酯薄膜表面的扫描电镜照片。
图12是实施例4制备的聚丙交酯-己内酯薄膜表面的台阶仪测试结果。
图13是雪旺细胞在实施例4制备的聚丙交酯-己内酯薄膜表面的迁移轨迹图。
具体实施方式
以下结合实施例进一步说明本发明。
实施例1:
称取0.1g聚丙交酯-己内酯于1.4mL三氯甲烷中充分搅拌溶解均匀,随后倒至底部放有1.1cm×1.3cm干净玻璃片的聚四氟乙烯模具中。常温条件下挥发除去溶剂使聚合物成膜,40℃烘箱放置一周除去溶剂三氯甲烷。将制备的聚丙交酯-己内酯薄膜置于恒温数显加热台上,升温至166℃并保持10min使薄膜完全熔融。将聚二甲基硅氧烷模板盖在聚丙交酯-己内酯薄膜上方并轻轻按压,1min后从加热台上取下盖有利用聚二甲基硅氧烷的聚丙交酯-己内酯薄膜,自然冷却到室温后用镊子取下聚二甲基硅氧烷模板,得到表面具有沟槽-嵴条带微图案的聚丙交酯-己内酯薄膜。将条带化的聚丙交酯-己内酯薄膜在丙酮:水=7:3(v/v)的混合液中溶胀20min,随后迅速置于2.5mg/mL的明胶水溶液中浸泡2h,整个实验过程中保持温度为37℃。最后,用镊子取出薄膜并用膜过滤水冲洗三次,置于40℃烘箱干燥。薄膜表面截留明胶的密度约为0.7μg/cm2。该实施例制备的聚丙交酯-己内酯薄膜表面形貌如图2、图3所示。由图可见,聚丙交酯-己内酯薄膜表面的沟槽-嵴条带尺寸为3/3μm、深度为3-4μm。固定明胶后条带表面变得粗糙,出现少许凸起,但整体形貌保持良好、沟槽-嵴排列规则有序。将雪旺细胞以1×104个/cm2的密度种植到该薄膜表面培养12h后,如图4所示,细胞沿条带方向定向地迁移。
实施例2:
方法同实施例1,区别在于改变明胶水溶液的浓度为15mg/mL。该实施例制备的聚丙交酯-己内酯薄膜表面截留明胶的密度约为1.9μg/cm2,形貌如图5、图6所示。由图可见,聚丙交酯-己内酯薄膜表面的沟槽-嵴条带尺寸为3/3μm、深度为3-4μm。固定明胶后条带表面变得更加粗糙,出现较多几百纳米的凸起,但条带的形貌依然保持良好。将雪旺细胞以1×104个/cm2的密度种植到该薄膜表面培养12h后,如图7所示,细胞沿条带方向快速、定向地迁移,而垂直于条带方向几乎不发生迁移。
实施例3:
方法同实施例1,区别在于改变聚丙交酯-己内酯的条带尺寸。该实施例制备的聚丙交酯-己内酯薄膜表面截留明胶的密度约为0.7μg/cm2,形貌如图8、图9所示。由图可见,聚丙交酯-己内酯薄膜表面的沟槽-嵴条带尺寸为10/10μm、深度为3-4μm。固定明胶后条带虽略粗糙,但沟槽-嵴依旧保持良好,排列规则有序。将雪旺细胞以1×104个/cm2的密度种植到该薄膜表面培养12h后,如图10所示,细胞较大程度地沿条带方向定向地迁移。
实施例4:
方法同实施例2,区别在于改变明胶水溶液的浓度为15mg/mL。该实施例制备的聚丙交酯-己内酯薄膜表面截留明胶的密度约为1.9μg/cm2,形貌如图11、图12所示。由图可见,聚丙交酯-己内酯薄膜表面的沟槽-嵴条带尺寸为10/10μm、深度为3-4μm。固定明胶后条带表面出现较多圆形凸起,但整体形貌保持良好。将雪旺细胞以1×104个/cm2的密度种植到该薄膜表面培养12h后,如图13所示,多数细胞沿平行于条带的方向做定向迁移。

Claims (8)

1.一种表面截留明胶的条带化聚丙交酯-己内酯薄膜的制备方法,其特征在于,包括如下步骤:
1)将聚丙交酯-己内酯溶于三氯甲烷,浇筑到聚四氟乙烯模具中待溶剂挥发后成膜;
2)利用具有条带化表面的聚二甲基硅氧烷模板置于步骤1)制备的聚丙交酯-己内酯薄膜表面,热压,自然冷却到室温后得到表面条带化的聚丙交酯-己内酯薄膜;
3)将步骤2)制备的条带化聚丙交酯-己内酯薄膜浸于丙酮和水的混合液中,溶胀20min后立即转移到明胶水溶液中静置至少2h;
4)将步骤3)制备的聚丙交酯-己内酯薄膜用膜过滤水冲洗多次,40℃烘箱干燥。
2.根据权利要求1所述的一种表面截留明胶的条带化聚丙交酯-己内酯薄膜的制备方法,其特征在于,步骤1)所述的聚丙交酯-己内酯其丙交酯与己内酯的投料比为75:25,重均分子量为50kDa。
3.根据权利要求1所述的一种表面截留明胶的条带化聚丙交酯-己内酯薄膜的制备方法,其特征在于,步骤1)所述的聚丙交酯-己内酯与三氯甲烷溶剂的质量体积比为7%-25%g/mL。
4.根据权利要求1所述的一种表面截留明胶的条带化聚丙交酯-己内酯薄膜的制备方法,其特征在于,步骤2)所利用的聚二甲基硅氧烷模板的条带化表面为:沟槽与嵴的宽度范围为3-40μm、深度为4-10μm。
5.根据权利要求1所述的一种表面截留明胶的条带化聚丙交酯-己内酯薄膜的制备方法,其特征在于,步骤2)所述的热压温度为166-180℃。
6.根据权利要求1所述的一种表面截留明胶的条带化聚丙交酯-己内酯薄膜的制备方法,其特征在于,步骤3)保持在37℃下完成,所述的丙酮与水以体积比为7:3混合。
7.根据权利要求1所述的一种表面截留明胶的条带化聚丙交酯-己内酯薄膜的制备方法,其特征在于,步骤3)所述的明胶水溶液的浓度为0.25-20mg/mL。
8.一种表面截留明胶的条带化聚丙交酯-己内酯薄膜,其特征在于,所述的聚丙交酯-己内酯薄膜表面具有条带化的沟槽和嵴结构,并修饰有明胶。
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