CN105641744A - 一种纳微米多尺度壳聚糖聚乳酸复合支架及其制备方法 - Google Patents
一种纳微米多尺度壳聚糖聚乳酸复合支架及其制备方法 Download PDFInfo
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Abstract
本发明涉及一种纳微米多尺度壳聚糖聚乳酸复合支架及其制备方法,它采用在一定条件下分别静电纺丝纳米和微米壳聚糖聚乳酸复合纤维膜,复合纤维膜经碱洗风干后按一定配比将纳米和微米壳聚糖聚乳酸复合纤维膜在水中高速剪切成短纤维,分样后,加入微米尺度的盐颗粒作为致孔剂,经冷冻干燥成型,水置换去除致孔剂,再经冷冻干燥得到纳微米多尺度壳聚糖聚乳酸复合三维支架。壳聚糖和聚乳酸复合克服两者的缺点,该制备方法工艺简单,支架在纤维直径和孔径同时具有微纳米尺度,赋予复合支架良好的生物学性能和力学性能。
Description
技术领域
本发明涉及一种纳微米多尺度壳聚糖聚乳酸复合支架及其制备方法,属于生物医用材料技术领域。
背景技术
壳聚糖作为一种阳离子碱性多糖,结构类似于人体内的硫酸软骨素和透明质酸等多糖类物质,在体内可被降解吸收,并具有止痛、止血、抑菌、促进伤口愈合、减小疤痕等优异特性,目前在生物医用材料领域有重要的应用。SundararajanV等人采用相分离技术首次制备了壳聚糖支架,其结构为数十到数百微米厚度的片状蜂窝结构,具有较好的生物学性能(Biomaterials,1999;20:1133-1142),但壳聚糖作为支架材料的主要缺点是易于溶胀。聚乳酸作为美国FDA最早批准的一种合成聚合物支架材料,具有无毒、与人体相容性好、在体内可完全降解吸收等优点,被广泛地应用于骨、软骨、血管和皮肤等组织和器官的修复(LeeEJ等,AnnBiomedEng,2014;42(2):323),聚乳酸憎水性强,细胞亲和性差,因此壳聚糖和聚乳酸复合可克服两者的缺点。李庚公开了一种壳聚糖-聚乳酸多孔复合材料及其制备方法,表明壳聚糖和聚乳酸的复合材料有较好的性能(CN102757625A)。
研究表明纳米纤维支架有较大的比表面积,可提供细胞粘附和分化的更好条件,但单纯的纳米纤维力学强度较低,无法承受支架植入过程的力学载荷。相比而言,微米纤维支架具有较好的力学性能。另外,支架要求具有三维贯通的孔结构,便于营养物质的传递、代谢物质的交换以及细胞的迁移。研究表明,对于营养物质的传递和代谢物质的交换,纳米尺度的孔具有较高的效率,并可促进支架的降解吸收和组织再生,但该尺度的孔阻碍细胞在支架内部的渗透和迁移,微米尺度的孔则对细胞的渗透、迁移以及随之的分化增殖、新血管和神经的长入是比较有利的。微纳米共存的孔结构可以保证细胞生长过程中营养物质的传递、细胞的迁移以及新血管和神经生长所需的空间。
综上,在纤维直径和孔径同时具有微纳米多尺度的三维支架具有良好的生物相容性和力学性能。目前研究者多采用相分离的方法制备三维支架,但无法控制其纤维直径和孔径,或采用静电纺丝的方法获得二维的纤维薄膜,但如何制备纤维直径和孔径同时具有微纳米多尺度的三维壳聚糖聚乳酸复合支架制备方法尚未见报导。
发明内容
本发明的主要设想是采用静电纺丝的方法分别制备纳米和微米壳聚糖聚乳酸复合纤维薄膜,按一定配比将纳米和微米复合纤维膜混合,并将复合薄膜高速剪切成短纤维,分样后,加入微米尺度的致孔剂,经冷冻干燥成型,水置换去除致孔剂,再经冷冻干燥得到纳微米多尺度壳聚糖聚乳酸复合支架三维支架。
本发明中纳微米多尺度壳聚糖三维支架的制备方法具体如下所述。
(1)静电纺丝纳米壳聚糖纤维膜:配制壳聚糖5.0%的溶液,溶剂为三氟乙酸和二噁烷的混合物,体积比为70:30;配制聚乳酸10.0%的溶液,溶剂为氯仿和二噁烷的混合物,体积比为70:30。将壳聚糖和聚乳酸溶液分别加入两个注射器中,用二合一针头将两个注射器连接,通以20~25kV的电压进行纺丝。
(2)静电纺丝微米壳聚糖纤维膜:配制壳聚糖7.5%的溶液,溶剂为三氟乙酸和二噁烷的混合物,体积比为90:10;配制聚乳酸20.0%的溶液,溶剂为氯仿和二噁烷的混合物,体积比为90:10。将壳聚糖和聚乳酸溶液分别加入两个注射器中,用二合一针头将两个注射器连接,通以12~15kV的电压进行纺丝。
(3)将静电纺丝得到的纳米和微米壳聚糖聚乳酸复合纤维膜浸泡在0.5mol/L的氢氧化钠溶液中去除未挥发的溶剂,水洗风干后待用。
(4)分别称取定量的纳米和微米壳聚糖聚乳酸复合纤维膜,加入到定量的水中,在水中高速剪切成短纤维。
(5)分样至称量瓶中,加入筛分后具有一定粒度分布的盐作为致孔剂,快速在液氮中冷冻成型并冻干。
(6)冻干后的样品经水置换去除盐,再经冷冻干燥可获得纳微米多尺度壳聚糖聚乳酸复合支架。
本发明制得的纳微米多尺度壳聚糖聚乳酸复合支架外观为白色圆盘,其形状和高度可在分样中采用不同的容器和分样体积调节。壳聚糖和聚乳酸的复合可克服两者的缺点,支架具有更好的综合性能。制备方法中主要通过静电纺丝获得纳微米尺度的纤维,制备中也可方便的调整纺丝条件得到不同尺度的纳米和微米壳聚糖聚乳酸复合纤维,通过纳米纤维之间的孔形成纳米尺度的孔,通过微米尺度的盐致孔剂获得微米尺度的孔。该支架在纤维直径和孔径同时具有微纳米尺度,赋予支架良好的生物相容性和力学强度,便于营养物质的输送和细胞的迁移。该制备方法工艺简单,易于得到不同直径的壳聚糖聚乳酸复合纤维,并可通过盐致孔获得微米尺度的孔,其孔径容易控制,相比于单纯的静电纺丝法,该法还可容易地制备三维结构的支架。
具体实施方式
下面结合具体实施例,对本发明内容作进一步的说明,但本发明的实现方式并不局限于此。
实施例1:分别取7ml的三氟乙酸和3ml的二噁烷,混合均匀,称取0.5g壳聚糖,搅拌溶解,静置12小时。分别取7ml的氯仿和3ml的二噁烷,混合均匀,称取1.0g聚乳酸,搅拌溶解,静置12小时。将壳聚糖和聚乳酸溶液分别加入到两个注射器中,用二合一针头将两个注射器连接,通以20kV的电压纺丝,收集在接地的铝箔上,可获得壳聚糖聚乳酸纳米复合纤维薄膜,纤维直径在50-500nm之间。分别取9ml的三氟乙酸和1ml的二噁烷,混合均匀,称取0.75g壳聚糖,搅拌溶解,静置12小时。分别取9ml的氯仿和1ml的二噁烷,混合均匀,称取2.0g聚乳酸,搅拌溶解,静置12小时。将壳聚糖和聚乳酸溶液分别加入到两个注射器中,用二合一针头将两个注射器连接,通以15kV的电压纺丝,收集在接地的铝箔上,可获得壳聚糖聚乳酸微米复合纤维薄膜,纤维直径在1-10μm之间。将静电纺丝得到的纳米和微米纤维膜浸泡在0.5mol/L的氢氧化钠溶液中去除未挥发的溶剂,水洗风干。分别称取0.2g和0.8g纳米和微米壳聚糖聚乳酸复合纤维膜,在100ml水中高速剪切成短纤维,分样至称量瓶中,加入筛分的氯化钠颗粒,其粒度分布在50-350μm之间,高度与分样的液面相同,快速在液氮中冷冻成型,在冻干机中冻干。冻干后的样品经水置换去除支架中的盐致孔剂,再经冷冻干燥可获得纳微米多尺度壳聚糖聚乳酸复合三维支架。
实施例2:分别取14ml的三氟乙酸和6ml的二噁烷,混合均匀,称取1.0g壳聚糖,搅拌溶解,静置12小时。分别取14ml的氯仿和6ml的二噁烷,混合均匀,称取2.0g聚乳酸,搅拌溶解,静置12小时。将壳聚糖和聚乳酸溶液分别加入到两个注射器中,用二合一针头将两个注射器连接,通以25kV的电压纺丝,收集在接地的铝箔上,可获得壳聚糖聚乳酸纳米复合纤维薄膜,纤维直径在50-500nm之间。分别取9ml的三氟乙酸和1ml的二噁烷,混合均匀,称取0.75g壳聚糖,搅拌溶解,静置12小时。分别取9ml的氯仿和1ml的二噁烷,混合均匀,称取2.0g聚乳酸,搅拌溶解,静置12小时。将壳聚糖和聚乳酸溶液分别加入到两个注射器中,用二合一针头将两个注射器连接,通以12kV的电压纺丝,收集在接地的铝箔上,可获得壳聚糖聚乳酸微米复合纤维薄膜,纤维直径在1-10μm之间。将静电纺丝得到的纳米和微米纤维膜浸泡在0.5mol/L的氢氧化钠溶液中去除未挥发的溶剂,水洗风干。分别称取2.4g和0.6g纳米和微米壳聚糖聚乳酸复合纤维膜,在100ml水中高速剪切成短纤维,分样至称量瓶中,加入筛分的氯化钠颗粒,其粒度分布在50-350μm之间,高度与分样的液面相同,快速在液氮中冷冻成型,在冻干机中冻干。冻干后的样品经水置换去除支架中的盐致孔剂,再经冷冻干燥可获得纳微米多尺度壳聚糖聚乳酸复合三维支架。
Claims (6)
1.一种纳微米多尺度壳聚糖聚乳酸复合支架及其制备方法,其特征在于在一定条件下分别静电纺丝纳米和微米壳聚糖聚乳酸复合纤维膜,在0.5mol/L的氢氧化钠溶液中去除未挥发的溶剂并水洗风干后,按一定配比将纳米和微米壳聚糖聚乳酸复合纤维膜在水中高速剪切成短纤维,分样后,加入具有一定粒度分布的微米氯化钠致孔剂,经冷冻干燥成型,水置换去除氯化钠致孔剂,再经冷冻干燥得到纳微米多尺度壳聚糖聚乳酸复合三维支架。
2.根据权利要求1所述的纳微米多尺度壳聚糖聚乳酸复合支架及其制备方法,其特征在于所使用的壳聚糖分子量在20~40万之间,聚乳酸分子量在5~10万之间。
3.根据权利要求1所述的纳微米多尺度壳聚糖聚乳酸复合支架及其制备方法,其特征在于静电纺丝纳米纤维膜时,壳聚糖浓度为5.0%,溶剂为三氟乙酸和二噁烷的混合物,体积比为70:30,聚乳酸浓度为10.0%,溶剂为氯仿和二噁烷的混合物,体积比为70:30,纺丝电压为20~25kV。
4.根据权利要求1所述的纳微米多尺度壳聚糖聚乳酸复合支架及其制备方法,其特征在于静电纺丝微米壳聚糖纤维膜时,壳聚糖浓度为7.5%,溶剂为三氟乙酸和二噁烷的混合物,体积比为90:10,聚乳酸浓度为20.0%,溶剂为氯仿和二噁烷的混合物,体积比为90:10,纺丝电压为12~15kV。
5.根据权利要求1所述的纳微米多尺度壳聚糖聚乳酸复合支架及其制备方法,其特征在于纳米壳聚糖聚乳酸复合纤维和微米壳聚糖聚乳酸复合纤维的质量比在20:80到80:20之间,壳聚糖聚乳酸纳米微米纤维在水中的总浓度在1~3%之间。
6.根据权利要求1所述的纳微米多尺度壳聚糖聚乳酸复合支架及其制备方法,其特征在于使用的氯化钠致孔剂的粒度范围在50~350μm之间。
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